Happy New Year to all and especially all who responded to my call for help. Thanks again. Best wishes for a great year. Sincerely, Peter A. Stolzenberg,Pesto Inc.
sounds like a research grant to me. I don't believe the salary of faculty in Chemistry departments of most major colleges would be "soft money" requiring grant support. I commend the Society of Analytical Chemists for trying to encourage this bridge. 20K of seed money for a new line of research can make an important difference to a junior faculty member.
} ---------------------------------------------------------------. } } } Hum....so for a 2000 hour year, this works out to be $10 per hour. } What a gift. And they probably want $30/hour of work in return. } } No matter how you package it, all of this babble does not measure } up to today's standards. Unless SEM, etc. is a obscure and } diminutive endeavor, I simply do not understand the cost-benefit } ratio. Maybe this is not an annual salary. OK. Is this in addition } to an existing income stream? Geeze, I hope it is the latter. But it } sounds like the position is on-site. So, the candidate gets a full time } job at McDonald's as well? } } All I can say is that I am glad and relieved that I do not have to } work and try to survive in this type of environment. Welcome to H-2 visas. } } gary g. } } } At 08:14 AM 12/31/99 , you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } I just received this notification from the Society for Analytical Chemists } } of Pittsburgh. Perhaps it will provide an important start for a new chem } } prof who has an interest in expanding the use of microscopy and/or in } } walking across the new bridge between microscopy and spectroscopy: } } } } "Twenty-first annual Analytical Chemistry Starter Grant Award" } } The society for Analytical Chemists of Pittsburgh will award one grant of } } $20,000 to an assistant professor in the field of analytical chemistry. } } The purpose of this grant is to encourage high-quality, innovative research } } by a new analytical chemistry professor and to promote the training and } } development of graduate students in this field. Assistant professors who } } have accepted a US college or university appoint since December 31, 1996 } } are eligible. Application forms available from: } } James Chadwick, Chairman } } Starter Grant Committee } } Society for Analytical Chemists of Pittsburgh } } 200 Penn Center Blvd., Suite 332 } } Pittsburgh, PA 15235 } } Ph: 1-800-825-3221, Xt. 208 } } Fx: 412-825-3224 } } } } Deadline for application receipt: February 29, 2000 } } Award winner announced: May 1, 2000 } } } } } } Best regards and welcome to the new millennium! } } Barbara Foster } } Consortium President } } Microscopy/Microscopy Education ...Educating microscopists for greater } } productivity. } } } } 125 Paridon Street Suite 102 Springfield, MA 01118 } } PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com } } Visit our web site {http://www.MME-Microscopy.com/education} } } ****************************************************** } } MME is America's first national consortium providing } } customized on-site workshops in all areas of } } microscopy, sample preparation, and image analysis.
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)
Thanks for the info. I understand what is going on now. sounds like a great opportunity.
I discussed compensation before and don't want to clutter up the list with a repeat. But I would appreciate an off-list communication which helps me better understand how the mechanics of SEM and its personnel work in the big picture. I get asked often how to get into the field. Since I use my SEM for personal research and photography only, I am not in academia or companys' formal research departments. If I sound ignorant about this, it is because I am. I work with some high school and upper level pre-high school students at times using my SEM and LMs. What can be done with the SEM and LMs turns them on as much as it still does me today. I'm hopelessly hooked.
But I lack facts and figures about this type of career. Rather than speculating about it, what do you careerists think? Is this a good career? What is involved in getting into the career at various levels? And what compensation is commensurate at these levels?
All responses will be kept confidential. Use PGP if you need to.
gary g.
At 12:39 PM 1/1/00 , you wrote: } sounds like a research grant to me. I don't believe the salary of faculty in Chemistry departments of most major colleges would be "soft money" requiring grant support. I commend the Society of Analytical Chemists for trying to encourage this bridge. 20K of seed money for a new line of research can make an important difference to a junior faculty member. } } } } ---------------------------------------------------------------. } } } } } } Hum....so for a 2000 hour year, this works out to be $10 per hour. } } What a gift. And they probably want $30/hour of work in return. } } } } No matter how you package it, all of this babble does not measure } } up to today's standards. Unless SEM, etc. is a obscure and } } diminutive endeavor, I simply do not understand the cost-benefit } } ratio. Maybe this is not an annual salary. OK. Is this in addition } } to an existing income stream? Geeze, I hope it is the latter. But it } } sounds like the position is on-site. So, the candidate gets a full time } } job at McDonald's as well? } } } } All I can say is that I am glad and relieved that I do not have to } } work and try to survive in this type of environment. Welcome to H-2 visas. } } } } gary g. } } } } } } At 08:14 AM 12/31/99 , you wrote: } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } I just received this notification from the Society for Analytical Chemists } } } of Pittsburgh. Perhaps it will provide an important start for a new chem } } } prof who has an interest in exd, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
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Thanks for the info. I understand what is going on now. sounds like a great opportunity.
I discussed compensation before and don't want to clutter up the list with a repeat. But I would appreciate an off-list communication which helps me better understand how the mechanics of SEM and its personnel work in the big picture. I get asked often how to get into the field. Since I use my SEM for personal research and photography only, I am not in academia or companys' formal research departments. If I sound ignorant about this, it is because I am. I work with some high school and upper level pre-high school students at times using my SEM and LMs. What can be done with the SEM and LMs turns them on as much as it still does me today. I'm hopelessly hooked.
But I lack facts and figures about this type of career. Rather than speculating about it, what do you careerists think? Is this a good career? What is involved in getting into the career at various levels? And what compensation is commensurate at these levels?
All responses will be kept confidential. Use PGP if you need to.
gary g.
At 12:39 PM 1/1/00 , you wrote: } sounds like a research grant to me. I don't believe the salary of faculty in Chemistry departments of most major colleges would be "soft money" requiring grant support. I commend the Society of Analytical Chemists for trying to encourage this bridge. 20K of seed money for a new line of research can make an important difference to a junior faculty member. } } } } ---------------------------------------------------------------. } } } } } } Hum....so for a 2000 hour year, this works out to be $10 per hour. } } What a gift. And they probably want $30/hour of work in return. } } } } No matter how you package it, all of this babble does not measure } } up to today's standards. Unless SEM, etc. is a obscure and } } diminutive endeavor, I simply do not understand the cost-benefit } } ratio. Maybe this is not an annual salary. OK. Is this in addition } } to an existing income stream? Geeze, I hope it is the latter. But it } } sounds like the position is on-site. So, the candidate gets a full time } } job at McDonald's as well? } } } } All I can say is that I am glad and relieved that I do not have to } } work and try to survive in this type of environment. Welcome to H-2 visas. } } } } gary g. } } } } } } At 08:14 AM 12/31/99 , you wrote: } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } I just received this notification from the Society for Analytical Chemists } } } of Pittsburgh. Perhaps it will provide an important start for a new chem } } } prof who has an interest in expanding the use of microscopy and/or in } } } walking across the new bridge between microscopy and spectroscopy: } } } } } } "Twenty-first annual Analytical Chemistry Starter Grant Award" } } } The society for Analytical Chemists of Pittsburgh will award one grant of } } } $20,000 to an assistant professor in the field of analytical chemistry. } } } The purpose of this grant is to encourage high-quality, innovative research } } } by a new analytical chemistry professor and to promote the training and } } } development of graduate students in this field. Assistant professors who } } } have accepted a US college or university appoint since December 31, 1996 } } } are eligible. Application forms available from: } } } James Chadwick, Chairman } } } Starter Grant Committee } } } Society for Analytical Chemists of Pittsburgh } } } 200 Penn Center Blvd., Suite 332 } } } Pittsburgh, PA 15235 } } } Ph: 1-800-825-3221, Xt. 208 } } } Fx: 412-825-3224 } } } } } } Deadline for application receipt: February 29, 2000 } } } Award winner announced: May 1, 2000 } } } } } } } } } Best regards and welcome to the new millennium! } } } Barbara Foster } } } Consortium President } } } Microscopy/Microscopy Education ...Educating microscopists for greater } } } productivity. } } } } } } 125 Paridon Street Suite 102 Springfield, MA 01118 } } } PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com } } } Visit our web site {http://www.MME-Microscopy.com/education} } } } ****************************************************** } } } MME is America's first national consortium providing } } } customized on-site workshops in all areas of } } } microscopy, sample preparation, and image analysis. } } 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)
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I am posting this for the NY Microscopical Society.
This is a Very Good Course, at a Great Price.
Best Regards
Joseph Passero mailto:jp-at-spacelab.net
New York Microscopical Society -- Course Announcement ====================================================
Bernard Friedman Memorial Workshop
Polarized Light Microscopy
April 8, 9, 15 & 16, 2000
An advanced course on polarized light microscopy which will cover the following topics:
The nature of polarized light
The origin and interpretation of interference colors
Birefringence and crystal orientation
The Indicatrix
Compensation and variable compensators
Interference figures and their interpretation
The workshop will consist of two consecutive weekends of lectures and hands on labs to cover the theoretical and practical aspects of polarized light microscopy.
The course instructors include;
Jan Hinsch of Leica, Inc.
John Reffner of Trace Consulting
N.Y.M.S. Instructor Donald O'Leary.
WHEN: April 8, 9, 15 & 16, 2000 from 10 A.M. to 4 P.M.
WHERE:
New York Microscopical Society Facility 1244 McBride Avenue West Paterson, NJ.
Phone (973) 812-8377
Web Site URL: http://www.nyms.org
(The facility has free parking and is accessible by public transportation, Information on car pools and transportation will be provided.)
COST: $275 for N.Y.M.S. members, $295 for non-members (includes membership) Lunch and course materials are included. Checks made out to N.Y.M.S.
WHO: advanced course for those who have completed "The Use of the Microscope" or are experienced in microscopy and familiar with the theory of its use.
HOW: Register using the form below. Limited to the first 12 registrants.
Thanks for your on-target observations ...and especially for getting Gary into a more positive perspective!
Best regards,
Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} At 02:39 PM 1/1/00 -0600, Tom Phillips wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am looking for any information (year of manufacture, company history, etc.) about a Vickers Instruments, M1500974C microscope. I would especially like to acquire a copy of the owners manual if possible. Please look at the following pictures for further identification.
American Chemical Society, "Applied Optical Microscopy"
3 days of exciting interactive lectures, lab, and demos on all aspects of Light Microscopy, with a touch of video imaging Learn how to -match optics to your application -interpret images from a variety of contrast techniques -troubleshoot for artifacts and misinformation -do simple measurement -put a camera system on your microscope
New Orleans Hyatt Regency, Mar 10-12,2000 Tuition, books, coffee breaks: $895 for ACS members, $995 for non-members
While many of the examples used will be from materials science, this course is NOT LIMITED TO CHEMISTS! Biologists are also welcome.
For a syllabus and enrollment information, visit MME's website: www.MME-Microscopy.com/education (B. Foster is course coordinator)
Please reserve early. This course is given in conjunction with Pittcon, the biggest analytical meeting in the country. Rooms fill up early.
Best regards .... and welcome to the positive side of Y2K!
Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education}
} I am forwarding this from someone who sent this to me. If anyone has any } ideas for this person, please contact him through his email address which } is listed. } } ML } } } From: "rwinn" {rwinn-at-mweb.co.za} } } To: {wong-at-msg.ucsf.edu} } } Subject: CHROMOSOME 5 (5P-) } } Date: Fri, 31 Dec 1999 07:21:48 +0200 } } MIME-Version: 1.0 } } X-Priority: 3 } } } } DEAR, MEI LEI WONG } } } } MY NAME IS RENEY WINN AND I AM MAILING YOU FROM SOUTH AFRICA. } } I AM 25 YRS OLD AND HAS A BABY BOY OF 14 MTHS. } } } } I FOUND OUT THAT HE HAS CRI-DU-CHAT SYNDROME WHICH IS ALSO CALLED CAT-CRY } } SYNDROME. HE HAS A DELETION ON THE SHORT ARM OF CHROMOSOME 5. HIS } } DELETION ON THE SHORT ARM IS FROM THE 14.2 MARK WHICH EXTENDS TO THE 15. } } MARK. } } } } I KNOW THAT THIS IS A VERY RARE SYNDROME AND HAVE BEEN IN TOUCH WITH THE } } SUPPORT GROUPS IN AUSTRALIA AND U.K. HOWEVER.. } } I STILL FEEL THAT I NEED MORE HELP FROM LABRORATRIES OR SCIENTISTS. I AM } } TRYING TO FIND OUT MORE ABOUT THE SEVERITY OF MY SON'S CONDITION. I AM } } TRYING TO FIND OUT HOW BADLY HE WILL BE AFFECTED BY HIS DELETION. I WOULD } } JUST LIKE TO KNOW HOW HIS DELETION SIVERITY WOULD BE CLASIFIED AS. MAYBE } } ON A SCALE OF 1-10 ONE AS BEING NONE SIVERE AND 10 AS MOST SEVERE. THIS } } WAY I WOULD BE ABLE TO UNDERSTAND +- HOW SEVERE MY CHILD'S CONDITION IS. I } } WOULD ALSO APPRECIATE IT IF YOU CAN TELL ME MORE OR LESS WHAT TO EXPECT } } WITH HIS DELETION. } } } } MEI, IF THIS IS NOT SOMETHING YOU CAN HELP ME WITH, COULD YOU PLSE } } FORWARD THIS MAIL TO SOMEONE WHO COULD PLSE. I AM VERY DESPERATE TO FIND } } OUT MORE ABOUT THE SEVERITY OF MY CHILDS DELETION. THIS WOULD BE MUCH } } APPRECIATED. } } } } BRGDS } } RENEY WINN. } } rwinn-at-mweb.co.z Mei Lie Wong Electron Microscope Laboratory Department of Biochemistry HHMI-UCSF Ph. 415-476-4441 Fax 415-476-1902 http://util.ucsf.edu/agard/wong/index.html email wong-at-msg.ucsf.edu
I know this is a little off the microscopy subject, but in the interest of encouraging scientific exploration in a young person. I hope you will indulge me.
I'm looking for materials for my daughter's science project. She is testing the effectiveness of different mouthwashes in killing bacteria found in the mouth. Her experiment matrix requires 12 petri dishes with a bacterial growth medium. I can probably scrounge something to use as petri dishes, but the growth medium is a problem. I'm not even sure what it is properly called or what its makeup is. Is it something we can easily purchase locally, or even better is there a recipe for making a suitable substitute from ingredients found in the home?
Any and all suggestions will be greatly appreciated.
The New York State Center for Advanced Thin Film Technology The University at Albany - State University at New York
The Center for Advanced Thin Film Technology at the University at Albany - SUNY seeks applicants for a postdoctoral position with established expertise in the area of analytical electron microscopy. Demonstrated experience in the development and application of SEM and/or TEM based analyses, including chemical analysis, is required to achieve targeted technique development goals and to support an expanding microscopy base at the Center. Experience with, or interest in, scanning probe microscopy techniques is also desirable. Candidates with a Ph.D. in applied physics, materials science and engineering, chemistry or related fields are encouraged to apply. This appointment is part of an ongoing multi-year investment in personnel and new laboratory facilities for thin film technology research areas at the Center.
The Center is a New York State sponsored high technology research and development center with a $60M state-of-the-art infrastructure, including class 1 clean rooms for back end IC manufacturing on 200 mm wafers, and advanced processing, patterning and characterization capabilities for single and multi-layered thin films. The Center is currently assembling a 200 mm wafer facility for MEMS integration. In addition, a new facility that will house a 25,000 ft2 class 1cleanroom for 300 mm wafer pilot manufacturing and workforce training is presently under construction.
Please send a letter of application including a summary of research interests, curriculum vitae, and three letters of recommendation to:
William Harris, Director of Analytical Sciences NYS Center for Advanced Thin Film Technology CESTM B110 251 Fuller Road Albany, NY 12203
The SUNY/Research Foundation is an Equal Opportunity/Affirmative Action Employer. All positions contingent upon availability of funding.
The New York State Center for Advanced Thin Film Technology The University at Albany - State University at New York
The New York State Center for Advanced Thin Film Technology at the University at Albany - SUNY is a fast growing, high technology research and development program with a mission of supporting industry and creating new jobs. This position will serve as the primary infrastructure support person for the Center's scientific and technical staff.
Job responsibilities include: performing routine maintenance and calibration of a variety of surface spectroscopic and electron microscopy instruments; performing analytical work with these instruments; assisting students with the operation of this equipment; diagnosing and repairing non-routine problems with the instrumentation; and maintaining an adequate store of spare parts and supplies.
The position requires: an associates degree in electronics, computer science, or related technology and two to four years of directly relevant experience; a working knowledge of high vacuum systems; proficiency with related computer hardware/software; demonstrated ability to work in a high energy, team oriented environment; and excellent communication and analytical skills. Experience with surface analysis and/or electron microscopy instrumentation is highly desirable.
Please send a letter of application including a summary of research interests, curriculum vitae, and three letters of recommendation to:
William Harris, Director of Analytical Sciences NYS Center for Advanced Thin Film Technology CESTM B110 251 Fuller Road Albany, NY 12203
The SUNY/Research Foundation is an Equal Opportunity/Affirmative Action Employer. All positions contingent upon availability of funding.
I am working on a spectra plotting program in Visual Basic 6 that is just about ready for general release. I was hoping to put it out there in time for the M&M MM meeting.
It started out just as a program that would take EMSA formatted EELS data from Gatan's ELP and convert them into an Excel compatible file so that I bring the file to a PC. Then I got carried away and used it to try to learn Visual Basic. Currently, the program will open and overlay up to five spectra, plot them in linear, log or rescale them and print them. They can also be copied to the clipboard and pasted in other applications. They can be re-colored and a couple other things as well. There is even a help file!
I am trying to make the program more general and make it completely compatible with the EMMFF standard for all spectra, EDS and EELS alike.I have a couple of requests for information.
1) I would like to have some EELS spectra of different elements in EMMFF format to test it out and include in a distribution packet that I can put on the MAS-LIB. I am particularly interested in transition metal oxides. Please send them to me if you have them and I could put them in the deployment file. Who knows, this could be a poor man's EELS atlas.
2) I would like to know how much interest there is in this program.
3) What features would you like in such a program. It doesn't do any analysis yet, but it may in the future.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
I have a multi-element standards mount which over time has deteriorated and become contaminated with salts. Three of the standards have fallen out of the holder. The mount was made by C.M. Taylor Corp., Sunnyvale, Calif. in 1982, and it appears that this company no longer exists. I am interested in finding someone to re-polish the mount and replace the lost standards. Does anyone know of such a company/person?
Thanks in advance,
Jean M. Howard Reynolds Metals Company Materials Characterization-Electron Microscopy E-mail: jmhoward-at-rmc.com Office: 804.751.2554
Tina, I have not been reading all posts so you may have been provided with all the help you needed, but I thought I would respond just in case... We do a lot of work with similarly embedded specimens. The problem you have is very familiar to me, and I believe you are right in your approach to "Hold the samples in a vacuum for some period of time before putting them into the scope" This is the best method that I have been able to come up with. Assuming: 1) the resin is fully cured, 2) you have vacuum infiltrated the resin into the specimen as thoroughly as practical, and 3) you have minimized the size of high surface area specimens prior to embedding, then there is not much more to try. In my experience the outgassing is most often the result of contaminants within the porosity of the specimen, and is not the result of the resin. If you still feel the need to check this out further, you might try an embedding procedure using a thermosetting resin (such as Bakelite) rather than a thermoplastic (epoxy).
If the sample prep involves polishing, then the source of the contamination is probably the water or oil lubricant in that procedure. Minimize these contaminants by preparing the smallest, and best vacuum infiltrated specimen with as little porosity as possible. I do this by repetitive evacuation/N2 back filling (with heat if possible) prior to embedding. This "clears the way" for better vacuum infiltration.
I have had polymerization problems with some resin/material combinations, and I have usually overcome these by choosing a different resin polymer. Epoxies and acrylics, for example, behave quite differently on various substrates, so I will try LR White if I am having trouble with epoxy, and visa versa. If you use LR White for the initial embedding, keep the reaction volume small, and re-embed in a larger mount if necessary. I do not know if there are any good references on the subject of vacuum infiltration for porous materials, but these are the approaches I have learned to take. Good Luck! Brad Huggins
} ---------- } From: Tina Carvalho[SMTP:tina-at-pbrc.hawaii.edu] } Sent: Tuesday, December 28, 1999 2:23 PM } To: Microscopy Listserver } Subject: SEM - epoxy mountants } } ------------------------------------------------------------------------ } 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- } } We have a Hitachi S-800 FESEM that is mostly used for biological } specimens. However, recently we have had a number of planetary geologists } looking at sections of meteorites mounted on epoxy resin. About the time } they started using the scope, we started having a number of problems that } suggest that we are getting some outgassing and contamination. I'm not } surprised; this happened before when looking at fish ear bones in similar } resin, despite the resin manufacturer's clain their product was completely } stable in the high vacuum and under the beam. } } My question is for those of you who routinely look at such samples: What } do you do to minimize the potential problems? Hold the samples in a } vacuum for some period of time before putting them into the scope? Paint } the exposed epoxy areas with carbon paint or similar? For the biological } samples we only require that the specimens be held over dessicant } overnight, but I suspect that more heroic measures must be taken for these } samples. } } Happy New Year to all! } } Tina } } ************************************************************************** } ** } * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu } * } * Biological Electron Microscope Facility * (808) 956-6251 } * } * University of Hawaii at Manoa * } http://www.pbrc.hawaii.edu/bemf* } ************************************************************************** } ** } }
Can someone suggest a reliable manufacturer for LaB6 filaments? I need the sturdiest, most poor-vacuum-tolerant variety for a Philips EM400T in a student lab! and I figure that users know best which filaments perform well in the real world.
Also, I am looking for a source for Philips bulk-sample mounts, the type used to carry SEM samples for the 6485/STEM system. (I need the mounts, not the specimen rod.) I need both low-background carbon (preferred) or beryllium for EDS, as well as standard copper carriers. My usual sources tell me they have not stocked these items for years. Maybe someone has some sitting unused in a cabinet somewhere?? or knows of a current supplier.
Offlist replies preferred, so as not to clog up the works... will summarize and send info to others who are interested.
Thanks for your help.
Ann Hein-Lehman Trinity College Hartford, CT 860-297-4289 ann.lehman-at-trincoll.edu
I have a side question spurred by Scott Walck's post. Is there a simple low cost program already available that would just allow me to view and print EMSA format spectra under a Windows PC environment? I found some info. in the archives regarding the EMMPDL site and the EMMFF software. I even got so far as to download the sourcecode. But, I don't have a compiler to turn it into an executable. Even so, I'm not sure from the documentation whether it will do what I want. Also I came across NIST's DTSA but that's only for Mac.
Any suggestions out there? And if not, then to Scott yes there is some interest from myself! Thanks, Karen Zaruba
"Walck. Scott 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 } -----------------------------------------------------------------------. } } Dear all, } } I am working on a spectra plotting program in Visual Basic 6 that is just } about ready for general release. I was hoping to put it out there in time } for the M&M MM meeting. } } It started out just as a program that would take EMSA formatted EELS data } from Gatan's ELP and convert them into an Excel compatible file so that I } bring the file to a PC. Then I got carried away and used it to try to learn } Visual Basic. Currently, the program will open and overlay up to five } spectra, plot them in linear, log or rescale them and print them. They can } also be copied to the clipboard and pasted in other applications. They can } be re-colored and a couple other things as well. There is even a help } file! } } I am trying to make the program more general and make it completely } compatible with the EMMFF standard for all spectra, EDS and EELS alike.I } have a couple of requests for information. } } 1) I would like to have some EELS spectra of different elements in EMMFF } format to test it out and include in a distribution packet that I can put on } the MAS-LIB. I am particularly interested in transition metal oxides. } Please send them to me if you have them and I could put them in the } deployment file. Who knows, this could be a poor man's EELS atlas. } } 2) I would like to know how much interest there is in this program. } } 3) What features would you like in such a program. It doesn't do any } analysis yet, but it may in the future. } } -Scott } } Scott D. Walck, Ph.D. } PPG Industries, Inc. } Glass Technology Center } Guys Run Rd. (packages) } P. O. Box 11472 (letters) } Pittsburgh, PA 15238-0472 } } Walck-at-PPG.com } } (412) 820-8651 (office) } (412) 820-8161 (fax)
-- Karen S. Zaruba kszaruba-at-mmm.com 3M Company, St. Paul, MN
As one of the co-authors of the MSA/MAS File format we had a goal to make the format simple enough to be useable in even a simple spreadsheet program.
A number of the major manufacturer's allow you to translate their data files into this format directly from their application programs. To use the data in a spreadsheet, simply specify dual column (x,y) format for the output file and then you can import the data files directly into a MS Excel spreadsheet, or even better a data graphing program like KaleidaGraph (which runs on both Mac's & PC's). Then plot to your hearts content.
Hi, I've been working on thin foils of CZT and I'm looking for some suggestions to help eliminate artifacts created by ion milling. So far I have polished side one and etched (to remove polishing damage) with bromine-methanol and then dimpled side two, ending with 0.1um alumina. I have been successful obtaining a final sample thickness of about 8 to 10 microns. However, when I put the samples into the PIPs (Gatan) to complete the thinning process I'm seeing beam damage. I was hoping that someone might have a different approach or suggestion that would eliminate the beam damage. Thanks for your help. Dorrance
I have an investigator who would like to stain just the cell wall of the yeast S.cerevisae to distinguish it from the cell membrane at the EM level. Any tips or references willbe appreciated. Many thanks
Manuela Palatsides Electron Microscopy Peter MacCallum Cancer Institute Locked Bag#1 A'Beckett Street Melbourne 3000
On Mon, 03 Jan 2000 18:44:05 -0800, Mike Southwell 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, } } I know this is a little off the microscopy subject, but in the interest } of encouraging scientific exploration in a young person. I hope you will } indulge me. } } I'm looking for materials for my daughter's science project. She is } testing the effectiveness of different mouthwashes in killing bacteria } found in the mouth. Her experiment matrix requires 12 petri dishes with } a bacterial growth medium. I can probably scrounge something to use as } petri dishes, but the growth medium is a problem. I'm not even sure } what it is properly called or what its makeup is. Is it something we } can easily purchase locally, or even better is there a recipe for making } a suitable substitute from ingredients found in the home? } } Any and all suggestions will be greatly appreciated. } } Michael Southwell } JEOL USA INC. } Austin, TX } } Michael - I'm sure that any bacterial growth medium requries agar or, at home, gelatin. If you were considering making it at home, then buy some clear gelatin. You'll need to boil water first, though, and then add the gelatin to make it liquid. Before it cools, pour the gelatin along with the apprpriate growth medium into the petri plates (typically, a plate holds about 10 ml of medium). This substance (err, the gelatin) acts primarily as a hardener for the actual medium. It is generally considered to be non-nutritive for most bacteria, although I believe that some may consider it to be nutritious (spelling ?). As for the main nutrients ... I've been dealing primarily with protozoa, but I'm pretty sure that bacteria can grow on an extract of boiled lettuce or even wheat grains, etc. I cannot tell you from my own experience, however, whether a boiled extract in combination with gelatin does indeed work as a suitable growth medium, only that I've heard that gelatin works and that, in my experience, a boiled extract works well for supporting bacterial growth for protozoa. If, instead, you have access to scientific catalogs, then it's fairly easy simply to order proteose peptone, glucose, and agar. The peptone is a standard component of typical "nutrient agar" plates that, I believe, can be bought commercially, and the glucose may be needed as a sugar source and carbon source (?). The agar serves to harden the medium much like gelatin is supposed to. I hope this helps. Nelson Conti [a graduate student formerly from San Francisco State University with a M.A. degree in microbiology & a B. A. degree from UC Davis in Bacteriology ]
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Dear tina and others, My experience with samples you have described makes me head for the nearest exit! However, I have looked at samples such as expanded teflon, and vascular castings mounted on nothing more than carbon sticky tabs. Frequently I have clientes who do not want samples to be gold coated and consequently, charging is an issue.
I frequently examine this specimens at an extremely reduced kV and adjust the spot size and working distance to help in obtaining the best resolution. The parameters will be different for a FESEM: I am using a tungsten system. I would think a coating of colloidal carbon or silver to ground would help under any circumstances. Any thoughts? Good luck!
Ken Tiekotter
On Tue, 28 Dec 1999, Tina Carvalho 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- } } We have a Hitachi S-800 FESEM that is mostly used for biological } specimens. However, recently we have had a number of planetary geologists } looking at sections of meteorites mounted on epoxy resin. About the time } they started using the scope, we started having a number of problems that } suggest that we are getting some outgassing and contamination. I'm not } surprised; this happened before when looking at fish ear bones in similar } resin, despite the resin manufacturer's clain their product was completely } stable in the high vacuum and under the beam. } } My question is for those of you who routinely look at such samples: What } do you do to minimize the potential problems? Hold the samples in a } vacuum for some period of time before putting them into the scope? Paint } the exposed epoxy areas with carbon paint or similar? For the biological } samples we only require that the specimens be held over dessicant } overnight, but I suspect that more heroic measures must be taken for these } samples. } } Happy New Year to all! } } Tina } } **************************************************************************** } * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * } * Biological Electron Microscope Facility * (808) 956-6251 * } * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* } **************************************************************************** } } }
I have a multi-element standards mount which over time has deteriorated and become contaminated with salts. Three of the standards have fallen out of the holder. The mount was made by C.M. Taylor Corp., Sunnyvale, Calif. in 1982, and it appears that this company no longer exists. I am interested in finding someone to re-polish the mount and replace the lost standards. Does anyone know of such a company/person?
Thanks in advance,
Jean M. Howard Reynolds Metals Company Materials Characterization-Electron Microscopy E-mail: jmhoward-at-rmc.com Office: 804.751.2554
Dear Jean,
Plano W. Plannet GmbH is a supplier for electron microscopy. This company offers refurbishment of SEM standards. The company is situated in Germany.
Address: Ernst-Befort-Str.12 D-35578 Wetzlar
e-mail: plano-at-t-online.de, or plano-at-plano-em.com http://www.plano-em.com
You might contact them to see if they can help you with your problem.
The New York State Center for Advanced Thin Film Technology The University at Albany - State University at New York
The Center for Advanced Thin Film Technology at the University at Albany - SUNY seeks applicants for a postdoctoral position with established expertise in the area of analytical electron microscopy. Demonstrated experience in the development and application of SEM and/or TEM based analyses, including chemical analysis, is required to achieve targeted technique development goals and to support an expanding microscopy base at the Center. Experience with, or interest in, scanning probe microscopy techniques is also desirable. Candidates with a Ph.D. in applied physics, materials science and engineering, chemistry or related fields are encouraged to apply. This appointment is part of an ongoing multi-year investment in personnel and new laboratory facilities for thin film technology research areas at the Center.
The Center is a New York State sponsored high technology research and development center with a $60M state-of-the-art infrastructure, including class 1 clean rooms for back end IC manufacturing on 200 mm wafers, and advanced processing, patterning and characterization capabilities for single and multi-layered thin films. The Center is currently assembling a 200 mm wafer facility for MEMS integration. In addition, a new facility that will house a 25,000 ft2 class 1cleanroom for 300 mm wafer pilot manufacturing and workforce training is presently under construction.
Please send a letter of application including a summary of research interests, curriculum vitae, and three letters of recommendation to:
William Harris, Director of Analytical Sciences NYS Center for Advanced Thin Film Technology CESTM B110 251 Fuller Road Albany, NY 12203
The SUNY/Research Foundation is an Equal Opportunity/Affirmative Action Employer. All positions contingent upon availability of funding.
The New York State Center for Advanced Thin Film Technology The University at Albany - State University at New York
The New York State Center for Advanced Thin Film Technology at the University at Albany - SUNY is a fast growing, high technology research and development program with a mission of supporting industry and creating new jobs. This position will serve as a primary infrastructure support person for the Center's scientific and technical staff.
Job responsibilities include: performing routine maintenance and calibration of a variety of surface spectroscopic and electron microscopy instruments; performing analytical work with these instruments; assisting students with the operation of this equipment; diagnosing and repairing non-routine problems with the instrumentation; and maintaining an adequate store of spare parts and supplies.
The position requires: an associates degree in electronics, computer science or related technology and two to four years of directly relevant experience; a working knowledge of high vacuum systems; proficiency with related computer hardware/software; demonstrated ability to work in a high energy, team oriented environment; and excellent communication and analytical skills. Experience with surface analysis and/or electron microscopy instrumentation is highly desirable.
Please send a letter of application including a summary of research interests, curriculum vitae, and three letters of recommendation to:
William Harris, Director of Analytical Sciences NYS Center for Advanced Thin Film Technology CESTM B110 251 Fuller Road Albany, NY 12203
The SUNY/Research Foundation is an Equal Opportunity/Affirmative Action Employer. All positions contingent upon availability of funding.
I have a question for all of the SEM microscope labs with EDS.
What is the percent of Biological use that the EDS gets in your facility?
This is going to be my first semester teaching a Biological EDS class. The facility here sees very little, if any, biological EDS projects. It is used here as in other places I am familiar with nearly 100% non biological applications.
I would like as many responses as possible so I can give a representative summary to the students in the class. A short reply directly to me ( Geoffrey.Lloyd.Williams-at-cmich.edu or willi1gl-at-cmich.edu ) would be much appreciated. A rough % bio use, most popular bio samples (plants or animals, type of organisms . . .) and most common analysis (Quantitative, Qualitative, Dot mapping. . .).
Thank you in advance. If there is sufficient interest I would be willing to put a summary up on the list for all.
-Geoff W.
-- Geoff Williams
Electron Microscope Facility Supervisor Biology Department Brooks Hall Central Michigan University Mt. Pleasant, MI 48859
Ken, If you have a vacuum evaporator Ken, I would try indirect carbon. Most metals will evaporate directionally only. Carbon will also evaporate indirectly (around corners) although at a reduced thickness. The advantage of this type of evaporation is the carbon will coat very convoluted surfaces which are not line of site. Another advantage is the carbon will add very little structure to your sample at high magnifications. Additionally heating of the sample can be reduced due to shading of the source from the sample. One caveat is, as you know, carbon is a very inefficient secondary producer and the carbon will not have the efficiency of gold. Indirect carbon, with a proper thickness, will prevent charging though. Just put a line of site shield between your source and sample while keeping it as small as possible. Good luck. Russ Gillmeister, Xerox
-----Original Message----- } From: Ken Tiekotter [mailto:tiekotte-at-up.edu] Sent: Wednesday, January 05, 2000 3:07 AM To: Tina Carvalho Cc: Microscopy Listserver
Dear tina and others, My experience with samples you have described makes me head for the nearest exit! However, I have looked at samples such as expanded teflon, and vascular castings mounted on nothing more than carbon sticky tabs. Frequently I have clientes who do not want samples to be gold coated and consequently, charging is an issue.
I frequently examine this specimens at an extremely reduced kV and adjust the spot size and working distance to help in obtaining the best resolution. The parameters will be different for a FESEM: I am using a tungsten system. I would think a coating of colloidal carbon or silver to ground would help under any circumstances. Any thoughts? Good luck!
Ken Tiekotter
On Tue, 28 Dec 1999, Tina Carvalho 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- } } We have a Hitachi S-800 FESEM that is mostly used for biological } specimens. However, recently we have had a number of planetary geologists } looking at sections of meteorites mounted on epoxy resin. About the time } they started using the scope, we started having a number of problems that } suggest that we are getting some outgassing and contamination. I'm not } surprised; this happened before when looking at fish ear bones in similar } resin, despite the resin manufacturer's clain their product was completely } stable in the high vacuum and under the beam. } } My question is for those of you who routinely look at such samples: What } do you do to minimize the potential problems? Hold the samples in a } vacuum for some period of time before putting them into the scope? Paint } the exposed epoxy areas with carbon paint or similar? For the biological } samples we only require that the specimens be held over dessicant } overnight, but I suspect that more heroic measures must be taken for these } samples. } } Happy New Year to all! } } Tina } } **************************************************************************** } * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * } * Biological Electron Microscope Facility * (808) 956-6251 * } * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* } **************************************************************************** } } }
Due to some recent high-resolution requirements in our lab, I find myself having to go back to Sputter Coating 101 (after years of just putting specimens in the coater and turning it on without a second thought!). We find ourselves in need of very thin coatings with as little structure as possible, in order to image samples down in the nanometer range.
We have a chromium coater, but often need to revisit samples days or weeks after the initial coating. The oxidation problem rears its ugly head. We intend to purchase a platinum target, but don't yet have one, so we're experimenting with our venerable Au/Pd coater.
My questions are:
1) I seem to remember a string on this listserver suggesting that lower deposition currents yield finer coating structure. Is this right? Does a low deposition current for a longer time yield a finer coating than a higher current for a shorter time? (I'm running some tests to check this, but would be very interested in others' experiences, too.)
2) Deposition current can be controlled by the initial current setting (i.e., the knob on the machine) and by the argon flow through the chamber. Is there any difference in the coating when adjusting the deposition current by either of these two ways?
3) Charts I have seen indicate that deposition current is directly proportional to coating rate. Is the same true for coating time? I.e., is a one minute coating twice as thick as a 30 sec. coating? It would seem so intuitively, but you know what they say about the word "assume".
My apologies if these are very basic questions, but, like I said, back to SC 101!
I'll be happy to summarize the responses for anyone who is interested.
Thanks!
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine Bldg. University of Missouri Columbia, MO 65211 (573)882-8304 tindallr-at-missouri.edu http://www.biotech.missouri.edu/emc/
You can probably use Agar, this is commonly used to grow yeast cultures for brewing. This is a vegetable based gelatin like substance. You can buy it at some organic type stores in the bulk section (you don't need much probably 10 tbsp) it costs about $100/ounce.
I don't remember the exact proportions but I think you use something like 1-3 tsp per 2cups of water and 1 tbsp of a complex sugar (brown sugar will probably be fine). You just get it al boiling for 5-10 min to sterilize it and pour into sterile glass dishes WITH LIDS. I use small Ball jars. Put it in the fridge and it should harden in 10-15 min. If not, then my proportions are off.
After it is hard, just wipe some saliva on the media and watch the green and red goo grow.
Good luck -andrew
----- Original Message ----- } From: Mike Southwell {mjsouth-at-flash.net} To: {microscopy-at-sparc5.microscopy.com} Sent: Monday, January 03, 2000 7:44 PM
Dear Dorrance, It seems to me that the etching step is redundant. The ion-milling should remove any polishing deformation and the etching will introduce surface profiling. I recall that the CZT is very soft and may require some modification to the ion beam voltage or current to reduce damage. Sounds like you are close to getting good results. At 06:34 PM 1/4/00 -0700, you wrote: } } Hi, } I've been working on thin foils of CZT and I'm looking for some suggestions } to help eliminate artifacts created by ion milling. So far I have polished } side one and etched (to remove polishing damage) with bromine-methanol and } then dimpled side two, ending with 0.1um alumina. I have been successful } obtaining a final sample thickness of about 8 to 10 microns. However, when } I put the samples into the PIPs (Gatan) to complete the thinning process I'm } seeing beam damage. I was hoping that someone might have a different } approach or suggestion that would eliminate the beam damage. } Thanks for your help. } Dorrance } 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
We have always kept our osmium solutions in glass containers. However, we are in the process of evaluating our safety procedures and discussed the idea of increasing the safety of the transport of osmium from the refrigerator to the fume hood by putting the osmium solution in plastic containers. (If they are dropped, they would not break and cause the danger of a spill outside the hood.) Does anyone have experience with osmium stored in plastic? Any comments about this particular subject or any of your safety with osmium procedures are welcomed. Thanks.
Donna Wagahoff SIU School of Medicine PO Box 19627 Springfield, IL 62794-9627 217-782-0898 fax217-524-3227
This experiment is very popular in the science fair circuit; it is only slightly less popular than "What kind of music do plants like best?". Moreover (as a topic RELEVANT TO THE LIST) van Leeuwenhoek showed more than 300 years ago, BY MICROSCOPY, that the experiment as usaully performed is invalid as a test of mouthwash efficacy in situ. You might want to consult his observations on the topic.
Prepared agar media can be purchased from educational scientific supply houses such as Carolina Biologicals or Ward's, both of which maintain web sites. They both offer "instant" formulations designed for preparation of Petri dishes without autoclaving. You would want a medium capable of supporting growth of Streptococcus species, which are an important component of the oral microflora. The streptococci are somewhat fastidious in their nutritional requirements. Therefore, select something rich in organic nutrients such as amino acids. Tryptic Soy Medium would probably work best. The proteose peptone/glucose composition suggested by a previous respondent would also probably work. Lettuce or wheat grain extract would probably not give satisfactory results.If you wish to make your own medium at home from local sources, I suggest table sugar, beef bullion (more concentrated than in culinary use) and agar (often available at health food stores). A pressure cooker would help to ensure sterility during preparation. Be advised that many of the bacteria in the oral cavity are obligate anaerobes; they will not grow on any medium if it is in contact with the atmosphere.
Best wishes for the success of your daughter's work! Mike Dalbey Biology Dept. University of California Santa Cruz, CA 95064
Randy Tindall wrote: ================================================= {snip} We find ourselves in need of very thin coatings with as little structure as possible, in order to image samples down in the nanometer range.
We have a chromium coater, but often need to revisit samples days or weeks after the initial coating. The oxidation problem rears its ugly head. We intend to purchase a platinum target, but don't yet have one, so we're experimenting with our venerable Au/Pd coater.
{snip} =================================================== One (maybe the only) solution to the problem is to coat with osmium metal in the osmium plasma coater. Pt grain size can be on the same size magnitude as your nanostructures. You can get information about this coater on our website at URL http://www.2spi.com/catalog/osmi-coat.html
One does not need to worry about grain size because it is an (apparently completely) amorphous layer and since it is a precious group metal, it has the intertness of Au and Pt and will essentially last forever.
The physics of the process is explained in the US Patent #5855682 which can be clicked to from the above URL.
We would be happy to coat some demo samples for you in our demo unit at any time, just let us know.
Disclaimer: SPI Supplies distributes the line of Osmium Plasma Coaters, manufactured by Nippon Laser and Electronics Ltd.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.spi.cc ######################## ============================================
Randy Tindall wrote: ================================================= {snip} We find ourselves in need of very thin coatings with as little structure as possible, in order to image samples down in the nanometer range.
We have a chromium coater, but often need to revisit samples days or weeks after the initial coating. The oxidation problem rears its ugly head. We intend to purchase a platinum target, but don't yet have one, so we're experimenting with our venerable Au/Pd coater.
{snip} =================================================== One (maybe the only) solution to the problem is to coat with osmium metal in the osmium plasma coater. Pt grain size can be on the same size magnitude as your nanostructures. You can get information about this coater on our website at URL http://www.2spi.com/catalog/osmi-coat.html
One does not need to worry about grain size because it is an (apparently completely) amorphous layer and since it is a precious group metal, it has the intertness of Au and Pt and will essentially last forever.
The physics of the process is explained in the US Patent #5855682 which can be clicked to from the above URL.
We would be happy to coat some demo samples for you in our demo unit at any time, just let us know.
Disclaimer: SPI Supplies distributes the line of Osmium Plasma Coaters, manufactured by Nippon Laser and Electronics Ltd.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.spi.cc ######################## ============================================
Agar can be purchased from some Asian stores at much cheaper rate if you can find them. It is made into strands, so, don't expect powder form.
Ann Fook Yang EM Unit, Eastern Cereal and Oilseed Research Centre, Rm 2091, K.W. Neatby Bldg., Central Experimental Farm, Ottawa, Ontario, Canada K1A 0C6
Phone: 613-759-1638 Fax; 613-759-1701
} } } "Andrew Cahill" {cahill-at-colorado.edu} 01/05 11:15 AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Mike,
You can probably use Agar, this is commonly used to grow yeast cultures for brewing. This is a vegetable based gelatin like substance. You can buy it at some organic type stores in the bulk section (you don't need much probably 10 tbsp) it costs about $100/ounce.
I don't remember the exact proportions but I think you use something like 1-3 tsp per 2cups of water and 1 tbsp of a complex sugar (brown sugar will probably be fine). You just get it al boiling for 5-10 min to sterilize it and pour into sterile glass dishes WITH LIDS. I use small Ball jars. Put it in the fridge and it should harden in 10-15 min. If not, then my proportions are off.
After it is hard, just wipe some saliva on the media and watch the green and red goo grow.
Good luck -andrew
----- Original Message ----- } From: Mike Southwell {mjsouth-at-flash.net} To: {microscopy-at-sparc5.microscopy.com} Sent: Monday, January 03, 2000 7:44 PM
Agar can be purchased from some Asian stores at much cheaper rate if you can find them. It is made into strands, so, don't expect powder form.
Ann Fook Yang EM Unit, Eastern Cereal and Oilseed Research Centre, Rm 2091, K.W. Neatby Bldg., Central Experimental Farm, Ottawa, Ontario, Canada K1A 0C6
Phone: 613-759-1638 Fax; 613-759-1701
} } } "Andrew Cahill" {cahill-at-colorado.edu} 01/05 11:15 AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Mike,
You can probably use Agar, this is commonly used to grow yeast cultures for brewing. This is a vegetable based gelatin like substance. You can buy it at some organic type stores in the bulk section (you don't need much probably 10 tbsp) it costs about $100/ounce.
I don't remember the exact proportions but I think you use something like 1-3 tsp per 2cups of water and 1 tbsp of a complex sugar (brown sugar will probably be fine). You just get it al boiling for 5-10 min to sterilize it and pour into sterile glass dishes WITH LIDS. I use small Ball jars. Put it in the fridge and it should harden in 10-15 min. If not, then my proportions are off.
After it is hard, just wipe some saliva on the media and watch the green and red goo grow.
Good luck -andrew
----- Original Message ----- } From: Mike Southwell {mjsouth-at-flash.net} To: {microscopy-at-sparc5.microscopy.com} Sent: Monday, January 03, 2000 7:44 PM
I store my osmium in a glass bottle with plastic cap (Schott bottle - orange cap - commonly used for tissue culture work). The inside portion of the cap turns black quite rapidly but the solution is stable for months at room temperature. I store this glass bottle inside an aluminum-foiled plastic container in my fume hood at room temp. The clear plastic of this outer container gets translucent black within weeks no matter how carefully I seal the glass bottle. I think the storage of osmium in any single container (except a sealed ampule) in the refrigerator is foolish and unnecessary. Due to University regulations, we are required to store our aldehyde fixatives and other toxic chemicals inside the refrigerator in "secondary containment" containers. We keep the glass bottles of aldehydes in small plastic containers and transport them this way to the fume hood for use.
} } } } We have always kept our osmium solutions in glass containers. However, we } are in the process of evaluating our safety procedures and discussed the } idea of increasing the safety of the transport of osmium from the } refrigerator to the fume hood by putting the osmium solution in plastic } containers. (If they are dropped, they would not break and cause the danger } of a spill outside the hood.) } Does anyone have experience with osmium stored in plastic? Any comments } about this particular subject or any of your safety with osmium procedures } are welcomed. } Thanks. } } Donna Wagahoff } SIU School of Medicine } PO Box 19627 } Springfield, IL 62794-9627 } 217-782-0898 } fax217-524-3227
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)
} We have always kept our osmium solutions in glass containers. However, we } are in the process of evaluating our safety procedures and discussed the } idea of increasing the safety of the transport of osmium from the } refrigerator to the fume hood by putting the osmium solution in plastic } containers. (If they are dropped, they would not break and cause the danger } of a spill outside the hood.) } Does anyone have experience with osmium stored in plastic? Any comments } about this particular subject or any of your safety with osmium procedures } are welcomed.
Dear Donna, How about putting the glass containers inside plastic containers? There are also padded and styrofoam containers which you could use for the transportation step. These can usually be made out of waste packaging material--better than sending it to a landfill. Yours, Bill Tivol
} I know this is a little off the microscopy subject, but in the interest } of encouraging scientific exploration in a young person. I hope you will } indulge me. } } I'm looking for materials for my daughter's science project. She is } testing the effectiveness of different mouthwashes in killing bacteria } found in the mouth. Her experiment matrix requires 12 petri dishes with } a bacterial growth medium. I can probably scrounge something to use as } petri dishes, but the growth medium is a problem. I'm not even sure } what it is properly called or what its makeup is. Is it something we } can easily purchase locally, or even better is there a recipe for making } a suitable substitute from ingredients found in the home? } } Any and all suggestions will be greatly appreciated. } Mike -
You're a supportive parent! I agree with you that purchasing prepared microbiological groth media doesn't have as much learning potential as starting from scratch, but it's undeniably easier. And probably cheaper. How old is she? Since you obviously don't know any microtechnique, I worry about the adequacy of her "experiment matrix". Some degree of sterile technique is required, implying the use of an alcohol lamp-sterilized wire loop.
If you want to cook your own, you'll need a pressure cooker and the instructions available in Zook et al., "The Microcosmos guide to exploring microbial space" (see the MICRO bibliography! URL below). I can send you photocopied pages, but if this is a major project you may want the book. Or you can buy prepared sterile plates (and the agarose that you'd need for home cooking) from any large biological supply house, such as Carolina Biological (800-334-5551). You may even have a medical supply source in town.
Caroline
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
Rick Felten-at-MACDERMID 01/05/2000 04:16 PM I have a couple of Ni/Au Samples that I need analyzed using AFM. The conductive samples would need to be scanned over a 10 X 10 micron region. Were are located in Waterbury Ct and the closer to us the better.
Is there a way to remove a moderate barrel distortion from an image? I have images taken with a 17 mm wide angle lens that are slightly distorted and wish to make some area measurements from them.
I have a picture of a grid of known size so it seems like there should be a way to 'stretch' the picture into shape in something like Photoshop, I just don't know where to look. Any help or ideas?
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
The drive belt on our Reichert (Leica) Ultracut E broke last night, leaving a number of people in a state of panic. Leica does not have any of these in the US, and the Vienna group is still on holiday.
I would appreciate and forever be indebted to anyone who could quickly send me the appropriate belt, which I could either pay for (list $38.34, plus shipping) or replace when ours come in.
Mahalo! Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Nestor, The Gatan ELP option for MSA formatted files outputs the files to 5 columns serial. There are other ASCII options, but they are not MSA format. I have found that Noran does the same thing in our new system. It is not easy to parse the data for a spreadsheet when it is in that format. I would have been happy if they had done it with the two column option that is available in the standard or if they simply used a single column. Once it is in that format, it's easy to do in a spreadsheet. With multiple columns, I had to go into the file with a word processor, take out the hard returns at the end of each row, and then replace all of the commas with hard returns. Then I could open them easily in the spreadsheet. That's how I got started with the Basic program -simply to read them in and output them to a two column text with the MSA format. Then I got carried away with VB.
-Scott
} -----Original Message----- } From: Nestor J. Zaluzec [mailto:zaluzec-at-Sparc5.Microscopy.Com] } Sent: Tuesday, January 04, 2000 8:14 PM } To: microscopy-at-Sparc5.Microscopy.Com } Subject: Spectra in MSA/MSA Format How to Plot... } } } -------------------------------------------------------------- } ---------- } 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 } } } -------------------------------------------------------------- } ---------. } } } } As one of the co-authors of the MSA/MAS File format we } had a goal to make the format simple enough to be } useable in even a simple spreadsheet program. } } A number of the major manufacturer's allow you to } translate their data files into this format directly } from their application programs. To use the data } in a spreadsheet, simply specify dual column (x,y) } format for the output file and } then you can import the data files directly into } a MS Excel spreadsheet, or even better a data } graphing program like KaleidaGraph (which runs } on both Mac's & PC's). Then plot to your hearts content. } } } } Nestor } Your Friendly Neighborhood SysOp. } } }
There is a Bacterial Growth Medium Kit for grades 3-6 in the Carolina K-6 Science Catalog ( I have the Fall 1998 Catalog). It contains 2 cans of Beef Browth, 2 Boxes of Unflavored Gelatin, Multivitamins, 20 sterile Petri Dishes and 10 sterile Cotton-Tipped Applicators. It recommends the use of a pressure cooker or autoclave for "complete" sterility. It costs $26.43 per kit (1998 price) Phone: 1-800-334-5551 Most of the items are available at the supermarket. The petri dishes and sterile applicators are more difficult to get. A local college microbiology department might give a hand or even some sterile Nutrient Agar Plates.
Whenever I tried storing osmium solutions in plastics they always reacted with the plastic causing it to blacken. This took place even in Teflon altho at a much slower rate.
Why not use a glass that has been coated with plastic and rendered breakproof. I see that many chemicals (like acids) come in such reagent bottles.
Hint: maybe one of the EM vendors may know about this.
John
} We have always kept our osmium solutions in glass containers. However, we } are in the process of evaluating our safety procedures and discussed the } idea of increasing the safety of the transport of osmium from the } refrigerator to the fume hood by putting the osmium solution in plastic } containers. (If they are dropped, they would not break and cause the danger } of a spill outside the hood.) } Does anyone have experience with osmium stored in plastic? Any comments } about this particular subject or any of your safety with osmium procedures } are welcomed.
#################################################################### John J. Bozzola, Ph.D., Director Micro-Imaging and Analysis Center 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 ####################################################################
The newer SEMs require software access to do some fundamental adjustments: magnification, high voltage, crt brightness, etc.
I am attempting to calibrate the magnification on a JEOL 5800 SEM. I am guessing this requires a sequence of keystrokes to access the computer in "service" mode. Does anyone have the keystroke sequence or know of the calibration procedure for the JEOL 5800?
We could quote on that and certainly, if it involved repolishing and re-coating only this would be worthwhile. In this case the separated standards would need to be identified, remounted and thoroughly polished. If much thickness is lost during the polishing there may be a problem with other standards. Certainly its possible but I expect that the cost will come close to our new standard blocks, complete with micro-engraving. I suggest that you check it out in our online. Disclaimer: ProSciTech supplies WDS/EDS standards.
Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Wednesday, January 05, 2000 1:48 AM, Howard, Jean M. [SMTP:JMHoward-at-rmc.com] wrote: } } Dear listers, } } I have a multi-element standards mount which over time has deteriorated and } become contaminated with salts. Three of the standards have fallen out of } the holder. The mount was made by C.M. Taylor Corp., Sunnyvale, Calif. in } 1982, and it appears that this company no longer exists. } I am interested in finding someone to re-polish the mount and replace the } lost standards. Does anyone know of such a company/person? } } Thanks in advance, } } Jean M. Howard } Reynolds Metals Company } Materials Characterization-Electron Microscopy } E-mail: jmhoward-at-rmc.com } Office: 804.751.2554 } } }
Some time ago I sent out a Kinney High Vacuum manual. There were 4 interested parties; one got the original; others, copies. I have now found a 2nd one of these antiques, and would be happy to send it to the highest bidder (i.e., free to the first to reply). The date in the front is listed as June, 1961.
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
Perhaps it is time to point out to some of the Manufacturers that they should implement the MSA/MAS standard with an option to specify the number of columns in the output file ( like the demo/test copy does). That would be in the spirit of how the format was originally designed so that "additional" programs code would not have to be written. They generally listen to customers so speak your mind!
In the mean time I'll look into also putting a version of the demo translator on-line.
(Grinning Devilishly)
Nestor Your Friendly Neighborhood SysOp
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Good Lord.......So this is what NAFTA brings us to.......Third world companies undermining real science with a New Brand of Burger King products and services? Care for an Enchilada while you wait for your SEM images?.....$5.95 please.......
Just a cynical comment that is much closer to the truth than I care to think about. -----Original Message----- } From: Paul Rennie (KIDDE) {Paul.Rennie-at-kidde-hq.com} To: microscopy-at-Sparc5.Microscopy.Com {microscopy-at-Sparc5.Microscopy.Com}
Dear All,
Having just purchased a new SEM, our old instrument, a Cambridge S200 is destined to be sent to one of our companies in Cuidad Victoria, Mexico (Although I'm trying to persuade management that it's not worth it). Although we will probably be installing the instrument ourselves, I would welcome details of any service companies in Mexico. Additionally, who sells consumables within Mexico, or is it best to purchase over the border in the USA.
Thanking you in advance.
Paul Rennie
Kidde International Research Mathisen Way Colnbrook Slough Berks SL3 0HB UK
Having just purchased a new SEM, our old instrument, a Cambridge S200 is destined to be sent to one of our companies in Cuidad Victoria, Mexico (Although I'm trying to persuade management that it's not worth it). Although we will probably be installing the instrument ourselves, I would welcome details of any service companies in Mexico. Additionally, who sells consumables within Mexico, or is it best to purchase over the border in the USA.
Thanking you in advance.
Paul Rennie
Kidde International Research Mathisen Way Colnbrook Slough Berks SL3 0HB UK
JEOL 100B - Disassembled, great source for Parts . $2,000 Cash and Carry.
PHILLIPS 201C - Will be taken out of service this month. Excellent working condition. Maintained on OEM Service Contracts since day one. Make an offer.
-- Ford M. Royer, MT(ASCP) Analytical Instruments, Ltd (Refurbished Histology, Cytology, & General Lab Equipment) 9921 13th Ave. N. Minneapolis, MN 55441-5004 800-565-1895 phone 612-929-1895 fax web site: http://www.aibltd.com
Received: from UICVM.UIC.EDU (UIC-VMNET.CC.UIC.EDU [128.248.2.49]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id KAA00322 for {zaluzec-at-sparc5.MICROSCOPY.COM} ; Thu, 6 Jan 2000 10:37:07 -0600 Received: by UICVM.UIC.EDU (IBM VM SMTP V2R4a) via spool with SMTP id 6780 ; Thu, 06 Jan 2000 10:15:16 CST Received: from UICVM (NJE origin SPAMPNS-at-UICVM) by UICVM.UIC.EDU (LMail V1.2a/1.8a) with BSMTP id 1309; Thu, 6 Jan 2000 10:15:16 -0600 Message-Id: {200001061637.KAA00322-at-Sparc5.Microscopy.Com}
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id {CF52CAJP} ; Thu, 6 Jan 2000 08:48:07 -0800 Message-ID: {EAF569A980E4D21185380090271F40EB125F35-at-EXCHANGE} {Microscopy-at-Sparc5.Microscopy.Com} , "',rfelten-at-Macdermid.com'" {,rfelten-at-Macdermid.com}
Rick,
While we are not too close to Connecticut (California to be exact), we have a great deal of experience in analyzing all types of samples with the AFM and overnight delivery services can make turnaround times very short. What information are you looking for from the samples? Please give me a call at 650-962-8767 or respond to this email so we can help you out.
-Rob
Robert J. Plano Staff Analyst, SPM Services Surface Science Labs (650)962-8767, ext. 742 http://www.cea.com http://www.surface-science.com
} -----Original Message----- } From: "rfelten-at-Macdermid.com"-at-Sparc5.Microscopy.Com } [mailto:"rfelten-at-Macdermid.com"-at-Sparc5.Microscopy.Com] } Sent: Wednesday, January 05, 2000 1:17 PM } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: Looking for a Contract laboratory that does AFM } } } -------------------------------------------------------------- } ---------- } 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/MicroscopyListser} ver/FAQ.html } } } -------------------------------------------------------------- } ---------. } } } } } Rick Felten-at-MACDERMID } 01/05/2000 04:16 PM } I have a couple of Ni/Au Samples that I need analyzed using AFM. The } conductive samples would need to be scanned over a 10 X 10 } micron region. } Were are located in Waterbury Ct and the closer to us the better. } } Any Help would be appreciated. } } Ric } } }
I have about 45 USED 8 inch 10 megabyte bernoulli flexible disk cartridges that I will ship to the first person that emails me at rgriffin-at-eng.uab.edu. Due to shipping charges, I will only ship to a continental U.S. address.
We have gotten rid of our computer that uses these disks and I thought someone out there that still needed them might appreciate them (since you can't buy them anymore!).
Robin Griffin Materials and Mechanical Engineering The University of Alabama at Birmingham
******************************************************* Corazon D. Bucana, Ph.D. Department of Cancer Biology U.T. M.D. Anderson Cancer Center 1515 Holcombe Blvd. Box 173 Houston, Texas 77030 Phone: (713) 792-8106 FAX: (713) 792-8747 Email:bucana-at-audumla.mdacc.tmc.edu FAX: (713) 792-8747
While they are in Florida, they deal extensively with Latin America. As for either Dr. Neddy Fookes or Dr. Barry Fookes... tell them I sent you.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
At 03:54 PM 1/6/00 +0000, Paul Rennie (KIDDE) wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Osmium vapors can penetrate and blacken plastics. The long term effects are uncertain. If you can find a Teflon bottle this might work.
Probably best to stay with glass bottles with Teflon colsures and do not hand carry the solution (i.e. use a lab cart for transport). Hope this is some help.
Charles Duvic, Ph.D. Chief Chemist
--
LADD RESEARCH 131 Dorset Lane Williston, VT 05495
TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net web site http://www.ladd.cc
This was sent to the MSA Education Committee. It seems to be a worthwhile endeavor, so I am passing it along to those on the listserver. Jay Jerome
Adam Fagen wrote:
} Dear Microscopy Community: } } The National Association of Graduate-Professional Students (NAGPS) has } recently received a grant from the Alfred P. Sloan Foundation to conduct a } survey of doctoral students on their graduate school experiences. The } survey will be completed on the Web {http://survey.nagps.org/} by current } and recent doctoral students from January - May 2000, and the results made } publicly available on the Web on a department specific basis in September. } } This effort is a follow-up to a more limited survey which occurred this } past spring, which was aimed at science and engineering doctoral students. } The aggregate results from that survey are available at } {http://www.phds.org/survey/results/} . } } The survey we are conducting is unique in at least two important ways: it } collects information on a department-specific basis, not only averaged } over entire institutions or disciplines (though discipline-level results } will also be available). So it will be possible to look at, for instance, } responses from individual biology programs, or to rank history departments } based on faculty mentoring. And it makes this data publicly available on } the Internet in Fall 2000. So we'll be opening the door about the } situation in individual departments for wider viewing by graduate } students, prospective students, faculty, administrators, etc. } } The survey is based upon best practices and covers issues in a number of } areas, including information for prospective students, curriculum breadth } and flexibility, career guidance and placement services, faculty } mentoring, time to degree, department climate, teaching, professionalism, } and overall satisfaction. In other words, the sort of best practices and } concerns outside of the reputation. The NAGPS survey itself will run from } January 18-May 1, 2000, and will be available on the Web at } {http://survey.nagps.org/} (which already has a number of resources). } } For this survey to be useful, it is vital that we reach as many current } and recent doctoral students (anyone who has been enrolled for at least } one semester in the past five years) as possible. We are hoping that we } can encourage a significant percentage of students to respond so that the } results will represent a broad range of experiences and a realistic } picture of department and institutional practices. } } In order to realize this level of participation, getting the word out is } obviously very important. One of the publicity strategies we're employing } is trying to reach current and recent doctoral students through the major } professional societies and organizations, such as the MSA. (Other } strategies include messages to relevant e-mail listservs, coverage in } campus and national media, and working through graduate student } organizations, department chairs and graduate deans, other organizations, } etc.) } } Since the MSA reaches so many current and recent graduate students, it is } certainly one of the organizations of prime importance for getting the } word out. We have thought of a few strategies for spreading the word } among your membership (and would welcome additional ideas and } suggestions): (1) distribution on e-mail listservs that reach a high } number of graduate students and recent graduates, those who have left } their programs, etc.; (2) notice in newsletters and other publications } that current and former students might see; and (3) publicity at meetings } and conferences. We are happy to provide whatever resources and materials } that would facilitate distribution (e.g., flyers, letters, posters, etc.). } We would certainly appreciate any insight you have on publicity within } (and outside of) the MSA. } } As a bit of background on our organization, NAGPS represents nearly } 900,000 graduate and professional students on 150 member campuses and is } dedicated to improving the quality of graduate and professional student } life and education by actively promoting the interests and welfare of } graduate- and professional-degree-seeking students. } } Of course, I'd be happy to answer any questions or provide any more info. } Thanks for your assistance in this important effort! } } --Adam Fagen } } Adam Fagen, Chair } Ad Hoc Committee on Faculty-Student Relations } National Association of Graduate-Professional Students (NAGPS) } } NAGPS Web: http://www.nagps.org/ } The National Doctoral Program Survey: http://survey.nagps.org/ } } Adam Fagen \ afagen-at-fas.harvard.edu } Molecular Biology/Education / http://www.fas.harvard.edu/~afagen/ } Harvard University GSAS \ http://mazur-www.harvard.edu/
-- Jay ---------------------------------------------- - AKA: W. Gray Jerome, Ph.D. - - Department of Pathology - - Wake Forest University School of Medicine - - Winston-Salem, NC 27157-1092 - - Ph: 336-716-4972, 336-716-2675 - - Fax: 336-716-6174 - - E-mail: jjerome-at-wfubmc.edu - ----------------------------------------------
Can anyone recommend image montaging software? That is, software that assists you in compositing several images into one larger image. I am familiar with the offerings from GATAN for their Digital Micrograph and QuickStitch from Enroute Software. Also, can anyone offer an evaluation of the QuickStitch software?
TIA
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 530 754 7536 fax http://katie.ucdavis.edu raharris-at-ucdavis.edu
Plastic is oxidized and so part of the osmium is exhausted in the vials. Even when frozen osmium diffuses through plastic containers. So you may save a rare breakage, but you are certain to have osmium in your refrigerator. It's just a bad idea to pack osmium in plastic. You could use a secondary container, be it plastic or glass, to increase overall safety. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Thursday, January 06, 2000 3:08 AM, Donna Wagahoff [SMTP:DWAGAHOFF-at-siumed.edu] wrote: } } We have always kept our osmium solutions in glass containers. However, we } are in the process of evaluating our safety procedures and discussed the } idea of increasing the safety of the transport of osmium from the } refrigerator to the fume hood by putting the osmium solution in plastic } containers. (If they are dropped, they would not break and cause the danger } of a spill outside the hood.) } Does anyone have experience with osmium stored in plastic? Any comments } about this particular subject or any of your safety with osmium procedures } are welcomed. } Thanks. } } Donna Wagahoff } SIU School of Medicine } PO Box 19627 } Springfield, IL 62794-9627 } 217-782-0898 } fax217-524-3227
I would not recommend storing osmium in ordinary laboratory plastics containers used on their own. Osmium penetrates polyolefin plastics (PE, PP) to some depth, and reacts with them, so containers made from these plastics or of polystyrene or polycarbonate with polyolefin closures cannot be recommended. Mostly, osmium is supplied in some protective packaging, but if you want to improve security still further I would consider placing the glass ampoules inside a polyethylene or polypropylene tube. That would give considerable shock-protection, and if the ampoule did break would protect against leakage, but only for the short period required to get it to a fume cupboard. The same principle can be applied to osmium solutions prepared in glass bottles - enclosure in an outer polyethylene bottle will give shock-protection and temporary containment. It will also indicate by its blackening how much leakage is occurring from your supposedly closed glass container. Speaking of which, we have a rule that osmium solutions are NEVER stored in the laboratory refrigerator. Why? because even when greatest care is taken, osmium blackening of the fridge walls and other objects in the fridge takes place, indicating that vapour is escaping into the fridge atmosphere. If you must store osmium at low temperatures, I recommend you consider installing a compact refrigerator in your fume hood or in a fan-ventilated cupboard.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } We have always kept our osmium solutions in glass containers. However, we } are in the process of evaluating our safety procedures and discussed the } idea of increasing the safety of the transport of osmium from the } refrigerator to the fume hood by putting the osmium solution in plastic } containers. (If they are dropped, they would not break and cause the danger } of a spill outside the hood.) } Does anyone have experience with osmium stored in plastic? Any comments } about this particular subject or any of your safety with osmium procedures } are welcomed. } Thanks. } } Donna Wagahoff } SIU School of Medicine } PO Box 19627 } Springfield, IL 62794-9627 } 217-782-0898 } fax217-524-3227 }
===================================================================== DR CHRIS JEFFREE BIOSEM - BIOLOGICAL SCIENCES EM FACILITY UNIVERSITY OF EDINBURGH Daniel Rutherford Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JH, Scotland, UK Tel. #44 131 650 5345 FAX. #44 131 650 6563 Mobile 0410 585 401 email c.jeffree-at-ed.ac.uk SEM / TEM bookings sem-at-ed.ac.uk =====================================================================
I just solved (with the help of some of you) my last problem with the carbon extraction replicas but the next specimen causing difficulties is on my desk...
I got a small semiconducor specimen with a layered structure that should be investigated. If I say small I mean it: it has a shape like a cube with a side length of about 250um. And now comes the real difficulty: We need to cut this one precious specimen into several slices in order to allow other investigations with other analytical methods. In fact we have five specimens to try with, but they are not completely identical to the one we have finally to investigate.
We are equipped with everything we need to do a TEM preparation of bulk materials and also of interfaces as long as the specimens are large enough (saw, disk cutter, dimpler, ion etcher, mounting tools, ...) but I don't see how I could be successful with such a small specimen.
Any ideas?
Petra
-------------------------------------------------------------- Dr. Petra Wahlbring Centre de Recherche Public - Gabriel Lippmann Laboratoire d'Analyse des Materiaux (LAM) 162a, av. de la Faiencerie L-1511 Luxembourg tel. +352-466644-402 fax +352-466644-400 e-mail: petra.wahlbring-at-crpgl.lu Visit our WWW site! http://www.crpgl.lu/~wahlbrin
I just solved (with the help of some of you) my last problem with the carbon extraction replicas but the next specimen causing difficulties is on my desk...
I got a small semiconducor specimen with a layered structure that should be investigated. If I say small I mean it: it has a shape like a cube with a side length of about 250um. And now comes the real difficulty: We need to cut this one precious specimen into several slices in order to allow other investigations with other analytical methods. In fact we have five specimens to try with, but they are not completely identical to the one we have finally to investigate.
We are equipped with everything we need to do a TEM preparation of bulk materials and also of interfaces as long as the specimens are large enough (saw, disk cutter, dimpler, ion etcher, mounting tools, ...) but I don't see how I could be successful with such a small specimen.
Any ideas?
Petra
-------------------------------------------------------------- Dr. Petra Wahlbring Centre de Recherche Public - Gabriel Lippmann Laboratoire d'Analyse des Materiaux (LAM) 162a, av. de la Faiencerie L-1511 Luxembourg tel. +352-466644-402 fax +352-466644-400 e-mail: petra.wahlbring-at-crpgl.lu Visit our WWW site! http://www.crpgl.lu/~wahlbrin
I think I can make several suggestions and point you toward several references that may help.
First, can you or have you tried dipping your ion milled (to perforation) samples into your bromine/methanol etch as the final step in your procedure? Yu et al (J.of Elec. Micro.Tech./18/315-324/1991) dipped their perforated samples in 0.5% Br2/Ch3OH for 1or 2 seconds followed by 20 seconds in HF/HCL of volume ratio 1:1. This technique worked well for them.
As you may know, reducing the energy during the final step(s) of ion milling can reduce the severity and/or the number of artifacts produced. If you have access to a mill that allows milling in the 100eV to 1keV range you may want to try this as the final step in your milling procedure.
The following techniques have also been used to prepare type II-VI compound semiconductors:
(1) Reactive ion milling. This can be done 2 different ways. In one method Iodine gas is used instead of Argon. The iodine gas is ionized to form I+, which is then used for milling (Cullis and Chew/ MRS symposium proceedings/ 115/ 3-14/1988). In the second method, Iodine gas is introduced into the Atmosphere of the milling chamber during Ar+ milling (Pecz and Barna /Vacuum/45/1-3/1994). The authors have applied these techniques to type II-VI compound semiconductors of CdTe, ZnS, and ZnSe in addition to other compound semiconductors. CAUTION must be used when introducing iodine gas into your ion mill. The gas is very corrosive and may damage mill parts and vacuum systems. I recommend that you check with the manufacturer of your ion mill before proceeding. (2) Chemomechanical polishing. Sabinina and Gutakovsky (Ultramicroscopy/45/411-415/1992) eliminated the need for ion milling completely by using this technique. They prepare samples of HgCdTe using a bromine-methanol etchant in a technique that is very similar to dimpling using padded tools.
Are you familiar with the Small Angle Cleavage Technique (SACT)? This technique may or may not work for your materials. I am not aware of any references for preparing II-VI materials using SACT but that certainly does not imply that it cant be done. I have used this technique to prepare samples of thin films on silicon substrates. It is quick and relatively easy to learn. The technique was developed by John McCaffrey and a good step by step procedure is given by Walck and McCaffrey (MRS symposium proceedings/480/149-171/1997).
The references listed above are meant to be a starting point and are by no means a complete listing.
Hope this helps.
E. Windsor
Eric Windsor Physical Scientist NIST 100 Bureau Dr. Stop 8371 Gaithersburg, MD 20899-8371 (301) 975-3930 fax: (301) 417-1321 eric.windsor-at-nist.gov
Dorrance McClean originally wrote:
Hi, I've been working on thin foils of CZT and I'm looking for some suggestions to help eliminate artifacts created by ion milling. So far I have polished side one and etched (to remove polishing damage) with bromine-methanol and then dimpled side two, ending with 0.1um alumina. I have been successful obtaining a final sample thickness of about 8 to 10 microns. However, when I put the samples into the PIPs (Gatan) to complete the thinning process I'm seeing beam damage. I was hoping that someone might have a different approach or suggestion that would eliminate the beam damage. Thanks for your help. Dorrance
Soft Imaging Systems has a program called analySIS that has a montage module called MIA. Have a look at their web site at: www.soft-imaging.de
David Rohde NORAN Instruments
-----Original Message----- } From: Rick Harris [mailto:raharris-at-ucdavis.edu] Sent: Thursday, January 06, 2000 6:47 PM To: microscopy-at-sparc5.microscopy.com
Can anyone recommend image montaging software? That is, software that assists you in compositing several images into one larger image. I am familiar with the offerings from GATAN for their Digital Micrograph and QuickStitch from Enroute Software. Also, can anyone offer an evaluation of the QuickStitch software?
TIA
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 530 754 7536 fax http://katie.ucdavis.edu raharris-at-ucdavis.edu
Dear Subscribers, I would be interested in hearing from anyone who has experience in in situ hybridization to mRNA. I am working with immature embryos of Arabidopsis and would like to recieve replies and comments to the following questions:
1. For embedding, Paraffin is mostly used. Why not use LR-White (London Resin)? or BMM (Butylmethylmethacrylate)? or another acrylic material? These hydrophilic materials should be easier to remove, or is it necessary to remove them for mRNA exposure? If it is necessary, what is best for removing them? Perhaps acetone?
2. About the probe itself, obviously RNA is best. Has anyone tried DNA oligos (~20 nucleotides) for less abundant mRNAs?
3. Has anyone used tailed oligos?
4. Is DIG significantly better to use than biotin labelled probes?
Many thanks to anyone who is prepared to take the time to help reveal the unknowns of plant embryogenesis. Maura ____________________ Dr. Maura C. Cannon Dept. Biochemistry & Molecular Biology Lederle Graduate Research Center University of Massachusetts Amherst, MA 01003
Ron Anderson has a technique that might do the trick for you. He took an ultrasonic drill and made a small cavity in a piece of Si at the surface. The drill was solid spherical end. He then epoxied his small sample into the hole. You might want to mount it on another piece of Si first and then put it in the hole. He then used the Tripod Polishing technique to examine it. The benefit is that he has the Si to gauge the thickness of the sample. I though that it was pretty slick.
Your other option and the one most easy to do if you have access to an instrument is to have the sample FIB'd.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies." --
} -----Original Message----- } From: Petra Wahlbring [mailto:wahlbrin-at-crpgl.lu] } Sent: Friday, January 07, 2000 8:17 AM } To: microscopy-at-Sparc5.Microscopy.Com } Subject: Mat: Cutting of small semiconductor specimen? } } } -------------------------------------------------------------- } ---------- } 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 } } } -------------------------------------------------------------- } ---------. } } } } A happy New Year to everybody! } } I just solved (with the help of some of you) my last problem with the } carbon extraction replicas but the next specimen causing } difficulties is on } my desk... } } I got a small semiconducor specimen with a layered structure } that should be } investigated. If I say small I mean it: it has a shape like a } cube with a } side length of about 250um. And now comes the real } difficulty: We need to } cut this one precious specimen into several slices in order } to allow other } investigations with other analytical methods. In fact we have five } specimens to try with, but they are not completely identical } to the one we } have finally to investigate. } } We are equipped with everything we need to do a TEM } preparation of bulk } materials and also of interfaces as long as the specimens are } large enough } (saw, disk cutter, dimpler, ion etcher, mounting tools, ...) } but I don't } see how I could be successful with such a small specimen. } } Any ideas? } } Petra } } } -------------------------------------------------------------- } Dr. Petra Wahlbring } Centre de Recherche Public - Gabriel Lippmann } Laboratoire d'Analyse des Materiaux (LAM) } 162a, av. de la Faiencerie L-1511 Luxembourg } tel. +352-466644-402 fax +352-466644-400 } e-mail: petra.wahlbring-at-crpgl.lu } Visit our WWW site! http://www.crpgl.lu/~wahlbrin }
Every plastic container we have seen turns black and some seals break down. Our current method is similar to Bill Trivol's. We use a 25ml Pyrex media bottle with the orange plastic lid (it does not seem affected). This is placed in a spare metal can (that the ampules of osmium crystals were shipped in) with paper padding. This new bottle (with parafilm around cap) also solved our osmium fumes in the refrig. problem. Everything is small enoug to transport from refrig. to hood.
Speaking of which, we have a rule that osmium solutions are NEVER stored in the laboratory refrigerator. Why? because even when greatest care is taken, osmium blackening of the fridge walls and other objects in the fridge takes place, indicating that vapour is escaping into the fridge atmosphere. If you must store osmium at low temperatures, I recommend you consider installing a compact refrigerator in your fume hood or in a fan-ventilated cupboard.
With all due respect Chris for over 20 years we have used our lab refrigerator to store Osmium with no blackening of the plastic walls. A 25ml 4% solution of Osmium is put into a 60 ml glass stopper bottle with the stopper tightly wrapped in parafilm. Next, that bottle is placed in a waxed 6" long x 3" diameter screw capped cardboard mailing tube. We prepare the waxed tubes by pouring hot paraffin inside and rotated quickly and evenly until the entire surface area is well coated. We also coat the inside of the metal screw cap but not the outside of the tube. Mailing tubes are available from the local shipping supply house and our waxed containers have lasted decades. Regards Jim
James S. Romanow The University of Connecticut Physiology and Neurobiology Department Electron Microscopy Facility U-2131 Storrs, CT 06269-2131 bsgphy3-at-uconnvm.uconn.edu 860 486-2914 voice 860 486-1936 fax
We have been using teflon bottles for many yeas with success. We got them from Fisher Sci
At 09:30 AM 1/7/00 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Gregory W. Erdos, Ph.D. Ph. 352-392-1295 Assistant Director, Biotechnology Program PO Box 110580 Fax: 352-846-0251 University of Florida Gainesville, FL 32611
} We have always kept our osmium solutions in glass containers. However, we } are in the process of evaluating our safety procedures and discussed the } idea of increasing the safety of the transport of osmium from the } refrigerator to the fume hood by putting the osmium solution in plastic } containers. (If they are dropped, they would not break and cause the danger } of a spill outside the hood.) } Does anyone have experience with osmium stored in plastic? Any comments } about this particular subject or any of your safety with osmium procedures } are welcomed.
Dear Donna,
In my point of view polypropylene and polyethylene are compatible with osmium tetroxide solutions. Only problem - to hold that nasty stuff inside the vial. Some plastics may partially be penetrable by osmium tetroxide.
I had some limited experience to store aqueous osmium tetroxide solutions in the polypropylene (?) NUNC 2 or 5 ml cryo-vials. I aliquot solution into that vials and store it at -40oC. For extra protection I stored cryo-vials in the 15 ml cell-culture tubes. Cryo-vial's seal may penetrate by osmium tetroxide vapors a little bit but second tube provides complete protection. 2-4% aqueous osmium tetroxide solutions (no buffer, please) are stable for at least 1-2 month in the dark at -20 - 40oC in that plastic containers.
Good luck.
Sergey.
_________________________________
Sergey Ryazantsev Ph. D. UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
There is a researcher at our Med. School who wants to do SEM on biofilms
on ear biopsies. He says the biopsies will be approximately 1mmX1mm. I proposed a standard glut.-osmium fix, Et-oh dehydration, CPD, mount, sput coat protocol. Is this sufficient? Do we need to affix the biopsies
to a substrate first? I would appreciate any suggestions. If anybody has reprints that contain a good protocol, I would certainly appreciate getting a copy. My address is in the footer and my FAX is 405-325-7619. Thanks for your help.
Bill Chissoe
-- ============================================================= Bill Chissoe III Electron Microscopist University of Oklahoma 770 Van Vleet Oval Norman, Ok. 73019 E-mail: wchiss-at-ou.edu Ph. (405)325-4391 =============================================================
A user of our facility wishes to do some special vacuum evaporation processes. He is looking for ceramic coated tungsten boats for the purpose, but has not been able to locate a source. Any help would be appreciated.
Replies offline would be welcome. Please reply to the list only if you have generally useful information.
Thanks,
John Chandler Colorado State University chandler-at-lamar.colostate.edu
For years, I have been using my diode GW BSE system with high beam currents and high gain (contrast) to yield electron channeling contrast on polished specimens.
FWIW: Contrary to some reports I have read, lower kV works better than higher. Since it is imperative to use high beam currents at the required resolution, I wonder if difficulty in achieving sufficient current at lower potentials may have been influencing opinions.
I have also been a very good customer for GW Electronics. When the detector is new, I generally have no problem, but as it ages, the same operating conditions will produce artifacts. A new detector every year or so is a bit expensive, but fixes the problem.
I now have a new detector (and SEM :) which produces this artifact and am now trying to find the reason.
The artifact can be described as video brightness overshoot from black to white when the black is strongly saturated. It occurs when the (very high gain) BSE signal goes from saturated black to some median gray. A low-Z inclusion or deep pore in the field of view will produce this artifact. When the beam leaves the inclusion/pore, it overshoots to white then settles back to the appropriate level. The effect is a black pore with a white "comet tail" pointing in the sweep direction. This problem does not manifest itself when using less extreme operating conditions.
I certainly cannot exclude the possibility that the detector is not the direct cause. The amount of signal gain (contrast) is not calibrated (nor have I paid much attention to the position of the adjustment) . If the detector simply loses some sensitivity, I would make up the difference in amplifier gain and not realize I was not at exactly the same operating conditions. ...Thus, the artifact could be amplifier gain related rather than simply a detector problem.
Paul Rennie wrote: =============================================== Having just purchased a new SEM, our old instrument, a Cambridge S200 is destined to be sent to one of our companies in Cuidad Victoria, Mexico (Although I'm trying to persuade management that it's not worth it). Although we will probably be installing the instrument ourselves, I would welcome details of any service companies in Mexico. Additionally, who sells consumables within Mexico, or is it best to purchase over the border in the USA. ================================================ With regard to the method of purchasing consumables from within Mexico, the situation is entirely analogous to that that exists in the UK vis a vis purchasing items from the USA directly or via a distributor in the UK. It is a matter of institutional policy and also, personal preference.
The main manufacturers of consumables all have local distributors in Mexico and those firms can be found on the websites of those respective firms. On the other hand, with the implimentation of NAFTA, making a shipment to a point in Mexico from the USA is not really all that different from making a shipment to some other point in the USA. So again it is a matter of institutional policy and personal preference.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.spi.cc ######################## ============================================
I have been trying for some time to find any information about a Vickers Instrument Company microscope. I had never heard of the Company but the scope looked interesting. I have searched hundreds of websites, posted messages on newsgroups and also on Microscopy, UK. So far I've had only two responses. The sum total of what I have learned is that the company was in York and that they closed in 1989. Cooke, Troughton and Simms were taken over by Vickers
The rather lengthy description of the scope follows:
Head: Binocular configuration, adjustable for interpupillary distance and dioptric differences. Interpupillary adjustment range, 50 to 72mm. Eyepieces(2): 10X, DIN standard. Objectives(4): Vickers microplan, Planachromatic objectives. Include 5X (0.15 N.A.), 10X (0.25 N.A.), 20X (0.50 N.A. spring loaded), 40X (0.65 N.A. spring-loaded), DIN standard objectives. All are parfocal, parcentered, and coated to resist reflection. Stage: Precision-machined mechanical stage with oversized, low-position, coaxial control knobs. Chemical-resistant finish with glass insert. This stage is exceptionally smooth in movement with a 3.25 x 2.75-inch range of motion. Focus Control: Low-position, coaxial, coarse/fine controls. . Condenser: Abbe condenser, fitted with an iris diaphragm. Illumination: Diascopic lower illumination features variable-intensity quartz halogen lamp ( 12V, 100 watt ) with metered solid-state control as well as field iris, condenser and centering adjustments. Episcopic illumination (30-watt lamp) is also of variable intensity via an independent control on front of the microscope base. Upper illuminator housing fitted with iris and condenser controls. As shown in the photos, the microscope can be separated from the 100-watt illuminator base Finish chemical-resistant paint with "hammertone" finish.
I have included some pictures that I have posted to help with the identification:
I am trying to find any information about the company and about the scope itself. I understand that this is rather difficult (impossible?) because of the practice of the company not to use serial numbers. I would especially like to be able to aquire a copy of the owners manual and a copy of the Vickers catalog.
When I began to attempt to collect information about the Vickers I never guessed that I would run into a blank wall. If you could assist me in any way at all it would be greatly appreciated.
I am looking for a simple method to indicate the presence of lipids /oils in plant tissue. Specifically palm fruit tissues. Ideally, if there is a procedure that I can used on fruits fixed in FAA that would be the best. I have tried Sudan Black B without success. Thank you in advance for your ideas or suggestions. Please reply via email to : mchapin-at-ntbg.org
The item you are looking for should be available from R.D. Matis. The following contact information is from www.vacuum.org. There are several vendor listed in the subsection for filaments if you care to browse. I have no financial interest in R.D. Matis.
R.D. Mathis Company 2840 Gundry Avenue Long Beach, CA 90806 Phone: 562-426-7049 FAX: 562-595-0907
Description
Specializes in the manufacture of hi-vacuum evaporation sources. We offer a comprehensive selection of tungsten, molybdenum and tantalum sources as well as custom fabrication to meet your specific needs. Display will be a variety of evaporation sources along with one of our "LV Series" low voltage high current power supplies and our "GP 100" inert gas purifier to compliment your evaporation process.
Product Categories Filaments
Good luck
Jim Campbell
James Campbell 36 Van Drive Bordentown, NJ 08505 Tel: 609-298-9206 Fax: 609-278-6969 e-mail campbell36-at-aol.com
In a message dated 1/7/00 9:56:09 PM Eastern Standard Time, chandler-at-lamar.ColoState.EDU writes:
} Subj: Vac Evap: Special needs request } Date: 1/7/00 9:56:09 PM Eastern Standard Time } From: chandler-at-lamar.ColoState.EDU (John Chandler) } To: Microscopy-at-Sparc5.Microscopy.Com } } ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } A user of our facility wishes to do some special vacuum evaporation } processes. He is looking for ceramic coated tungsten boats for the } purpose, but has not been able to locate a source. Any help would be } appreciated. } } Replies offline would be welcome. Please reply to the list only if you } have generally useful information. } } Thanks, } } John Chandler } Colorado State University } chandler-at-lamar.colostate.edu
James Campbell 36 Van Drive Bordentown, NJ 08505 Tel: 609-298-9206 Fax: 609-278-6969 e-mail campbell36-at-aol.com
Dana: This may not help, but I have a book I purchased about 1970 that was published by Vickers entitled: The Polarizing Microscope by A.F. Hallimond. It is an excellent work and has many pictures of the Vickers line of polarizing scopes and accessories from the late 60s. They did buy out Cooke, Troughton and Simms and marketed a number of innovative and competitively priced (cheaper than Leitz and Zeiss) scopes. Dr Hallimond was one of their consultants on design. I am sure you can find the book in a large University Geology department library ( I hope). Or try interlibrary loan. It is still an excellent and definitive reference for polarizing light microscopes and their use.
Michael L. Boucher Sr. mboucher-at-isd.net http://www.isd.net/mboucher
I am trying to quantify the alloy amount of Al/Si. The standard alloy ratio is 1-3 wt % Si/Al. However, with Z of each only 1 unit apart, and low Z at that, EDAX and AES cannot detect the Si. My next attempt is to try dynamic SIMS and then time of flight SIMS.
The specimen is a microcircuit die. I am analyzing the bonding pad metalization.
Has anyone done this sort of thing before and had success? If so, how did you do it?
Vickers was a well known British manufacturer of microscopes. I have had the privilege of using some of their more interesting measuring equipment. Several contacts come to mind, most of them from the UK: Clive Cowan, Micro Instruments in Oxford (UK) 199-388-9616 and Gerard Turner, who is a curator in microscopy at the Museum of Science at Oxford University, Dr. Savile Bradbury, retired from Oxford (but could probably be reached by letter there; also, if you are interested, I can probably get a more recent address), and Dr. Cecile Fox at Molecular Histology in Gaithersburg (301-216-1564). Cecile put together major microscopy exhibits for both the Smithsonian and the Am. Mus. of Natural History in the mid- 1980's. Please give my regards to them (Gerard may only remember me as an RMS student from the distant past) and best of luck on your search.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
At 12:41 PM 1/8/00 EST, Lugosi1936-at-aol.com"-at-Sparc5.Microscopy.Com wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Do you have an evaporator in the lab? Simultaneous evaporation of Pt (wire) and carbon gives a very fine and permanent coating. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Thursday, January 06, 2000 2:15 AM, Tindall, Randy D. [SMTP:TindallR-at-missouri.edu] wrote: } } } Hi, } } Due to some recent high-resolution requirements in our lab, I find myself } having to go back to Sputter Coating 101 (after years of just putting } specimens in the coater and turning it on without a second thought!). We } find ourselves in need of very thin coatings with as little structure as } possible, in order to image samples down in the nanometer range. } } We have a chromium coater, but often need to revisit samples days or weeks } after the initial coating. The oxidation problem rears its ugly head. We } intend to purchase a platinum target, but don't yet have one, so we're } experimenting with our venerable Au/Pd coater. } } My questions are: } } 1) I seem to remember a string on this listserver suggesting that lower } deposition currents yield finer coating structure. Is this right? Does a } low deposition current for a longer time yield a finer coating than a higher } current for a shorter time? (I'm running some tests to check this, but } would be very interested in others' experiences, too.) } } 2) Deposition current can be controlled by the initial current setting } (i.e., the knob on the machine) and by the argon flow through the chamber. } Is there any difference in the coating when adjusting the deposition current } by either of these two ways? } } 3) Charts I have seen indicate that deposition current is directly } proportional to coating rate. Is the same true for coating time? I.e., is a } one minute coating twice as thick as a 30 sec. coating? It would seem so } intuitively, but you know what they say about the word "assume". } } My apologies if these are very basic questions, but, like I said, back to SC } 101! } } I'll be happy to summarize the responses for anyone who is interested. } } Thanks! } } Randy } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility } W122 Veterinary Medicine Bldg. } University of Missouri } Columbia, MO 65211 } (573)882-8304 } tindallr-at-missouri.edu } http://www.biotech.missouri.edu/emc/ }
I'm not 100% sure but I thought that the Microscience Division of Bio-Rad bought Vickers some years ago. You might want to give them a shot.
best regards, Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chapel Hill, NC 27599 Phone office 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
Don't go around saying the world owes you a living; the world owes you nothing; it was here first. Mark Twain [Samuel Langhornne Clemens] (1835-1910)
-- Begin original message -- } -----------------------------------------------------------------------. } } } } I have been trying for some time to find any information about a Vickers } Instrument Company microscope. I had never heard of the Company but the scope } looked interesting. I have searched hundreds of websites, posted messages on } newsgroups and also on Microscopy, UK. So far I've had only two responses. } The sum total of what I have learned is that the company was in York and that } they closed in 1989. Cooke, Troughton and Simms were taken over by Vickers } } The rather lengthy description of the scope follows: } } Head: Binocular configuration, adjustable for interpupillary distance and } dioptric differences. Interpupillary adjustment range, 50 to 72mm. } Eyepieces(2): 10X, DIN standard. Objectives(4): Vickers microplan, } Planachromatic objectives. Include 5X (0.15 N.A.), 10X (0.25 N.A.), 20X (0.50 } N.A. spring loaded), 40X (0.65 N.A. spring-loaded), DIN standard objectives. } All are parfocal, parcentered, and coated to resist reflection. Stage: } Precision-machined mechanical stage with oversized, low-position, coaxial } control knobs. Chemical-resistant finish with glass insert. This stage is } exceptionally smooth in movement with a 3.25 x 2.75-inch range of motion. } Focus Control: Low-position, coaxial, coarse/fine controls. . Condenser: Abbe } condenser, fitted with an iris diaphragm. Illumination: Diascopic lower } illumination features variable-intensity quartz halogen lamp ( 12V, 100 watt } ) with metered solid-state control as well as field iris, condenser and } centering adjustments. Episcopic illumination (30-watt lamp) is also of } variable intensity via an independent control on front of the microscope } base. Upper illuminator housing fitted with iris and condenser controls. As } shown in the photos, the microscope can be separated from the 100-watt } illuminator base Finish chemical-resistant paint with "hammertone" finish. } } I have included some pictures that I have posted to help with the } identification: } } {A HREF="http://members.aol.com/lugosi1936/vic1a.jpg"} http://members.aol.com/l } ugosi1936/vic1a.jpg {/A} } {A HREF="http://members.aol.com/lugosi1936/vic2.jpg"} http://members.aol.com/lu } gosi1936/vic2.jpg {/A} } {A HREF="http://members.aol.com/lugosi1936/vic3.jpg"} http://members.aol.com/lu } gosi1936/vic3.jpg {/A} } } I am trying to find any information about the company and about the scope } itself. I understand that this is rather difficult (impossible?) because of } the practice of the company not to use serial numbers. I would especially } like to be able to aquire a copy of the owners manual and a copy of the } Vickers catalog. } } When I began to attempt to collect information about the Vickers I never } guessed that I would run into a blank wall. If you could assist me in any way } at all it would be greatly appreciated. } } Best regards, } Dana } }
first, I would like to wish you all the best for the new year. Now my question: Does anyone know the shelf-life/usability of LR-Gold stored in a fridge? Is it that delicate as LR-White (Meanwhile I don«t use LR-W older than half a year). My bottle which was not opened many times, could be roundabout three years old.
Thanks a lot, Michael
Michael Reiner Department of Anatomy I University of Cologne Germany michael.reiner-at-smail.uni-koeln.de
Please send me (if possible) a brief explanation of gamma correction in image analysis and a good reference source (book, papers, etc) to get the details... Thanks
Dear Randy, There was a good series of articles in the Scanning Electron Microscopy, 1980, volume I (pp. 143--218), that examined a number of thin-film deposition methods and measured the films for feature size, mostly using TEM. They found that lower kV made for finer films, using Mo, W or Ta made a more featureless coating, Pt was a little finer than Au/Pd and that lowering the temperature of the substrate also made the films finer. In my own experience I found that, on smoother specimens, even a two second coating would sometimes be enough to stop charging. Try very short times and turn the specimen and coat again for a very short time, if the first one isn't enough. I used 700V, if your coater can change voltage. If you can find the Scanning Electron Microscopy articles, they have other good suggestions. At 10:15 AM 1/5/00 -0600, you wrote:
} Hi, } } Due to some recent high-resolution requirements in our lab, I find myself } having to go back to Sputter Coating 101 (after years of just putting } specimens in the coater and turning it on without a second thought!). We } find ourselves in need of very thin coatings with as little structure as } possible, in order to image samples down in the nanometer range. } } We have a chromium coater, but often need to revisit samples days or weeks } after the initial coating. The oxidation problem rears its ugly head. We } intend to purchase a platinum target, but don't yet have one, so we're } experimenting with our venerable Au/Pd coater. } } My questions are: } } 1) I seem to remember a string on this listserver suggesting that lower } deposition currents yield finer coating structure. Is this right? Does a } low deposition current for a longer time yield a finer coating than a higher } current for a shorter time? (I'm running some tests to check this, but } would be very interested in others' experiences, too.) } } 2) Deposition current can be controlled by the initial current setting } (i.e., the knob on the machine) and by the argon flow through the chamber. } Is there any difference in the coating when adjusting the deposition current } by either of these two ways? } } 3) Charts I have seen indicate that deposition current is directly } proportional to coating rate. Is the same true for coating time? I.e., is a } one minute coating twice as thick as a 30 sec. coating? It would seem so } intuitively, but you know what they say about the word "assume". } } My apologies if these are very basic questions, but, like I said, back to SC } 101! } } I'll be happy to summarize the responses for anyone who is interested. } } Thanks! } } Randy } 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-interchg.ubc.ca
Is anyone willing to do some contract cryoSEM or TEM in the Chicagoland area? I have a primary emulsion with particle size less than 1.0 micrometers. 2 samples. Please contact me via the server. Joanne Crudele-Unilever-Rolling Meadows Il. Give a price estimate per sample.
You are corrrect in that the time constant of diode BSE detectors is relatively slow. Unfortunatly, I am already sweeping about as slowly as possible to minimize the effect and help the signal to noise ratio.
Woody, Is the problem independent of scan speed? Slower scan speeds often overcome distortion using solid state BEI detectors.
At 05:41 PM 1/7/00 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
id AA30574; Mon, 10 Jan 2000 19:01:49 +0800 Message-Id: {200001101101.AA30574-at-aphy.iphy.ac.cn}
Hi, Microscopist,
International Kunming Symposium on Microscopy (IKSM) will be held on July 2-5, 2000, in Kunming, one of the most attractive tourist destinations in China.
Call-for-paper circular and pre-registration form for International Kunming Symposium on Microscopy (IKSM) are available on request by email. For more information, pls visit http://www.iphy.ac.cn/microsc/IKSM.html .
Rick Felten-at-MACDERMID 01/10/2000 04:20 PM Whenever I wanted to merge images I used corel draw 8. I inserted the images where I wanted them and exported the entire image as a tiff. Seem to work ok. Not sure if it is the most efficient way. In fac, it is the only thing that I liked about corel draw over MS publisher.
To the Board, We have a problem with spotty,black,amorphous artifacts on our SEM samples visible at 200X and up. The problem seems to arise only with one sample type, epoxy resin casts made with amine-blush resistant hardener. We use professional dental impression molds,a two part process, base and catalyst to make the flexible mold (President, polyvinylsiloxane). Then Araldite 506 epoxy with HY 356 hardener is poured and cured at room temp overnight. Sputter coating glass slides with gold palladium does not seem to cause the artifact, but another question, is gold palladium more susceptible to oxide formation than pure gold, and do oxides look like the above described artifact. Also, etching the sample does not help. Any suggesstions will be greatly appreciated. Thanks.
We store Os in a glass bottle with Parafilm around the top, inside a larger plastic jar with a corn oil-soaked paper towel taped to the inside of the lid, and then the outer container is Parafilmed around the top. No black frig. (Change the oil-soaked towel periodically.)
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
I use Denton spatter coater with Au/Pd target for magnifications up to 100,000 without any problems. I can observe collagen striations and other fine details and do not see coating structure. Parameters for coating: current 15 ma, time 5-10 sec. Some charging can occur for samples with "multilayer" surface, but it is tough case for almost any coating anyway. I have also magnetron Cr coater, but do not use it often because it's much more time consuming.
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} -----Original Message----- } From: Tindall, Randy D. [mailto:TindallR-at-missouri.edu] } Sent: Wednesday, January 05, 2000 10:15 AM } To: 'microscopy-at-sparc5.microscopy.com' } Subject: SEM: Sputter coating } } } -------------------------------------------------------------- } ---------- } 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, } } Due to some recent high-resolution requirements in our lab, I } find myself } having to go back to Sputter Coating 101 (after years of just putting } specimens in the coater and turning it on without a second } thought!). We } find ourselves in need of very thin coatings with as little } structure as } possible, in order to image samples down in the nanometer range. } } We have a chromium coater, but often need to revisit samples } days or weeks } after the initial coating. The oxidation problem rears its } ugly head. We } intend to purchase a platinum target, but don't yet have one, so we're } experimenting with our venerable Au/Pd coater. } } My questions are: } } 1) I seem to remember a string on this listserver suggesting } that lower } deposition currents yield finer coating structure. Is this } right? Does a } low deposition current for a longer time yield a finer } coating than a higher } current for a shorter time? (I'm running some tests to check } this, but } would be very interested in others' experiences, too.) } } 2) Deposition current can be controlled by the initial current setting } (i.e., the knob on the machine) and by the argon flow through } the chamber. } Is there any difference in the coating when adjusting the } deposition current } by either of these two ways? } } 3) Charts I have seen indicate that deposition current is directly } proportional to coating rate. Is the same true for coating } time? I.e., is a } one minute coating twice as thick as a 30 sec. coating? It } would seem so } intuitively, but you know what they say about the word "assume". } } My apologies if these are very basic questions, but, like I } said, back to SC } 101! } } I'll be happy to summarize the responses for anyone who is interested. } } Thanks! } } Randy } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility } W122 Veterinary Medicine Bldg. } University of Missouri } Columbia, MO 65211 } (573)882-8304 } tindallr-at-missouri.edu } http://www.biotech.missouri.edu/emc/ } }
Posted at author's request to remain anonymous....
} To: zaluzec-at-Sparc5.Microscopy.Com } Date: Mon, 10 Jan 2000 01:20:59 +0530 } Subject: Nestor, would you post this for me? } } Osmium Vapor- A Safety Question } } The last time I mentioned a safety concern about the rumor of an possible } "explosive" warning about some Ruthenium compound , I got "Battered" by } certain vendors(for quite a while too I might add..certainly didn't help } my previously supportive, pro-vendor position-since I used to be one!) . } I was warned and pestered frequently and asked to retract any suggestion } of such a possibility, but I refused. So here we go again.... } } On approximately January 6, 2000 I noticed a request under the } title: } } : Donna Wagahoff {DWAGAHOFF-at-siumed.edu} } } "....Does anyone have experience with osmium stored in } plastic?..." } } and the only responses on the list server were: } } " How about putting the glass containers inside plastic containers? } There are also padded and styrofoam containers which you could use for } the transportation step." } } and } } ......" I store this glass bottle inside an aluminum-foiled plastic } container in my fume hood at room } temp. The clear plastic of this outer container gets translucent black } within weeks no matter how carefully I seal the glass bottle"....... } } ( I'm sure these were very fine suggestions..but..) } } This concerns me for several reasons: } } 1. I believe there is a considerably more dangerous situation here than } many of us may realize, or..will discuss. We must check, and we may find } that Osmium and Ruthenium compounds(and others?) may penetrate all } plastics and we may only see this at a macroscopic level long after the } heavy metal compounds or there derivatives have penetrated many layers } and possible contaminated at some lower and less visible level( but } undetermined danger level that may be a health risk) a much larger area. } Many of us may have seen the effect in EM refrigerators as the interior } blackens over a long period of storage of Osmium. } } 2. There doesn't seem to be an acceptable safe level of these compounds } or known affects, but probably fixation or contamination and loss of } function at some level occurs(microscopic?). } } 3.Also venting these vapors through hoods should be scrubbed or } absorbed(?), reduced,etc.( "I'm no chemist") and the level of the output } probably should be carefully and technically monitored. } } 4. The safety level must be obvious to those who receive bottles of } solutions in sealed glass(or Pirex?) and the factories that manufactor } them.(What glass type materials are safe?) } } Does anyone know of the level of danger(or safety) or know of some good } research sites or periodicals that have dealt with these issues? Any } personal experience, or are we all afraid of retribution and retaliation? } The MSDS's seem a little less than thorough to me. I'm sure I'm again not } alone here in this concern and only wish to continue this difficult and } frequently dangerous craft(EM) with more care for myself , my associates } and my community. } } Anonymous (chicken....) } (...and Vendors and manufacturers are our backbone and our support! } Thank you all!) } } } }
Does anyone have an idea of a good negative film scanner to use for TEM negatives? I am trying to eliminate the darkroom printing process. These are the stipulations: 1. Must be able to scan 3 1/4 x 4 inch negatives. (Kodak 4489 film) 2. Must be compatible to a PC. 3. Need to know what software is needed. 4. Must be priced under $3,000.00 5. Must provided the best quality resolution for diagnostic results.
It is our experience, at South Bay Technology, that Cr films deposited by ion beam sputtering remain conductive for a short time. The amount of time depends on the film thickness and sample surface topography. Because Cr deposition requires a water vapor free environment, usually not possible in a sputter coater, you may have more success with Ir target. Under 200Kx, ion beam deposited Ir (better than Pt since Ir films minimize beam damage) does not display any artifacts (grains cannot be seen) or specimen damage. We recommend using Cr (or Ti, W or Ta) only when looking at mag } 200Kx.
An Ir target in your sputter coater may work well and an Ir target in your "Cr coater" may solve the oxidation problem more effectively.
Some advantages of Ion Beam Sputtering: - Controlled thickness on angstrom level since the average deposition rates are 10A/min - Precise thickness measurements reported by quartz thickness monitor as result of low energy sputterant energy striking crystal - No damage or artifacts as a result of 30ev sputterant energy - Any material can be deposited although Cr is suggested for } 200Kx mags, Ir for {200Kx - C like metal films are amorphous. C ddoes not display grains or create damage - X-ray production from 10A films is lost in the noise - Image improvement results from increased signal to noise as well as conductivity
Disclosure: South Bay Technology is the manufacturer of the IBS/E Ion Beam Sputtering and Etching System and therefore has a vested interested interest in promoting it use.
Regards, Mike Mizell
************************************************************************* Michael K. Mizell Tele: 949-492-2600 VP Sales & Marketing Fax: 949-492-1499 South Bay Technology 1120 Via Callejon mizell-at-southbaytech.com San Clemente, Ca 92673 USA ************************************************************************** South Bay Technology is an American manufacturer of precision sample preparation equipment and supplies for metallography crystallography and electron microscopy.
} } } } } } } Please visit us at http://www.southbaytech.com { { { { { { { {
I would like to know what is a nomarski disc. What it is used for? Is there any difference between this and the routine phase contrast condenser we use in light microscopy. Regards, M. Angou SUMKA SONS INSTRUMENTATION1137-B, THADAGAM ROAD,R S PURAM, COIMBATOREINDIA - 641002 FAX: 91-422-473227TEL:91-422-474378 URL:www.sumka.com
first, I would like to wish you all the best for the new year. Now my question: Does anyone know the shelf-life/usability of LR-Gold stored in a fridge?
Is it that delicate as LR-White (Meanwhile I don«t use LR-W older than half a year). My bottle which was not opened many times, could be roundabout three years old.
Thanks a lot, Michael
Michael Reiner Department of Anatomy I University of Cologne Germany michael.reiner-at-smail.uni-koeln.de
We scan our 4489 negs with a UMAX PowerLook III regularly on a Mac G3. The UMAX is compatable with PCs too, cost about 1100 dollars last summer with the transparancy adaptor and has 1200x2400 dpi actual resolution. We had to modify the 4x5" transparancy holder (easy) to fit the smaller EM negs. UMAX tech help could improve but the scanner works well. Stand alone or PhotoShop plug-in software. Connect via SCSI.
Other units to check out are the Agfa DuoScan T2500 and Microtek ScanMaker 5.
The major electronic trade show meetings are soon so all of the new 'improved' equipment will be ordered and available for summer/fall 2000. The remaining stock of last year's discontinued models will be discount priced by Spring. Good Luck
} Date: Mon, 10 Jan 2000 18:44:54 -0600 } To: Microscopy-at-Sparc5.Microscopy.Com } From: "ELMA Cortinas" {ECORTI-at-childmed.dallas.tx.us} } Subject: Negative film scanner }
} } Does anyone have an idea of a good negative film scanner to use for } TEM negatives? I am trying to eliminate the darkroom printing } process. These are the stipulations: } 1. Must be able to scan 3 1/4 x 4 inch negatives. (Kodak 4489 } film) } 2. Must be compatible to a PC. } 3. Need to know what software is needed. } 4. Must be priced under $3,000.00 } 5. Must provided the best quality resolution for diagnostic } results. } } Thanks in advance! }
James S. Romanow The University of Connecticut Physiology and Neurobiology Department Electron Microscopy Facility U-2131 Storrs, CT 06269-2131 bsgphy3-at-uconnvm.uconn.edu 860 486-2914 voice 860 486-1936 fax
We are using an Epson Expression 800 and have been very happy with it. We routinely scan in at 600 dpi, which has been more than enough for publication quality, in our experience. The unit is capable of higher resolutions than that. I don't remember what we paid, but it was considerably less than $3000. It came with a transparency adapter and all necessary software, including SilverFast, text recognition software, calibration software, etc. Ours works through Adobe Photoshop's Import functions, but may be usable in other programs, too.
Best wishes, Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine Bldg. University of Missouri Columbia, MO 65211 (573)882-8304 tindallr-at-missouri.edu http://www.biotech.missouri.edu/emc/
-----Original Message----- } From: ELMA Cortinas [mailto:ECORTI-at-childmed.dallas.tx.us] Sent: Monday, January 10, 2000 6:45 PM To: Microscopy-at-Sparc5.Microscopy.Com
Does anyone have an idea of a good negative film scanner to use for TEM negatives? I am trying to eliminate the darkroom printing process. These are the stipulations: 1. Must be able to scan 3 1/4 x 4 inch negatives. (Kodak 4489 film) 2. Must be compatible to a PC. 3. Need to know what software is needed. 4. Must be priced under $3,000.00 5. Must provided the best quality resolution for diagnostic results.
{fontfamily} {param} Times_New_Roman {/param} {smaller} Postdoctoral position available immediately to study the 3-D structure of insect flight muscle. The research project, recently renewed for a 4 year period, involves several experimental and theoretical approaches to studying crossbridge structure in different states. Approaches include electron microscope tomography, alignment and classification of 3-D crossbridge structures (see recent publication (Winkler & Taylor, Ultramicroscopy 77:141-152 (1999) for details) and fitting of atomic coordinates of actin and myosin S1 into the envelope of the 3-D images. Emphasis is on the study of quick-frozen, contracting muscle, freeze-substituted for thin section electron microscopy and 3-D image reconstruction. We use stretch activated muscle as well as an isometrically contracting state dubbed stretch activation. Specimens are mechanically monitored right up the point of freezing to facilitate the interpretation of the structures in terms of muscle force and stiffness. Parallel X-ray diffraction experiments make for thoroughly characterized specimens. Please see recent publication (Taylor et al., Cell 99:421-431 (1999)) for current status of this project. This project is a collaboration with Michael and Mary Reedy (Duke Univ. Med. Center), Yale Goldman and Clara Franzini-Armstrong (U. Penn. Med. School) and Richard Tregear (MRC, Cambridge, UK). Successful applicants can work on any of several aspects of the problem of identifying structural features and relating them to the generation of tension in muscle. The project has evolved to the point that most of the effort needs to be put on classification and averaging, model building, model refinement and interpretation of X-ray data. An individual with experience in either image reconstruction or protein structure and function who is interested in gaining further experience in the interpretation and correlation of diverse experimental data on a topical biophysical problem would be ideal for this position. The experimental emphasis of correlating high resolution structures with lower resolution EM data is expected to become a growth area for structural biology. Our laboratory is equipped with Silicon Graphics workstations, one of which is dedicated to this position, a cluster of 3 DEC Alpha compute servers, a Perkin-Elmer PDS 1010M microdensitometer and a Philips CM300-FEG electron microscope. Salary is commensurate with relevant experience. Successful candidates will join a strong Program in Structural Biology with 4 groups using primarily X-ray diffraction, 3 using NMR, 2 using EPR and one using EM. The Program enjoys close ties with the National High Field Magnetic Laboratory and the Supercomputer Computation Research Institute on campus. Additional information about the Structural Biology Program can be found at http://www.sb.fsu.edu/. Interested applicants should send their CV and names, addresses and phone numbers of 3 references to Dr. Kenneth A. Taylor, Institute of Molecular Biophysics, Florida State University, Tallahassee, FL 32306, USA. E-mail address is taylor-at-bio.fsu.edu. Phone number 1-850-644-3357, FAX 1-850-561-1406.
The Institute of Molecular Biophysics and Florida State University are located in Tallahassee, the capitol of Florida. The city has a population of ~200,000. The city is surrounded by rolling hills and pine forests and is 35 miles from pristine beaches on the coast of Gulf of Mexico. Tallahassee residents enjoy many cultural and sporting events.
Posted at author's request to remain anonymous....
Dear Anon,
} Osmium Vapor- A Safety Question } } The last time I mentioned a safety concern about the rumor of an possible } "explosive" warning about some Ruthenium compound , I got "Battered"... } The EM Safety Handbook also warns of the risk of explosion from RuO4.
} On approximately January 6, 2000 I noticed a request under the } title: } } : Donna Wagahoff {DWAGAHOFF-at-siumed.edu} } } "....Does anyone have experience with osmium stored in } plastic?..." } } and the only responses on the list server were: } } " How about putting the glass containers inside plastic containers? } There are also padded and styrofoam containers which you could use for } the transportation step." } } and } } ......" I store this glass bottle inside an aluminum-foiled plastic } container in my fume hood at room } temp. The clear plastic of this outer container gets translucent black } within weeks no matter how carefully I seal the glass bottle"....... } } ( I'm sure these were very fine suggestions..but..) } } This concerns me for several reasons: } } 1. I believe there is a considerably more dangerous situation here than } many of us may realize, or..will discuss. We must check, and we may find } that Osmium and Ruthenium compounds(and others?) may penetrate all } plastics... } This is very likely.
} 2. There doesn't seem to be an acceptable safe level of these compounds } or known affects, but probably fixation or contamination and loss of } function at some level occurs(microscopic?). } The EM Safety Handbook gives a TLV--threshold limit value, defined as a concentration "to which *it is believed* (emphasis is mine) *nearly* all workers may be repeatedly exposed day after day without adverse effects."--of 2parts in 10^10. Note the very small value and the cautionary wording.
} 3.Also venting these vapors through hoods should be scrubbed or } absorbed(?), reduced,etc.( "I'm no chemist") and the level of the output } probably should be carefully and technically monitored. } Since OsO4 is a powerful oxidant, and since it reacts with unsaturated lipids, I had heard the recommendation that corn oil be used to clean up spills; however, the EM Safety Handbook recommends ascorbate powder "as it reacts quickly", and, come to think of it, is more suited for treating an aqueous solution than is an immiscible oil.
} 4. The safety level must be obvious to those who receive bottles of } solutions in sealed glass(or Pirex?) and the factories that manufactor } them.(What glass type materials are safe?) } I doubt that any particular glass is less safe (but would not be surprised to be corrected). Any glass which doesn't oxidize shouldn't react with OsO4.
} Does anyone know of the level of danger(or safety) or know of some good } research sites or periodicals that have dealt with these issues? Any } personal experience, or are we all afraid of retribution and retaliation?
The extensively-quoted EM Safety Handbook is a good source.
} The MSDS's seem a little less than thorough to me.
Although, the existance of an MSDS for di-hydrogen oxide (flamed here some time ago) would seem to argue otherwise. There is further info available from company web sites and phone lines (listed in some instances on the MSDS), and your safety office should have other relevant info.
} I'm sure I'm again not } alone here in this concern and only wish to continue this difficult and } frequently dangerous craft(EM) with more care for myself , my associates } and my community. } } Anonymous (chicken....) } (...and Vendors and manufacturers are our backbone and our support! } Thank you all!) } I'd say the first order of safety is to be concerned, next is to get all the info available. Yours in chickenhood, Bill Tivol
This technology needs more thorough discussion, I believe. There are several rather serious safety issues that, I believe, we might find some better solutions to here, if we open this area for discussion . For example vapor level and detection, penetration, permeability, scrubbing, reactive absorption, isolation, etc.. Some plastics stain easily and some do not, but all are apparently permeable. On the productive side the commercially astute might find several new product ideas valuable to this community. 'JD'
previously:
Plastic is oxidized and so part of the osmium is exhausted in the vials. Even when frozen osmium diffuses through plastic containers. So you may save a rare breakage, but you are certain to have osmium in your refrigerator. It's just a bad idea to pack osmium in plastic. You could use a secondary container, be it plastic or glass, to increase overall safety. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Thursday, January 06, 2000 3:08 AM, Donna Wagahoff [SMTP:DWAGAHOFF-at-siumed.edu] wrote: } } We have always kept our osmium solutions in glass containers. However, we } are in the process of evaluating our safety procedures and discussed the } idea of increasing the safety of the transport of osmium from the } refrigerator to the fume hood by putting the osmium solution in plastic } containers. (If they are dropped, they would not break and cause the danger } of a spill outside the hood.) } Does anyone have experience with osmium stored in plastic? Any comments } about this particular subject or any of your safety with osmium procedures } are welcomed. } Thanks. } } Donna Wagahoff } SIU School of Medicine } PO Box 19627 } Springfield, IL 62794-9627 } 217-782-0898 } fax217-524-3227
Recently an E3 Electroscan has become available and I'm becoming aware of a much different SEM technology than I'm used to seeing over the last 30 years. It obviously operates only at vacuum levels in the torr range, right? I am still a little confused how resolution can be maintained in these vacuum levels. And dispersive X-ray spectroscopy, does this broaden the peaks and what happens to spectral resolution? It also appears that this particular design cannot pump down below 10-4(?). There are some complex gas background issues that are different. Are there ways of using these design parameters to the benefit of the materials imaging analysis in samples that are not hydrated or partially volatile? 'JD'/Texas
Continuing in the line of ICPIC '95 held at IIT, Kharagpur and ICVGIP '98 held at IIT, Delhi, ICVGIP 2000 will be organized by the Centre for Artificial Intelligence & Robotics at Bangalore during December 20-22, 2000. The conference is intended to bring together the Vision, Graphics, and Image Processing communities together with a special emphasis on India. A high quality technical track will be augmented by presentations from various R&D institutions in the country and the industry.
Important Dates: ----------------
Submissions due: May 15, 2000 Notifiation of acceptance: Sep 01, 2000 Final papers due: Oct 15, 2000 Conference dates: Dec 20-22, 2000
Topics: -------
We strive to host a high quality conference in India. An additional goal his to bring the community of Indian practitioners of these areas together at a single forum. We encourage papers related to system development, innovative applications etc., in addition to research papers. We especially encourage papers by student. The topics of interest include, but are not limited to, the following:
Electronic submissions are highly encouraged. Acceptable formats are: Acrobat PDF, standard PostScript, self-contained LaTeX with psfig, and Word 7.0. Check the official web page for details on electronic submission. Manuscripts should not exceed 20 double-spaced pages including figures and tables. The submission should include a cover page with the title, the authors' names, abstract and keywords. Those submitting hard-copy manuscripts should send four copies to the following address:
ICVGIP 2000 Secretariat Centre for Artificial Intelligence & Robotics (CAIR) Raj Bhavan Circle, High Grounds Bangalore, 560 001. INDIA
-------------------------------------------------------------------- Patrons: -------- Prof. M. Vidyasagar, CAIR Prof. R. Narasimha, NIAS
General Chair: -------------- Dr. P. J. Narayanan, CAIR pjn-at-cair.res.in
Program Co-Chairs: ------------------ Prof. Ramakant Nevatia, USC nevatia-at-usc.edu Prof. Jayanta Mukherjee, IIT, KGP jay-at-cse.iitkgp.ernet.in
Organizing Chair: ----------------- Prof. Swamy Manohar, IISc manohar-at-csa.iisc.ernet.in
Plenary Chair: -------------- Dr. P. Anandan, Microsoft anandan-at-microsoft.com
Publications Chair: ------------------- Dr. C. V. Jawahar, CAIR jawahar-at-cair.res.in
Treasurer: ---------- Dr. Subrata Rakshit, CAIR subrata-at-cair.res.in
-------------------------------------------------------------------- Organized by Centre for Artificial Intelligence and Robotics (CAIR) --------------------------------------------------------------------
Hi all- I am looking for someone to provide in house service for a Denton Sputter Coater in the Rhode Island area. Please feel free to contact me offline.
---------- } From: c j day {wa5ekh-at-juno.com} } To: Microscopy-at-sparc5.microscopy.com } Subject: ESEM } Date: January 11, 2000 10:24 PM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Recently an E3 Electroscan has become available and I'm becoming aware } of a much different SEM technology than I'm used to seeing over the last } 30 years. It obviously operates only at vacuum levels in the torr range, } right? I am still a little confused how resolution can be maintained in } these vacuum levels. And dispersive X-ray spectroscopy, does this } broaden the peaks and what happens to spectral resolution? It also } appears that this particular design cannot pump down below 10-4(?). } There are some complex gas background issues that are different. Are } there ways of using these design parameters to the benefit of the } materials imaging analysis in samples that are not hydrated or partially } volatile? } 'JD'/Texas } } This is exactly the model we've been using here for the past 7 or so years. In fact, the instrument can be operated in "normal" high vacuum mode, too (if your samples don't mind it). The resolution is not bad, even in "wet" mode - say, 2 - 10 torr. There is a certain amount of beam diffusion, of course, in a wet atmosphere, but we've been doing EDS for the past 5 or 6 years with pretty good, reproducible, results. (We have a NORAN ultra-thin window detector and Voyager 3 software.) Since we have a LaB6 gun in ours, there is also an ion pump, and the gun vacuum is maintained at a little better than 10 -6 torr. As you know, the instrument can be used with, instead of water vapour, inert gases as an atmosphere in the chamber. As it happens, we don't do that with our instrument, so I couldn't really comment. I suspect that there probably aren't any major advantages in using an instrument like this for materials studies, except that it has a very large specimen chamber that can accomodate several types of stage. And, of course, the fact that samples generally require no coating before examination. Much of our usage is earth sciences, and it's nice not to have to coat type fossils with carbon or metals. There are two major disadvantages with the E3's. One is that the field of view is very small - you can't really image anything larger than about 1 mm in length or diameter, because of the configuration of the ESD detector/final aperture assembly. This can be a major pain. Another is that, with the standard stage, samples can not be thicker than about 25 mm or so, especially if you want to do EDS. FWIW, our instrument has been dead reliable since it's installation. Other than biannual column cleanings, the odd hose leak, and an occasional glitch with some miscellaneous part, the machine is hardly ever down. (Kind of like my Harley - there may even be a few shared parts :-). If you do wind up acquiring the E3, the Philips service rep for the American southeast is (or at least was a couple years ago) Steve Booth. (You probably know that Philips bought out ElectroScan a few years ago, so they handle the service contracts now). Steve was here twice to do service calls on ours, because both times, the regular US Northeast guy was unavailable. Steve runs a horse ranch somewhere in Texas, I believe, but knows his E3's pretty well, too. This might be a whole lot more than you wanted to know, but I'll admit to being a bit of a fan of our instrument - like my bike, it's ruggedly built, but is no more complex than it has to be. Just last week, myself and a local SEM service tech who'd never seen an ESEM before completed a biannual column cleaning, and it all went very well - takes a day or two to get the gun vacuum down to operating levels again, though.
No connection with Philips Electron Optics, etc......
F.C. Thomas MicroAnalysis Facility Geological Survey of Canada Bedford Institute of Oceanography Dartmouth, Nova Scotia Canada B3L 4C8
Dear List, can you help me once again. Can anyone recommend a sub $1500 video camera for video microscopy. It will be generally used for relatively high intensity fluorescence microscopy (imaging GFP bacteria) and basic microscopy. At present we have a Sony XC-999P (752x582 pixels) and are looking to upgrade - preferably in resolution and sensitivity - but resolution is the most important factor. If people wish they can respond off=list and I will produce a prcis of the information I receive. Thanks. -------------------------------------------------------------------------------- -----------------------Dr. Giles SandersZeneca / SmithKline Beecham Centre for Analytical SciencesChemistry DepartmentImperial College of Science, Technology and MedicineLondonSW7 2AY (44) - 0171-594-5749 Never express yourself more clearly than you think. -- Niels Bohr (1885-1962) Danish physicist -------------------------------------------------------------------------------- --------------------
Hello All! I guess National Geographic Explorer Magazine will be showing some of David Scharf's work in the next "Explorer" show on TBS (Turner Broadcasting System-yes it's not the Braves or Clint Eastwood!) on January 16th. They will show some of his work imaging parasites with the modified SEM. Should be cool! I love it when they show electron microscopy on TV. Just thought you might want to know!
Tracey Pepper Supervisor Bessey Microscopy Facility Iowa State University ph: 515-294-3872 fax: 515.294.1337
A few of the simpler advantages of an ESEM (we have a model E3): No need to coat a sample saves a few minutes (at least) allows for back-and-forth work with a light microscope No need to pre-pump to remove volatiles - the differential pumping system handles this (although your rough pump oil becomes your trap) Gas evolution (degradation on heating, for example)is not a problem (see above) Aging/dynamic studies don't get compromised due to sample coating. near-atmospheric pressure minimizes sublimation without cryogenics You can study influence of water (swelling, for example)or other sample/gas interactions ESD detector is light-insensitive, so you can watch the sample as you position it. Also as you poke/prod it with the micromanipulator option.
As for EDX, yes, there is a significant beam spread from the imaging gas. I typically line up the region-of-interest, then dial the chamber pressure to zero before collecting spectra. The spread is significantly reduced.
Without trying to touch off arguments, my practical experience is that above about 20,000X I don't collect images worth writing home about. They are useful, but not beautiful.
It's an instrument that fills an interesting niche, even for a materials scientist. (The real forte' is biological/wet stuff. That's where the fun really begins!)
Bill Heeschen The Dow Chemical Company
-----Original Message----- } From: c j day [mailto:wa5ekh-at-juno.com] Sent: Tuesday, January 11, 2000 9:24 PM To: Microscopy-at-Sparc5.Microscopy.Com
Recently an E3 Electroscan has become available and I'm becoming aware of a much different SEM technology than I'm used to seeing over the last 30 years. It obviously operates only at vacuum levels in the torr range, right? I am still a little confused how resolution can be maintained in these vacuum levels. And dispersive X-ray spectroscopy, does this broaden the peaks and what happens to spectral resolution? It also appears that this particular design cannot pump down below 10-4(?). There are some complex gas background issues that are different. Are there ways of using these design parameters to the benefit of the materials imaging analysis in samples that are not hydrated or partially volatile? 'JD'/Texas
Does anyone have a method for flattening 0.5 um thick epoxy (Spurr) sections for LM? We are using water pickup and mild heat drying onto glass slides and that doesn't seem to be doing the trick.
TIA
Bob Dr. Robert R. Wise Department of Biology University of Wisconsin Oshkosh Oshkosh, WI 54901 (920) 424-3404 wise-at-uwosh.edu http://www.uwosh.edu/departments/biology/wise/wise.html
I want to look at what I hypothesize is a densely packed array of membranes in TEM. In routine osmium fixed, LR White and Epon embedded specimens, the image is not overwhelming. I plan to try ruthenium tetroxide ala the skin people looking at lamellar bodies. I am wondering whether I am missing another obvious approach. Any ideas gratefully accepted. 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)
I am currently working on preparing TEM foils of single crystalline gold. The electropolishing was completely fruitless, until I tried Bernie Kestel's solution "BK-2". This has worked wonders, giving a very smooth and shiny surface. FYI, I use a Struers Tenupol twin-jet electropolisher, so my conditions are slightly different than with the South Bay polisher. One problem remains: the electron-transparent edges of the foil are very prone to bending, and thus I have regions that are full of bend contours. This isn't terribly surprising, since the gold is from a grown single crystal and has been subjected only to about 1% plastic strain. Any ideas on reducing the amount of bending, so that I can see the dislocations more easily? Are there any specific profiles for foil perforations that will help keep the edges rigid (other than a smooth, small hole)? Any ideas, either for improving specimen prep, or for "fixing" foils I already have, would be greatly appreciated.
Regards,
John -- ____________________________________ John Balk 200 Latrobe Hall Johns Hopkins University 3400 N. Charles St. Baltimore, MD 21218 ph: (410) 516-8284 fax: (410) 516-4316 e-mail: balk-at-kjhsgi.me.jhu.edu
We are in the market for a CCD camera for a JEOL 1200CX TEM. I would appreciate any information regarding to this type of products available on the market. Thank you.
Those of you who know me can understand that I agree completely with JDs comment below. The whole issue of safety in handling laboratory chemicals is one that, I believe, has still not been completely addressed and accepted by everyone. We all know the short term effect that exposure to toxic levels of OsO4 can have on a person, eyes is one that comes to mind. Formaldehyde and glutaraldehyde are others that I and a pathologist, who I just met yesterday, are now well aware of its possible long term effects. But what about long term, synergistic effects of some chemicals that are by themselves relatively innocuous? Combinations of two, three, four? The only answer is to develop and follow safely procedures for handling each chemical as if it were very toxic. There is no reason not to, you already do it for some chemical, and there are many reasons to do so.
For those who don't know me and need a little motivation, think "bilateral, single sequential lung transplant" .
Damian
At 07:55 PM 1/11/00 -0600, c j day wrote:
} This technology needs more thorough discussion, I believe. There are } several rather serious safety issues that, I believe, we might find some } better solutions to here, if we open this area for discussion . For } example vapor level and detection, penetration, permeability, scrubbing, } reactive absorption, isolation, etc.. Some plastics stain easily and some } do not, but all are apparently permeable. } On the productive side the commercially astute might find several } new product ideas valuable to this community. } 'JD'
What is the longterm effect of exposure to osmium on the brittleness and permeability of common plastics? We leave exposed polymer block faces in osmium vapour overnight before sectioning.
Hello all! We are looking for an ESEM in the greater Washington DC area for a limited amount of work (maybe a dozen samples over a two-month period). This could be a contractual situation, although we are rather hoping for the owner's generosity.... Please reply off-list to: rjpalmer-at-dir.nidcr.nih.gov Robert J. Palmer Jr., Ph.D. Natl Inst Dental Craniofacial Res - Natl Insts Health Oral Infection and Immunity Branch Bldg 30, Room 310 30 Convent Drive Bethesda MD 20892 ph 301-496-2088 fax 301-402-0396
I have a Reichert compound microscope that needs servicing. The number I have for the company who used to service my microscope is no longer valid. Is there anyone in the RTP, NC area who could recommend a service vendor? Please respond to gail.harrison-at-reichhold.com
Does anyone have a solution for this problem? I managed to get a spot ( {3mm around) of Toluidine Blue on the cuff of my new shirt (the only part extending beyond my lab coat). Is there any way to get rid of all (or most) of the stain without it spreading? The shirt is 100% cotton. (I don't care if it gets on my lab coat, but this is the first time in over fourteen years that I've gotten it on my clothes.)
If I don't get any response, my inclination is to try a sodium tetraborate paste applied with a cotton swab.
Jaclynn Lett, Research Assistant jmlett-at-cid.wustl.edu
Fay and Carl Simons Center for the Biology of Hearing and Deafness Central Institute for the Deaf 818 S. Euclid Ave. St. Louis, MO 63110
I am having great difficulty extracting large carbides from a steel specimen using the standard single stage carbon extraction technique. The film is either not releasing or it is breaking up into unusable pieces. My standard practice is as follows:
1) Etch polished surface in 2% Nital for 15 to 45 seconds 2) Release sections in 10% Nital 3) Float sections in Dist. water and retrieve
I have used an electrolytic process to aid in releasing the sections but often this process tends to create somewhat dirty replicas.
I also am going to try a two-stage replication technique, but would prefer to have sucessful single stage replicas.
Does anyone have any suggestions that would be of assistance in my single stage replication technique? Thank you for your consideration.
Kevin Downey Research Analyst Bethlehem Steel Corp. e-mail: rkedo-at-bsco.com
} Does anyone have a solution for this problem? I managed to get a spot ( {3mm } around) of Toluidine Blue on the cuff of my new shirt (the only part } extending beyond my lab coat). Is there any way to get rid of all (or most) } of the stain without it spreading? The shirt is 100% cotton. (I don't care } if it gets on my lab coat, but this is the first time in over fourteen years } that I've gotten it on my clothes.)
We have some stuff called Erada-Stain. Its made for histological stain removal from hands, glassware, etc. We've had our tube of it for forever(15 years +) but it seems like it would be one of those things you should still be able to find. Its always worked for me when I had a clothes splash. Good Luck
Paula Moore Wake Forest Univ. Baptist Medical Center EM Lab pmoore-at-wfubmc.edu
Embeded in TIFF images is data describing the "print" size, X inches by Y inches at N dpi. Of course, this is directly related to the pixel matrix size.
My problem is that my digital imaging equipment (Hitachi S-3500 and IXRF EDS) saves the image data at 72 (or less) dpi. The pixel array size is correct, but at {= 72 dpi, many programs (like PhotoShop) want to print a huge image.
I can overcome this by (in P.S. for example) resetting the image to 300 dpi and adjusting the print size so that the pixel array is not altered. At the very least, this is cumbersome and time consuming when a large number of images must be printed.
I would like to find a way to change the file saving default value for the dpi to avoid image resizing for most print applications.
I've received a lot of inquiries about the education potential of the $100 (or less, in some stores) toy digital microscope introduced by Mattel just before Christmas; it's a plastic-bodied scope with simple image processing software which requires a wired connection to a Windows 98 computer (I'm a Mac user and I don't often feel envious, but...). It's been hard to get good information, but Jim Harper has just posted an excellent article on the web. He describes its capabilities well - far better than any of the other reviews that I've read. And he gives detailed instructions on how to mount it on ANY light microscope! Don't miss the hotlinks at the end of his piece.
Educationally, it's no substitute for the one student - one microscope approach of Project MICRO and "Microscopic Explorations", its manual. But it has exciting potential for classroom demonstrations and science fair projects. And listserver readers who are looking for low cost digital recording of LM may find that it's adequate for a lot of applications. Please let us know if it works for you.
The site is http://www.microscopy-uk.org.uk/mag/artjan00/jhqx3.html
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/PCI/pci.html
I am soliciting contributors (or names of potential contributors) for a symposium for the natiional Microscopy & Microanalysis Annual Meeting to be held on August 13-17, 2000 in Philadelphia, Pa.
Talks may range in length from 25 to 45 minutes. Deadline for receipt of 2 page absracts is Feb 15, 2000.
A description of the symposium follows.
SYMPOSIUM: MICROORGANISMS: THE GOOD, THE BAD, THE UNUSUAL
This symposium will deal with microorganisms (viruses, bacteria, parasites, prions) found in the environment as well as in higher life forms (animals and plants). Newly discovered pathogens or organisms with unique capabilities (detoxification, invasiveness, resistance to antibiotics) are of interest in this symposium. Of particular interest are those orgamisms that represent extremes, as for example: the ability to grow in extreme environments, having extreme virulence or invasiveness, or being difficult to visualize using conventional prepartory procedures. Hopefully, the participants shall describe some of the features of extreme organisms that give rise to these capabilities. Finally, many of these organisms are often difficult to visualize using standard preparatory procedures. Papers describing procedures to prepare the specimens for visualization would be germane to this symposium.
==============================================
#################################################################### John J. Bozzola, Ph.D., Director Micro-Imaging and Analysis Center 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 ####################################################################
} Hello All, } } Embeded in TIFF images is data describing the "print" size, X inches by Y } inches at N dpi. Of course, this is directly related to the pixel matrix } size. } } My problem is that my digital imaging equipment (Hitachi S-3500 and IXRF } EDS) } saves the image data at 72 (or less) dpi. The pixel array size is correct, } but } at {= 72 dpi, many programs (like PhotoShop) want to print a huge image. } } I can overcome this by (in P.S. for example) resetting the image to 300 dpi } and } adjusting the print size so that the pixel array is not altered. At the } very } least, this is cumbersome and time consuming when a large number of images } must } be printed. } } I would like to find a way to change the file saving default value for the } dpi } to avoid image resizing for most print applications. }
I also have a Hitachi S-3500N. Use the Action Palette in PhotoShop to change the print size and adjust the pixel array with a single key stroke.
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 530 754 7536 fax http://katie.ucdavis.edu raharris-at-ucdavis.edu
I suggest getting a hold of ThumbsPlus. I print everything from it. It can stretch an image to full scale and can print annotation text with the image. It is a graphics database program that works very well. In addition, it can convert from almost any format to any other format.
What it can do for you is to convert your image from 72 dpi -big to 300 -small and keep it in the same format, e.g. Tiff. You can do a batch convert easily.
Find out more at www.cerious.com
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
} -----Original Message----- } From: White, Woody N [mailto:Woody.N.White-at-mcdermott.com] } Sent: Thursday, January 13, 2000 1:56 PM } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: TIFF image dpi format ? } } } -------------------------------------------------------------- } ---------- } 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, } } Embeded in TIFF images is data describing the "print" size, } X inches by Y } inches at N dpi. Of course, this is directly related to the } pixel matrix } size. } } My problem is that my digital imaging equipment (Hitachi } S-3500 and IXRF } EDS) } saves the image data at 72 (or less) dpi. The pixel array } size is correct, } but } at {= 72 dpi, many programs (like PhotoShop) want to print a } huge image. } } I can overcome this by (in P.S. for example) resetting the } image to 300 dpi } and } adjusting the print size so that the pixel array is not } altered. At the } very } least, this is cumbersome and time consuming when a large } number of images } must } be printed. } } I would like to find a way to change the file saving default } value for the } dpi } to avoid image resizing for most print applications. } } Any suggestions??? } } Thanks, } Woody White } McDermott Technology }
I have a problem doing x-ray mapping on my Philips XL30 (W) with an attached Oxford ISIS 200 EDS system. I'm running the ISIS software on the same pc that controls the XL30, a 133 Pentium with 32Mb memory, running NT4 with service pack 3. I have ISIS v3.32 and XL v5.5 software. When I try to collect say 6 elemental maps using the SPEEDMAP software the system locks after 2 or 3 scans and the computer has to be re-booted. I have tried increasing the memory up to 80Mb but this had no effect. I did not have this problem when running Windows 3.1, although the system would give 'out of memory' errors which is why I upgraded to NT. The problem also occurs when collecting an image using the ISIS AUTOBEAM software and integrating several frames. Single frame acquisitions are ok. If you have a similar system configuration would you please let me know it you experience this problem? I know of stand alone NT systems that are ok, so can only assume it is some conflict with my particular software/hardware combination.
Thanks
Dave
Dave Phelan EM/X-Ray Unit University of Newcastle NSW 2308 AUSTRALIA Ph 02 4921 5667 Fax 02 4921 7019 emudp-at-mail.newcastle.edu.au
We are having antifield systems from Lindgren RF Enclosures , fitted on our two Hitachi FESEMs, an S-4500II and an S-900.
It would be very advisable for the installing engineer to inspect columns of these models before they set out for Sydney.
Could any operator of these models around New York who would allow inspection of their microscopes please mail me?
thanks,
Dr. Mel Dickson, Deputy Director, The Electron Microscope Unit, Adjunct Associate Professor, School of Microbiology & Immunology The University of New South Wales Sydney, Australia. Phone +612 9385 6383 Fax +612 9385 6400
Revisiting an old chestnut - safety and aldehyde fixatives.
In the UK, the Health & Safety Commission have just lowered exposure limits for glutaraldehyde.
Formalin/formaldehyde has a MEL (maximum exposure limit) of 2 ppm or 2.5 mg/cu m - this is measurable and legally enforcible. Glutaraldehyde used to have an OES (occupational exposure standard) of 0.2 ppm or 0.83 mg/cu m. over a 15 minute period. OES is a standard to aim for, but not prosecutable if you weren't achieving it.
Now, glutaraldehyde suddenly has a MEL of 0.05 ppm, both for 15 minute short term reference limit AND THE 8 HOUR TIME WEIGHTED AVERAGE EXPOSURE!! This is a quarter of what it previously was and 40 times lower than that for formaldehyde! Does anyone know why or have evidence or anecdotes of glutaradehyde-related health problems? Is it so nasty??
I appreciate that it is used in bulk as a bacteriocide in hospitals and possibly in horticulture/agriculture.
I am trying to contact HSC specialist committees for a response and will post anything that I receive. I suspect that may be very little.
Regards - Keith (reincarnated after "early retirement")
PS - Hello, Daniele! And those who know her! _______________________________ Dr. Keith Ryan Marine Biological Association of the UK The Laboratory Citadel Hill Plymouth Devon PL1 2PB England
Tel. ++44 (0)1752 633249 Tel. ++44 (0)1752 633279 The 279 number has an answering machine
There may be some confusion about my question. Perhaps I can clarify....
I can resize/fix the image size parameters ok. Either totally manually or with a macro from PhotoShop, etc.
My goal is to NOT have to do that. If the original images are SAVED at the appropriate print dpi this would not be required.... That is my goal. That is... Is there any way to modify the SAVING software (Hitachi PC-SEM/IXRF Iridium) so that the images are, for example, 300 dpi rather than 72 or 26 which is what the software(s) does now.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Does anyone have a solution for this problem? I managed to get a spot ( {3mm } } around) of Toluidine Blue on the cuff of my new shirt (the only part } } extending beyond my lab coat). Is there any way to get rid of all (or most) } } of the stain without it spreading? The shirt is 100% cotton. (I don't care } } if it gets on my lab coat, but this is the first time in over fourteen years } } that I've gotten it on my clothes.) } } We have some stuff called Erada-Stain. Its made for histological stain removal } from hands, glassware, etc. We've had our tube of it for forever(15 years +) } but it seems like it would be one of those things you should still be able to } find. } Its always worked for me when I had a clothes splash. } Good Luck } } Paula Moore } Wake Forest Univ. Baptist Medical Center } EM Lab } pmoore-at-wfubmc.edu } } } Hi,
To destain a slide which has been contrasted with toluidine blue (or any of the other blues), soak the slide in acid alcohol. To one liter of 70% ethanol, add 10ml of conc hydrochloric acid. Soak the cuff in that. If that does not do it, paint your cuff. I have done this frequently. Use laundry marking stick or magic marker or whatever. If I have a lot of staining to do, I simply wear a multicolor blue blouse. No problem.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Does anyone have a solution for this problem? I managed to get a spot ( {3mm } } around) of Toluidine Blue on the cuff of my new shirt (the only part } } extending beyond my lab coat). Is there any way to get rid of all (or most) } } of the stain without it spreading? The shirt is 100% cotton. (I don't care } } if it gets on my lab coat, but this is the first time in over fourteen years } } that I've gotten it on my clothes.) } } We have some stuff called Erada-Stain. Its made for histological stain removal } from hands, glassware, etc. We've had our tube of it for forever(15 years +) } but it seems like it would be one of those things you should still be able to } find. } Its always worked for me when I had a clothes splash. } Good Luck } } Paula Moore } Wake Forest Univ. Baptist Medical Center } EM Lab } pmoore-at-wfubmc.edu } } } Hi,
To destain a slide which has been contrasted with toluidine blue (or any of the other blues), soak the slide in acid alcohol. To one liter of 70% ethanol, add 10ml of conc hydrochloric acid. Soak the cuff in that. If that does not do it, paint your cuff. I have done this frequently. Use laundry marking stick or magic marker or whatever. If I have a lot of staining to do, I simply wear a multicolor blue blouse. No problem.
Greetings friends, A researcher wants to know how to stain for, or find by TEM, Barr bodies that are already prepped and embedded in resin for TEM. Any advise that you can supply will be most appreciated. Thanks, Linda M. Fox Loyola University Stritch School of Medicine Core Imaging Facility 2160 S. First Ave. Maywood, Il 60153 Bld. 102 Room 0617 1-708-216-3395 lfox1-at-wpo.it.luc.edu
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I spent a lot of time many years ago working on 316 stainless. Many of the specimens I made were single stage carbon extraction replicas. The technique I found most effective was to use dilute hydrochloric acid and electrolytic activation. I used this both to etch the surface prior to carbon coating and also to release the carbon replica. I also used the same process on ferritic steels. It was very effective it even released large sheets of M23C6 carbides from grain boundaries. The only 'precipitate' it would not work on was ferrite in austenite.
regards, -- Larry Stoter 17, Rocks Park Road, Uckfield, E. Sussex, TN22 2AT, UK email: LPS-at-teknesis.demon.co.uk, Home Phone/Fax: +44 (0)1825 767967
Position open at the Australian National University , Canberra www.anu.edu.au/hr/jobs
RESEARCH SCHOOL OF BIOLOGICAL SCIENCES ELECTRON MICROSCOPY UNIT ELECTRON MICROSCOPIST ANU OFFICER GRADE 7 (TECHNICAL) $43,506 - $47,010 per annum (Plus generous superannuation provisions)
Reference No: G000011. The ANU Electron Microscopy Unit, a multidisciplinary research and teaching support facility with four SEMs, three TEMS and ancillary equipment, requires a skilled and experienced person to join its team of 5-6 staff. The successful applicant will have a history of work in electron microscopy in a diversified research-oriented environment, preferably with some administrative experience, and areas of expertise that support and complement those of existing staff. They will have up-to-date expertise in a number of areas of electron microscopy and image analysis. Among these, experience with quantitative energy-dispersive X-ray analysis, research projects in plant or animal cell biology, and cryopreparation techniques is a necessity.
The ANU EMU website is http://online.anu.edu.au/EMU
Contact for selection documentation: Ms Susan Toscan , ph (02) 6249 4752, email: susan.toscan-at-rsbs.anu.edu.au For further information contact: Dr Sally Stowe, email: stowe-at-rsbs.anu.edu.au Closing date: 31st January 2000.
Tenure-Track Faculty Position in Condensed Matter: Electron Microscopist. The candidate should have a strong background in transmission electron microscopy and diffraction, and an interest in the applications of advanced TEM techniques to materials physics. The candidate would be expected to have a broad knowledge of electron diffraction theory, of high resolution microscopy and electron spectroscopy and of materials physics. Possible areas of interest include (but are not limited to), microscopy of magnetic and/or superconducting materials, including holography; defects and interfaces in materials; ferroelectrics; diamond films and film growth; nanoscale materials; amorphous materials; quantitative microscopy; and 3-D tomography. Although not essential, an interest in electron optics would be valuable. The candidate will have the opportunity to establish a joint program with the Electron Microscopy Center in The Materials Science Division at Argonne National Laboratory. Send curriculum vitae and references by March 17, 2000 to: Physics Dept., NIU, DeKalb, IL 60115, Attn: J. C. Shaffer, Chairman. NIU is an AA/EEO Institution.
Scripps Institution of Oceanography Analytical Facility
http://sioaf.ucsd.edu/flyer/
----- Original Message ----- } From: {"rfelten-at-Macdermid.com"-at-sparc5.microscopy.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, January 05, 2000 2:16 PM
Dear colleagues,
Would you please give us some information about a beam blanking device and a cryostat which can be set inside the microscope chamber. In fact, we would like to modify our old Phillips microscope (SEM 505) with these two devices. In particularly, could you give us the quotation for these two devices,
Thanking you in advance, Cordially,
email adresse for the answer : abdelillah.elhdiy-at-univ-reims.fr
I've received a lot of inquiries about the education potential of the $100 (or less, in some stores) toy digital microscope introduced by Mattel just before Christmas; it's a plastic-bodied scope with simple image processing software which requires a wired connection to a Windows 98 computer (I'm a Mac user and I don't often feel envious, but...). It's been hard to get good information, but Jim Harper has just posted an excellent article on the web. He describes its capabilities well - far better than any of the other reviews that I've read. And he gives detailed instructions on how to mount it on ANY light microscope! Don't miss the hotlinks at the end of his piece.
Educationally, it's no substitute for the one student - one microscope approach of Project MICRO and "Microscopic Explorations", its manual. But it has exciting potential for classroom demonstrations and science fair projects. And listserver readers who are looking for low cost digital recording of LM may find that it's adequate for a lot of applications. Please let us know if it works for you.
The site is http://www.microscopy-uk.org.uk/mag/artjan00/jhqx3.html
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/PCI/pci.html
FIXED TERM - Total Remuneration: Level 3 (38 hours): A$35,824 - A$41,251 per year. (Salary Level 3: A$30,272 - A$34,858 per year plus up to 17% employer superannuation plus leave loading.)
The Electron Microscope Unit is a central infrastructural research facility, containing nine principal instruments, which support a wide range of projects. The Unit seeks a self-motivated, enthusiastic person to offer technical support to assist in the smooth running of the Unit. The successful applicant will be required to perform a range of routine laboratory tasks such as film processing, specimen preparation and assist users of the Unit with operation of microscopes.
Essential criteria: familiarity with the operation of both scanning and transmission electron microscopes and with microscope specimen preparation techniques; previous experience in research laboratory environment; familiarity with common windows-based software packages; good interpersonal skills and a knowledge of EEO/AA principles.
Desirable criteria: experience with microscopy of biomedical specimens, experience with cryomicroscopy techniques; ability to use image processing and analysis software. This is a fixed term position to 31 December 2000
Information about the Unit can be found on its website: http://srv.emunit.unsw.edu.au
Enquiries may be directed to Associate Professor Paul Munroe on telephone (02) 9385 4435, facsimile (02) 9385 6400 or email: p.munroe-at-unsw.edu.au.
Applications close 28 January 2000.
Dr. Mel Dickson, Deputy Director, The Electron Microscope Unit, Adjunct Associate Professor, School of Microbiology & Immunology The University of New South Wales Sydney, Australia. Phone +612 9385 6383 Fax +612 9385 6400
I have been looking for some relevant references to put in my PhD thesis which are applicable to the work undertaken.
I was trying to immunogold label (using 5nm gold conjugated to Fab) an epitope on the giant muscle protein titin within muscle fibre bundles (all Ab labelling was done prior to sample fixation). However, the labelling seen was low and inconsistent.
Labelling with FITC conjugated Ab or unconjugated Ab labelled samples which were then stained was fine though.
Does anyone one know of similar work where immunogold labelling has failed and possible reasons for this has been explicitly mentioned within the paper.
I have inherited a Leitz Aristoplan microscope for integration into a digital imaging system for histopathology. Unfortunately, the mid-range objectives (10, 25, 40x) are all fluors, although the scope is not equipped for fluorescence. I would like to acquire planapochromats for each of these magnifications. The Aristoplan is a fixed tube length (160mm) instrument that is excellent optically, but the fluotars cause significant vignetting at all magnifications, even with the correct C mount. If you can provide any or all of these lenses, please contact me off-list with pricing information and purchasing details.
Roger Moretz Dept of Toxicology
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Not having a lot of microscopy experience, I was wondering if anyone could recommend some solutions for effectively clearing Arabidopsis seed coats. I would like to use as non-toxic a solution as possible (no Xylene!) and also one which works quickly. Can anyone give me some handy hints?
Thanks for you help!
Stephen Evans Wheat Improvement Centre Norwich Research Park Colney Norwich NR4 7UH stephen.evans-at-aguk.zeneca.com
Hi Roy: Sorry I haven't gotten back with you sooner, but I was off for the holidays. Do you still need equipment? Give me a call so we can discuss what you need. Regrards, Mike Coviello 817 272-5496
I received this question from a co-worker. Please respond directly to me.
thanks in advance, Marisa Ahmad
--------------------
Small question. We have a Leybold mechanical pump hooked to a Reactive Ion Etcher. It is running 24 hrs/ day, and pumps actively on the chamber about 35 times per day. How often should a pump under these conditions be rebuilt? The reason I am asking is the pump seems to be blowing seals and requires rebuilding about once a year, the manufacturer says this is normal...is it? If not, what should we do to improve the time between rebuilds?
Michael Davidson at Florida State University has an excellent demonstration of the QX3 microscope at: http://microscopy.fsu.edu
Don O'Leary ----- Original Message ----- } From: "Caroline Schooley" {schooley-at-mcn.org} To: {Microscopy-at-Sparc5.Microscopy.Com} Sent: Sunday, January 16, 2000 12:44 PM
11th International Congress of Histochemistry and Cytochemistry (ICHC 2000)
ON-LINE REGISTRATION NOW AVAILABLE
Dear Collegues,
I have mentioned before the upcoming meeting ICHC 2000 3-8 Sept. 2000 York, UK
There will a range of sessions that will I am sure be of great interest to this group. Please take a look at our web-site at http://www.med.ic.ac.uk/external/ichc_2000
Hope to see you their
Best wishes
Gary Coulton Organiser ICHC 2000
Dr. Gary Coulton Molecular Pathology Division of Biomedical Sciences Imperial College School of Medicine The Sir Alexander Fleming Building South Kensington London SW7 2AZ
tel 0044 (0)171 594 3190 fax 0044 (0)171 594 3022
e-mail g.coulton-at-ic.ac.uk
------------------------------------- Announcing the 11th International Congress of Histochemistry and Cytochemistry (ICHC 2000)
"Understanding Biocomplexity: The Post-Genome Challenge"
September 3-8, 2000, York, United Kingdom
ICHC 2000 will comprise 27 symposia addressing the latest developments and applications of histochemistry and cytochemistry in the life sciences including medicine.
Many leading experts to speak
On-line registration now open!!!!!!!!!!
For further details of the meeting and how to pre-register please visit our web-site at http://www.med.ic.ac.uk/external/ichc_2000
} Fellow Microscopists, } } I "found" a Sony Mavigraph UP-D7000 Digital Color Printer with 2 cases } of consumables. What I could not find is any of the software to run the } little beast; and besides it was suppose to go on a Unix system, so } even if I found the software it would be useless {I think}. } } I went to the Sony Web site and found no downloads, any other } suggestions or even some discs etc... would be a great help. BTW, the } Sony will be on a PC. } } Thanks, } } John Grazul } Lucent
Dear Listers, I thought I'd beat Tina to the weather report. Here in Albany, NY (where cryo-microscopy means working with the windows open) we are having Martian summer--yesterday's high was -15 C. Yours, Bill Tivol
Sorry to bug everyone, but I'm trying to reach Robert Derby. If you are out there, please e-mail me! Or if anyone has a contact address for him, could you send it to me?
{/bigger} {/fontfamily} {bigger} {bold} {fontfamily} {param} Times {/param} {bigger} RESEARCH ASSOCIATE
Electron Microscopy Facility
University of Kentucky
{/bigger} {/fontfamily} {/bold} {fontfamily} {param} Times {/param} {bigger} The University of Kentucky invites applications for a Research Associate in the area of electron microscopy. The Electron Microscopy Facility houses two SEMs, two TEMs one AFM and extensive sample preparation equipment that are used by undergraduate and graduate students in support of research and education. The primary responsibility of the Research Associate will be to oversee the JEOL 2010F Field Emission TEM and auxiliary equipment, including an Oxford energy dispersive x-ray spectrometer (EDS), Gatan electron energy loss imaging filter and acquisition hardware and software. S/he will be responsible for coordinating and administering maintenance, access, training and use of the equipment by internal and external users. S/he will also be encouraged to develop sponsored research programs and collaborate with university faculty members. The qualified candidate will have a Ph.D. and practical experience in analytical electron microscopy and demonstrate good communication skills. The salary will be commensurate with qualifications and experience. The University of Kentucky offers comprehensive insurance and benefits packages and is an equal opportunity employer. Please send applications to:
Hi, I am trying to get frozen sections of rat skin about 10 um thick. The tissue was perfusion fixed w/ 4% para in PBS then cryoprotected in sucrose/ PBS and mounted with OTC compound. More often then not my sections are sticking to the knife edge and are hard to remove even with a brush. I'm a bit knew to cryo-sectioning so any tips would be greatly appreciated. Thanks, Andy
} Dear Listers, } I thought I'd beat Tina to the weather report. Here in } Albany, NY (where cryo-microscopy means working with the } windows open) we are having Martian summer--yesterday's } high was -15 C. } Yours, } Bill Tivol
OK, OK, it's in the 70sF, raining with enough sun for spectacular rainbows, and there's a break in the winter North Shore surf season. But I'm stuck in a windowless lab just like the rest of you guys!
http://wavetrak.surfline.com/pipecam.asp
My condolences on your winter blues.
Aloha, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Hello, I have version 1.1 of a plugin for Adobe Photoshop (Macintosh). I can send you a copy to try. You'll be printing "pink" images in no time.
John Bonevich
} john grazul wrote: } } } Fellow Microscopists, } } } } I "found" a Sony Mavigraph UP-D7000 Digital Color Printer with 2 cases } } of consumables. What I could not find is any of the software to run the } } little beast; and besides it was suppose to go on a Unix system, so } } even if I found the software it would be useless {I think}. } } } } I went to the Sony Web site and found no downloads, any other } } suggestions or even some discs etc... would be a great help. BTW, the } } Sony will be on a PC. } } } } Thanks, } } } } John Grazul } } Lucent
-------------------------- John Bonevich, Ph.D. NIST, Metallurgy, Stop 8554 100 Bureau Drive Gaithersburg, MD 20899 USA TEL: (301) 975-5428 FAX: (301) 975-4553
Please don't consider this a "commercial" posting; my goal is to encourage lots of experimentation. The Mattel microscope (http://www.microscopy-uk.org.uk/mag/artjan00/jhqx3.html )lists for $100, but Toys-R-Us is now selling it for $69.95, in both its retail stores and website. And if you go to www.etoys.com, you can download a Mattel $30 rebate certificate, good till 4/29. So it looks like you can get one for ~$40. Have fun, folks!
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/PCI/pci.html
We have a Micro-g anti-vibration platform that our JEOL 1200EX TEM was sitting on until recently. This allowed us to do decent microscopy on a vibration prone second floor with very good results. We no longer have a need for this unit. It could be used with a TEM or SEM.
What we would like to do is trade it for a new Mac. We can not buy Macs. If you need a platform, let's talk trade. I think that this would be a good deal.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Hi All, We have a Balzars 301 Freeze Fracture with quartz thin film monitor, double replica accessory, and electron guns for both platinum and carbon up for grabs. New seals on the mechanical pump. Original equipment bought circa 1977. Last used 1998, and was perfectly fine. Free, as is. Recipient will pay for moving. Kristen
Kristen A. Lennon Cell, Molecular & Developmental Biology Group Department of Botany & Plant Sciences University of California Riverside, CA 92521 kalen-at-citrus.ucr.edu
Dear Listers, We're investigating both confocal microscopy and epi-fluorescence combined with de-convolution software for a multi-user facility. I'm requesting input from directors and managers of other shared technology laboratories regarding systems chosen and whether the system has met their expectations. Please include websites which include FAQs and clarification of terms. Thanks in advance for your input. Rosemary Walsh EM Facility for the Life Sciences Life Science Consortium & Biotechnology Institute Penn State University University Park, PA. 16802 (814) 865-0212
Dear ALL, there is a lot of good news about the QX3 microscope (toy-)attachment, and I would like to try it under Windows95 or Windows NT4. Is there any driver for this systems? any thanks Lajos Pogany
I have the task to analyse (by EELS technique) precipitates in steel. I would greatly appreciate some hints coming from people who are already familiar with the difficulties connected with: 1. carbon analysis. It is obvius that a carbon film support for the precipitates extracted from the steel is to be avoided. Has anyone experince on the use of other kind of supporting film (silicium monoxide ?, beryllium ?). I have the same problem with using copper grids, because the precipitates are supposed to contain copper too. What kind of grids is most appropriate?
2. would it be a better solution to perform precipitate analysis on thinned steel specimens ? Can it occurr an annoying interference originating in the material surrounding the precipitate ?
3. last but not least, I would greatly appreciate some bibliographical hints on that very specific topics.
Recently, an interesting medical school project has appeared at my lab door. I've been asked to quantify Ca and P in scar tissue found in sections of hamster hearts. Obtaining appropriate standards is critical. Are there commercially available biological standards or procedures to create such standards out there? Any suggestions would be appreciated and thanks for your time.
Dan
===================================================== Dan Kremser Washington University Department of Earth and Planetary Sciences/Campus Box 1169 (Wilson Hall, Room 108-----for packages) One Brookings Dr. St. Louis, MOÊ 63130-4899
Dear Colleagues What is the best glue for making cross-section samples to study ~ 30 micron reaction layer formed by a coating on a Ni-base superalloy? The M-bond 610 which used to work great for Si and ceramic samples does not seem to work here. Though the glue I have is pretty old. TIA Anita
Anyone have any recommendations for organizing digital images and associated data in electronic form in a GLP environment? I'm going crazy trying to find something that will be easy to use, will link the image with associated data information, and will have restriction or audit capabilities to track any changes to the data that may be made.
Any input is greatly appreciated.
Gregg Sobocinski Parke-Davis Pharmaceuticals Ann Arbor, Michigan USA Gregg.Sobocinski-at-wl.com
} What about the Mac? :-) } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
John Mansfield PhD CPhys MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 763-2282 Cellular Phone: (734) 358-7555 (Leaving a phone message at 936-3352 is preferable to 358-7555) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42¡ 16' 48" Long. 83¡ 43' 48"
I need to make some ~100nm thick oxide layers on silicon to align an ion gun in an auger system. I assume that most people use thermal oxidation to grow the films.
Does anyone have a recipe for making these films? (time, temperature, atmosphere) It would be really great if the recipe allowed me to get an exact thickness too!
Thanks, Henk
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://web.ceof.ohio-state.edu An optimist believes that we live in the best of all possible worlds. A pessimist fears that this is true.
According to the data sheet packed with M-Bond 610, the room temp pot life, after mixing, is six weeks.
We note that it fails quickly, i.e. it works fine one day and doesn't the next. As we can't abide samples coming unglued, we toss mixed M-Bond 610 after 30 days and mix fresh.
Also, M-Bond 610 works best bonding smooth surfaces. If we have rough surfaces we use GATAN G-1 (Epo-Tec 353ND).
Ron
Ron Anderson, IBM, Hopewell Jct., New York, USA. anderron-at-us.ibm.com
IBM Analytical Services; http://www.chips.ibm.com/services/asg
Anita Garg {Anita.Garg-at-lerc.nasa.gov} on 01/20/2000 10:51:15 AM
To: Microscopy-at-sparc5.microscopy.com cc:
Dear Colleagues What is the best glue for making cross-section samples to study ~ 30 micron reaction layer formed by a coating on a Ni-base superalloy? The M-bond 610 which used to work great for Si and ceramic samples does not seem to work here. Though the glue I have is pretty old. TIA Anita
It's raining cats and dogs, and it's cold and miserable. Feel better, now?
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Does anyone have technical manuals or documentation for a B&L Research II Metallograph which I could copy. MCCC just received one as a donation. Thanks!
Michael Mohn Assistant Professor of Materials Technology Monroe County Community College Phone: 734-384-4122 Fax: 734-242-9711 http://www.monroe.cc.mi.us/mmohn
} } Aloha, } Tina } } It's raining cats and dogs, and it's cold and miserable. Feel better, } now? } } **************************************************************************** } * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * } * Biological Electron Microscope Facility * (808) 956-6251 * } * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* } ****************************************************************************
But what you call cold and what we call cold are two very different animals! Its been snowing all day in the Northeast, and the temps haven't nosed above (or even near) freezing) in days!
Lee
Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Confocal Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
I attempted to do this many years ago. I made replicas using aluminium. This was written up in the proceedings of a conference on microanalysis ("Measurement of carbon in V(C,N) precipitates extracted from HSLA steels in aluminum replicas," by A.J. Garratt-Reed, in "Quantitative Microanalysis with High Spatial Resolution," The Metals Society, London, Book No. 277, 1981, p. 165.) Then someone else tried to replicate my work (for the moment I can't remember his name, but he was at Strathclyde University in Glasgow) and could not get consistent results. He concluded that the aluminum was somehow catalysing the oxidation of the carbon in the carbides. I expect he published the results, but I can't now remember where. I have some memory that he used silicon monoxide to make replicas, but when I tried that, the replicas would break up because of beam-induced charging. Of course, it would have defeated the point to have put a carbon coat on the films!
When copper has been an issue, I have often used nickel, which are very satisfactory. You can also get grids of many other materials, including Ti, Al, Au, Mo, etc., etc. - check your EM suppies catalogues.
Tony G-R.
At 10:33 AM 01/20/2000 +0100, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
** Anthony J. Garratt-Reed ** MIT Room # 13-1027 ** 77 Massachusetts Avenue ** Cambridge, MA 02139-4307 ** USA ** ** Phone: (+) 1-617-253-4622 ** Fax: (+) 1-617-258-6479 **
Henk, Ta is usually used for measuring sputter rates and you can see where the beam was put. You electrochemically grow a specific thickness and measure the sputter rate. If you want to pursue this, I can give you a referral to a couple of surface scientists who have done this. They are in your neck of the woods.
Another thing that you might want to try is something that I wrote up a number of years ago in JVST as a shop note for aligning an ion gun system where I could not see the beam. Take Double sticky tape and put it on your sample holder. It works in a UHV system well enough, trust me. Then push the sample holder into yellow WO3 powder to cover the sticky tape. Where the beam hits the sample, the WO3 will turn blue. A couple of sample transfers and you have it.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies." --
} -----Original Message----- } From: Hendrik O. Colijn [mailto:colijn.1-at-osu.edu] } Sent: Thursday, January 20, 2000 12:54 PM } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: MAT: making silicon oxide layers } } } -------------------------------------------------------------- } ---------- } 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 all, } } I need to make some ~100nm thick oxide layers on silicon to } align an ion } gun in an auger system. I assume that most people use } thermal oxidation to } grow the films. } } Does anyone have a recipe for making these films? (time, } temperature, } atmosphere) It would be really great if the recipe allowed } me to get an } exact thickness too! } } Thanks, } Henk } } Hendrik O. Colijn colijn.1-at-osu.edu } Campus Electron Optics Facility Ohio State University } (614) 292-0674 http://web.ceof.ohio-state.edu } An optimist believes that we live in the best of all possible worlds. } A pessimist fears that this is true. } }
I read Tony's answer to your question and I can't add anything to that. However, I just have a minor point. If you are using EELS, why are you worried about the grids? The grids would not show up in the EELS spectrum. Of course it would interfere if you are using EDS.
You can get your grids in almost any material you want nowadays. Ni, Mo, Be, Al, Cu, C, etc. Just pick one that doesn't interfere with your EDS spectrum.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies." --
} -----Original Message----- } From: Corneliu Sarbu [mailto:Corneliu.Sarbu-at-mtm.kuleuven.ac.be] } Sent: Thursday, January 20, 2000 4:33 AM } To: microscopy-at-Sparc5.Microscopy.Com } Subject: steel precipitates analysis by EELS } } } -------------------------------------------------------------- } ---------- } 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 fellow microscopists, } } I have the task to analyse (by EELS technique) precipitates in steel. } I would greatly appreciate some hints coming from people who are } already familiar with the difficulties connected with: } 1. carbon analysis. It is obvius that a carbon film support for the } precipitates extracted from the steel is to be avoided. Has anyone } experince on the use of other kind of supporting film } (silicium monoxide ?, beryllium ?). I have the same problem with } using copper grids, because the precipitates are supposed to } contain copper too. What kind of grids is most appropriate? } } 2. would it be a better solution to perform precipitate analysis on } thinned steel specimens ? Can it occurr an annoying interference } originating in the material surrounding the precipitate ? } } 3. last but not least, I would greatly appreciate some bibliographical } hints on that very specific topics. } } Thank you in advance. } } Corneliu Sarbu } MTM Dept. of KULeuven, Belgium }
I am celating information about current TEM and Confocal technologies with the intention of setting up both a confocal suite and an EM suite at a new facility. However, I am new to this field and would like some advice on the pros. and cons. of different systems based on other user experiences, to give me an idea of what I should be looking out for. Specifically, the confocal suite would allow the study of both live (EGFP) and fixed samples. Thus, the type of confocal needed should have good resolution for both live and fixed samples as well as good phase contrast. Importantly, this will probably be a multi-user facility, which needs reliable lasers that do not need to be realigned often. It would also be important to have good quality microscope, filters and objectives as well as extras such as a heated stage, video and CCD cameras, appropriate computer workstations and software. Any advice that you could offer on such equipment would be very much appreciated.
The results of the recent election of officers to the Society are now official The current list of elected officers is below.
The new officers elected this year are:
President-Elect RON ANDERSON
Secretary JANET H. WOODWARD (2000-2002)
Director, Physical Sciences THOMAS F. KELLY (2000-2002)
Director, Biological Sciences SARA E. MILLER (2000-2002)
------------------------------------------- The complete list of officers is given below and on the MSA WWW site http://www.msa.microscopy.com/MSADocs/MSAOfficers.html --------------------------------------------
MSA COUNCIL 2000
President KEN DOWNING 326 Donner Lab Lawrence Berkeley Lab Berkeley, CA 94720 (510) 486-5941; Fax (510) 486-6488 Email: khdowning-at-lbl.gov
President-Elect RON ANDERSON IBM Analytical Services IBM Zip-41E Hopewell Junction, NY 12533 (914) 892-2225; Fax (914) 892-2555 E-mail: ron-anderson-at-vnet.ibm.com
Past-President DAVID JOY Rm.232 Science and Engineering Research Facility Univ. of Tennessee Knoxville, TN 37996-0810 (865) 974-3642, Fax (865) 974-9449 and Rm. S189, High Temperature Materials Laboratory Oak Ridge National Laboratory Oak Ridge, TN 37831-6064 (865) 574-6799 E-mail: djoy-at- utk.edu
Secretary JANET H. WOODWARD (2000-2002) Buckman Laboratories, Inc. 1256 N. McLean Blvd. Memphis, TN 38108-1241 (901) 272-6408; Fax (901) 272-6451 E-mail: jhwoodward-at-buckman.com
Treasurer KATHI ALEXANDER (1999-2001) Los Alamos National Lab MST-8 G755 P. O. Box 1663 Los Alamos, NM 87545 (505) 665-4750, Fax (505) 667-8021 Email: kbalexander-at-lanl.gov
Directors, Physical Sciences
J. MURRAY GIBSON (1998-2000) Argonne National Laboratory Materials Science Division Argonne, Ill 60439 {p} Email:gibson-at-anl.gov
THOMAS F. KELLY (2000-2002) 2021 Chamberlain Ave. Madison, WI 53705-4076 (608) 263-1073; Fax (608) 262-8353 E-mail: tfkelly-at-engr.wisc.edu
MIKE KERSKER (1999-2001) JEOL USA 11 Dearborn Rd Peabody MA 01960 (508) 535-5900 Fax (508) 536-2205 E-mail: kersker-at-jeol.com
Directors, Biological Sciences SARA E. MILLER (2000-2002) Duke Medical Center, Pathology Box 3712 Durham, NC 27710 (919) 684-3452; Fax (919) 684-8735 E-mail: saram-at-acpub.duke.edu
AVRIL SOMLYO (1998-2000) Dept. of Molecular Physiol. & Bio. Phys. Box 10011, Health Sciences Center Univ. of Virginia Charlottesville, VA 22906-0011 (804) 982-0825; Fax (804) 982-1616 E-mail: avs5u-at-virginia.edu
JOHN BOZZOLA (1999-2001) EM Ctr - Mailcode 4402 Southern Illinois Univ Carbondale IL 62901 (618) 453-3730, Fax (618) 453-2665 E-mail: bozzola-at-siu.edu
Director, Local Affiliated Societies EV OSTEN (2000-2002) 3M Company 3M Center, Bldg. 201-BE-16 St. Paul, MN 55144-1000 (651) 736-0104; Fax (651) 733-0648 E-mail: efosten-at-mmm.com
============================================== Nestor Your Friendly Neighborhood SysOp
I was wondering if anyone could tell me how to properly install a reticle in an Olympus microscope eyepiece. The eyepiece housing does not seem to come apart, although there are two tiny round holes on either site of the lens. Is there a special tool needed?
I have to admit that until I read Scott's posting, I had missed the point about the elemental interference from the grid not being a problem with EELS analysis.
However, in the case of copper, there is another effect to worry about when making replicas, if the process involves any sort of flotation on, or picking the sample up from, water. Because of its electronegativity, copper can dissolve in the water and then ion-exchange with other elements from the sample, replacing them. Then you really do have copper in the sample. The problem isn't particularly severe with extraction replicas from steels (but then, I have never used extraction replicas from steels to try to analyze for small amounts of copper in the precipitates), but has been terrible, for example, when looking at iron sulphides, which actually had a visible shell of copper sulphide (but I was also fishing those samples out of brine, not distilled water!). The problem doesn't seem to occur with nickel grids, which I use in preference to copper for this type of application, just to be on the safe side.
BTW, I'm glad you got my posting, Scott - I haven't seen it come back to myself yet. I guess the poor mail redirector gets indigestion with the volume it has to deal with!
Tony.
** Anthony J. Garratt-Reed ** MIT Room # 13-1027 ** 77 Massachusetts Avenue ** Cambridge, MA 02139-4307 ** USA ** ** Phone: (+) 1-617-253-4622 ** Fax: (+) 1-617-258-6479 **
************************************************ Robert J. Derby New Mexico Institute of Technology Socorro, N.M. Phone - 505-835-5866 E-mail - rjderby-at-excite.com ************************************************ I have heard through the grapevine that Tamara Howard from Cold Spring Harbor is looking for me. I was told it was posted here but I never got it. Tamara call me at 505-835-5866(Iam 2 hours behind you) Robert
_______________________________________________________ Get 100% FREE Internet Access powered by Excite Visit http://freeworld.excite.com
Just a quick question, I was wondering if anyone had ever tried using a microwave oven to harden their resin??? And if so what sort of results were achieved. Another question I have is how to remove / dissolve hardened resin from an aluminium surface???
I would appreciate any suggestions or advice. Thank you
Many thanks to all who responded on and off-line to my request for information. I hope I acknowledged each one individually.
My quest was to find the real reason for lowering the exposure limit. I won't summarise the replies but: (1) pulmonologists at one hospital are thinking that long term, low level exposure to formaldehyde and glutaraldehyde may cause idiopathic pulmonary fibrosis, (2) I was told of a serious skin burn caused by a splash.
I have now heard from the UK Health & Safety Commission's Advisory Committee on the Toxicity of Substances (ACTS). I have copy from: HSE Review 1997, published 1999, section C58 (consisting of 4 pages). Quotes from this are below.
The official reason for lowering the exposure limit is that it has not been possible to set a no-observed adverse effect level for glutaraldehyde with regard to the induction of asthma.
Carcinogenicity is not supported by the available good-quality evidence to date. ________________________________________
Glutaraldehyde must be labelled under the Chemicals (Hazard Information and Packaging for supply) Regulations 1994 (CHIP) as Toxic, Corrosive, Sensitising and Dangerous for the Environment.
Several thousand tonnes are imported into the UK each year. It is primarily used as a biocide and disinfectant in the health care, off-shore, paper-making and agricultural sectors.
.. it is estimated that a considerable number of (health care) workers are intermittently exposed, given the widespread use in that sector. Similarly, ....... for several hundred workers in the manufacture of glutaraldehyde solutions.
..... available exposure data relates mainly to use in the health care sector....... suggests that under normal operational conditions short term exposures are generally less than 0.2 ppm. This can be exceeded during the cleaning of endoscopes ...... or the wiping of surfaces.
HEALTH EFFECTS - ANIMAL STUDIES. Glutaraldehyde is acutely toxic to rats by inhalation, oral and dermal routes. The principal effects are due to its irritant properties.
Glutaraldehyde is clearly a skin sensitiser in rabbits, guinea pigs, rats and mice.
Glutaraldehyde is clearly mutagenic in vitro, in bacterial and mammalian cells. ............. No firm conclusions can be drawn from the available evidence on chromosomal aberrations, but glutaraldehyde clearly causes sister chromatid exchange (SCE) and unscheduled DNA synthesis (UDS) in mammalian cells.
......... However, the clearly negative results of recent, good-quality bone marrow cytogenetics and peripheral blood micronucleus tests, together with those of the liver UDS assay, provide reassurance that the genotoxic effects shown in vitro are unlikely to be expressed in vivo.
No reports of carcinogenicity studies of glutaraldehyde by the inhalation or dermal routes of administration are currently available. A recent oral study provided no convincing evidence ... in rats ..... drinking water for up to two years.
No significant effects on reproduction were reported in a modern two generation study ........... There were no indications of significant gonadal effects in 13 weeks inhalation studies carried out in rats or mice, or in a lethal assay in mice.
HUMAN DATA Glutaraldehyde is irritant to to human skin at concentrations of 2-10%, but not 0.5%. Higher concentrations have not been investigated.
There is substantial evidence that glutaraldehyde is a skin sensitiser in humans. Concentrations as low as 0.13% have induced allergic contact dermatitis. The majority of cases have been reported in health or funeral workers.
A fair body of evidence ............. indicates that glutaraldehyde has the potential to cause occupational asthma.
.......... several workplace studies with exposure data in which no cases of asthma have been found among contemporary workers. ..... However, superimposed on these data are other reports of sensory irritation and / or asthma in endoscopy nurses where the reported levels of exposure overlap with those in the above studies. .................. From the data available, it has not been possible to determine a NOAEL (no-observed adverse effect level) for the induction of asthma.
No information is available in genotoxicity in humans.
The only available mortality study is of limited value, but does not provide any evidence that glutaraldehyde caused cancer or increased mortality in glutaraldehyde production workers.
On the basis of the very limited information available, it does not appear that glutaraldehyde causes reproductive toxicity in humans.
REFERENCE: Glutaraldehyde: Criteria document for an occupational exposure limit EH/65/32. HSE Books ISBN 0 7176 1443 3. _________________________________________________
Dr. Keith Ryan Marine Biological Association of the UK The Laboratory Citadel Hill Plymouth Devon PL1 2PB England
Tel. ++44 (0)1752 633249 Tel. ++44 (0)1752 633279 The 279 number has an answering machine
Fax ++44 (0)1752 633102
e-mail: kpr-at-wpo.nerc.ac.uk
PS - Daniele, you're still here! Keep smiling!! That was a nice evening in Strasbourg. Do you know which Gewurztraminer we all had as an aperatif? Now the world will wonder?!
************************************************ Robert J. Derby New Mexico Institute of Technology Socorro, N.M. Phone - 505-835-5866 E-mail - rjderby-at-excite.com ************************************************ Does anyone or any company know of a ccd that will capture single photon at the 852-872 wavelength? This is needed for a special project. Any help would be of great help Thanks,
Robert
_______________________________________________________ Get 100% FREE Internet Access powered by Excite Visit http://freeworld.excite.com
Treasurer KATHI ALEXANDER (1999-2001) Los Alamos National Lab MST-8 G755 P. O. Box 1663 Los Alamos, NM 87545 (505) 665-4750, Fax (505) 667-8021 Email: kbalexander-at-lanl.gov
Directors, Physical Sciences
J. MURRAY GIBSON (1998-2000) Argonne National Laboratory Materials Science Division Argonne, Ill 60439 {p} Email:gibson-at-anl.gov
THOMAS F. KELLY (2000-2002) 2021 Chamberlain Ave. Madison, WI 53705-4076 (608) 263-1073; Fax (608) 262-8353 E-mail: tfkelly-at-engr.wisc.edu
MIKE KERSKER (1999-2001) JEOL USA 11 Dearborn Rd Peabody MA 01960 (508) 535-5900 Fax (508) 536-2205 E-mail: kersker-at-jeol.com
Directors, Biological Sciences SARA E. MILLER (2000-2002) Duke Medical Center, Pathology Box 3712 Durham, NC 27710 (919) 684-3452; Fax (919) 684-8735 E-mail: saram-at-acpub.duke.edu
AVRIL SOMLYO (1998-2000) Dept. of Molecular Physiol. & Bio. Phys. Box 10011, Health Sciences Center Univ. of Virginia Charlottesville, VA 22906-0011 (804) 982-0825; Fax (804) 982-1616 E-mail: avs5u-at-virginia.edu
JOHN BOZZOLA (1999-2001) EM Ctr - Mailcode 4402 Southern Illinois Univ Carbondale IL 62901 (618) 453-3730, Fax (618) 453-2665 E-mail: bozzola-at-siu.edu
Director, Local Affiliated Societies EV OSTEN (2000-2002) 3M Company 3M Center, Bldg. 201-BE-16 St. Paul, MN 55144-1000 (651) 736-0104; Fax (651) 733-0648 E-mail: efosten-at-mmm.com
============================================== Nestor Your Friendly Neighborhood SysOp
--CAB19553.948442177/styx.services.ou.edu--
} From MAILER-DAEMON Fri Jan 21 02:09 CST 2000 Received: from localhost (localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with internal id CAA00349; Fri, 21 Jan 2000 02:09:34 -0600
Hello, all-
I'm hoping to hear from those of you who have worked out the optimum size and resolution to make images in e.g., Photoshop that are destined for PowerPoint to be made into transparencies. An image that is 4 x 5 inches and 400-600 dpi seems to be overkill for a 35mm slide.
Your opinions?
Mahalo, Tina **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
I found the following recipe from Bernie Kestel. He states that this is used for surface polishing using the beaker method. Although he did not use it for "jet polishing", it may be a good starting point.
Inconel 718 (annealed) 10% HClO4 90% Ethanol Temperature = -60C Current: 275mA Volts: as required
There is some more information about stir rate, sample orientation etc. that is only relevant for the beaker method. If you'd like to try this approach, I would be happy to send you the entire reference.
David Henriks Vice President 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
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by Schryvers Dominique } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I'm looking for a good electropolishing solution + conditions for as-received and annealed Inconel 718 for use with a Tenupol 3 system to produce well thinned matrix + precipitates for TEM work. Any suggestions?
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
A minor observation ...........
While the issue of overlaps is generally not a problem with EELS (as opposed to EDS), if can arise with steel precipates. The vanadium L and Oxygen K edges are really quite close, so if you have vanadium precipatates, a silicon oxide support film will cause problems for quantitation.
regards, -- Larry Stoter 17, Rocks Park Road, Uckfield, E. Sussex, TN22 2AT, UK email: LPS-at-teknesis.demon.co.uk, Home Phone/Fax: +44 (0)1825 767967
Yikes! Great! set the microscope near the window and get those snow flake pictures! It has been close to 80 degrees a few days this week here in Arizona! I have a great batch of infusion brewing on the back patio.
Got the happage win TV card to add to the computer....Have the c-mount ccd camera I found for $100.... now... I should be able to share pictures soon!
Ed Sharpe archivist for SMECC
{ { Subj: Martian summer Date: 1/22/00 7:50:37 AM Pacific Standard Time From: tivol-at-wadsworth.org (William Tivol) Sender: tivol-at-wadsworth.org To: microscopy-at-sparc5.microscopy.com
------------------------------------------------------------------------ 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, I thought I'd beat Tina to the weather report. Here in Albany, NY (where cryo-microscopy means working with the windows open) we are having Martian summer--yesterday's high was -15 C. Yours, Bill Tivol } }
The main Microscopy Server was replaced this weekend (read that as many blurry eyed hours in front of a monitor, and lots of cursing at new formats of configuration files for DNS and SENDMAIL).
The old beast was growing increasing unreliable with hardware crashes nearly daily. I believe that most of the services have been restored however until all databases are reconfigured and tested there will be some glitches. Please be patient.
I think I have been able to capture the few messages that were sent over the weekend. If those of you that posted items over the weekend don't see things in the next day please repost them.
Dear Tina, Our medical photography dept. requests that files submitted for slides are at least 1 Mb and no more than 3 Mb in size. Less than this doesn't have enough pixels to adequately fill the image (similar to a thin negative), and more is unneeded. I create the slide figure, set to 300 dpi and adjust dimensions to put the file size into that range.
Regards, Glen
Glen MacDonald Research Scientist Hearing Research Laboratories of the Virginia Merrill Bloedel Hearing Research Center Box 35-7923 University of Washington Seattle, WA 98195-7923 (206) 616-4156 glenmac-at-u.washington.edu
On Fri, 21 Jan 2000, Tina Carvalho 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- } } I'm hoping to hear from those of you who have worked out the optimum size } and resolution to make images in e.g., Photoshop that are destined for } PowerPoint to be made into transparencies. An image that is 4 x 5 inches } and 400-600 dpi seems to be overkill for a 35mm slide. } } Your opinions? } } Mahalo, } Tina } **************************************************************************** } * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * } * Biological Electron Microscope Facility * (808) 956-6251 * } * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* } **************************************************************************** } } }
Position: Full/Associate/Assistant Professor of Materials Science and Engineering. This is a full time, tenure track teaching and research position beginning Fall 2000.
Rank and Salary: Rank will depend upon background and experience; salary competitive.
Responsibilities: Teach undergraduate and graduate courses in materials science and engineering, advise undergraduate and graduate students, seek and conduct funded research, supervise theses and projects, and serve on university, college and department committees.
Academic Qualifications: Earned Ph.D. in materials science and/or engineering or a closely related field is required. Demonstrated experience in research and grant seeking is highly desired. Prior teaching experience a plus.
Experience Qualifications: Demonstrated experience in research and grant seeking is highly desired. Prior teaching experience a plus.
Department: The Department of Construction Engineering, Materials Engineering and Industrial Design at Western Michigan University currently offers two undergraduate BSE -- Materials Engineering and Construction Engineering and Management, and BS in Industrial Design. The Department also offers two Master of Science programs - Materials Science and Engineering and Construction Management.
University: Western Michigan University, with a student body of approximately 28,000 is located in southwest Michigan. Kalamazoo is halfway between Chicago and Detroit and 45 miles south of Grand Rapids. The population of the greater Kalamazoo area is approximately 200,000. Its industry is highly diversified and it is the center of many cultural and sporting events.
Applications: Review of applications will begin on January 10, 2000, and will continue until the position is filled. Please send the following credentials: Letter of Application addressing qualifications, Vitae, Transcripts from all institutions, and the names/addresses, telephone/fax numbers of three references
Contact: Please send credentials to:
Dr. Roman Rabiej, Chair Western Michigan University Department of Construction, Materials, & Industrial Design 2007 Kohrman Hall Kalamazoo, MI 49008
AN EQUAL OPPORTUNITY/AFFIRMATIVE ACTION EMPLOYER Western Michigan University is an Equal Opportunity Employer. In addition, it has embarked upon a vigorous affirmative action program and encourages the applications of women and members of minority groups. ============================== Pnina Ari-Gur, D.Sc., Professor of Materials Science and Engineering Western Michigan University Kalamazoo, MI 49008 (616) 387-3372 FAX: (616) 387-6517 email: pnina.ari-gur-at-wmich.edu http://www.wmich.edu/cmd/arigur.htm ==============================
Does anyone have a Probe Current Detector accessory for a JEOL 840 surplus to requirements and available for purchase? I think its designation is PCD40, it's the pneumatically-operated thing which mounts on the column opposite to the objective aperture holder, and shoots a Faraday cup across into the beam.
thanks
Ritchie Sims
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Dear Dr. Fashing, it is a pleasure receiving news from you. I have to apologize for the trouble you have reported in your letter and I hope to help you.
First of all, the website is going to be updated because of the second circular is ready to be sent.
The second point is that you have enough time to submit your paper or poster. The deadline for submission has been established on 30 April 2000. I hope you answered to the first circular (my database has not been recently updated - Migliorini is working on this and he has the complete and updated archive), so in this case you will receive the second circular directly on your computer. Otherwise I suggest you to fill the application on the web site.
Let me know if I could help you more. I will be glad to do that.
My best wishes and see you in Siena
Enrico
} Dear Dr. De Lillo, } } I am interested in attending the EURACC symposium in Siena this July, } and would greatly appreciate receiving information concerning that } meeting. I keep checking the meeting homepage on the web site } (http://www.unisi.it/ricerca/dip/bio_evol/sitoeuraac/siena2000.html), } but very little information is given. I also contacted the e-mail } address (euraac2000-at-unisi.it) a few months back and have not yet } received a reply. I wrote to Dr. Leo P. S. van der Geest, the EURAAC } Secretary, yesterday and he gave me your e-mail address and thought } perhaps you could help. } } I would like to present a paper at the meeting (probably a poster) } and need to know when titles and abstracts are due and to whom they } should be sent. Also the format that should be used for submitting } the paper. } } Many thanks for your help; it is greatly appreciated. I look forward } to hearing from you. } } Sincerely, } } Norm } } Norm Fashing } Professor of Biology } Department of Biology } College of William and Mary } P.O. Box 8795 } Williamsburg, VA 23187-8795 } 757 221-2221 (Office) } 757 221-6483 (FAX) } njfash-at-facstaff.wm.edu } http://www.wm.edu/biology/Fashing.html
dr Enrico de Lillo Istituto di Entomologia agraria - Universit Bari - Italy via Amendola, 165/A - 70126 Bari - Italy tel. +39 080 5443105 fax +39 080 5442876 email: delillo-at-agr.uniba.it http://193.204.185.103/de_lillo.htm
The power supply in our cryo microtome is having problems which might be related to the transformer. I was told by the service engineer that the transformer is no longer supported by Reichert. Does any one know where I can get a replacement ?
Dear Nestor, Thanks for taking such good care of us. Lee
Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Confocal Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
Anyone have any recommendations for organizing digital images and associated data in electronic form in a GLP environment? I'm going crazy trying to find something that will be easy to use, will link the image with associated data information, and will have restriction or audit capabilities to track any changes to the data that may be made.
Any input is greatly appreciated.
Gregg Sobocinski Parke-Davis Pharmaceuticals Ann Arbor, Michigan USA Gregg.Sobocinski-at-wl.com
Contact Robin Griffin at UAB. I bought that unit there and we worked on 718 and developed the polishing conditions for it. I believe that we used the butyl cellusolve(SP?)/perchloric acid solution at low temp to do it. She should have the recipe or know who to contact. Her Email address is rgriffin-at-eng.uab.edu or you might get a hold of Ray Thompson whose student did the work. As I recall, the as-cast material was very difficult to do and had very narrow conditions. The annealed samples were a little easier. To save time, we had the samples initially cut out of the bulk samples using an EDM machine.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies." --
} -----Original Message----- } From: Schryvers Dominique [mailto:schryver-at-ruca.ua.ac.be] } Sent: Friday, January 21, 2000 2:53 AM } To: Microscopy MAIL } Subject: Inconel 718 polish } } } -------------------------------------------------------------- } ---------- } 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'm looking for a good electropolishing solution + conditions for } as-received and annealed Inconel 718 for use with a Tenupol 3 } system to } produce well thinned matrix + precipitates for TEM work. Any } suggestions? } } Nick Schryvers } } } *=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=* } *=* *=* } *=* Dr. D. Schryvers *=* } *=* Electron Microscopy for Materials Research (EMAT) *=* } *=* University of Antwerp, RUCA *=* } *=* Groenenborgerlaan 171 *=* } *=* B-2020 ANTWERP *=* } *=* Belgium *=* } *=* tel: 32-3-2180247 *=* } *=* fax: 32-3-2180257 *=* } *=* e-mail: schryver-at-ruca.ua.ac.be *=* } *=* homepage: http://www.ruca.ua.ac.be/EMAT *=* } *=* *=* } *=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=* } }
The short answer to you question would be an image of about 1024 pixels across should be adequate for a slide used for a presentation. The slide will appear to most people as a 8x10"-print held at arm's length, or as a computer screen at 4 to 6 feet. If you cannot make out the pixels in those images, then you probably have enough pixels in your image for PowerPoint.
I think I start seeing the pixels when the resolution drops to 800 or 640 pixels across. I might see a benefit in raising the pixels to 1280 across, but I am not able to see improvement beyond that point. This would give you an image of about 1 million pixels.
Now if you are shooting your slide with a 35-mm camera, it might be good to use 24-bit color on the image if your output device (e.g., printer) can well render it. However, since the slide is only for a presentation, you can probably get by with much less color depth if file size per slide is an issue. I venture to say that 8-bit color at 1024 pixels across is plenty adequate for most presentations.
Remember, the above considerations are only for images for slide presentations. If the slides are meant to archive the images for other purposes, then you probably want every bit of resolution and color depth that your technology allows and justifies.
Warren
At 02:18 PM 1/21/2000 -1000, you wrote: } Hello, all- } } I'm hoping to hear from those of you who have worked out the optimum size } and resolution to make images in e.g., Photoshop that are destined for } PowerPoint to be made into transparencies. An image that is 4 x 5 inches } and 400-600 dpi seems to be overkill for a 35mm slide. } } Your opinions? } } Mahalo, } Tina
Periodically questions appear on this list concerning the use of cryostats for cutting histological sections. I usually reply to the sender off line and offer a copy of a handout that I got from a workshop at a Histochem Meeting some years ago. Even though it is old, cryostat sectioning has not changed a lot. I think there is some valuable info there, especially for beginners. I thought it might be helpful to make this available on the net for whomsoever might want to take a look. It can be found at : http://www.biotech.ufl.edu/sems/
Look for the snowflake
It was written by Bruce Quinn, then of MIT. I hope he has no objections to my posting it.
Greg Erdos Gregory W. Erdos, Ph.D. Ph. 352-392-1295 Assistant Director, Biotechnology Program PO Box 110580 Fax: 352-846-0251 University of Florida Gainesville, FL 32611
A full time position is available immediately for a highly motivated individual to work in the Center for C. elegans Anatomy at the Albert Einstein College of Medicine, located in the Bronx, New York. The candidate should have a Bachelor's degree in Biology or some related science, and some previous laboratory experience. We are particularly looking for an individual with training in transmission electron microscopy and thin section microtomy. Experience with immunocytochemistry and/or computerized image analysis is helpful but not required.
The College offers a generous compensation package including 4 weeks vacation and tuition reimbursement. Qualified candidates should submit a resume and a list of references to:
Dr. David Hall Department of Neuroscience Albert Eistein College of Medicine 1410 Pelham Parkway Bronx, NY 10461
Qualifications (education, certification, language, etc.) and Experience required: A candidate with a BS or MS or PHD degree in physical science is preferred. Prior experience in electron probe microanalysis is essential.
Job Overview: The Surface/Microscopy/X-ray (SMX) Group in Dow's Corporate R&D Analytical Science Laboratory has one professional level full time opening for an electron microprobe analyst. The candidate should have theoretical and practical experience in electron beam techniques, including quantitative x-ray microanalysis, digital imaging, digital x-ray imaging, electron beam/solid interactions, scanning electron microscopy and material science. Good computer skills are very desirable. Good written and oral communication skills and the ability to work both independently and in a team environment are extremely important.
Key responsibilities will include: 1. Extensive problem solving on a wide variety of Dow materials and processes 2. Operation and routine maintenance of a CAMECA SX-50 electron microprobe. 3. Some sample preparation including microtomy and metallography 4. Operation of light microscopes. 5. Operation of scanning and transmission microscopes as needed. 6. Interpretation of images. 7. Documentation of work. 8. Compliance with safety and quality systems
Interested: Please e-mail or send your resume and cover letter, with reference to this ad to: Email: R&D-at-Dow.com or The Dow Chemical Company, Workforce Planning, P. O. Box 150, Plaquemine, Louisiana 70765-0150. E-mail respondents must list Job 006145 and their last name as the first and second items on the Subject line. Only those selected for an interview will be contacted. Only U.S. citizens or aliens who are authorized to work in the United States will be considered for employment.
We are an equal opportunity employer and offer a competitive compensation and benefits package including 401k, stock purchase, tuition reimbursement and performance incentives. The Dow Chemical Company is the fifth largest chemical company in the world with annual sales of US$20billion. Dow manufactures and supplies chemicals, plastics and agricultural products for customers in 164 countries and employs approx. 43,000 people worldwide. For more news and information about Dow, please visit our web site at www.dow.com.
Robert C. Cieslinski The Dow Chemical Company Microscopy & Microanalysis (517) 636-6875 email: rccieslinski-at-dow.com
We successfully import our digital images into a PDF file using Adobe Acrobat.
Harry Ekstrom
-----Original Message----- } From: Sobocinski, Gregg [mailto:Gregg.Sobocinski-at-WL.com] Sent: Monday, January 24, 2000 8:35 AM To: 'Microscopy-at-MSA.Microscopy.Com'
Anyone have any recommendations for organizing digital images and associated data in electronic form in a GLP environment? I'm going crazy trying to find something that will be easy to use, will link the image with associated data information, and will have restriction or audit capabilities to track any changes to the data that may be made.
Any input is greatly appreciated.
Gregg Sobocinski Parke-Davis Pharmaceuticals Ann Arbor, Michigan USA Gregg.Sobocinski-at-wl.com
Does anyone or any Company know of a CCD that will do single photon detection at 852 wavelength? This is for a very specialized app. Thanks in advance
************************************************ Robert J. Derby New Mexico Institute of Technology Socorro, N.M. Phone - 505-835-5866 E-mail - rjderby-at-excite.com ************************************************
_______________________________________________________ Get 100% FREE Internet Access powered by Excite Visit http://freeworld.excite.com
first, I would like to wish you all the best for the new year. Now my question: Does anyone know the shelf-life/usability of LR-Gold stored in a fridge?
Is it that delicate as LR-White (Meanwhile I don«t use LR-W older than half a year). My bottle which was not opened many times, could be roundabout three years old.
Thanks a lot, Michael
Michael Reiner Department of Anatomy I University of Cologne Germany michael.reiner-at-smail.uni-koeln.de
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Dear all,
we're starting a project of HRTEM on small (nanoscale) colouring metal particles in glass. Does anyone have experience with sample preparation for this type of material. We prefer ion-milling as this retains the relative positions of the particles with respect to the surface. Any suggestions, also on literature, are welcome.
Many thanks in advance,
Nick Schryvers
*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=* *=* *=* *=* Dr. D. Schryvers *=* *=* Electron Microscopy for Materials Research (EMAT) *=* *=* University of Antwerp, RUCA *=* *=* Groenenborgerlaan 171 *=* *=* B-2020 ANTWERP *=* *=* Belgium *=* *=* tel: 32-3-2180247 *=* *=* fax: 32-3-2180257 *=* *=* e-mail: schryver-at-ruca.ua.ac.be *=* *=* homepage: http://www.ruca.ua.ac.be/EMAT *=* *=* *=* *=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*
by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id IAA06203 for dist-Microscopy; Tue, 25 Jan 2000 08:53:13 -0600 (CST) Received: from no_more_spam.com (sparc5.microscopy.com [206.69.208.10]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id IAA06200 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Tue, 25 Jan 2000 08:52:42 -0600 (CST) Received: from snarl.biotech.ufl.edu (snarl.biotech.ufl.edu [128.227.60.109]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id IAA06193 for {Microscopy-at-sparc5.microscopy.com} ; Tue, 25 Jan 2000 08:52:31 -0600 (CST) Received: from empc1 by snarl.biotech.ufl.edu (8.8.5/4.09) id JAA02445; Tue, 25 Jan 2000 09:47:10 -0500 (EST) Message-Id: {4.2.0.58.20000125094650.00a545f0-at-biotech} X-Sender: gwe-at-biotech X-Mailer: QUALCOMM Windows Eudora Pro Version 4.2.0.58
Some people have trouble and some do not. It is an Acrobat . After you get the blank screen, try hitting the reload button on your browser menu. This has worked for some people. I need to consult a web expert to see why my PDF files cause trouble.
Nestor, are you out there?????
If you still have trouble, let me know and I will put it into an HTML file. My apologies to anyone having trouble.
Greg Erdos
At 09:39 AM 01/25/2000 -0500, you wrote: } Dear Greg; } I was most interested to look at your tips etc. for cryo sectioning, } but when I clicked on the snowflake, all I ended up with was a blank } screen. Any idea what I did wrong (or is my computer system to blame?) } } thanks in advance } shea } } } } Dr. S. Shea Miller } Agriculture and Agri-Food Canada } Eastern Cereal and Oilseed Research Centre } 2068 K.W. Neatby Bldg } Central Experimental Farm } Ottawa, Ontario } Canada K1A 0C6 } email: millers-at-em.agr.ca } phone: 613-759-1760 } fax: 613-759-1701
Gregory W. Erdos, Ph.D. Ph. 352-392-1295 Assistant Director, Biotechnology Program PO Box 110580 Fax: 352-846-0251 University of Florida Gainesville, FL 32611
Your best source of advice would be Scott Walck, at these contacts:
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
Scott has been in the 'TEM of glass' business for years and has developed a series of techniques relevant to the preparation of cross-sections of this material, which I assume you require when you say, "we prefer ion-milling as this retains the relative positions of the particles with respect to the surface." As Scott will probably tell you, there is a small-angle cleaving technique that you may find preferable to ion milling.
Cheers John
John P. McCaffrey National Research Council of Canada M-50, Montreal Rd. Ottawa, Ontario K1A 0R6 CANADA
-----Original Message----- } From: Schryvers Dominique [mailto:schryver-at-ruca.ua.ac.be] Sent: Tuesday, January 25, 2000 9:51 AM To: Microscopy MAIL
Dear all,
we're starting a project of HRTEM on small (nanoscale) colouring metal particles in glass. Does anyone have experience with sample preparation for this type of material. We prefer ion-milling as this retains the relative positions of the particles with respect to the surface. Any suggestions, also on literature, are welcome.
Many thanks in advance,
Nick Schryvers
*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=* *=* *=* *=* Dr. D. Schryvers *=* *=* Electron Microscopy for Materials Research (EMAT) *=* *=* University of Antwerp, RUCA *=* *=* Groenenborgerlaan 171 *=* *=* B-2020 ANTWERP *=* *=* Belgium *=* *=* tel: 32-3-2180247 *=* *=* fax: 32-3-2180257 *=* *=* e-mail: schryver-at-ruca.ua.ac.be *=* *=* homepage: http://www.ruca.ua.ac.be/EMAT *=* *=* *=* *=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*
we're starting a project of patterning on BG of YBCO films. I got difficulties to find the BG by using opital microscope, because we can not etch the sample before patterning. Does anyone have experience with checking the GB by OM? We appraciate any suggestions or references.
University of Connecticut Institute for Materials Science
Postdoctoral Research Position in Electron Microscopy
The Institute for Materials Science (IMS) at UConn is an interdisciplinary center with the threefold mission of fostering education, research and outreach in all areas of the materials sciences. Due to a re-organization of the IMS Microscopy Unit, a Postdoctoral Position has become available in the area of transmission electron microscopy. The appointee will be involved in a range of academic and industrial projects, and will assist in developing the TEM facilities. Candidates should hold a PhD in Materials Science, Physics or a related discipline and must have extensive hands-on experience in a broad range of electron microscopy techniques. Experience in instrument development and/or computer image processing/simulation would be beneficial. The appointment is for one year in the first instance and is available immediately. Screening of the applications will begin immediately and will continue until the post is filled. Applications from under-represented groups, including minorities, women and people with disabilities are encouraged.
Interested candidates should send a curriculum vitae, including publication list, and the names of at least three referees with postal addresses, telephone numbers and Email addresses to: Prof. M. Aindow, Institute for Materials Science, University of Connecticut, 97 North Eagleville Road, U-3136, Storrs, CT 06269-3136 USA. Email: maindow-at-ims.uconn.edu
I recently asked Michael Davidson of FSU (http://microscopy.fsu.edu/) if he had any success running the QX3 on a Mac w/ USB and he said "It works with a windows 98 emulator, but very very slowly."
Several people have had success running the QX3 with twain drivers from other programs such as Paint Shop Pro and Photoshop (http://clubs.yahoo.com/clubs/qx3). Does anybody know of a twain driver that would run the QX3 from a Mac?
Position Title: (Technical-Level) Scientist-Electron Microscopy
Company: Alcon Laboratories, Inc., a wholly owned subsidiary of Nestle, S.A., has 10,000 employees and worldwide sales of $2.4 billion. It is the global leader in the discovery, development, manufacture and marketing of ophthalmic pharmaceuticals and medical devices. Alcon expects to double its sales over the next five years and has achieved a profit growth rate of approximately 11% over the last several years. Historically, the company commits 10% of sales to research and development. Products developed in the last ten years generate 50% of current sales. The current product pipeline is strong. Alcon was just renamed to the Fortune List of the 100 Best Companies to Work for in America.
Location: Fort Worth was recognized by USA Today as one of the 20 best cities in which to live and work.
Position Responsibilities: The EM Unit is a core resource for R&D, witnessed by the fact that the staff of three generated 9,350 electron micrographs from 1,160 processed specimens in 1998 alone. The successful candidate will be a key member of the EM Unit who processes, examines and provides preliminary interpretation of ophthalmic devices as well as human and animal tissue specimens from a wide variety of R&D groups. S/he will provide electron microscopic research and method development directed towards the discovery of new drug candidates and unique ophthalmic devices, the understanding of pathogenic mechanisms and the identification of new therapeutic agents. S/he will handle the EM Unit commitment to several groups: Surgical-Intraocular Lens, Toxicology, and Degenerative Diseases, as well as contributing to Glaucoma Therapeutic Research, Surgical, Formulations, Consumer Technical Support, Physical Characterization and Pharmacokinetics.
Responsibilities include preparing ophthalmic devices for SEM and x-ray analysis and human and animal tissue for TEM and SEM; use and daily maintenance of Zeiss CEM-902, Cambridge Stereoscan-120 and Zeiss DSM-940, and PGT System 4+ x-ray system; developing and implementing EM techniques for various research projects; assisting with human and animal tissue procedures; and providing preliminary interpretation of EM data.
Preferred Qualifications: Candidates should possess a Bachelor of Science degree in a related discipline plus at least seven years of significant EM experience related exclusively to human and animal tissue. Collaborative and problem solving skills are essential for this position. The successful candidate will also demonstrate highly refined interpersonal and technical writing skills. Certification by or eligibility to be certified by the MSA is a plus.
Alcon professionals enjoy state-of-the-art facilities in a year-round business casual environment. Our company offers competitive salaries and a wide array of excellent benefits: a very generous retirement plan and dollar-for-dollar matching (up to 5%) 401K, medical, dental, vision, life, and accident insurance, death, dismemberment, illness and disability benefits, tuition reimbursement, employee credit union, adoption assistance, dependent care, and wellness programs, on-site fitness center, running track, cafeteria, and company store, innovative paid time off and holidays, and retiree medical coverage.
An Equal Opportunity and Affirmative Action Employer. Pre-employment drug testing.
Please email your resume and salary requirements to: Job28_1261-at-careers.alconlabs.com Reference Code: EM
Apparently several were unable to read the cryostat technique pdf file that Dr. Greg Erdos had generously posted at his website. The answer to the problem could be the version of Acrobat used. I had the "blank page" problem with version 3.0 but no problem at all with version 4.0 (Mac).
} } Aloha, } Tina } } It's raining cats and dogs, and it's cold and miserable. Feel better, } now? }
But what you call cold and what we call cold are two very different animals! Its been snowing all day in the Northeast, and the temps haven't nosed above (or even near) freezing) in days!
Dear Lee, Fair is fair. Often wind and rain at slightly above freezing is more miserable than snow at a few degrees below--having been both in San Francisco for the first condition and Albany NY for the second, I can say this with first-hand authority. Yours, Bill Tivol
Appropriate vendors, I have a Kevex detector which has been giving peak widths larger than specs. It probably just needs an overhaul, but there may be some repair necessary for the pre-amp and FET. Could anyone who can undertake this please respond to me off-list with estimates for various contingencies? TIA. Yours, Bill Tivol
Our Ohio company is seeking an engineer / scientist to research the relationships between the chemistry and microstructure of solid lubricant and hard coatings and their performance in the lubrication of aerospace systems. Research will involve a variety of surface analytical tools (XPS, Raman, etc.) so that fundamental mechanisms of lubrication can be elucidated. Emphasis on microstructure will require expertise with TEM and SEM including preparation of SEM & TEM specimens of thin films and wear scars on steel and ceramic substrates. Research will also involve correlating thin film properties with deposition plasma characteristics and making recommendations for improving lifetime and performance of such materials in different environments: e.g., vacuum, moist air, high temperature, etc.
It is important that candidates have capabilities in cross-section TEM, analytical TEM, analysis of unique microstructures; and understand TEM of thin films on a fundamental level. It is desirable that the candidate have knowledge of tribological materials and experience with TEM/XTEM of wear tracks.
Contact Ronald Decker - mailto:decker-at-utcdayton.com
{} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} Ronald C. Decker Program Manager Universal Technology Corporation 1270 N FAIRFIELD RD DAYTON OH 45432-2600
Voice (937) 426-8530, Fax (937) 426-7753 (Voice mail is available at my extension, 270)
While browsing news from the latest MacWorld Expo I came across a brief comment about a USB video microscope for the Mac. Further searches at the MacWorld Expo website or at Apple's web site have not turned up anything more about it, although there were announcements that Data Translation and National instruments have released additional PCI and USB I/O boards for the Mac. Has anyone heard more of this?
Regards, Glen
Glen MacDonald Research Scientist Hearing Research Laboratories of the Virginia Merrill Bloedel Hearing Research Center Box 35-7923 University of Washington Seattle, WA 98195-7923 (206) 616-4156 glenmac-at-u.washington.edu
High Resolution Scanning Electron Microscopist/Engineer
United Technologies Research Center is seeking an engineer to fill the HR-SEM operator/engineer position at the United Technologies Research Center in East Hartford, CT. This position will provide support to the United Technologies Corporation Business Unites including Pratt & Whitney, Carrier, Sikorsky, Hamilton Sundstrand and Otis. The SEM operator/engineer will be responsible for the full utilization of both high-resolution secondary and back-scattered imaging to characterize a wide range of metallic and non-metallic materials, including surface coatings and advanced structural materials including metals and ceramics. In addition, the ability to recognize fracture modes and origins of fractures is strongly desired. The candidate should be experienced in the use of EDS for both qualitative and quantitative analyses, including compositional mapping and line profiles. The qualified candidate must be capable of judging the optimal combinations of imaging and EDS to yield t! he most informative characterization of a particular specimen. Good communication and interpersonal skills are essential. Experience with electron backscatter diffraction (EBSD) is a plus.
Qualified candidates will have BS in Materials Science or an equivalent discipline, with a minimum of 2 years SEM experience. U.S. citizenship or permanent resident status is required.
Please visit our web site at http://www.utrc.utc.com for additional general information. Interested parties should send a letter of application and a resume to Employment Opportunities, Code MATS-2050-9049, United Technologies Research Center, 411 Silver Lane, East Hartford, CT 06108 or e-mail employment-at-utrc.utc.com. United Technologies Research Center is an equal opportunity employer.
I tried to open the file with Acrobat 4.0 in Windows 98. No luck.
Damian Neuberger etc., etc.
} Apparently several were unable to read the cryostat technique pdf file that } Dr. Greg Erdos had generously posted at his website. The answer to the } problem could be the version of Acrobat used. I had the "blank page" } problem with version 3.0 but no problem at all with version 4.0 (Mac).
We have an ISI SS40 SEM that is in need of a discontinued part. It is a NEC transistor, number D588. It is used in the filament current control circuit. I'm having trouble locating the part because it has not been manufactured by NEC since 1984. Does anyone know of a source for this part or a substitute transistor?
Thanks in advance,
Bill Carmichael
______________________________________ Bill Carmichael Electron Microscopy Faculty Madison Area Technical College 3550 Anderson St. Madison, WI 53704 608-243-4309 wcarmichael-at-madison.tec.wi.us
Dominique, Glass is readily microtomed with a diamond knife and may be a suitable inexpensive technique for you to consider. Particularly if the materials of your sample are sufficiently dissimilar in reaction to the chemical and ion etching of some techniques. I've been embedding and sectioning coated glass, optics, and other hard materials for 18 years (even diamond coated silicon in Microscopy Research and Technique, vol. 31, p. 308, 1995). The imaging and analysis of nano-structures in micron sized areas near the surface of glass is routine, fast, and inexpensive for physical microstructure and chemistry. Mechanical artifacts generated in ultramicrotomy tend to be quite large, readily visible, and easily ignored but may interfere with the analysis of naturally occurring deformation features (i.e. twinning, slip, etc.). Any good diamond knife will work with meticulous and careful technique, but experience has shown that 35 degree knives yield the best results with hard and ultra-hard materials.
The critical elements for microtomy of hard, non-porous materials include: 1. Minimize the cross-sectional area to be sectioned. An easy way is to do this is to pop concoidal micro-chips from the surface. These tend to be very thin at the edges and may be further broken to form very pointed thin samples. [Time = ~20 minutes] 2. Optimize sample orientation for sectioning and preferred orientation. Some physical microstructures are anisotropic and are difficult to interpret when viewed in the wrong orientation. [Time = ~10 minutes] 3. Maximize adhesion to the resin through the selection of an appropriate resin (low viscosity and non-reactive with your sample), meticulous and contamination-free sample prep, and the addition of adhesion promoters (such as Dow Corning Z-6040). [Time = ~1 hour, with an over night epoxy cure] 4. Section using standard procedures, but minimize the sectioning speed (optimize cutting speed). [Time = ~1 hour]
These times are approximate for 1 sample, and there could be economy in numbers. As always, each case will require individual attention. Cheers,
Phil Swab Engineering Development Deposition Sciences Inc. Santa Rosa, CA 707-566-3718 phil.swab-at-depsci.com
-----Original Message----- } From: Schryvers Dominique [SMTP:schryver-at-ruca.ua.ac.be] Sent: Tuesday, January 25, 2000 6:51 AM To: Microscopy MAIL
Dear all,
we're starting a project of HRTEM on small (nanoscale) colouring metal particles in glass. Does anyone have experience with sample preparation for this type of material. We prefer ion-milling as this retains the relative positions of the particles with respect to the surface. Any suggestions, also on literature, are welcome.
Many thanks in advance,
Nick Schryvers
*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=* *=* *=* *=* Dr. D. Schryvers *=* *=* Electron Microscopy for Materials Research (EMAT) *=* *=* University of Antwerp, RUCA *=* *=* Groenenborgerlaan 171 *=* *=* B-2020 ANTWERP *=* *=* Belgium *=* *=* tel: 32-3-2180247 *=* *=* fax: 32-3-2180257 *=* *=* e-mail: schryver-at-ruca.ua.ac.be *=* *=* homepage: http://www.ruca.ua.ac.be/EMAT *=* *=* *=* *=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*
we are analyzing frequently this kind of specimens. In our case the particles are silver. Depending on density and size distribution they are changing the color of the glass. Since we are interested in the depth distribution starting at the interface with a silver containging layer on the glass, we need to do a cross-sectional preparation. Therefore, we use several steps of preparation as described below:
á gluing of two coated glass surfaces face to face
á cutting and polishing of the obtained block to a size of 2 x 1 x 10 mmÒ with the interface plane running parallel to the 2 mm x 10 mm face and in the middle of the block
á gluing of this block in a steel-cylinder of 3 mm diameter and 10 mm length with a 1 mm x 2 mm rectangular hole along the cylinder axis
á cutting of 0.5 - 1 mm thick slices perpendicular to the cylinder axis
á fine polishing of one face of the obtained disk (lowest grain size: 1µm)
á grinding of the disk from the opposite face down to 100 µm thickness with subsequent fine polishing
á dimpling of a crater into one face of the disk with a residual thickness in the middle of the disk of about 10 µm
á ion etching of the flat side of the sample with argon ions of 4 kV under an angle of 2 - 4 degrees with a current of about 12 µA until electron transparency is reached in the region of the interface.
Some of the results were presented on the FEMMS-Meeting in Irsee, Germany, 1998. For more details, you might contact me directly.
Hope this helps,
Petra
At 15:50 25.01.00 +0100, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-------------------------------------------------------------- Dr. Petra Wahlbring Centre de Recherche Public - Gabriel Lippmann Laboratoire d'Analyse des Materiaux (LAM) 162a, av. de la Faiencerie L-1511 Luxembourg tel. +352-466644-402 fax +352-466644-400 e-mail: petra.wahlbring-at-crpgl.lu Visit our WWW site! http://www.crpgl.lu/~wahlbrin
Can anybody offer a method for the preparation of Halophilic bacteria for 'standard' SEM observation? post fixation washing appears to produce cell lysis!.
For those who are unable to view the PDF file of Cryostat information that I posted, I have also posted it in ugly HTML. I am still trying to find out why some can read the PDF and others cannot. The Version of Acrobat does not seem to be the answer.
My apologies to anyone who got frustrated. Once again the site is: www.biotech.ufl.edu/sems/
Greg Erdos Gregory W. Erdos, Ph.D. Ph. 352-392-1295 Assistant Director, Biotechnology Program PO Box 110580 Fax: 352-846-0251 University of Florida Gainesville, FL 32611
} } } {"wft03-at-health.state.ny.us"-at-sparc5.Microscopy.Com} 01/25 3:14 PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } Aloha, } Tina } } It's raining cats and dogs, and it's cold and miserable. Feel better, } now? }
But what you call cold and what we call cold are two very different animals! Its been snowing all day in the Northeast, and the temps haven't nosed above (or even near) freezing) in days!
Dear Lee, Fair is fair. Often wind and rain at slightly above freezing is more miserable than snow at a few degrees below--having been both in San Francisco for the first condition and Albany NY for the second, I can say this with first-hand authority. Yours, Bill Tivol
} } } {"wft03-at-health.state.ny.us"-at-sparc5.Microscopy.Com} 01/25 3:14 PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } Aloha, } Tina } } It's raining cats and dogs, and it's cold and miserable. Feel better, } now? }
But what you call cold and what we call cold are two very different animals! Its been snowing all day in the Northeast, and the temps haven't nosed above (or even near) freezing) in days!
Dear Lee, Fair is fair. Often wind and rain at slightly above freezing is more miserable than snow at a few degrees below--having been both in San Francisco for the first condition and Albany NY for the second, I can say this with first-hand authority. Yours, Bill Tivol
It would be convenient to be able to bring a video camera into an elementary school classroom, hook it up to a microscope (with an eyepiece adaptor) and display the microscope image for an entire classroom to see. If there is a video monitor (or a TV with a video input), this is fairly easy. If there is not an available monitor, I should be able to bring in a laptop and display the image on the laptop screen.
I am looking for an inexpensive lightweight video camera (with a C-mount) with either a firewire or USB linkage.
Does anyone have any suggestions?
Thanks
Peter Guthrie Department of Neurobiology & Anatomy University of Utah School of Medicine 50 N Medical Drive Salt Lake City, UT 84132 (801) 581-8336 (801) 581-4233 fax Peter.Guthrie-at-hsc.utah.edu
DearBill, When my Kevex detector showed degraded resolution, I turned off the bias and grounded the BNC plug with a paper clip, then warmed it up completely to get rid of ice and frost in the detector. This brought back my resolution, but degraded my LN2 holding time. Then I had a friend in Physics pump out the dewar and now I'm back to peak performance. 04:03 PM 1/25/00 -0500, you wrote: } Appropriate vendors, } I have a Kevex detector which has been giving peak widths } larger than specs. It probably just needs an overhaul, but there may } be some repair necessary for the pre-amp and FET. Could anyone } who can undertake this please respond to me off-list with estimates } for various contingencies? TIA. } Yours, } Bill Tivol 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
Recently, an interesting medical school project has appeared at my lab door. I've been asked to quantify Ca and P in scar tissue found in sections of hamster hearts. Obtaining appropriate standards is critical. Are there commercially available biological standards or procedures to create such standards out there? Any suggestions would be appreciated and thanks for your time.
Dan
===================================================== Dan Kremser Washington University Department of Earth and Planetary Sciences/Campus Box 1169 (Wilson Hall, Room 108-----for packages) One Brookings Dr. St. Louis, MOÊ 63130-4899
How about using a device like Dazzle or Snappy to display the video stream from your video camera on a laptop? It would require a little software setup but should work. The models I am familiar with used parallel port connections, but there ought to be models around that would use USB.
At 09:28 AM 1/26/2000 -0700, you wrote: } It would be convenient to be able to bring a video camera into an } elementary school classroom, hook it up to a microscope (with an eyepiece } adaptor) and display the microscope image for an entire classroom to see. } If there is a video monitor (or a TV with a video input), this is fairly } easy. If there is not an available monitor, I should be able to bring in a } laptop and display the image on the laptop screen. } } I am looking for an inexpensive lightweight video camera (with a C-mount) } with either a firewire or USB linkage. } } Does anyone have any suggestions? } } Thanks
Listservers, Can anyone lead me to a good reference for processing human sperm for TEM? Or if you have a procedure can you please forward the details. TIA Hank Adams Integrated Microscopy Core Molecular and Cell Biology Baylor College of Medicine Houston, TX
Stefanini M, De Martino C, Zamboni L. 1967. Fixation of ejaculated spermatozoa for electron microscopy. Nature 216:173-174.
Meschede D, Keck C, Zander M, Cooper TG, Yeung CH, Nieschlag E. 1993. Influence of three different preparation techniques on the results of human sperm morphology analysis. Int J Androl 16:362-369.
Phillips DM. 1995. Fixation of mammalian spermatozoa for electron microscopy. In: Dentler W, Witman G (Eds.), Methods in Cell Biology, Vol 47, vol 47. Academic Press Inc (San Diego), pp 199-204.
Kent
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ A. Kent Christensen, Professor Emeritus Department of Cell and Developmental Biology, Medical Science II Building University of Michigan Medical School, Ann Arbor, MI 48109-0616 Office: 2703A Medical Science II Building Tel (work) (734) 763-1287, Fax (work) (734) 763-1166 E-mail: akc-at-umich.edu http://www.umich.edu/~akc/ ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
} Listservers, } Can anyone lead me to a good reference for processing human sperm for } TEM? Or if you have a procedure can you please forward the details. TIA } Hank Adams } Integrated Microscopy Core } Molecular and Cell Biology } Baylor College of Medicine } Houston, TX
If you intend to use a "video" camera, you may need a capture card rather than, or in addition to, a USB or Firewire interface. Most video cams are analog (NTSC in the US, SECAM or PAL elsewhere in the world). Others can certainly have digital circuits internal (like the new digital camcorders or the PCcams used for video conferencing on the net). ...You first need to determine the source format.
Warren's suggestion implements an inexpensive external NTSC* video frame grabber (Snappy). Which will "grab" an analog video frame and digitize it. *May do PAL too??
Parallel port I/O is a bit slow (or is that a byte slow :) for pictures containing lots of data, but is cheap and works since the inherent resolution of typical NTSC video is { 640x480. USB is very much faster and Firewire faster yet (and usually a lot more $$ for the interface card). For applications other than full frame rate/resolution streaming video, USB is fine.
They have a USB version and the software interface is great. It is easy to use and has a street price of about $180!
Good Luck,
Lawrence Kordon Nikon, Inc. Columbia, Maryland nikon-at-jagunet.com
***I have no affiliation with Dazzle, Inc.***
"White, Woody N" wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Peter, } } If you intend to use a "video" camera, you may need a capture card rather } than, } or in addition to, a USB or Firewire interface. Most video cams are analog } (NTSC in the US, SECAM or PAL elsewhere in the world). Others can certainly } have digital circuits internal (like the new digital camcorders or the } PCcams } used for video conferencing on the net). ...You first need to determine } the } source format. } } Warren's suggestion implements an inexpensive external NTSC* video frame } grabber } (Snappy). Which will "grab" an analog video frame and digitize it. *May do } PAL } too?? } } Parallel port I/O is a bit slow (or is that a byte slow :) for pictures } containing lots of data, but is cheap and works since the inherent } resolution of } typical NTSC video is { 640x480. USB is very much faster and Firewire } faster } yet (and usually a lot more $$ for the interface card). For applications } other } than full frame rate/resolution streaming video, USB is fine. } } Woody White
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} Please keep weather reports private. } } Ann Fook
Why? A little light heartedness never hurt anyone. For the record, LA was sunny as usual today. Happy I don't live in CT anymore.
Regards,
Paul Webster, Ph.D. Associate Scientist & Director Ahmanson Advanced Electron Microscopy & Imaging Center House Ear Institute 2100 West Third St. Los Angeles, CA 90057
by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id UAA11239 for dist-Microscopy; Wed, 26 Jan 2000 20:43:46 -0600 (CST) Received: from no_more_spam.com (sparc5.microscopy.com [206.69.208.10]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id UAA11236 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Wed, 26 Jan 2000 20:43:16 -0600 (CST) Received: from staff2.cso.uiuc.edu (staff2.cso.uiuc.edu [128.174.5.53]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id UAA11229 for {Microscopy-at-sparc5.Microscopy.Com} ; Wed, 26 Jan 2000 20:43:05 -0600 (CST) Received: from [130.126.25.46] (rochester-46.slip.uiuc.edu [130.126.25.46]) by staff2.cso.uiuc.edu (8.9.3/8.9.3) with ESMTP id UAA27358 for {Microscopy-at-sparc5.Microscopy.Com} ; Wed, 26 Jan 2000 20:38:21 -0600 (CST) Mime-Version: 1.0 X-Sender: lamiller-at-ux1.cso.uiuc.edu Message-Id: {v04210100b4b55eb12196-at-[130.126.26.199]}
I currently video with a Sony DV Video camera, fire wire connect to my computer with no capture board.
It does require special software however, We bought Final Cut Pro, But from trying out the demo and reading, It appears Adobe Priemier also will allow firewire capture without a board. Though Adobe's is one I have not tried, I'd call first.
This works fine for video, and I can pull off individual frames for low res images for the web, and ok small images to print if very small.
But, in emailing to and from Sony, It was my understanding that if I were to firewire images, I WOULD need a board.
My video camera will do both images and video. FinalCut Pro pulls off the video, but I can't seem to get it to recognize the individual image shots. So I believe Sony, though I may just not have selected the right buttons etc.
If pulling in video by fire wire, especially pulling it onto a firewire hard drive ( ie VST) It is pretty close to real time video.
Lou Ann *************************** Lou Ann Miller Service Supervisor Microscopic Imaging Lab College of Vet. Medicine University of Illinois 2001 S Lincoln Ave Urbana,Illinois 61801 217-244-1566 lamiller-at-ux1.cso.uiuc.edu
Microscopy Home Page: http://treefrog.cvm.uiuc.edu
Central States Microscopy Society http://treefrog.cvm.uiuc.edu/csmms
Personal Home Page: http://treefrog.cvm.uiuc.edu/lam
I did some TEM work on the extreme holophile bacterias (they grow in saturated NaCl solutions) some 30 years ago. Very difficult specimens, it seems no fixation is complete and can prevent osmotic shock.
I have not tried SEM on halophiles, but I suggest this: You could try excessive fixation, using 2 hours at room 20 degrees. I would use a several molar solution of ammonium acetate to rinse the specimen after fixation. Ammonium acetate is a volatile salt solution and leaves no crystals after evaporation. The still wet sample (mounted on a 10mm coverslip) could then be placed in a glass Petrie dish which has a double layer of filter paper, saturated with chloroform. Place the closed Petrie dish in the fridge for a day or two. Warm the dish before opening (to avoid condensation) and metal coat prior to SEM.
Ah, your first problem could be the fixation. The osmium (I'd forget GA), would need to go into the bacteria's growth medium, or use vapour fixation only. Even then, I expect much damage before any further processing. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Thursday, January 27, 2000 12:01 AM, Hyman, S.C. [SMTP:sch10-at-leicester.ac.uk] wrote: } } } Can anybody offer a method for the preparation of Halophilic bacteria for } 'standard' SEM observation? post fixation washing appears to produce cell } lysis!.
Moreover, it is well known that weather may affect the quality of EM samples. For instance, humidity is very critical for many EM techniques. I utilized very unusual technique for holey-film preparation with calcium-rhodanide. This technique is extremely sensitive for humidity/temperature combination. When I was working in Russia (without conditioner in the room), I was able predict the weather changes using that technique. Again, it was tricky to work when temperature in the room was around 7oC (at winter). The guys from East Coast may have something like that. Why not to share experience how to work at different weather conditions?
Have a good weather!
Sergey
} Date: Wed, 26 Jan 2000 17:52:26 -0800 } From: Paul Webster {pwebster-at-hei.org} } Subject: Re:No weather report please } To: MSA listserver submission {Microscopy-at-sparc5.Microscopy.Com} } Reply-to: Paul Webster {pwebster-at-hei.org} } X-Mailer: QuickMail Pro 1.5.4 (Mac) } X-MIME-Autoconverted: from quoted-printable to 8bit by ultra5.microscopy.com id } TAA11085 } } ------------------------------------------------------------------------ } 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
Dear Hank and others, I have had good success this Stefanini's buffered picric acid paraformaldehyde (PAF) for spermatozoan. I do not have the fixative formula at my fingertips, but the reference is: Nature Vol. 216, OCT 14. 1967.
I will look it up if you are interested and get back to you or you can email me-at- tiekotte-at-up.edu. -Ken
Ken Tiekotter Dept. of Biology The University of Portland 5000 N Willamette Blvd. Portland, OR 97303
On Wed, 26 Jan 2000, hank adams 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 } -----------------------------------------------------------------------. } } } Listservers, } Can anyone lead me to a good reference for processing human sperm for TEM? Or if you have a procedure can you please forward the details. } TIA } Hank Adams } Integrated Microscopy Core } Molecular and Cell Biology } Baylor College of Medicine } Houston, TX } }
} From: Greg Erdos {gwe-at-biotech.ufl.edu} } To: Microscopy-at-sparc5.microscopy.com } Subject: Cryostat info. } Date: Wed, 26 Jan 2000 09:19:58 -0500 } } ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } For those who are unable to view the PDF file of Cryostat information that } I posted, I have also posted it in ugly HTML. I am still trying to find } out why some can read the PDF and others cannot. The Version of Acrobat } does not seem to be the answer. } } My apologies to anyone who got frustrated. } Once again the site is: } www.biotech.ufl.edu/sems/ } } Greg Erdos } Gregory W. Erdos, Ph.D. Ph. } 352-392-1295 } Assistant Director, Biotechnology Program } PO Box 110580 Fax: } 352-846-0251 } University of Florida } Gainesville, FL 32611 }
I think that we are proceeding slightly tongue-in-cheek here - but to continue the 'serious' note;
The most weather sensitive task I have ever tackled is the production of formvar films. With the extremely high humidities we experience here there are times when the formvar will simply NOT separate from the substrate, despite 'air-conditioning'.
Tony
Tony Bruton Head, Centre for Electron Microscopy University of Natal, Pietermaritzburg Tel +27 (0) 33 260 5155 Fax +27 (0) 33 260 5776 website via:http:www.nu.ac.za Email:bruton-at-emu.unp.ac.za postal address; Private Bag X01, Scottsville, 3209 KwaZulu-Natal South Africa
} } } Sergey Ryazantsev {sryazant-at-ucla.edu} 01/27/00 10:31AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Moreover, it is well known that weather may affect the quality of EM samples. For instance, humidity is very critical for many EM techniques. I utilized very unusual technique for holey-film preparation with calcium-rhodanide. This technique is extremely sensitive for humidity/temperature combination. When I was working in Russia (without conditioner in the room), I was able predict the weather changes using that technique. Again, it was tricky to work when temperature in the room was around 7oC (at winter). The guys from East Coast may have something like that. Why not to share experience how to work at different weather conditions?
Have a good weather!
Sergey
} Date: Wed, 26 Jan 2000 17:52:26 -0800 } From: Paul Webster {pwebster-at-hei.org} } Subject: Re:No weather report please } To: MSA listserver submission {Microscopy-at-sparc5.Microscopy.Com} } Reply-to: Paul Webster {pwebster-at-hei.org} } X-Mailer: QuickMail Pro 1.5.4 (Mac) } X-MIME-Autoconverted: from quoted-printable to 8bit by ultra5.microscopy.com id } TAA11085 } } ------------------------------------------------------------------------ } 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
Can anyone tell me who currently makes and sells the retrograde neuronal tracer, Fluorogold? We have a customer who is confusing it with our FluoroNanogold products (not the first time this has happenned) and I would lkke to point them to the right source!
Thanks,
Rick Powell
********************************************************************** * NANOPROBES, Incorporated | US Toll-free: (877) 808-2101 * * 25 East Loop Road, Suite 113 | Tel: (919) 510-0590 * * Stony Brook, NY 11790-3350, | Fax: (919) 510-0590 * * USA | rpowell-at-lihti.org * * * * NOW EASY TO FIND ON THE WEB: http://www.nanoprobes.com * **********************************************************************
The main reference is: Stefanini et al., 1967, Nature 216:173.
Ramin Rahbari PARKE-DAVIS Pharmaceutical Research Worldwide Preclinical Safety 2800 Plymouth Road Ann Arbor, MI 48105 Voice (734) 622-3383 Fax (734) 622-5001 Ramin.Rahbari-at-WL.COM
-----Original Message----- } From: hank adams [mailto:hpadams-at-bcm.tmc.edu] Sent: Wednesday, January 26, 2000 2:39 PM To: 'microscopy-at-msa.microscopy.com'
Listservers, Can anyone lead me to a good reference for processing human sperm for TEM? Or if you have a procedure can you please forward the details. TIA Hank Adams Integrated Microscopy Core Molecular and Cell Biology Baylor College of Medicine Houston, TX
If anyone remembers I use to end all my e-mails to the listees with a quite sarcastic weather and/or olfactory report from the garden state. Either no one read my posts or they just didn't get the East Coast thing.
Cold, windy and something smells real odd under the Pulaski Skyway...Aloha!
John Grazul Lucent
Sergey Ryazantsev wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Moreover, it is well known that weather may affect the quality of EM } samples. For instance, humidity is very critical for many EM techniques. I } utilized very unusual technique for holey-film preparation with } calcium-rhodanide. This technique is extremely sensitive for } humidity/temperature combination. When I was working in Russia (without } conditioner in the room), I was able predict the weather changes using that } technique. Again, it was tricky to work when temperature in the room was } around 7oC (at winter). The guys from East Coast may have something like } that. Why not to share experience how to work at different weather } conditions? } } Have a good weather! } } Sergey } } } Date: Wed, 26 Jan 2000 17:52:26 -0800 } } From: Paul Webster {pwebster-at-hei.org} } } Subject: Re:No weather report please } } To: MSA listserver submission {Microscopy-at-sparc5.Microscopy.Com} } } Reply-to: Paul Webster {pwebster-at-hei.org} } } X-Mailer: QuickMail Pro 1.5.4 (Mac) } } X-MIME-Autoconverted: from quoted-printable to 8bit by } ultra5.microscopy.com id } } TAA11085 } } } } ------------------------------------------------------------------------ } } 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 } } -----------------------------------------------------------------------. } } } } } } } Please keep weather reports private. } } } } } } Ann Fook } } } } Why? A little light heartedness never hurt anyone. } } For the record, LA was sunny as usual today. Happy I don't live in CT } anymore. } } } } Regards, } } } } Paul Webster, Ph.D. } } Associate Scientist & Director } } Ahmanson Advanced Electron Microscopy & Imaging Center } } House Ear Institute } } 2100 West Third St. } } Los Angeles, CA 90057 } } } } Phone: (213) 273-8026 } } Fax: (213) 413-6739 } } e-mail: pwebster-at-hei.org } } http://www.hei.org/htm/aemi.htm } } } } } } } _____________________________________ } } Sergey Ryazantsev Ph. D. } Electron Microscopy } UCLA School of Medicine } Department of Biological Chemistry } Box 951737 } Los Angeles, CA 90095-1737 } } Phone: (310) 825-1144 } Pager: (310) 845-0248 } FAX (departmental): (310) 206-5272 } mailto:sryazant-at-ucla.edu } http://www.bol.ucla.edu/~sryazant
Molecular Probes sells hydroxystilbamidine, methanesulfonate (cat. # H-7599) which I beleive is the active chemical in Fluorogold. (see paper by Martin W. Wessendorf in Brain Res 553(1): 135-48. Jul 1991).
Karen Zaruba
P.S. I have no interest in Molecular Probes other than a satisfied customer.
Rick Powell at Nanoprobes 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 Light Microscopists: } } Can anyone tell me who currently makes and sells the retrograde neuronal } tracer, Fluorogold? We have a customer who is confusing it with our } FluoroNanogold products (not the first time this has happenned) and I would } lkke to point them to the right source! } } Thanks, } } Rick Powell } } ********************************************************************** } * NANOPROBES, Incorporated | US Toll-free: (877) 808-2101 * } * 25 East Loop Road, Suite 113 | Tel: (919) 510-0590 * } * Stony Brook, NY 11790-3350, | Fax: (919) 510-0590 * } * USA | rpowell-at-lihti.org * } * * } * NOW EASY TO FIND ON THE WEB: http://www.nanoprobes.com * } **********************************************************************
-- Karen S. Zaruba kszaruba-at-mmm.com 3M Company, St. Paul, MN
Tried to respond to a message posted here from Cynthia Shannon re: a used TEM, it keeps bouncing. Cynthia, if you are out there, how can I contact you?
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
ELECTRON MICROSCOPY TECHNICIAN The Integrated Microscopy Core, Department of Molecular and Cell Biology, Baylor College of Medicine is expanding and has an immediate full-time opening for an electron microscopy technician. The Integrated Microscopy Core is a growing, state-of-the-art facility with 2 TEMs, deconvolution, laser scanning confocal, 2 CCD-based fluorescent microscopes and multiple PC and Silicon Graphics workstations for imaging software. The applicant should have at least one year of experience in various aspects of sample preparation for biological TEM including fixation, embedding, ultrathin sectioning and staining. The applicant should have darkroom experience and experience in the operation of TEMs. Other duties include preparation of solutions, embedding media and the maintaining of records. The position offers excellent opportunities for training in advanced light and electron microscopy techniques, including immunofluorescence and immunogold labeling, laser scanning confocal and deconvolution microscopy, as well as live imaging of GFP-tagged proteins. Training in several image computer-based imaging programs will also be available (PhotoShop, MetaMorph, SoftWorks). The position requires a minimum of a Bachelors degree and will start as a Lab Technician II; salary will be commensurate with experience, and includes the standard Baylor benefits package.
Send CV and letter of research/technical interests to:
Hank Adams Laboratory Manager Integrated Microscopy Core Department of Molecular and Cell Biology Baylor College of Medicine One Baylor Plaza Houston, TX 77030 Email submissions to: hpadams-at-bcm.tmc.edu Fax submissions to: 713 790 0545
Baylor College of Medicine is an Equal Opportunity, Affirmative Action and Equal Access Employer.
I can't see how it can be netscape vs IE, since I got the blank page using IE4. I haven't tried netscape or IE5 (have both at home--but not here at work). Acrobat seems to work on every other PDF file I have opened (the intranet and internet standard here for public documents in a read-only setting), so I don't think the version of Acrobat is the problem either. I tried Dr. Erdos' original work-around, but still got the blank page. ????
On Thu, 27 Jan 2000 11:36:12 +0100, Shu-You Li wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Dr. Erdos, } } Perheps it is due to the difference between Netscape and IE. I got a blank } page with netscape but read it correctly with IE4.0. } } Shu-You Li } ************************************************** } Shu-You Li, Dr. } Institut fuer Physikalische Chemie } Johannes Guttenberg Universitaet } Jakob-Welder-Weg 11 } D-55099 Mainz, Germany } } E-mail: syli-at-mail.uni-mainz.de; syli16-at-hotmail.com } Fax: +49-6131-3923768 } Tel: +49-6131-3923148(O) } ************************************************** } } } } } From: Greg Erdos {gwe-at-biotech.ufl.edu} } } To: Microscopy-at-sparc5.microscopy.com } } Subject: Cryostat info. } } Date: Wed, 26 Jan 2000 09:19:58 -0500 } } } } ------------------------------------------------------------------------ } } 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 } } -----------------------------------------------------------------------. } } } } } } For those who are unable to view the PDF file of Cryostat information that } } I posted, I have also posted it in ugly HTML. I am still trying to find } } out why some can read the PDF and others cannot. The Version of Acrobat } } does not seem to be the answer. } } } } My apologies to anyone who got frustrated. } } Once again the site is: } } www.biotech.ufl.edu/sems/ } } } } Greg Erdos } } Gregory W. Erdos, Ph.D. Ph. } } 352-392-1295 } } Assistant Director, Biotechnology Program } } PO Box 110580 Fax: } } 352-846-0251 } } University of Florida } } Gainesville, FL 32611 } } }
Roger C Moretz, Ph.D.
_______________________________________________________ Get 100% FREE Internet Access powered by Excite Visit http://freeworld.excite.com
I think, there is some confusion out there about digital and analog cameras and different protocols and interfaces. Woody's posting is correct, but perhaps a look at some of the current implementations might help:
1) Cameras come either with a digital output or analog output. As Woody mentions, there are several different analog standards (PAL, NTSC, RS-170,...) and formats for transmitting the data (RGB, S-VHS, composite, ...)
2) Regardless of what the signal is, there must be some "device" that translates the incoming signals into "numbers" that the computer can understand. Sometimes this is implemented on the motherboards (USB), or needs an additional card (frame grabber). It depends on the age of the computer and its make and Operating system which of the different options are supported.
3) Currently there are 3 ways of getting the signals into a PC (other than serial RS-232 and parallel ports which are way to slow):
a) USB b) 1394 or firewire c) PCI boards
USB
USB is a serial interface that is now supported on most computers out of the box. The bandwidth of a USB connection is a maximum of 12 MegaBIT/second, which translates of course into 1.5 Mega-BYTE per second. This is too slow for video (about 4-5 MegaByte per second), but enough for still images, unless the video is compressed. Compression is OK for "consumer" video, but not acceptable for "scientific" video unless it is lossless (JPEG and MPEG is usually not). Several devices on the USB bus can compete for bandwidth.
1394
1394 (or firewire) is also a serial interface with a maximum bandwidth of 400 Mega-BIT per second (or 50 Mega-BYTES per second). This is fast enough for video. However, I believe that again more than one device can be attached to a 1394 port which will again compete for bandwidth. Firewire is not generally available on PCs (I believe the newer Macs have it), so that for a firewire implementation on a PC normally a card is necessary that fits into a PCI slot.
PCI
PCI boards, while a bit more cumbersome to install, have the highest throughput. I think, they are now implementing a new PCI-X specification that allows up to 1 GigaBYTE per second, i.e., about 20 times faster than firewire. The reason is of course that PCI is a parallel standard and not a serial like USB or firewire.
So, there are various ways to attach a camera:
Analog camera: There is no other way than to use a board to transform the analog signals into digital signals and then send them to the computer. This can be through a PCI or other card or other electronics for example on the video card, but the translation is necessary.
Digital cameras: Digital cameras essentially put the digitizer into the camera and then transmit digital signals. They can then be transferred through USB (slow but available everywhere), firewire (faster, currently on Macs (I believe) and PCs with additional card), or through boards for the PCI bus (fastest, widely available, require board installation).
So, for most users (of PCs at least) there is currently no real difference between using a firewire or other camera, as they either have to install a PCI-} firewire card, or another PCI card for image acquisition. That may change if the motherboard manufacturers start building firewire circuitry into the motherboards and the operating systems start supporting this option. For highest performance, however, I believe that we will see PCI boards for some time to come. Firewire may run into a performance problem in the future for image streams with large images. A 1600x1200 image stream with 24 bit color and 30 frames per second requires a bandwidth of about 170 MBytes/second uncompressed.
I hope I have not confused anybody with this.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
} ---------- } From: White, Woody N[SMTP:WOODY.N.WHITE-at-MCDERMOTT.COM] } Sent: Wednesday, January 26, 2000 4:13:00 PM } To: "Peter Guthrie" ; Microscopy-at-sparc5.Microscopy.Com } Subject: Re:USB or firewire cameras } Auto forwarded by a Rule } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Peter,
If you intend to use a "video" camera, you may need a capture card rather than, or in addition to, a USB or Firewire interface. Most video cams are analog (NTSC in the US, SECAM or PAL elsewhere in the world). Others can certainly have digital circuits internal (like the new digital camcorders or the PCcams used for video conferencing on the net). ...You first need to determine the source format.
Warren's suggestion implements an inexpensive external NTSC* video frame grabber (Snappy). Which will "grab" an analog video frame and digitize it. *May do PAL too??
Parallel port I/O is a bit slow (or is that a byte slow :) for pictures containing lots of data, but is cheap and works since the inherent resolution of typical NTSC video is { 640x480. USB is very much faster and Firewire faster yet (and usually a lot more $$ for the interface card). For applications other than full frame rate/resolution streaming video, USB is fine.
Woody White
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
The IMAC DV comes with a firewire port standard and a firewire cable. Connection to a Sony DV camcorder is easy and it gives you complete control from the computer. See http://www.apple.com/firewire/ for more information.
To get firewire into a PCI (mac or windows) machine Sony sells this card http://www.sel.sony.com/SEL/consumer/ss5/office/digitalvideo/minidvcamcorderspro ducts/dvbk-2000_specs.shtml for around $350 which gives similar controls for live video and digital stills.
No interest in either company except as a satisfied customer. Scott
} } 1394 (or firewire) is also a serial interface with a maximum bandwidth } of 400 Mega-BIT per second (or 50 Mega-BYTES per second). This is fast } enough for video. However, I believe that again more than one device can } be attached to a 1394 port which will again compete for bandwidth. } Firewire is not generally available on PCs (I believe the newer Macs } have it), so that for a firewire implementation on a PC normally a card } is necessary that fits into a PCI slot. } ..snip... } So, for most users (of PCs at least) there is currently no real } difference between using a firewire or other camera, as they either have } to install a PCI-} firewire card, or another PCI card for image } acquisition. That may change if the motherboard manufacturers start } building firewire circuitry into the motherboards and the operating } systems start supporting this option. For highest performance, however, } I believe that we will see PCI boards for some time to come. Firewire } may run into a performance problem in the future for image streams with } large images. A 1600x1200 image stream with 24 bit color and 30 frames } per second requires a bandwidth of about 170 MBytes/second uncompressed. } } Michael Bode, Ph.D.
-------------------note: new mailing address---------------------- Scott Wight e-mail: scott.wight-at-nist.gov NIST 222/A113 W voice: 301-975-3949 100 Bureau Dr STOP 8371 | fax: 301-417-1321 Gaithersburg, MD 20899-8371 \|/ disclaimer: Any opinion expressed is my own and does not represent those of my employer.
-----Original Message----- } From: Paul Webster [mailto:pwebster-at-hei.org] Sent: Wednesday, January 26, 2000 6:52 PM To: MSA listserver submission
} Please keep weather reports private. } } Ann Fook
Why? A little light heartedness never hurt anyone. For the record, LA was sunny as usual today. Happy I don't live in CT anymore.
Regards,
Paul Webster, Ph.D. Associate Scientist & Director Ahmanson Advanced Electron Microscopy & Imaging Center House Ear Institute 2100 West Third St. Los Angeles, CA 90057
We would apreciate to inform interested young researcher of the following available positions in our research project. Thank you very much for getting this circulated. Pierre
Pierre Ruterana Laboratoire d'Etude et de Recherche sur les Materiaux (LERMAT) Unite associee CNRS No 6004 Institut Superieur de la Matiere et du Rayonnement(ISMRA) 6, Bd Marechal Juin 14050 Caen Cedex France Tel: (33 2) 31 45 26 53 Fax:(33 2) 31 45 26 60 e-mail: ruterana-at-lermat8.ismra.fr
Research Training Network EC Contract N¡: HPRN-CT-1999-00040 Interface analysis at atomic level and Properties of Advanced Materials (IPAM)
Eight positions are immediately available, eligible young ( { 35 years) researchers must be citizens of EC or associated countries (Norway, Island, Israel, Lichtenstein, Bulgaria, the Czech Republic, Estonia, Hungary, Lithuania, Poland, Romania, Slovakia, Slovenia and Letonia), however any foreigner who has spent five years in an EC country may apply. Women candidates are particularly encouraged to apply and equal opportunity between women and men will govern our choice. Following the mobility criteria, the young researchers will not apply for a position in their native country.
1. POSTDOCTORAL POSITION at Fritz Haber Institute, Max Planck Society, Berlin, Germany A postdoctoral position at the Fritz-Haber-Institut in Berlin (Germany) is available in the group "Surface morphology and growth of semiconductors" under the supervision of Dr. Joerg Neugebauer. The research will be mainly focused on the theoretical modeling of electronic properties and atomic structure of interfaces and interfacial defects employing first principles total energy calculations. The basic materials for this project will be gallium nitride based semiconducting layers where extended defects are well known to occur in large concentrations. The research will be performed in close collaboration with experimental, industrial, and theoretical partners within the EC. Strong interaction with the other groups is therefore expected. The successful candidate should have a PhD in Physics, Chemistry or Materials Science, and have a strong interest on microscopic simulations. Preference will be given to candidates with strong background in any (or several) of these fields: electronic structure calculations, molecular modeling, density functional theory, empirical potentials, and analysis of transmission electron microscopy measurements. Interested candidates should send immediately a CV, list of publications, name and address (including email) of three references, preferably by email or fax, to: Dr Joerg Neugebauer E-mail: neugebauer-at-fhi-berlin.mpg.de, Phone: ++49 30 8413 4826, Fax: ++49 30 8413 4701, www: http://www.fhi-berlin.mpg.de/th/JG
2. POSTDOCTORAL POSITION at Universitat Politcnica de Catalunya, Barcelona, Spain A postdoctoral position is available at the department of Applied Mathematics in the UPC. We are seeking a computational materials scientist interested in modeling the atomic structure of interfaces and defects in crystals, mainly gallium nitride based materials. A PhD in Physics, Materials Science or related discipline and having experience with atomic simulations is required. It is highly desirable an ability to develop empirical interatomic potentials. It is intended that he/she visits the other laboratories working in the project to learn how to interpret the experimental observations and how to use the theoretical concepts. Interested candidates should send immediately a CV, list of publications, name and address (including email) of three references, preferably by email or fax, to: Prof. Anna Serra E-Mail: a.serra-at-upc.es, Phone: ++34 93 401 68 86, Fax: ++ 34 93 401 18 25
3. A FULL TIME PhD STUDENT at CRHEA, Valbonne, France These last years, CRHEA-CNRS has implemented an expertise in the growth of heteroepitaxial GaN layers on different substrates: sapphire, SiC and Si by different techniques, MBE, MOVPE and HVPE. The group has developed a proprietary Epitaxial Lateral Overgrowth (ELO) technology which allows to decrease by orders of magnitude the density of dislocations in GaN heteroepitaxial layers on sapphire, SiC or Si. Therefore, a great interest in the procurement of high quality GaN substrates currently exists. The successful candidate will strongly support our present effort to produce self-supported GaN of ELO quality by combining ELO-MOVPE and HVPE. Parallel to the growth, he will contribute to the development of in depth analysis of basis mechanisms linked to the generation and propagation of threading dislocations(TDs). More precisely, it is planned to determine the core structure of defects in ELO GaN, their electronic structures (by EELS), the mechanism of bending of these TDs, to implement new ways of further decreasing the density of dislocations. He will be able to use two MOVPE, one HVPE reactor and all basic characterisation tools (double X-ray diffraction, magnetotransport, low temperature photoluminescence, HRTEM,.). Candidates should send immediately a CV, name and address (including email) of two references, preferably by email or fax, to: Dr Pierre Gibart, email: Pierre.Gibart-at-crhea.cnrs.fr, Tel: ++33 4 93 95 42 27, Fax: ++33 4 93 95 83 61 CHREA-CNRS is located at the French Riviera near Nice ( see: http://www.crhea.cnrs.fr/)
4. POSTDOCTORAL POSITION at ISMRA Caen, France Candidates should preferably have a Phd with experience in electron microscopy and/or atomic structure modeling. The project will involve experimental high resolution electron microscopy and image analysis. In parallel, atomic structure modeling of defects and interfaces will use empirical and tight binding methods. A connection will be established with ab initio techniques developed in partner groups and the successful candidate will undertake quantitative comparison of experimental and simulated images. The overall aim is the understanding of the role of defects and interfaces on the optoelectronic properties in the Ga based nitride semiconductors. Candidates should send immediately a CV, list of publications, name and address (including email) of three references, preferably by email or fax, to: Dr Gerard Nouet on ++33 2 31 45 26 47, email:gerard.nouet-at-labolermat.ismra.fr or Dr Pierre Ruterana on ++33 2 31 45 26 53 email: ruterana-at-lermat8.ismra.fr
5. POSTDOCTORAL POSITION at University of Liverpool, Great Britain A three year full-time appointment funded by the European Commission is available to study defect mechanisms in gallium nitride based electronic device structures within the III-V semiconductor materials group. Candidates should preferably have postgraduate experience in the growth or processing of semiconductor device materials. The project will involve the chemical beam epitaxy of GaN based materials and the fabrication of model device structures. The influence of processing parameters on defect propagation will be investigated using analytical methods such as electron microscopy, Raman spectroscopy and surface analytical techniques. This appointment is part of a Research Training Network and eligible candidates must be citizens of EC member countries other than the United Kingdom. Candidates should send immediately a CV, list of publications, name and address (including email) of three references, preferably by email or fax, to: Dr Paul Chalker: ++44 151 794 4313, email pchalker-at-liv.ac.uk or Prof Robert Pond: ++44 151 794 43 13 / 46 60, email R.C.Pond-at-liverpool.ac.uk
6. POSTDOCTORAL POSITION at Universitt Erlangen-Nrnberg, Germany Focus of the work in Erlangen university will be on direct correlation of structural, optical and electrical properties of extended defects in (i) group-III nitrides (ii) group III-nitride based heterostructures. Experimental work is based on (scanning) transmission electron microscopy ((S)TEM) at all levels of resolution. These comprise high resolution imaging with atomic resolution, optical characterisation by cathodoluminescence in the STEM and analysis of electrical properties (electrical activity of extended defects, diffusion length of minority carriers) by the electron beam induced current (EBIC) both in the SEM and the STEM. Theoretical analyses are based on TEM contrast simulation for defect analysis, analysis of the tetragonal distortion from high resolution TEM images and finite element simulations of the strained state of heterostructures. Candidates should send immediately a CV, list of publications, name and address (including email) of three references, preferably by email or fax, to: Prof Horst Strunk, Tel: ++49 9131 85 2 8601, Fax: ++49 9131 85 2 8602, email: strunk-at-cmp03ww7.ww.uni-erlangen.de, Dr Martin Albrecht, Tel.: ++49 9131 85 2 8613, Fax: ++49 9131 85 2 8602, e-mail: albrecht-at-cmp04ww7.ww.uni-erlangen.de,
7. POSTDOCTORAL POSITION at the Aristotle University of Thessaloniki, Greece A research opportunity is available for postdoctoral candidates with a background in Electron Microscopy, Crystal Growth, Materials Science. The primary responsibility of the candidate will be the study of the structure and properties of the heterophase interfaces between thin films on gallium nitride based materials. The project will offer the necessary training for the specific skills to meet the requirements of the job. Candidates should send immediately a CV, list of publications, and name and address (including email) of three references, preferably by email or fax, to: Prof Philomela Komninou, Tel: ++30 31 99 81 95 / Fax: ++30 31 99 80 61 e-mail: komnhnoy-at-auth.gr
8. A FULL TIME PhD STUDENT at The University of Cambridge, Great Britain A three year research studentship position leading to a PhD degree is available to study the microscopy and analysis of defects in gallium nitride layers and device structures. This exciting project will use a wide range of state-of-the-art electron microscopy and analysis techniques to study the atomic structure of defects (using high resolution electron microscopy), their chemical composition and electronic properties (using x-ray spectroscopy and electron energy loss spectroscopy), including which defects give rise to states in the band gap. This project is part of a European Research Training Network aimed at optimising devices in GaN-based materials. The research student will form strong links with the other European partners in this project. Eligible candidates should have a top quality degree in physics, chemistry, materials science or electrical engineering. Candidates should send immediately a CV, name and address (including email) of two references, preferably by email or fax, to: Prof Colin Humphreys on +44 1223 334457, email {colin.humphreys-at-msm.cam.ac.uk} , or Dr Dave Tricker on +44 1223 334469, email {dmt1000-at-cus.cam.ac.uk}
The Candidates will gain time by sending a copy of their CV also to Prof. Philomela Komninou, leader of the Training Programme. Tel: ++30 31 99 81 95 / Fax: ++30 31 99 80 61 e-mail: komnhnoy-at-auth.gr; Please indicate the position of interest.
Caen, January 27 2000 Dr. Pierre Ruterana Coordinator
It seems nobody replied to Michaels question. Perhaps because there is no single answer. LR White and LR Gold I would expect to have a very similar shelf-life - under similar conditions.
The trouble is to know the starting point. LR White slowly "goes off" from when the catalyst is added. At room temperature or higher this happens at a much faster rate than when it's kept refrigerated. Catalyzed LR White could be kept at the room temperatures for some months. Because the shipping time (even by air) to our home-market (Australia) from the UK is too long and often at high temperatures, much and an indeterminable part of the shelf-life would be expired prior to sale.
Consequently we only procure uncatalysed LR White and the end-user must add and thoroughly mix the catalyst prior to first use. Our users expect a full year of refrigerated shelf-life. Shelf-life is really a "when, how long and at what temperature" question. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Tuesday, January 25, 2000 4:52 PM, Michael Reiner [SMTP:michael.reiner-at-Smail.Uni-Koeln.de] wrote: } } } Dear members of the list, } } first, I would like to wish you all the best for the new year. } Now my question: } Does anyone know the shelf-life/usability of LR-Gold stored in a fridge? } } Is it that delicate as LR-White (Meanwhile I don?t use LR-W older than } half a year). My bottle which was not opened many times, could be } roundabout three years old. } } Thanks a lot, } Michael } } Michael Reiner } Department of Anatomy I } University of Cologne } Germany } michael.reiner-at-smail.uni-koeln.de
Dear list members, The intent of this note is to formally announce a call for papers for the upcoming Spring 2000 AReMS (Appalachian Regional Microscopy Society) meeting to be held in Raleigh, NC.The meeting dates are March 30 and 31. The theme for this meeting is "Recent Advances in Microscopy for 2000",specifically we would like to focus on recent (but not limited to) advances in microscopical instrumentation and applications therefrom.
Please forward any abstracts, papers or related items to me at mailto:\\rlmcgill-at-eastman.com or you may contact me at the number below.
The most recent meeting info is -at- http://www.wise.virginia.edu/cvc/arems/raleigh.html
Thanks in advance for your interest!
Rick McGill Microscopy Research Eastman Chemical Company - - - Phone: (423) 229-5473 - - - Fax: (423) 229-4558 - - - e-mail: rlmcgill-at-eastman.com
You should logon to Histonet - they're much friendlier!
Had a beautiful drive from old Plymouth to Reading, Berkshire, with my retired boss yesterday to collect x-ray microanalysis equipment. Clear blue sky, -2 to +5 degrees. Saw the most gorgeous scenes of trees covered by thick hoar frost - photographers' dream. Met two nice ladies - Hello, Jill and Hilary! Thanks for the help!
Keith Ryan Marine Biological Association of the UK PS - Don't read this if you don't like the weather ! PPS - See, Paul, I did get there! PPS - Hello, Daniele - time to write!
You ought to logon to "Histonet" - they're much friendlier!
I agree about the weather being important to EM. 20-30 years ago we had some seaweed hanging in the microtome room (about 100 m from the sea). If it was damp we didn't even try cutting some resins!
Had a car trip yesterday from old Plymouth to Reading, Berkshire, collect EM equipment., -2 to +5 degrees, clear blue sky, gorgeous scenes of thick hoar frost on the winter trees - a photographers' dream. Bonus - met two nice girls - Hello, Jill and Hilary, thanks for the help!
Keith Ryan Marine Biological Association of the UK PS - Don't read this if you don't like weather! PPS - Paul, I made it! PPPS - Hello, Daniele, its time you wrote!
Neither rain nor sleet, snow nor heat, humidity, hell nor highwater can keep me from getting Formvar films off glass slides. Only time can...see below.
My secret? Just rinse glass slides - both sides & all around edges, except for the end you are holding onto - with 95% ethanol and air dry. Then use right away - dunk into Formvar solution (.25 to .5% w/v in ethylene dichloride), drain and air dry. Score around edges to break film and float off onto clean water surface. I like to score the edge by using the corner of a razor blade to score on the top surface of the slide, near the edge, in addition to scoring the actual corner edge of the slide with the blade held perpendicular to the edge - know what I mean? Also, score across the slide near the "top" edge of the Formvar film, near the end you are holding on to, for clean release of the end of the film.
Second point, Formvar film solutions older that 3 months tend to stick to glass. I've been tracking that for years. Put mix date on bottle of fresh solution, after 3 months expect poor release effects to appear.
Gib
P.S. I tend to agree with Fook that this forum should not be used to discuss the weather ONLY, but if someome wants to put a current local weather "tag" at the end, AFTER the Microscopy stuff, with their signature, thats OK with me. Everyone is entitled to their (short) bit of poetry, sage sayings, or weather commentary there.
For example: "The weather here is unremarkable at this time."
Gib Ahlstrand Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN. USA. 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.cdl.umn.edu http://biosci.umn.edu/MIC/consortium.html
Dear All, I have a user who is trying to make TEM samples from nanophase metals. They come out of the process as a fine (10 to 100um) powder. The study is to compare the microstructure at different processing temperatures (77K to 400K). The current sample preparation technique is to embed the powder in epoxy, slice and polish, and finish with l-N2 ion milling (BTW, direct dispersion of the powder does not work as the edges are too thick). There are several problems with this technique, the worst being that we have found these materials age rapidly even at room temperature. The epoxy cure and ion milling thermal budgets may be a problem. I am considering ultramicrotomy for these samples, but I have zero experience in this field. McMahon and Malis (1995 Micro Res & Tech v31 267) worked on a similar system and outline the use of thermally cured LR-White as the embedding material, so I think it is an appropriate option, but I am worried about the thermal budget. My question is: Does any one have experience with low-temperature, UV cured resins for materials of these type? If so I would greatly appreciate any advice.
Thanks in advance, Ray
************************* Ray D. Twesten, PhD Center for Microanalysis of Materials University of Illinois (217) 244-6177 fax:(217) 244-2278
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Please see request below. If you have any suggestions, either send to the list and I will forward them to the investigator, or send them directly to Dale. Thanks for the input. Debby Sherman
--------------------------------------
Greetings, I can make two suggestions.
1) Ignore the staining in the cell wall. Since you know that your protein of interest is not there and you know also that secondary cell wall must be outside of the cell, this should not interfere with staning inside the cell.
2) Try to pre-absorb with "wood". The idea here would be to incubate your serum with some sort of wood pulp, and then spin out the wood pulp presumably taking with it all the ab's that bind wood, but leaving behind the ones of interst. I am not sure how best to make a suitable pulp of wood. Maybe take a pencil sharpener and grind a dowell, and then grind the shavings further in a mortar and pestle or maybe use a homogenizer of some sort. I am guessing wildly here. Certainly the wood bits should be easy to spin after absorption.
Hope this helps, Tobias.
} } Date: Thursday, January 27, 2000 } } From: Dale Karlson {dtk-at-omni.cc.purdue.edu} } } } } QUESTION: How to minimize artifact labelling with secondary cell walls } } } To whom it may concern: } } I am attempting to localize protein in a woody plant and consistently } observe an interaction with secondary cell walls. This is an artifact, we } know that the protein does not exist in the cell wall. We are using a } polyclonal antibody that was raised against a protein that was excised } from an SDS gel, suspended in Freund's complete adjuvant and used for the } immunization (in chicken). Chicken antibodies were purified by an } ammonium sulfate precipiation method described by Song et al. (Song, C.S., } J.H. Yu, D.H. Bai, P.Y. Hester, and K.H. Kim. 1985. Antibodies to the } 5-subunit of insulin receptor from eggs of immunized hens. Journal of } Immunology 135: 3354-3359). I have tried Western blot affinity } purification of the antigen and antibody and it has not solved this } problem. We do not have access to a "purified" form of the protein, so } affinity purification with a purified protein is not an option. } } It is obvious that the chicken had an "allergy" that was not visible } during our screening process (with western blots) to select the host } animal. The chicken obviously has specific antibodies to some secondary } wall component, and I would like to know what it might be and how I could } remove this artifact. } } Any input would be greatly appreciated. } } } ------------------------------------------- } } Debbie, } } let me know if this is suitable..or if it is way too long etc. } } Thanks, } } -Dale } } } } } } _______________________________________________________________________ } } Dale Karlson .***. .***. .***. } 1165 Horticulture Building * | | | * | | | * * | | | * } Purdue University * | | | * * | | | * * | | | * } W.Lafayette. IN 47907-1165 * | | | * * | | | * | | | * } '***' '***' '***' } } Home Phone: (765) 742-8379 } Lab Phone: (765) 494-1345 } _______________________________________________________________________
The decision has been made to get rid of the following piece of equipment. We obtained the unit approximately five years ago when an outside Contractor brought the system very near operational state. Numerous distractions and circumstances have prevented the TEM from becoming the valuable research investigative tool we had planned.
HITACHI H-600 TEM
1) Model H-600-1 Analytical Electron Microscope 2) Model H-6015 EDX Interface Kit 3) Model H-6012 Micro-Diffraction Unit 4) Spot Scan 5) Model H-6006 Auto Data Display Unit 6) Reduced Area Scan Unit 7) Polaroid Camera w/ Adaptor 8) Model H-6007 High Resolution CRT 9) Model H-5001-C Cobling Holder 10) 2 ea. Overhauled Mechanical Vacuum Pumps & 1 ea. Diffusion Pump
Anyone interested in requesting a Bid Form should contact me at following address:
Jim Goodman University of Tennessee Space Institute (UTSI) 411 B. H. Goethert Pkwy. Tullahoma, TN 37388 TEL: (931) 393-7494 FAX: (931) 393-7543 e-mail: jgoodman-at-utsi.edu
Yes, I also agree. Today in San Diego it is about 68 degrees with 76% humidity. Great for cryoultramicrotomy and regular cutting! Take care all, Jo Dee
Witold Zielinski 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 } -----------------------------------------------------------------------. } } *Date sent: 26 Jan 00 17:52:26 -0800 } *From: Paul Webster {pwebster-at-hei.org} } *Subject: Re:No weather report please } *To: MSA listserver submission {Microscopy-at-sparc5.microscopy.com} } *Send reply to: Paul Webster {pwebster-at-hei.org} } } *------------------------------------------------------------------------ } *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 } *-----------------------------------------------------------------------. } * } * } *} Please keep weather reports private. } *} } *} Ann Fook } * } *Why? A little light heartedness never hurt anyone. } *For the record, LA was sunny as usual today. Happy I don't live in CT anymore. } * } *Regards, } * } *Paul Webster, Ph.D. } *Associate Scientist & Director } *Ahmanson Advanced Electron Microscopy & Imaging Center } *House Ear Institute } *2100 West Third St. } *Los Angeles, CA 90057 } * } *Phone: (213) 273-8026 } *Fax: (213) 413-6739 } *e-mail: pwebster-at-hei.org } *http://www.hei.org/htm/aemi.htm } * } * } I agree with you Paul. } In Warsaw, Poland yesterday was snow on the ground today is } rain and no chance for sunshine. } Stay cool, } Witold
-- Jo Dee Fish Electron Microscopy Assistant The Burnham Institute 10901 N. Torrey Pines Rd. La Jolla, CA 92037 858-646-3100 ext.3620
This request is posted on behalf of an associate who does not subscribe to this list. He has assumed responsibility for the following equipment and is looking for service providers. Equipment is located in south central Massachusetts.
Jeol 840A sem Kevex delta 5 EDS & XRF with Syquest drive upgrade
He should be contacted off line by e-mail at LapradeB-at-burle-eo.com
FYI, I just tested an Optronics digital camera that had a on board firewire connection to a Gateway 366MHz notebook computer. One can also get PCMCIA card to connect to most notebook computers. Nice camera but should be for $13k, without notebook computer!
Damian
} 1394 (or firewire) is also a serial interface with a maximum bandwidth } of 400 Mega-BIT per second (or 50 Mega-BYTES per second). This is fast } enough for video. However, I believe that again more than one device can } be attached to a 1394 port which will again compete for bandwidth. } Firewire is not generally available on PCs (I believe the newer Macs } have it), so that for a firewire implementation on a PC normally a card } is necessary that fits into a PCI slot.
I'm trying to find a used ion beam sputter coater or something similar... The system I am used to is the old VCR group ion beam sputter coater. Any leads that you might have would be greatly appreciated...
Thanks,
Raj
********************************************* Raj Lartius, Ph.D. NovaScan Technologies Iowa State University Research Park 2501 North Loop Drive Ames, Iowa USA 50010
We are interested in acquiring an EDS detector for our JEOL JEM-1200EXII. Will trade a Kevex Be-windowed 30mm2 that was formerly attached to an EM400 for a Kevex/TN/Noran 10mm2/30mm2 Be or UTW. Need detector only (no MCA, P. Processor, etc). Will purchase or trade.
If you have a suitable detector and are in a position to trade/sell immediately, please reply off line to sender.
Bob Roberts EM Lab Services, Inc 2409 S. Rural Rd Tempe, Arizona 85282 (480) 967-3946
I have been reading about FRET and have a question about dual label imaging with probes like FITC/RHo or Cy3/Cy5. We have to worry about excitation of the long wavelength probe at the shorter probe wavelength, and FRET as two ways in which we can be mislead about the co-localization of two probes. I am wondering to what extent one also has to worry about non-FRET energy transfer. It seems that there is some possibility that, for example, Cy5 could become excited by absorbing photons from Cy3 emission. My presumption is that the density of photons is low, and this would limit the effect, but it seems that as proximity gets closer, the chances of this radiative exchange (rather than resonance exchange) would become greater. Are there any experimental guidelines as to when to worry about this? Thanks- Dave --
Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 75 N. Eagleville Rd. U-125 Storrs, CT 06269 Knecht-at-uconnvm.uconn.edu 860-486-2200 860-486-4331 (fax) home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html ************************************************************
On Mon, 24 Jan 2000 13:33:28 -0500, Greg Erdos 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 } -----------------------------------------------------------------------. } } } Periodically questions appear on this list concerning the use of cryostats } for cutting histological sections. I usually reply to the sender off line } and offer a copy of a handout that I got from a workshop at a Histochem } Meeting some years ago. Even though it is old, cryostat sectioning has not } changed a lot. I think there is some valuable info there, especially for
} beginners. I thought it might be helpful to make this available on the net } for whomsoever might want to take a look. } It can be found at : } http://www.biotech.ufl.edu/sems/ } } Look for the snowflake } } It was written by Bruce Quinn, then of MIT. I hope he has no objections } to my posting it. } } Greg Erdos } Gregory W. Erdos, Ph.D. Ph. 352-392-1295 } Assistant Director, Biotechnology Program } PO Box 110580 Fax: } 352-846-0251 } University of Florida } Gainesville, FL 32611 } } } Finally got onto things here at home, and using Netscape 4.5 and Acrobat 3.0, the document opens up the way it should. Thanks for the info and link, Greg.
Roger C Moretz, Ph.D.
_______________________________________________________ Get 100% FREE Internet Access powered by Excite Visit http://freeworld.excite.com
A method I've used for controlling the position of the cleave in silicon wafers is to use focused ion beams (FIBs) to mill micro-cleaving grooves into the silicon. I found that the grooves can determine the position of a cleave to within 200 nm.
If you want any more details I can forward you a pre-print on the technique.
Richard
-------------------------------------------------------------- Richard M Langford
Department of Materials, University of Oxford Parks Road, Oxford, OX1 3PH, UK
----- Original Message ----- } From: Timothy Dimitri {tdimitri-at-us.ibm.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Friday, January 28, 2000 12:56 PM
Folks: I thought I should let you all know about the Second annual course in Quantitative Fluorescence Microscopy to be taught between june 19 and 24th 2000 at the Mount Desert Island Marine Biology Laboratories in Arcadia National Park in Maine. This team taught course led by Dave Piston (Vanderbilt), Mike Nathanson (Yale), Guillermo Romero (Pittsburgh) and myself focusses specifically on the development and application of modern fluorescence microscopic methods. This intensive course covers all aspects of the technology from microscope and dye design, cameras, confocal microscopy, live cell microscopy, multiphoton microscopy and GFP. Considerable attention is also given to quantitative analysis in 2 and 3 dimensions and time. The specific focus of the course allows an in depth treatment of these methods. The goal of the course it to teach students how to best implement these methods within their labs, using either their own cells and tissues or using material supplied by the course. An extensive array instrumentation, provided by all the major microscope and associated software, hardware and camera manufacturers will be available for students to use. Last year it was a very successful event and we were encouraged to give the course again. A full description of the course lectures together with lecture outlines, registration etc. is available at http://sbic6.sbic.pitt.edu/microscopy. I would encourage you to spread the word, or sign up if you are interested. The total number of students is limited to 20, enrollment is decided by the course faculty. If you have any further questions please feel free to contact me Thanks Simon
----------------------------------------- Simon C. Watkins Ph.D. MRCPath Associate Professor Director: Center for Biologic Imaging University of Pittsburgh Pittsburgh PA 15261 tel:412-648-3051 fax:412-648-8330 URL: http://sbic6.sbic.pitt.edu ----------------------------------------- Simon C. Watkins Ph.D. MRCPath Associate Professor Director: Center for Biologic Imaging University of Pittsburgh Pittsburgh PA 15261 tel:412-648-3051 fax:412-648-8330 URL: http://sbic6.sbic.pitt.edu
At 1:09 PM -0500 1/29/0, "IMZartTchr-at-aol.com"-at-sparc5.Microscopy.Com wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
*********** If you don't get an answer from MSA people, try the listserver for the histologists out there: "HistoNet Server" {HistoNet-at-Pathology.swmed.edu}
Lee
Lee Cohen-Gould EM & Confocal Facilities Weill Medical College of Cornell U. (212)746-6146 Rms A-105, LC-207
The Cape Town weather is great today, beautiful clear blue skies and a nice warm 25 C.
We have a Philips TEM 420 which has an EDAX system attached (which is non-functional at the moment). We are looking for video / digital image grabbing system that we can use to grab images for prints and possibly Image analysis. Is this possible on the 420 ? Can anyone suggest a system?
A graduate student here (Rita Ware) had some success fixing halophilic bacteria for TEM several years ago. She used the growth medium as a buffer. She made a poster presentation at an MSA meeting.
} Date: Mon, 24 Jan 2000 06:46:26 -0700 } From: Marti, Jordi {jordi.marti-at-honeywell.com} } To: 'Microscopy' {Microscopy-at-sparc5.Microscopy.Com} } Subject: Reichert Ultracut E with cryo FC 4D } } ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } The power supply in our cryo microtome is having problems which might be } related to the transformer. I was told by the service engineer that the } transformer is no longer supported by Reichert. Does any one know where I } can get a replacement ? } } Thanks } } Jordi Marti } }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
South Bay Technology, Inc. manufactures an SEM Cleaving System which provides a means to precisely and quickly cleave a wafer while in the inspection mode. A wafer is mounted to a vacuum chuck which is positioned under an optical microscope. The exact area of interest is located visually and the sample is cleaved at that point. SEM compatible versions of the cleaving system are under development which will allow the user to image and cleave while mounted in the SEM. The Cleaving System is quick, easy to operate and precise. It allows anyone to quickly and repeatably prepare SEM cross sections.
If you have an interest, please let me know and we can discuss your requirements in detail.
David Henriks Vice President 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
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by Timothy Dimitri } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello,
I am looking for the names of any manufactures (other than SELA) that have a product that can cleave sub micron features on silicon wafers...
Thank you
Timothy Dimitri ASTC Failure Analysis Laboratory IBM Microelectronics Division
I forget to mention in my original posting that South Bay Technology also produces the MicroCleave kit which is designed for TEM cross sectioning. Again, if you have an interest, please let me know and I'll get you additional information.
David Henriks Vice President 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
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy. Message text written by Timothy Dimitri } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello,
I am looking for the names of any manufactures (other than SELA) that have a product that can cleave sub micron features on silicon wafers...
Thank you
Timothy Dimitri ASTC Failure Analysis Laboratory IBM Microelectronics Division
While I don't have any leads on a used IBS system, I wanted to let you know that we at South Bay Technology, Inc. are continuing the manufacture of the IBS system formerly produced by VCR Group. Actually, we have updated the system and are now offering the IBS/E. The IBS/E now adds the capability of etching samples as well as coating and will accommodate samples up to 2" in diameter.
If you would like additional information, please feel free to contact me.
David Henriks Vice President 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
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by "Dr. Raj Lartius" } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I'm trying to find a used ion beam sputter coater or something similar... The system I am used to is the old VCR group ion beam sputter coater. Any leads that you might have would be greatly appreciated...
Thanks,
Raj
********************************************* Raj Lartius, Ph.D. NovaScan Technologies Iowa State University Research Park 2501 North Loop Drive Ames, Iowa USA 50010
Are we like kids sitting in a sandbox and squabbling? Yes weather, no weather! He said it first! She started it! I don't want weather on the listserver. I want to read about weather in the boondocks. Who cares about you, anyway? Sort of funny when you think of it that way, isn't it?
A friend has the chance to buy a used JEOL JSM840 - AM1000 with EDX and all the bells and whistles. He has asked if this is a good instrument, and worth the price (whatever that is - he won't tell me). Since I haven't seen the instrument and I don't know JEOLs at all, I told him I would ask the experts.
If anyone can tell me a little about the instrument and what it might be worth (the second part of the question being more difficult), I would appreciate it.
In Honolulu it is gloriously clear and sunny, with temperatures near 80F during the day and about 60F at night, which is unusually cold but really nice.
Aloha, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
You are invited to participate in a symposium of remote access microscopy, and /or teaching microscopy to take place at the Microscopy & Microanalysis Annual Meeting August 13-17, 2000 in Philadelphia, Pa.
Platform and poster contributions are welcome. Please contact me directly for more information about the symposium.
Deadline for receipt of a 2page abstract is Feb 15, 2000. For registrationand abstract forms, see http://www.microscopy.com/MSAMeetings/MMMeeting.html
ADVANCES IN INSTRUMENTATION AND TECHNIQUES SYMPOSIUM 19: TEACHING MICROSCOPY IN THE NEW MILLENNIUM
Organizer: Steve Barlow
The use of computers to control microscope operations, the ability to control microscopes remotely over the Internet, and the creation of microscope computer simulations allow researchers and students to access microscopes in new ways. These developments mean changes in the way microscopy can be taught to students and researchers. This symposium will examine different ways to teach microscopy and microscope theory and operation to reseachers and students of all levels, in the context of new laboratory configurations, computer simulations, remote access usage, and classroom exercises.
Dr. Steven Barlow, Associate Director EM Facility/Biology Department San Diego State University 5500 Campanile Drive San Diego CA 92182-4614 phone: (619)594-4523 fax: (619) 594-5676 email: sbarlow-at-sunstroke.sdsu.edu website: http://www.sci.sdsu.edu/emfacility/
Chairman, Educational Outreach subcommittee promoting access to microscopes Microscopy Society of America http://www.msa.microscopy.com/
We have a new (to us) Philips CM-12 TEM in our lab and are wondering how to get the intermediate lens focused on the diffraction aperture (for making the first image plane and the diffraction aperture coincident prior to obtaining a SAED pattern). It doesn't seem to be covered in the manual.
Any help would be appreciated as this is a completely new microscope to us.
The JEOL 840 is an excellent instrument: very reliable & very easy to use. Enjoy with confidence.
Earl Weltmer
P.S.: Does your friend need someone to install the SEM? I would love to install it assuming it is in Hawaii.
Tina Carvalho 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, microscopists- } } A friend has the chance to buy a used JEOL JSM840 - AM1000 with EDX and } all the bells and whistles. He has asked if this is a good instrument, } and worth the price (whatever that is - he won't tell me). Since I } haven't seen the instrument and I don't know JEOLs at all, I told him I } would ask the experts. } } If anyone can tell me a little about the instrument and what it might be } worth (the second part of the question being more difficult), I would } appreciate it. } } In Honolulu it is gloriously clear and sunny, with temperatures near 80F } during the day and about 60F at night, which is unusually cold but really } nice. } } Aloha, } Tina } } **************************************************************************** } * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * } * Biological Electron Microscope Facility * (808) 956-6251 * } * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* } ****************************************************************************
Hello All, Does anyone know of a supplier for Historesin, formerly Cambridge, Leica, LKB? And, the weather in SoCal is quite lovely today - it finally rained. Thanks, Kristen Kristen A. Lennon Cell, Molecular & Developmental Biology Group Department of Botany & Plant Sciences University of California Riverside, CA 92521 kalen-at-citrus.ucr.edu
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Hi,
Are we like kids sitting in a sandbox and squabbling? Yes weather, no weather! He said it first! She started it! I don't want weather on the listserver. I want to read about weather in the boondocks. Who cares about you, anyway? Sort of funny when you think of it that way, isn't it?
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Would you be interested in a Zeiss 9s2 TEM? 60k is the top magnification. It has been a nifty scope as we have upgraded to a Zeiss 109. If you are interested let me know. Cheers! -Ken ------------ Ken Tiekotter Dept. of Biol. The University of Portland 5000 Willamette Blvd. Portland, OR 97303
On Wed, 26 Jan 2000, Cynthia Shannon 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 } -----------------------------------------------------------------------. } } } Subject: Wanted: Used TEM for virus work } Date: Tues, 26 Jan 2000 } } From: cshannon-at-nctimes.net } To: Microscopy-at-sparc5.microscopy.com } } Does anyone have an old TEM for virus work? } I am the electron microscopist for the county veterinarian. We are short } of funds. Please contact me by email. } Thanks. } Cindy Shannon } } }
-Obviously an advantage for many bacteria species, but a nightmare for the microscopist who wants to count them. For several reasons we want to split the aggregates of bacteria into single cells before counting. We have tried mild detergent treatment and ultrasound though with limited success. The samples are initially taken, as filter samples in working atmospheres were bacteria could be airborne, e.g. farm work. Afterwards the bacteria are washed off the filter, resuspended, stained (AO), refiltered on black pc-filter, mounted and counted. Any suggestions for a treatment, which will de-aggregate the bacteria, are most welcome.
Asbjorn Skogstad National Inst. of Occup. Health, Oslo, Norway asbjorn.skogstad-at-stami.no
That's why I tried to bring in a useful element to it by mentioning, a little tongue in cheek, my humidity vs formvar films question. But thanks to the silly weather thread I had some useful and interesting responses. May I be forgiven for saying that there's a silver lining..............
Tony Bruton University of Natal South Africa
} } } "Chuck Butterick" {cbutte-at-ameripol.com} 02/01/00 04:06AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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Hi,
Are we like kids sitting in a sandbox and squabbling? Yes weather, no weather! He said it first! She started it! I don't want weather on the listserver. I want to read about weather in the boondocks. Who cares about you, anyway? Sort of funny when you think of it that way, isn't it?
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Does anyone have the geometry settings for a Kevex Quantum detector on a JEOL 2000FX? I am using DTSA and need the values. I would like to have the sample to detector distance and the azimuthal angle. I am using +45 for the azimuthal angle and 90 for the detector angle. Are these correct? I know that the takeoff angle is 70. -Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Hi all. I have just joined this list and am posting without the usual "lurking time" due to time constraints - please forgive if recently covered. I am also a novice in all microscopy techniques, especially fluorescence.
I am trying to detect GFP-containing cells in liver tissue and having some problems.
1. I can't determine the spectra for the filter blocks I am using (on a Nikon Eclipse TE300 with TE-FM Epifluorescence attachment). The blocks are marked "UV", "B-2" (blue from source, green in field) and "G" (green from source, red in field). I have e-mailed Nikon and to be fair they've only had a few days but I'm under some pressure. No help from the manual. I have been using B-2 for GFP.
2. I get quite a lot of background fluorescence even with frozen sections (fixed in neutral buffered formalin). Can anyone suggest a way to reduce this? (eg any extra filters?)
3. I have read conflicting opinions on fixation. Most previous work has used thick sections (50 microns cut eg with Vibratome, ? to avoid having to embed tissue blocks). GFP is interfered with by acetone (and probably other organic solvents) but I would have thought that after fixation with formalin it should be stabilized and resistant to the xylene/alcohol used in paraffin embedding. I have seen fluorescence retained after this treatment but perhaps it can be improved.
4. For those with liver fluoro experience - on "UV" setting I see bright fluorescence which photobleaches. I believe I am looking at retinoids in stellate cells, although the bleaching is incomplete and a bit slower than I have seen before. Can anyone confirm this?
Apologies for long post. Any help much appreciated.
David Lockwood University of Qld Dept of Surgery PA Hospital Brisbane Australia
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A colleague would like your responses to these questions. I believe that I say something on the first question about a year ago and apologize for asking again.
} Here's what I/we need to know. } } 1) Are Microstar diamond knives equal to Diatome. We have an offer that } will save us about $1200 on three knives ( one 3mm ultra with 45 degree } angle, one 3mm ultra with 35 degree angle, one 10 mm histo knife). } } } 2) Is the power controller feature on the Pelco Microwave Tissue processor } (model #3451) worth it? (costs about $1300 additional compared to the } Pelco 3450 without the power controller). I plan to do immunolocalization } work, but am not sure this power controller feature is necessary or worth } the cost. } } Casey } } } Thanks for your time } Marty Reed Equipment Technician Biology Department Humboldt State University Arcata CA 95521 707-826-3234 707-826-3201 FAX mmr7001-at-axe.humboldt.edu
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Hi! Could anybody tell me what would be the sale price for a Hitachi H600? Thanks Dorota
I don't use a CM series microscope on a regular basis, but what is important is that you set it up the same way every time.
You can not make the back focal plane of the objective lens coincident with the objective aperture. The back focal plane is well above the location of the objective aperture plane.
There are two ways that I use to set up diffraction patterns reproducibly depending on whether I am using CBED or SAD. Both are set up after the sample has been made eucentric and the image focussed.
CBED: This method can be done for both CBED and SAD. The shadow of the condenser aperture defines the diameter of the diffraction disk. When the intermediate lens is adjusted properly, the edges of the diffraction disks will be in focus. You are grabbing the back focal plane for your diffraction pattern in the projector lens system. You will note that all of the HOLZ lines (if you can see them) are the sharpest at this condition. If you have a highly polycrystalline sample with continuous diffraction rings, this method is difficult to do.
SAD. Spread the beam with the condenser all the way. (clockwise in the CM-12 will go to more parallel beam faster than CCW -I think.) Then focus the spot to the smallest that you can. You can take a really long exposure or cheat a little and put some intensity back into the pattern with the condenser lens. You will note that the objective aperture is not focused in this method.
The most important thing to remember is to make the sample eucentric and focus before during either of these methods and to do the diffraction focussing consistently from sample to sample. -Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies." --
} -----Original Message----- } From: Valerie Leppert [mailto:vjleppert-at-ucdavis.edu] } Sent: Monday, January 31, 2000 6:44 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Question on Philips CM-12 SAD Alignment } } } -------------------------------------------------------------- } ---------- } 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 there; } } We have a new (to us) Philips CM-12 TEM in our lab and are } wondering how } to get the intermediate lens focused on the diffraction aperture (for } making the first image plane and the diffraction aperture coincident } prior to obtaining a SAED pattern). It doesn't seem to be } covered in the } manual. } } Any help would be appreciated as this is a completely new } microscope to } us. } } Thanks, } Valerie Leppert } }
I received Simon Watkins' note about the microscopy class in Maine and wonder if anyone knows about similar courses closer to California in the near future? We've got a number of technicians working on fluorescence microscopy in our lab, and I think it would be great to have some more formal training for us!
Sincerely, Laurie Wallin UCSD Department of Anesthesiology 9500 Gilman Drive, 0629, La Jolla, CA 92093
We are disposing of our old Philips 410 transmission electron microscope. It is in pretty good shape but is not fully functioning. At minimum it needs its ion getter reconditioned. Does anyone know of someone who might want to buy it and fix it up or use it for parts? -Robert
____________________________________
Robert S. Dotson, Ph.D., Laboratory Supervisor, Microscopy
Coordinated Instrumentation Facility 605 Lindy Boggs Building Tulane University 6823 St. Charles Avenue New Orleans, LA 70118-5698
I am thankful to those who took this weather thread and imparted some information that I can really use. The retired professor that taught me how to release forvar films had no idea why sometimes it worked and sometimes it didn't. Now at least I have a couple of likely variables to check.
Thanks! Chris Best Mol. Biol. Juniata College Huntingdon, PA 16652
PS - I can't help but be amused that even on a forum for scientists, the petty &/or silly items get the most responses. Tell the truth, do you stay up at night watching Jerry Springer? (For heaven sake, don't answer that!)
I am afraid that I do not agree with Scott Walck about back focal planes. I do not have a CM 12 in the lab here. So I can not check that I am not confusing the CM 12 with other Philips/FEI instruments. However, when Scott says You can not make the back focal plane of the objective lens coincident with the objective aperture. The back focal plane is well above the location of the objective aperture plane. he is wrong. On all microscopes except the very few which have very small pole-piece gaps for high resolution, the objective aperture should coincide with the back focal plane.
There is an easy and accurate way to find the true back focal plane on a CM 12 (or any other microscope with an immersion lens). Use a crystalline sample, go to convergent-beam diffraction then use the diffraction focus to make the Kikuchi lines as sharp as you can. That is the back focal plane.
If the microscope is set up properly, the image of the objective aperture should be sharply in focus at nearly the same setting of the diffraction focus. If the diffraction focus to give a sharp image of the objective aperture is very different from the diffraction focus to make the Kikuchi lines sharp, get your service engineer to reset the height of the objective aperture until they agree.
If the sample is at the correct eucentric height, a selected-area diffraction pattern in the true back focal plane will have sharp spots for a C2 setting almost but not quite to the maximum (almost fully clockwise). Again, if it does not, ask the service engineer to fix it.
Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvannia 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
Alwyn, We have a CM-12 at our other facility and I will check it out in a day or so.
I thought that I did and it worked like our JEOL 2000FX. In the 2000FX, because the lens is highly excited, there are 3 cross overs in the objective lens after the sample. That means the true back focal plane is inside the objective lens and it is not possible to put the aperture at that plane. In the 2000FX, I have focused the CBED pattern and have found the objective aperture not in focus. I have also found that the two methods that I outlined do not agree with the camera constants. The 2000FX has a condenser mini lens to make the beam parallel, but the highly excited lens allows the small probes. Since the CM-12 can form the small probes, I thought that the lens system was working in a similar manner.
I will try it out unless someone beats me to it. How about it CM owners?
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
} -----Original Message----- } From: Alwyn Eades [mailto:jae5-at-lehigh.edu] } Sent: Tuesday, February 01, 2000 3:02 PM } To: microscopy-at-sparc5.microscopy.com } Subject: SAD and the back focal plane } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } } I am afraid that I do not agree with Scott Walck about back } focal planes. } I do not have a CM 12 in the lab here. So I can not check } that I am not } confusing the CM 12 with other Philips/FEI instruments. } However, when } Scott says "You can not make the back focal plane of the } objective lens } coincident with the objective aperture. The back focal plane } is well above } the location of the objective aperture plane." he is wrong. On all } microscopes except the very few which have very small } pole-piece gaps for } high resolution, the objective aperture should coincide with } the back focal } plane. } } There is an easy and accurate way to find the true back focal } plane on a CM } 12 (or any other microscope with an immersion lens). Use a } crystalline } sample, go to convergent-beam diffraction then use the } diffraction focus to } make the Kikuchi lines as sharp as you can. That is the } back focal plane. } } If the microscope is set up properly, the image of the } objective aperture } should be sharply in focus at nearly the same setting of the } diffraction } focus. If the diffraction focus to give a sharp image of } the objective } aperture is very different from the diffraction focus to make } the Kikuchi } lines sharp, get your service engineer to reset the height of } the objective } aperture until they agree. } } If the sample is at the correct eucentric height, a selected-area } diffraction pattern in the true back focal plane will have } sharp spots for } a C2 setting almost but not quite to the maximum (almost } fully clockwise). } Again, if it does not, ask the service engineer to fix it. } } } } Alwyn Eades } Department of Materials Science and Engineering } Lehigh University } 5 East Packer Avenue } Bethlehem } Pennsylvannia 18015-3195 } Phone 610 758 4231 } Fax 610 758 4244 } jae5-at-lehigh.edu } }
Jordi: For Reichert repair and parts I would suggest Helmut Patzig of MOC (Microscopical Optical Consulting Inc.), phone (914) 268-6450 or e-mail MOCLeica-at-Aol.com Helmut is known to carry spare parts for Reichert and LKB ultramicrotomes. I have no vested interest in MOC other than as a satisfied customer. If he cannot help, you could ask him what your Reichert transformer voltage output should be and using a voltage meter adjust the voltage of a variable voltage transformer to the correct amount. Make sure to incorporate a "stop" on the dial so the voltage can't be accidentally moved above the correct amount. Henry
Henry Eichelberger, Manager Electron Microscopy Facility Department of Biological Sciences Binghamton University Binghamton, NY 13902-6000
-----Original Message----- ------------------------------------------------------ On Mon, 24 Jan 2000, Marti, Jordi wrote:
} The power supply in our cryo microtome is having problems which might be } related to the transformer. I was told by the service engineer that the } transformer is no longer supported by Reichert. Does any one know where I } can get a replacement ? } } Thanks } } Jordi Marti
I apologize in advance if this posting offends anyone. However there are a good many people who use silver membranes in their work and to have the supply from the worlds only manufacturer (to my knowledge) come to a halt, has the potential of being highly disruptive at least to some programs: ================================================= Osmonics has announced the closing of our Phoenix, AZ manufacturing facility effective 1 May 2000. Since our silver membranes are manufactured in this facility, Osmonics has decided to end production of this membrane because of declining sales and the very expensive costs associated with moving the mfg. . plant to another location. All orders placed before 1 March, 2000 will be honored and filled. ================================================== We plan to make a "last buy" before the cut off date of March 1, 2000. I would advise anyone depending on these silver membranes for their work to take stock of their future requirements because after the cut off date, sales will be possible only from remaining stocks.
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I have someone here who wants to freeze and cut cryostat sections of marine larvae (5 microns?) for immuno/light microscopy. The specimens are 100-200 microns in length.
1. What would be the best way of handling these small items? 2. What would be the best support/medium e.g TissueTek? 3. What about cryoprotection? I am familiar with 2.3 molar sucrose for EM. 4. Any general tips for immuno (protein/amino peptidases)?
Thanks - Keith _______________________ Keith Ryan (Dr) Marine Biological Association Citadel Hill Plymouth Devon PL1 2PB England
Like the other respondees I am not a CM12 user (where are they?), however, as an ex CM12 user of some 10 years ago I think that I remember the alignment that Valerie is asking about. In the basic alignment of the machine there was a step during which the SAD aperture was focussed. I think that it was in the `service calibrations' page. This may have changed in later software revisions.
The alignment of this microscope was usually carried out by the engineer when the instrument was installed. They would set up the objective lens current and adjust the specimen goniometer height to ensure that when it was at the eucentric position it was correctly in focus. Following this a complete column alignment was carried out including focussing the SAD aperture. The manufacturers then assumed that the operator would set up the specimen to the eucentric height, it should focus in the same position and the aperture should be in focus.
If the aperture is not in focus when in the SA range (as noted next to the mag readout) then check you are correctly at the eucentric position, if you are then either it was not aligned properly after installation or something has changed. NOTE: DO NOT ADJUST THE SERVICE ALIGNMENTS UNLESS YOU KNOW WHAT YOU ARE DOING AS YOU CAN LOSE THE INSTRUMENT ALIGNMENTS.
Good luck, Ron
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
I got a tip for finding the condenser lens settings required for parallel illumination from Angus Kirkland (Cambridge).
Make sure you are in microprobe mode and set up for SAED and the specimen is out of the way. Spread the beam, put in the SA Aperture, go to diffraction and then sweep the SA aperture (SAA) from side to side and minimise the deflection of the diffraction spot by tweaking the illumination control (C2 or second condenser lens). A little bit of thought will convince you that anything other than a parallel beam will give you a side to side motion (tilt) when the SAA is swept.
I have found it useful to keep a table of C2 condenser lens current settings for each spot size (C1 excitation) in microprobe mode. Setting the C2 lens for parallel illumination and adjusting the diffraction focus to get the sharpest spot will then give you near perfect SAD conditions.
Anyone contemplating electron holography for example, will have to start from the parallel illumination to get the best spatial coherence for their holograms.
******************************************************** Dr Jonathan Barnard
Analytical Materials Physics The Angstrom Laboratory, Uppsala University P O Box 534, SE-751 21 Uppsala, Sweden Phone: +46-(0)18-4716838 Fax: +46-(0)18-500131 http://www.angstrom.uu.se/analytical/home.html
I am working on fluorescent imaging of thin sections cut from samples embedded in TEM embedding resin, and am having a significant problem with resin fluorescence. Does anyone know of an embedding resin that doesn't autofluoresce?
Matt Plantinga Amway Corporation matt_plantinga-at-amway.com
We have a Zeiss Axiophot microscope and a Dvorak-Stotler Chamber that we would like to assemble into a temperature-controlled environment for digital live-cell imaging. I have several questions about the best way to combine these elements:
i) Can anyone recommend a heating stage compatible with both the microscope and the chamber?
ii) Would the best position for a temperature sensor be on the top (near the objective?) or on the bottom of the chamber, or both (two sensors)?
iii) Is it better to heat the media/fluids in their containers, or to have an in-line heater ( brand recommendations?)
iv) Are there any fuid flow or temperature considerations specific to this chamber that need to be addressed (gravity vs. pump feed, shear forces, heating stage redundant, etc.)?
v) Are there any problems related to objective lens mag/NA that need to be overcome when using this D- S apparatus?
Any references, anecdotes, or words of wisdom would be appreciated. Dealers, if you have a product that you think I could use, please don't hesitate to contact me. Thank you in advance. {FontFamily} {param} BrushScript BT {/param} {bigger}
{nofill} Regards,
Andrew Ochalski, Microscopy Technician, Dept. of Biology, University of Ottawa Room 108, Gendron Bldg. 130 Louis Pasteur, Ottawa, Ontario CANADA K1N 6N5 613-562-5800 x6343 FAX: 613-562 5486
I would like to get some information from some of the top companies on microscopes with photographic and possibly fluourescent capabilities. Please email me or call 313-993-4195 Thank you
Not really a reply but another question!! Anybody know of a similar course a little further east - England (Great Britain) to be exact !!? Thanks in Advance,
Baz
Barry Shaw Senior Imaging Technician Molecular & Cell Biology E Floor School Of Biomedical Sciences University of Nottingham Medical School NG7 2UH
We are a company that has specialized on extracting 3D information from stereoscopic SEM images. Currently we try to figure out the potential applications and the future prospects of such techniques for the microscopy community.
What we do exactly is that we take two images from a sample on the SEM from slightly different tilt angles and use digital image processing
to compute a 3D elevation model. From there you can then perform various analysis steps like extraction of 3D profiles, determination of surface roughness etc.
My questions now are:
1.) How would you judge the importance of obtaining 3D data from a SEM image? 2.) What would be the major requirements for such a data set to be useful (like density, accuracy, number of false alarms, etc.)? 3.) Is it of importance to you that you get the 3D data and the image data together and not separated (like it is the case with AFM)? 4.) How would you judge the future prospects and influence of such a technique on your personal field of work? 5.) Do you generally trust the measurement of 3D data via stereoscopic images? 6.) Do you already have experience with stereoscopic depth measurments and what are your conclusions?
Best regards, Manfred Prantl R&D Alicona GmbH, Germany www.alicona.com
Irene Piscopo of FEI/Philips probably can answer your question. I'm copying this message to her although if she's in the "field" a response may take a few days. Also, Max Otten of FEI/Philips is another good source for that type of information.
We have a CM-120 so we're aware of your problems in deciphering the operator's manual. Good luck!
Bruce F. Ingber, Biologist USDA-ARS, SRRC 1100 Robert E. Lee Blvd. New Orleans, LA 70124
-----Original Message----- } From: Valerie Leppert [mailto:vjleppert-at-ucdavis.edu] Sent: Monday, January 31, 2000 5:44 PM To: Microscopy-at-sparc5.Microscopy.Com
I can't speak about Philips CM12's, but the back focal plane of the objective coincides with the objective aperture in the CM30 here.
---------------------------------------------------------------------- Russell E. Cook, Ph.D. Electron Microscopy Center for Materials Research Argonne National Laboratory Materials Science Division, Building 212 9700 South Cass Avenue Argonne, IL 60439-4838 (630)252-7194 FAX: (630)252-4289 cook-at-horus.msd.anl.gov
Hello all: Valerie Leppart, you can reach me at 203-853-3256. I cannot call you since there is no phone number to contact. Irene Piscopo
The CM 12 microscope has four mag ranges: LM, M, SA, Mh
As long as you are eucentric and using the SA range the image plane and the diffraction aperture plane will be in focus. (It is done automatically by the instrument.) This will give you accurate SAD down to 1um. The objective mag in the plane of the diffraction aperture is approxmately 27X. If you wish to obtain diffraction from areas smaller than one micron, use uD, or uuD. If you call me I will discuss these methods with you and send you detailed instructions on using these methods.
Irene Piscopo
} -----Original Message----- } From: Ingber, Bruce F. [SMTP:bingber-at-commserver.srrc.usda.gov] } Sent: Wednesday, February 02, 2000 10:14 AM } To: Microscopy-at-MSA. Microscopy. com (E-mail) } Cc: Piscopo Irene (CS) (E-mail); Max T. Otten (E-mail) } Subject: FW: Question on Philips CM-12 SAD Alignment } } Irene Piscopo of FEI/Philips probably can answer your question. I'm } copying } this message to her although if she's in the "field" a response may take a } few days. Also, Max Otten of FEI/Philips is another good source for that } type of information. } } We have a CM-120 so we're aware of your problems in deciphering the } operator's manual. Good luck! } } Bruce F. Ingber, Biologist } USDA-ARS, SRRC } 1100 Robert E. Lee Blvd. } New Orleans, LA 70124 } } (504) 286-4270 phone } (504) 286-4419 fax } bingber-at-nola.srrc.usda.gov } } -----Original Message----- } From: Valerie Leppert [mailto:vjleppert-at-ucdavis.edu] } Sent: Monday, January 31, 2000 5:44 PM } To: Microscopy-at-sparc5.Microscopy.Com } Subject: Question on Philips CM-12 SAD Alignment } } ------------------------------------------------------------------------ } 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 there; } We have a new (to us) Philips CM-12 TEM in our lab and are wondering how } to } get the intermediate lens focused on the diffraction aperture (for making } the first image plane and the diffraction aperture coincident prior to } obtaining a SAED pattern). It doesn't seem to be covered in the manual. } Any help would be appreciated as this is a completely new microscope to } us. } Thanks, } Valerie Leppert
I don't have the reference right in front of me, but I believe that Gonzalez and Wintz say that
1. The eye can respond to an intensity range of ~10^10 in light intensity, but this is the adaptive response. The perceived brightness is a log (some will argue power law) function of the incident intensity.
2. In any one point in an image, the eye can distinguish at most ~20-30 gray levels... But... in a complex image you need at least 100 gray levels for the eye to see it as smooth. In other words, the eye seems to adapt as it scans the image.
Cheers, Henk
At 02:15 PM 2/2/00 -0600, rgriffin-at-eng.uab.edu"-at-sparc5.Microscopy.Com wrote:
} How many gray levels can the human eye distinguish? We've found several } references that disagree. } If anyone knows of a reference - that would be best. } } } Robin Griffin } UAB
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://web.ceof.ohio-state.edu Murphy's Law: "If anything can go wrong, it will." Commentary: "Murphy was an optimist."
Thanks for your reply and everyone else's reply regarding the SAD alignment. There seem to be a lot of different opinions about how this done!
We did set the sample at the eucentric and did notice that the SAD aperture is not quite in focus when in image mode. Of course this will cause the diffraction information to come from an area that is displaced with respect to the aperture position.
I am accustomed to being able to independently focus the intermediate lens on the SAD aperture, and then bring the image into focus using the objective lens - making the image plane and the SAD aperture coincident. This is on a much older Philips model and a much older Hitachi model.
However, on the CM-12, there doesn't seem to be a way for the user to do this in normal operation. I've tried a few of the suggestions (haven't worked my way through them all yet!) with no luck yet.
It does seem that this adjustment has been grouped under the category of "service alignment" in newer models.
We have a sony DXC-760 MD ccd camera that we have mounted to a standard camera lens with a bayonet adapter. Unfortunately, we have borrowed the adapter and have to return it. Is there a standard size (i.e. Nikon, Minolta, Canon) lens adapter that works with this camera mount? We can't find a part number or description for the adapter, and the book doesn't describe the size. We have already ordered and returned one that didn't fit.
Thanks in advance.
Karl Hagglund Proctor and Gamble Food and Beverage Analytical Microbiology
Fellow Listers, Does anyone know of a (preferrably free) software package which simulates ray paths, including the diffraction mode, in the TEM? We would like to use such a package to teach physics students a bit of optics.
Thanks, Eric
---------------------- Dr Eric Lachowski Department of Chemistry University of Aberdeen Meston Walk Old Aberdeen AB24 3UE Scotland +44 (0)1224 272934 fax 272921 e.lachowski-at-abdn.ac.uk
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Reply to: RE: LSCM Need help with non-fluorescing resin Dear Matt,
Try any of the immunocytochemical resins (Lowicryl, LR White or Gold, Unicryl etc). When specific antibodies are used on embedded tissue, they produce very beautiful fluorescent patterns that look like line drawings.
Paul Webster
Matt_Plantinga-at-amway.com wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I am working on fluorescent imaging of thin sections cut from samples } embedded in TEM embedding resin, and am having a significant problem with } resin fluorescence. Does anyone know of an embedding resin that doesn't } autofluoresce? } } Matt Plantinga } Amway Corporation } matt_plantinga-at-amway.com } } } } } RFC822 header } ----------------------------------- } } Received: from ultra5.microscopy.com [206.69.208.10] by mailhouse.hei.org } with ESMTP } (SMTPD32-5.05) id A77A9A2E007C; Wed, 02 Feb 2000 16:10:34 -0800 } Received: (from daemon-at-localhost) } by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id HAA02782 } for dist-Microscopy; Wed, 2 Feb 2000 07:22:38 -0600 (CST) } Received: from no_more_spam.com (sparc5.microscopy.com [206.69.208.10]) } by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id HAA02778 } for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Wed, 2 Feb 2000 } 07:21:41 -0600 (CST) } From: "Matt_Plantinga-at-amway.com"-at-sparc5.Microscopy.Com } Received: from mail1.mailrouter.net ([167.23.241.35]) } by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id HAA02771 } for {Microscopy-at-sparc5.microscopy.com} ; Wed, 2 Feb 2000 07:21:30 -0600 (CST) } Received: from lnot21.mailrouter.net ([10.10.16.153]) } by mail1.mailrouter.net (Pro-8.9.3/Pro-8.9.3) with ESMTP id IAA16955 } for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 2 Feb 2000 08:11:43 -0500 (EST) } Subject: LSCM Need help with non-fluorescing resin } To: Microscopy-at-sparc5.microscopy.com } Date: Wed, 2 Feb 2000 08:15:48 -0500 } Message-ID: {OF23F7A00C.431C958F-ON85256879.00486259-at-mailrouter.net} } X-MIMETrack: Serialize by Router on LNOT21/ANet(Release 5.0.2 |November 4, } 1999) at 02/02/2000 08:16:41 AM } MIME-Version: 1.0 } Content-type: text/plain; charset=us-ascii } Errors-to: Microscopy-request-at-sparc5.Microscopy.Com } X-RCPT-TO: {pwebster-at-hei.org} } X-UIDL: 242867701 } Status: U } Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm
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In a message dated 2/2/00 4:00:26 PM, rgriffin-at-eng.uab.edu-at-sparc5.Microscopy.Com writes:
} How many gray levels can the human eye distinguish? We've found several } } references that disagree. } } If anyone knows of a reference - that would be best.
Most of the books on image processing say the same thing (probably all copied from each other and other non-prime sources), but I don't know of an original reference based on real research. You can find the same ideas in books like Frisby "Seeing: illusion, brain and mind", Oxford, 1980; Marr "Vision", Freeman, 1982; and Rock "Perception", Freeman, 1984 which deal with the human visual system not in the context of computer graphics. These are probably closer to the original research.
The basic idea is that it takes about a 2-2.5% change in absolute brightness (i.e. a ratio of 1.025) to be just detectable, and only if it is a fairly sudden change spatially (or temporally for that matter). That is where the 20-30 distinct brightness levels over the full range of brightness that can be seen at one time (i.e. without accommodations like varying pupil size) comes from. The requirement that an entire scene have about 100 levels to avoid banding is more controversial, based on the idea that the eye can perform some accommodation as it traverses from one part of a scene to another. This probably doesn't apply when (e.g.) looking through a microscope, so the 20-30 number would be more applicable. It also ignores color, which complicates things further.
} From what is my understanding of optics - this will not give you parallel illumination at the specimen plane (if this what you meant?). By tweaking C2 you are moving the crossover plane (which is actually the diffraction plane). You can in the same way make the spot not to move by changing the diffraction focus. For each setting of C2 you can find the crossover by changing the diff. focus. Try this - spread the beam, go to diffraction, focus by diffraction focus for smallest spot, go back to imaging mode put the SAA, go to diffraction and try the procedure you described. I think the spot will not move. Ofcourse if you expand the beam too much you can no longer produce small diffraction spot (without the use of SAA) because you go out of paraxial mode. The diffraction plane is always the crossover plane not the theoretical back focal plane of the objective. Its z-position changes with the change of the C2 excitement. The spatial coherency of the electron waves is very high (actually the electrons are flying one by one through the microscope). The limiting factors are the source size, the illumination angle and the energy spread.
Please correct me if I'm wrong ... I'm still "green" in electron microscopy .. I'll be happy to learn from experienced users.
Best wishes,
Rado
--------------------------------------------------------------------- Radostin Danev Laboratory of Ultrastructure Research National Institute for Physiological Sciences Myodaiji-cho, Okazaki 444-8585, JAPAN e-mail: rado-at-nips.ac.jp --------------------------------------------------------------------- ----- Original Message ----- } From: Jonathan Barnard {Jonathan.Barnard-at-Angstrom.UU.SE} To: {microscopy-at-sparc5.microscopy.com} Sent: Wednesday, February 02, 2000 7:20 PM
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We are making preparations for experiments on determination of the chemical composition of semiconductor oxides, formed locally. The size of the modified area should be about 1 micron, and we are not sure yet if it is possible to make mentioned chemical analysis with Auger microprobe at all.
Another problem is that our newly came Auger spectroscope JAMP-7810 has no English manual with it. And believe me, it is barely possible to read that in Japanese!
We would be very grateful for your suggestions on both my questions: - how to measure the best chemical bonding on the spot with 1x1 sq. micrometer dimensions - where to get the manual (or copy of it) from. Remark: Mails to USA and Japanese representatives of JEOL gave no result!
Best regards. Dmitri
__________________________________________ Dmitri V. Sokolov, Doctor Course student Research Center for Interface Quantum Electronics, Hokkaido University, North 13, West 8, Kitaku, Sapporo 060, Hokkaido, Japan Phone 81-11-706-7174 Fax 81-11-716-6004 http://www.geocities.com/SiliconValley/Campus/1314 AOL Instant Messenger FalconDot ICQ 9418072 __________________________________________
Dear listers, I have started a research project on ammonium -bearing mineral (zeolites), and now I'm trying to quantify N using electron microprobe analysis. My problems of dealing with N are:
a) The thickness of the coating (in the literateur = approximately 150 A¡). Does anyone have information on the model of coater to making these films? Does anyone have experience with preparation of such samples?
b) Obtaining appropriate standards. (I have Si3N4, but it contains too much N (=39.96%) towards my samples (5%). Does anyone known any good nitrogen standards for such a sample ?
Any other suggestions about quantitative nitrogen analysis would be greatly appreciated !! Thanks for your time.
My best regards
Eugenia
Eugenia Marchi Universit degli Studi di Modena Dip. Scienze della Terra P.le S.Eufemia n¡.19 41100 Modena (Italy) Tel. +39-059-417289 Fax. +39-059-417399 e-mail: bengi-at-unimo.it
It's unfortunate that this manufacturer is stopping production. Aren't there other manufacturers?
Dennis B. Barr (dennbarr-at-eastman.com) Physical Chemistry Research Laboratory Physical & Analytical Chemistry Research Division Eastman Chemical Company Kingsport, TN 37662-5150
B-150B, R-132E, (423) 229-2188
} -----Original Message----- } From: Garber, Charles A. [SMTP:cgarber-at-2spi.com] } Sent: Wednesday, February 02, 2000 2:37 AM } To: MICROSCOPY BB } Subject: Silver membranes being discontinued } } ------------------------------------------------------------------------ } 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: Garber, Charles A. * EMC.Ver #3.1 ] -- } } Hello, } } I apologize in advance if this posting offends anyone. However there are a } good many people who use silver membranes in their work and to have the } supply from the worlds only manufacturer (to my knowledge) come to a halt, } has the potential of being highly disruptive at least to some programs: } ================================================= } Osmonics has announced the closing of our Phoenix, AZ manufacturing } facility } effective 1 May 2000. Since our silver membranes are manufactured in this } facility, Osmonics has decided to end production of this membrane because } of } declining sales and the very expensive costs associated with moving the } mfg. } .. plant to another location. All orders placed before 1 March, 2000 will } be } honored and filled. } ================================================== } We plan to make a "last buy" before the cut off date of March 1, 2000. I } would advise anyone depending on these silver membranes for their work to } take stock of their future requirements because after the cut off date, } sales will be possible only from remaining stocks. } } For those who do run out, and are in a bind, we are prepared help them } find } alternative filtration media, of which there are indeed some alternatives } for some applications. } } Chuck } } ============================================ } } Charles A. Garber, Ph. D. Ph: 1-610-436-5400 } President 1-800-2424-SPI } SPI SUPPLIES FAX: 1-610-436-5755 } PO BOX 656 e-mail:cgarber-at-2spi.com } West Chester, PA 19381-0656 USA } Cust.Service: spi2spi-at-2spi.com } } Look for us! } ######################## } WWW: http://www.spi.cc } ######################## } ============================================
Yes, I agree with John. A better question than "how many gray levels can one see" is probably "how many gray levels can one see under what circumstances". The answer is probably very different for a static image and a dynamic image, with edges or without, in low light conditions or bright light conditions.
The human eye (and brain) has had Millions of years to evolve and develop some rather sophisticated algorithms to deal with different conditions and the answer is probably also very complex. For example, our vision is extremely good at detecting movement. On a completely static image this tends to depress differences, while on a dynamic image it tends to enhance differences (we automatically focus on the changes).
Maybe you can test that yourself:
take the same image on a computer at different bit depths (=levels of gray). Then randomly put them on the screen (without watching the change) and try to decide, which image it is. Then do the same but watch the changes and try to decide how many gray levels you need before the changeover becomes invisible. You could do this with a "noise" image, an image that shows some object, and a smooth gray wedge. I for one would be interested to hear about the results.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
} ---------- } From: "DrJohnRuss-at-aol.com"-at-sparc5.Microscopy.Com[SMTP:"DRJOHNRUSS-at-AOL.COM"-at-SPA RC5.MICROSCOPY.COM] } Sent: Wednesday, February 02, 2000 5:56:04 PM } To: Microscopy-at-sparc5.Microscopy.Com; rgriffin-at-eng.uab.edu } Subject: Re: Gray level } Auto forwarded by a Rule } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
In a message dated 2/2/00 4:00:26 PM, rgriffin-at-eng.uab.edu-at-sparc5.Microscopy.Com writes:
} How many gray levels can the human eye distinguish? We've found several } } references that disagree. } } If anyone knows of a reference - that would be best.
Most of the books on image processing say the same thing (probably all copied from each other and other non-prime sources), but I don't know of an original reference based on real research. You can find the same ideas in books like Frisby "Seeing: illusion, brain and mind", Oxford, 1980; Marr "Vision",
Freeman, 1982; and Rock "Perception", Freeman, 1984 which deal with the
human visual system not in the context of computer graphics. These are probably closer to the original research.
The basic idea is that it takes about a 2-2.5% change in absolute brightness (i.e. a ratio of 1.025) to be just detectable, and only if it is a fairly sudden change spatially (or temporally for that matter). That is where the 20-30 distinct brightness levels over the full range of brightness that can be seen at one time (i.e. without accommodations like varying pupil size) comes from. The requirement that an entire scene have about 100 levels to avoid banding is more controversial, based on the idea that the eye can perform some accommodation as it traverses from one part of a scene to another. This probably doesn't apply when (e.g.) looking through a microscope, so the 20-30 number would be more applicable. It also ignores color, which complicates things further.
Most industrial cameras use C-mounts. C-mount to camera lens adapters are available from Edmund Industrial Optics (http://www.edmundoptics.com phone:800-363-1992) on page 165 of their 2000 catalog. They have mounts for several types of cameras, and list for $62 each. If your connecting the camera to a 'scope, Edmund sells a relay lens for $240 which mounts the camera to a 23mm eyepiece tube. For connecting to trinocular and CCD ports on a scope, Diagnostic Instruments sells relay lenses for many types of 'scopes (http://www.diaginc.com) for $299.
Michael Mohn Assistant Professor of Materials Technology Monroe County Community College Phone: 734-384-4122 Fax: 734-242-9711 http://www.monroe.cc.mi.us/mmohn
} } } "Karl Hagglund-KW" by way of Nestor J. Zaluzec {hagglund.kw-at-pg.com} 02/02 6:06 PM } } } We have a sony DXC-760 MD ccd camera that we have mounted to a standard camera lens with a bayonet adapter. Unfortunately, we have borrowed the adapter and have to return it. Is there a standard size (i.e. Nikon, Minolta, Canon) lens adapter that works with this camera mount? We can't find a part number or description for the adapter, and the book doesn't describe the size. We have already ordered and returned one that didn't fit.
Thanks in advance.
Karl Hagglund Proctor and Gamble Food and Beverage Analytical Microbiology
Are you equating brightness levels with grey levels here? I was told something like this years ago when we were buying our first digital B&W camera, when I asked the vendor about the competitors thousand plus grey levels vs his 256 grey levels and was told that the eye could not distinguish greater levels. Now, four or five years later everyone is pitching 10, 12 and 16? bit cameras, and yet the images from these cameras do look better. Is it the bits and increased grey levels? I know that if your doing image analysis at the pixel level that more bits are to your advantage but what about just photgraphic quality?
Sorry, but I think we are getting a little confused here.
1) The diffraction plane is (by definition) the objective lens focal length below the sample - it is the focus of parallel directions coming from the sample. The focal length (and parameters such as Cs, Cc) change rapidly with the objective lens current so an eucentric position makes life simple but is not necessarily the best condition to use. (For HREM you generaly want to have the objective lens stronger and reduce Cs.)
2) In general C2 does not change the focal length at all. You can have some effect from coupling of the magnetic fields, but all C2 is supposed to do is change the convergence angle.
3) The objective aperature is (approximately) in the back-focal plane, given that it is finite and height adjustments are only done coarsely (for a specific value of the objective current only).
4) The selected area aperature is (approximately) in the same plane as the object for the same reasons as above. Remember that there are positional errors in SA diffraction due to Cs (see Hirsch et al for instance).
5) You should focus Kikuchi-lines, not go for the smallest spot, since these are real object in the diffraction pattern. What you will see is then representative of you incident beam, sometimes a Gaussian range of directions.
6) The simplest way to get more parallel illumination in general is to reduce the condensor aperture size.
7) All bets are off if you have a FEG. Rather than having incoherent illumination things like the Cs of the prefield (above the sample) and postfield (below) add coherently making it much more complicated. For instance, the illumination angle varies across the field of view. While this is also there with LaB6, it is a weak effect (except for certain classes of diffracted beams, e.g. those with a screw-axis). With a FEG you can get a "spot" pattern many ways, and most of these will have large aberrations (e.g. pincushion).
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University fax: (847) 491-7820 mailto:l-marks-at-nwu.edu http://www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
Eugenia, the microprobe is not the best tool for the analysis of ammonia in zeolites because ammonia is very volatile in the electron beam. If your zeolite is phase pure it would be far better to dissolve it and do a classical chemical analysis.
regards, Eric On Thu, 3 Feb 2000 14:54:20 +0100 bengi {bengi-at-unimo.it} 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, } I have started a research project on ammonium -bearing mineral (zeolites), } and now I'm trying to quantify N using electron microprobe analysis. My } problems of dealing with N are: } } a) The thickness of the coating (in the literateur = approximately 150 A°). } Does anyone have information on the model of coater to making these films? } Does anyone have experience with preparation of such } samples? } } b) Obtaining appropriate standards. (I have Si3N4, but it contains too much } N (=39.96%) towards my samples (5%). Does anyone known any good nitrogen } standards for such a sample ? } } Any other suggestions about quantitative nitrogen analysis would be greatly } appreciated !! } Thanks for your time. } } My best regards } } Eugenia } } } Eugenia Marchi } Università degli Studi di Modena } Dip. Scienze della Terra } P.le S.Eufemia n°.19 } 41100 Modena (Italy) } Tel. +39-059-417289 } Fax. +39-059-417399 } e-mail: bengi-at-unimo.it } } }
---------------------- Dr Eric Lachowski Department of Chemistry University of Aberdeen Meston Walk Old Aberdeen AB24 3UE Scotland +44 (0)1224 272934 fax 272921 e.lachowski-at-abdn.ac.uk
I need somebody to repair a Hitachi SEM S-2700. Please contact me individually not the list. Thanks for help, -- L.Paul Bédard, ing. Ph.D. Resp. Lab. Géochimie | Lab. Manager Sciences Appliquées ; Université du Québec à Chicoutimi Canada Tél. : 418/545-5011 x 2276 | Alt. E-mail : pbedard-at-saglac.qc.ca {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} La période humaine, depuis la création d'Adam jusqu'à nos jours, n'est qu'une moisissure dans l'histoire de notre planète JCK Laflamme 1886
Let me offer my guess that the improved appearance of the images is be due largely to the improved signal-to-noise ratio in the newer cameras.
Of course there is also something to be gained in the dynamic range by the extra bits of resolution. They are much more forgiving and allow substantial changes in contrast and brightness without sacrificing the information contained in the data.
Warren S.
At 09:39 AM 2/3/2000 -0600, you wrote:
} Are you equating brightness levels with grey levels here? } I was told something like this years ago when we were buying our first } digital B&W camera, when I asked the vendor about the competitors thousand } plus grey levels vs his 256 grey levels and was told that the eye could not } distinguish greater levels. Now, four or five years later everyone is } pitching 10, 12 and 16? bit cameras, and yet the images from these cameras } do look better. Is it the bits and increased grey levels? I know that if } your doing image analysis at the pixel level that more bits are to your } advantage but what about just photgraphic quality? } } Rick Vaughn
Dear Colleagues, Has anyone used gold enhancement reagents from Nanoprobes, Inc. (Stonybrook, NY - USA) as an alternative to silver enhancement for enlarging 1 nm immunogold probes?
Advertisements state that gold enhancement has lower background and is compatible with osmium. Can anyone verify this from personal experience?
In addition to these advantages, I am hoping that milder pH conditions will be less damaging to ultrastructure in cultured neurons (using a preembedment protocol).
} The diffraction plane is always the crossover plane not the theoretical back } focal plane of the objective. Its z-position changes with the change of the } C2 excitement.
Your statement above is not wrong, but isn't the standard TEM viewpoint (or terminology). Yes, the plane at which the spot is sharpest (the in focus plane for the demagnified filament image) is the crossover plane. However to generally call this the "diffraction plane" is confusing. For example in the case of CBED, the crossover plane has moved (up or down, depending on whether C2 was initially under or overfocused) to the specimen plane. This doesn't make the specimen plane the "diffraction plane". Rather, the "diffraction plane" is now considered the in-focus plane for the C2 aperture. This is at the objective lens back-focal plane - the plane at which points on the specimen are magnified to infinity and where the position variable corresponds to the illumination angle. In collecting an SADP you can correct for slightly non-parallel illumination by correcting diffraction focus slightly off of the true back focal plane, but this should be only slight.
What was proposed in the original mail was a technique of determining how parallel the incident beam is, and of improving things by tweaking C2. If the beam isn't very parallel, you'll see the diffraction spots shift when you move the SA aperture. This is because the incident beam inclination depends on the position on the sample.
I would think the "tweak" to C2 for obtaining more parallel illumination would always in the direction of greater beam spread (unless I am misunderstanding something?).
Regards, Wharton ++++++++++++++++++++++++++++++++++++++++++++++++++ Wharton Sinkler Argonne National Laboratory West P. O. Box 2528 Idaho Falls, ID 83403
} Rado } } --------------------------------------------------------------------- } Radostin Danev } Laboratory of Ultrastructure Research } National Institute for Physiological Sciences } Myodaiji-cho, Okazaki 444-8585, JAPAN } e-mail: rado-at-nips.ac.jp } --------------------------------------------------------------------- } ----- Original Message ----- } } From: Jonathan Barnard {Jonathan.Barnard-at-Angstrom.UU.SE} } To: {microscopy-at-sparc5.microscopy.com} } Sent: Wednesday, February 02, 2000 7:20 PM } Subject: TEM:finding the parallel beam } } } } ------------------------------------------------------------------------ } } 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 got a tip for finding the condenser lens settings required for parallel } } illumination from Angus Kirkland (Cambridge). } } } } Make sure you are in microprobe mode and set up for SAED and the specimen } } is out of the way. Spread the beam, put in the SA Aperture, go to } } diffraction and then sweep the SA aperture (SAA) from side to side and } } minimise the deflection of the diffraction spot by tweaking the } } illumination control (C2 or second condenser lens). A little bit of } thought } } will convince you that anything other than a parallel beam will give you a } } side to side motion (tilt) when the SAA is swept. } } } } I have found it useful to keep a table of C2 condenser lens current } } settings for each spot size (C1 excitation) in microprobe mode. Setting } the } } C2 lens for parallel illumination and adjusting the diffraction focus to } } get the sharpest spot will then give you near perfect SAD conditions. } } } } Anyone contemplating electron holography for example, will have to start } } from the parallel illumination to get the best spatial coherence for their } } holograms. } } } } ******************************************************** } } Dr Jonathan Barnard } } } } Analytical Materials Physics } } The Angstrom Laboratory, Uppsala University } } P O Box 534, SE-751 21 Uppsala, Sweden } } Phone: +46-(0)18-4716838 Fax: +46-(0)18-500131 } } http://www.angstrom.uu.se/analytical/home.html } } } } ******************************************************** } } } } }
You can avoid your concern with C coating by coating your standards and unknowns at the same time. You should use the minimum coating thickness--just enough to prevent beam charging.
However, as stated below by Dr. Lachowski, ammonia volatility would be a big problem, especially since you will need help from high beam intensities and/or long counting times to get enough N counts to give decent precision. The expected intensity measured with a 10kV beam from a zeolite nominally of analcime composition (NaAlSi2O6.H2O) with 5 wt% N would be about 2.5% that of the intensity from Si3N4. High N intensity is difficult to achieve even with the best spectrometer and crystal designs. And to add insult to injury, the addition of the C coating absorbs even more of the N x-rays from the sample. The mass absorption coefficient for N K-alpha emission with a C absorber is huge (greater than 23,000).
Plus, you should check for possible N peak shape differences between Si3N4 (or other standards) and your ammonium-bearing zeolite.
All in all, it is a very difficult analytical problem.
Good luck!
Carl
At 4:24 PM +0000 2/3/00, Dr Eric Lachowski wrote: } Eugenia, } the microprobe is not the best tool for the analysis of } ammonia in zeolites because ammonia is very volatile in the } electron beam. If your zeolite is phase pure it would be } far better to dissolve it and do a classical chemical } analysis. } } regards, } Eric } On Thu, 3 Feb 2000 14:54:20 +0100 bengi {bengi-at-unimo.it} } wrote: } } } } } Dear listers, } } I have started a research project on ammonium -bearing mineral (zeolites), } } and now I'm trying to quantify N using electron microprobe analysis. My } } problems of dealing with N are: } } } } a) The thickness of the coating (in the literateur = } approximately 150 A°). } } Does anyone have information on the model of coater to making these films? } } Does anyone have experience with preparation of such } } samples? } } } } b) Obtaining appropriate standards. (I have Si3N4, but it contains too much } } N (=39.96%) towards my samples (5%). Does anyone known any good nitrogen } } standards for such a sample ? } } } } Any other suggestions about quantitative nitrogen analysis would be greatly } } appreciated !! } } Thanks for your time. } } } } My best regards } } } } Eugenia } } } } } } Eugenia Marchi } } Università degli Studi di Modena } } Dip. Scienze della Terra } } P.le S.Eufemia n°.19 } } 41100 Modena (Italy) } } Tel. +39-059-417289 } } Fax. +39-059-417399 } } e-mail: bengi-at-unimo.it } } } } } } } } ---------------------- } Dr Eric Lachowski } Department of Chemistry } University of Aberdeen } Meston Walk } Old Aberdeen AB24 3UE } Scotland } +44 (0)1224 272934 fax 272921 } e.lachowski-at-abdn.ac.uk
====================================== Carl Henderson Electron Microbeam Analysis Laboratory University of Michigan 2501 C.C. Little Bldg. Ann Arbor, MI 48109-1063 USA (734) 936-1550 FAX (734) 763-4690 ======================================
Does anyone out there have experience with the Technoorg-Linda IV3H ion beam thinner. We have their instruction manual but it has been a long time since we had a training session and are having some problem with the set up. Does anyone have a revised instruction manual or perhaps some tips on the alignment procedure of the guns. (please please please please).
Thanks Paul Nolan
P.S.. It is cold in Canada..everyday...all year. hehe
Now I'm really confused. I used to think I understood this somewhat.
1. In the JEOL 2010F FEG (the "all bets are off" case in your item#7), changing the illumination spread by changing C2 profoundly moves the "diffraction plane" relative to the objective aperture. [There are actually two objective apertures at different heights in this microscope, but that's beside the point]. By diffraction plane, I mean the plane below the objective lens where the diffraction spots are in sharp focus. In other words (assuming fixed OL and condenser minilens excitations), in this microscope there is only one C2 value that allows the diffraction spot pattern and OL aperture to be in focus simultaneously. If you focus on the OL aperture (with the intermediate lens), the spot pattern can be focused at only this one C2 setting. At any other C2 setting, you can focus the diffraction pattern with the intermediate lens ("diffraction focus") but the OL aperture will then be defocused. Changing the condenser minilens excitation changes the diffraction plane (relative to the OL aperture) to a different C2 setting. This is actually quite a useful feature of the microscope, e.g., for getting very intense focused diffraction patterns, but a different story.
2. The reason for choosing to focus the diffraction spots rather than the K-lines is --to put it simply-- that's where the intensity is. In amplitude contrast imaging (conventional brightfield and darkfield imaging), the intensity contribution to the image is limited by the OL aperture. It's really important to have the defining aperture and diffraction spot pattern in focus simultaneously. In addition, if I aim to measure diffraction spot patterns there is little incentive to focus the K-lines instead of the spots.
3. Textbooks such as Hirsch et al. often discuss the positional error in SA diffraction due to Cs. What readers often miss is the consequence of the fact that the error depends strongly on the diffraction angle. If measurements are confined to the low-index (low-angle) reflections, the selecting area can be reduced (to 200 nm or so at 200 kV) without big positional errors. Maybe that's an example of differences in experimental and theoretical viewpoints.
4. Reducing the condensor aperture size will give less illumination, but it won't give parallel illumination. It will change the size of the diffraction spots, but not their focus.
5. I'm not quite sure why the illumination spread has such a strong effect on the apparent position of the diffraction plane relative to the OL aperture, but suspect it arises from the action of the strong condensor-objective in this microscope. I would also question that it has much to do with coherent/incoherent beams in a FEG or with prefield/postfield Cs effects.
Larry
Larry Thomas Pacific Northwest National Laboratory MSIN P8-16 P.O. Box 999 Richland, WA 99352 Phone: (509)372-0793 Fax: (509)376-6308 Email: mailto: Larry.Thomas-at-pnl.gov
---------- From: L. D. Marks Reply To: L-marks-at-nwu.edu Sent: Thursday, February 3, 2000 8:06 AM To: Radostin Danev Cc: MSA listserver Subject: Re: TEM:finding the parallel beam
------------------------------------------------------------------------ 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 -----------------------------------------------------------------------.
Sorry, but I think we are getting a little confused here.
1) The diffraction plane is (by definition) the objective lens focal length below the sample - it is the focus of parallel directions coming from the sample. The focal length (and parameters such as Cs, Cc) change rapidly with the objective lens current so an eucentric position makes life simple but is not necessarily the best condition to use. (For HREM you generaly want to have the objective lens stronger and reduce Cs.)
2) In general C2 does not change the focal length at all. You can have some effect from coupling of the magnetic fields, but all C2 is supposed to do is change the convergence angle.
3) The objective aperature is (approximately) in the back-focal plane, given that it is finite and height adjustments are only done coarsely (for a specific value of the objective current only).
4) The selected area aperature is (approximately) in the same plane as the object for the same reasons as above. Remember that there are positional errors in SA diffraction due to Cs (see Hirsch et al for instance).
5) You should focus Kikuchi-lines, not go for the smallest spot, since these are real object in the diffraction pattern. What you will see is then representative of you incident beam, sometimes a Gaussian range of directions.
6) The simplest way to get more parallel illumination in general is to reduce the condensor aperture size.
7) All bets are off if you have a FEG. Rather than having incoherent illumination things like the Cs of the prefield (above the sample) and postfield (below) add coherently making it much more complicated. For instance, the illumination angle varies across the field of view. While this is also there with LaB6, it is a weak effect (except for certain classes of diffracted beams, e.g. those with a screw-axis). With a FEG you can get a "spot" pattern many ways, and most of these will have large aberrations (e.g. pincushion).
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University fax: (847) 491-7820 mailto:l-marks-at-nwu.edu http://www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
South Bay Technology does have quite a bit of experience with the TL IV3 Ion Mill and we are happy to offer our assistance. We market the IV3 system throughout the world for Technoorg-Linda and offer technical support as well. I will send you the most up to date IV3 manual that we have produced and will include by separate e-mail the section on the gun alignment.
I will follow up with you to be sure that you have all of the information you need.
David Henriks Vice President 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
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by INTERNET:"Paul.Nolan-at-Alcan.Com"-at-sparc5.Microscopy.Com } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Does anyone out there have experience with the Technoorg-Linda IV3H ion beam thinner. We have their instruction manual but it has been a long time since we had a training session and are having some problem with the set up. Does anyone have a revised instruction manual or perhaps some tips on the alignment procedure of the guns. (please please please please).
Thanks Paul Nolan
P.S.. It is cold in Canada..everyday...all year. hehe {
} Laurie, } } Now I'm really confused. I used to think I understood this somewhat. } } 1. In the JEOL 2010F FEG (the "all bets are off" case in your item#7), } changing the illumination spread by changing C2 profoundly moves the } "diffraction plane" relative to the objective aperture. [There are actually two } objective apertures at different heights in this microscope, but that's beside } the point]. By diffraction plane, I mean the plane below the objective lens } where the diffraction spots are in sharp focus. In other words (assuming fixed } OL and condenser minilens excitations), in this microscope there is only one C2 } value that allows the diffraction spot pattern and OL aperture to be in focus } simultaneously. If you focus on the OL aperture (with the intermediate lens), } the spot pattern can be focused at only this one C2 setting. At any other C2 } setting, you can focus the diffraction pattern with the intermediate lens } ("diffraction focus") but the OL aperture will then be defocused. Changing the } condenser minilens excitation changes the diffraction plane (relative to the OL } aperture) to a different C2 setting. This is actually quite a useful feature of } the microscope, e.g., for getting very intense focused diffraction patterns, but } a different story. } OK, the bets off case. In a FEG you have a somewhat coherent range of incident directions. The "diffraction pattern" is therefore something which is effected by the coherent aberrations of the microscope, unlike the case with incoherent illumination. Consider the case with focussed illumination, when the diffraction pattern shows an image of the condensor aperture. In a FEG As a you can "focus" this (coherent) image to a small spot by going out of focus in a true sense for the diffraction pattern. This gives you a spot-like pattern, but you will also see severe distortions at higher angles. With incoherent illumination you cannot do this and going out of focus (in the diffraction pattern) will give in general a blurred image of the condensor aperture - life is simple. What you need to do is focus "real diffraction" features, e.g. Kikuchi lines, then not play with the diffraction focus at all. As you change C2 the size of the spots will grow or shrink, this is fine.
} 2. The reason for choosing to focus the diffraction spots rather than } the K-lines is --to put it simply-- that's where the intensity is. In amplitude } contrast imaging (conventional brightfield and darkfield imaging), the intensity } contribution to the image is limited by the OL aperture. It's really important } to have the defining aperture and diffraction spot pattern in focus } simultaneously. In addition, if I aim to measure diffraction spot patterns } there is little incentive to focus the K-lines instead of the spots. } Yes, you can always get a "spot" pattern with a FEG, and it may be prettier. However, beware the distortions which make doing measurements from it very dangerous.
} 3. Textbooks such as Hirsch et al. often discuss the positional error } in SA diffraction due to Cs. What readers often miss is the consequence of the } fact that the error depends strongly on the diffraction angle. If measurements } are confined to the low-index (low-angle) reflections, the selecting area can be } reduced (to 200 nm or so at 200 kV) without big positional errors. Maybe that's } an example of differences in experimental and theoretical viewpoints. } Hirsch et al is just a good reference to read, about this as well as many other things!
} 4. Reducing the condensor aperture size will give less illumination, } but it won't give parallel illumination. It will change the size of the } diffraction spots, but not their focus. } It makes it closer to parallel. Of course, with really parallel illumination there is no intensity!
} 5. I'm not quite sure why the illumination spread has such a strong } effect on the apparent position of the diffraction plane relative to the OL } aperture, but suspect it arises from the action of the strong } condensor-objective in this microscope. I would also question that it has much } to do with coherent/incoherent beams in a FEG or with prefield/postfield Cs } effects. } Sorry, I don't think that is correct. I would suggested that people look at the distortions with a standard sample and see how bad they can be.
} Larry } } } Larry Thomas } Pacific Northwest National Laboratory } MSIN P8-16 } P.O. Box 999 } Richland, WA 99352 } Phone: (509)372-0793 Fax: (509)376-6308 } Email: mailto: Larry.Thomas-at-pnl.gov } } } } ---------- } From: L. D. Marks } Reply To: L-marks-at-nwu.edu } Sent: Thursday, February 3, 2000 8:06 AM } To: Radostin Danev } Cc: MSA listserver } Subject: Re: TEM:finding the parallel beam } } ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } Sorry, but I think we are getting a little confused here. } } 1) The diffraction plane is (by definition) the objective lens } focal length below the sample - it is the focus of parallel } directions coming from the sample. The focal length (and } parameters such as Cs, Cc) change rapidly with the objective lens } current so an eucentric position makes life simple but is not } necessarily the best condition to use. (For HREM you generaly } want to have the objective lens stronger and reduce Cs.) } } 2) In general C2 does not change the focal length at all. You } can have some effect from coupling of the magnetic fields, but } all C2 is supposed to do is change the convergence angle. } } 3) The objective aperature is (approximately) in the back-focal } plane, given that it is finite and height adjustments are only } done coarsely (for a specific value of the objective current } only). } } 4) The selected area aperature is (approximately) in the same } plane as the object for the same reasons as above. Remember that } there are positional errors in SA diffraction due to Cs (see } Hirsch et al for instance). } } 5) You should focus Kikuchi-lines, not go for the smallest spot, } since these are real object in the diffraction pattern. What you } will see is then representative of you incident beam, sometimes } a Gaussian range of directions. } } 6) The simplest way to get more parallel illumination in general is } to reduce the condensor aperture size. } } 7) All bets are off if you have a FEG. Rather than having incoherent } illumination things like the Cs of the prefield (above the sample) } and postfield (below) add coherently making it much more complicated. } For instance, the illumination angle varies across the field of view. } While this is also there with LaB6, it is a weak effect (except for } certain classes of diffracted beams, e.g. those with a screw-axis). } With } a FEG you can get a "spot" pattern many ways, and most of these will } have } large aberrations (e.g. pincushion). } } ++++++++++++++++++++++++++++++++++++++++++++++++ } Laurence Marks } Department of Materials Science and Engineering } Northwestern University } fax: (847) 491-7820 } mailto:l-marks-at-nwu.edu } http://www.numis.nwu.edu } ++++++++++++++++++++++++++++++++++++++++++++++++ } } }
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University fax: (847) 491-7820 mailto:L-marks-at-nwu.edu http://www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
Greetings, Butyl methyl methacrylate is in our hands non fluorescent totally. You can use it for TEM, although it is not as good as epoxies. Hope this helps. Tobias } } } I am working on fluorescent imaging of thin sections cut from samples } embedded in TEM embedding resin, and am having a significant problem with } resin fluorescence. Does anyone know of an embedding resin that doesn't } autofluoresce? } } Matt Plantinga } Amway Corporation } matt_plantinga-at-amway.com
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Does anyone know of a current source for this chemical? It's an ingredient of an embedding medium made up by our group. We have a supply on hand but have to reserve it if it's no longer commercially available.
} } We are very excited about the possibilities of HACH as a solution for a } } problem } } that has troubled us for a couple of years with our PU grafts. We have, } } unfortunately, one serious problem in that we cannot find a source for HA. } } We have tried Sigma Aldrich and they informed us that they withdrew } } 2-Hydroxyhexane-dial from their range in 1990. We have tried a web search } } and are contacting our local suppliers, so far without success. Do you have } } any ideas where we can find it? We are very anxious to try HACH on our PU } } samples and will gratefully appreciate any additional help that you can give } } us.
Have I been cut off, or is nobody talking anymore?
-- Dr MP Roberts Phone: [+27](0)46 603 8313 Dept of Geology Fax: [+27](0)46 622 9715 Rhodes University e-mail: malc-at-rock.ru.ac.za 6140 Grahamstown SOUTH AFRICA
I have tried GoldEnhance for preembedding protocol to label intracellular structures. It is marvelous. Buy it and use it - you will not be unsatisfied. I have no any interest in Nanoprobes, I just like these dense gold particles. Practically all they claim is true: - light insensitive - no self-nucleation - do not react with buffer ions - is not dissolved by uranil and osmium (I have treat for 1 h)
Sincerely, Dr. Alexander A. Mironov Jr., MD, PhD Unit of Morphology Dept. of Cell Biology and Oncology Consorzio Mario Negri Sud Via Nazionale, S.Maria Imbaro (Ch) 66030 Italy
} ------------------------------------------------------------------------ } 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, } Has anyone used gold enhancement reagents from Nanoprobes, Inc. } (Stonybrook, NY - USA) as an alternative to silver enhancement for } enlarging 1 nm immunogold probes? } } Advertisements state that gold enhancement has lower background and is } compatible with osmium. Can anyone verify this from personal experience? } } In addition to these advantages, I am hoping that milder pH conditions will } be less damaging to ultrastructure in cultured neurons (using a } preembedment protocol). } } Thank you, } Michael } }
The comparison between Zeiss and Olympus '100x' objective lenses is impossible unless one knows the type of lens be compared.
Catagories used to determine the quality of such a lens include: aberration correction (spherical and chromatic), correction of flatness of field, numerical aperture, the presence of an iris diaphram, whether the system is based on a 160mm tube length or infinity correction, and if the lens is used for brightfield or phase contrast for instance.
Resolution is still basically related to half the wavelength utilized. Hey, let's look at the advantages of oblique illumination for contrast enhancement and a differerent formula for resolution!!
Is your client looking at a system or an indivdual lens? Do they have a Zeiss or an Olympus system? One does not mixed and match objectives from different systems merely for the convenience of pricing.
An additional consideration is the type and correction of the condenser lens. Check the numerical aperture of not only the objective, but the condenser lens as well.
More questions than answers...Let me know if I can further complicate the answers!
Cheers! Ken
------------ Ken Tiekotter Dept. of Biology The University of Portland 5000 N Willamette Blvd. Portland, OR 97303
On Thu, 3 Feb 2000, Steve Niemela 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 } -----------------------------------------------------------------------. } } } To: {Microscopy-at-MSA.Microscopy.Com} } Subject: A comparison of optics } Date: Thu, 3 Feb 2000 10:57:19 -0600 } MIME-Version: 1.0 } Content-Type: multipart/alternative; } boundary="----=_NextPart_000_001F_01BF6E35.70E8F2E0" } X-Priority: 3 } X-MSMail-Priority: Normal } X-Mailer: Microsoft Outlook Express 5.00.2615.200 } X-MimeOLE: Produced By Microsoft MimeOLE V5.00.2615.200 } } A client of ours wonders about a comparison between Zeiss and Olympus } optics. The Zeiss 100X lense cost $10,000, and the Olympus about $4,000: } does that mean the Olympus is inferior? That's what our client's } colleagues imply. What measure of lense function is relevant? Does Olympus } (or Zeiss) have a specification, like 'resolves particles to 0.1 micron' } or something like that? Is there a measure of distortion? Best } regards, Steve NiemelaVayTek, Inc.305 West Lowe AvenueFairfield, IA } 52556email: sniemela-at-vaytek.com Phone: 515.472.2227Fax: 515.472.8131web: } www.vaytek.comgeneral email: vaytek-at-vaytek.com } } } }
} We are making preparations for experiments on determination of the chemical } composition of semiconductor oxides, formed locally. The size of the } modified area should be about 1 micron, and we are not sure yet if it is } possible to make mentioned chemical analysis with Auger microprobe at all.
I can recommend you some former colleagues of mine performing auger spectroscopy and many other semiconductor surface analytics. Please contact Prof. Wolfgang Richter: richter-at-pinet.uni-jena.de. The URL is: http://www.physik.uni-jena.de/~layer/
W'ere all waiting to hear news of MSSA Conference 2000 { :-)
Tony
} } } Dr Malcolm Roberts {malc-at-rock.ru.ac.za} 02/04/00 09:04AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Have I been cut off, or is nobody talking anymore?
-- Dr MP Roberts Phone: [+27](0)46 603 8313 Dept of Geology Fax: [+27](0)46 622 9715 Rhodes University e-mail: malc-at-rock.ru.ac.za 6140 Grahamstown SOUTH AFRICA
} Does anyone out there have experience with the Technoorg-Linda IV3H ion beam } thinner. } We have their instruction manual but it has been a long time since we had a } training session and are having some problem with the set up. } Does anyone have a revised instruction manual or perhaps some tips on the } alignment procedure of the guns. (please please please please).
We have the IV3 here in Sheffield - I'll try and help you if I can. You need to be able to actually see the beams (cover your head and the viewing glass with a black sheet if you cannot darken the room). Get one gun to fire through the center of the sample holder (tilt it at right angles to the beam) and to hit the opposite gun where it's beam would exit. Do the same for the other gun. Now keep this gun orientation, load the specimen and tilt the holder to the desired milling angle.
Hope this helps,
Alan Walker
********************************************* Alan Walker Dept of Electronic and Electrical Engineering University of Sheffield Mappin Street Sheffield S1 3JD United Kingdom
A Kevex 8000 (with huge 8 or 10" disk drives), attached to an old Amray 1000, has been donated to us. All components were fully functional when they were crated up a year ago. (They are sitting in our basement until we move into our new building this June).
A colleague in Physics (I'm a biologist) wishes to use the system to quantify elemental composition (beryllium, gallium, etc) in semi-conductor ceramics. We already have (up and running) an Hitachi HS-510 (SEM) and an Hitachi H-7100 (STEM).
1) Is this (Kevex 8000) an appropriate instrument for her to do the analyses?
2) Which would be wiser: Trying to get the Amray installed and running or getting an adapter to use the Kevex in one of the Hitachi's? (Space is a consideration, so I'd prefer not to have to install (and keep in running condition) another scope.)
3) The Kevex is pretty old. Are we wasting money trying to get or keep running something that is this old? I realize that we may need to replace the detector, since it has been at room temp for about a year. Is there anything else we should automatically consider replacing or upgrading as part of the installation?
4) She also has been sending her samples out to do x-ray analysis of the material to determine it's crystalline structure. The place she sends it to has a special x-ray diffraction instrument. Could the x-ray diffraction on the H-7100 be used to get equal results, or does this other instrument have capabilities that the TEM doesn't.
Thanks for any advice.
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
The cost of an objective depends on a number of issues, not the least of which are the corrections for chromatic and spherical aberration as well as the size of the numerical aperture. I'd need more information to really compare. "Optimizing Light Microscopy" has a detailed discussion of all of this which might be helpful. Ordering information is available on our website.
The best test is to try each system with your applications. Since much of Vaytek's work centers on deconvolution of fluorescence images, fluorescent beads of varying sizes would be a good test object for you and high throughput and crisp imaging would probably be two key benchmarks. Since fluorescence detects objects beyond the resolution limits, the normal resolution tests are meaningless. By the way, the numerical aperture of the objective is the major deciding factor on a microscope's ability to resolve fine detail (as well as providing good edge information).
If you'd like to write or call me directly, I may be able to give you further guidance.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education}
Caveat: MME does have a commercial interest in the sale of "Optimizing Light MIcroscopy:"
At 04:04 PM 2/3/00 -0600, Steve Niemela wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
If your Kevex has a Quantum thin window detector you should be able to see about half of a Boron peak, but there is no commercially available detector (that I know of) that reliably sees all of the beryllium peak. And if you have a standard window, you will only be able to see sodium and above.
I suppose the detector may still be functional even after storage. I recall that it is not a trivial thing to switch detectors between scope models. The adapters can be quite different and can get pricey. But I have not priced one in over 10 years.
I suppose with rigorous use of standards, you might be able to analyze Be by difference, but it would be a trick.
I think the Kevex 8000 may be worth installing if you are primarily interested in x-ray analysis. We used a Kevex Delta for many years and found it quite adequate. I might be able to talk you through some of the technical issues.
However, if you are interested in digital imaging or x-ray mapping, the improvements in the new equipment is fantastic compared to the old systems. We run an IXRF Systems EDS on a JEOL 840A (having replaced our Kevex) and a Link ISIS on a Hitachi 2460N. I could hardly imagine giving up their capabilities and going back to the Delta or an 8000. Disk storage, the operating environment, digital imaging can hardly be compared between the old and the new.
FWIW, Warren
At 09:17 AM 2/4/2000 -0500, Donald L. Lovett wrote: } A Kevex 8000 (with huge 8 or 10" disk drives), attached to an old Amray } 1000, has been donated to us. All components were fully functional when } they were crated up a year ago. (They are sitting in our basement until we } move into our new building this June). } } A colleague in Physics (I'm a biologist) wishes to use the system to } quantify elemental composition (beryllium, gallium, etc) in semi-conductor } ceramics. We already have (up and running) an Hitachi HS-510 (SEM) and an } Hitachi H-7100 (STEM). } } 1) Is this (Kevex 8000) an appropriate instrument for her to do the } analyses? } } 2) Which would be wiser: Trying to get the Amray installed and running } or getting an adapter to use the Kevex in one of the Hitachi's? (Space is } a consideration, so I'd prefer not to have to install (and keep in running } condition) another scope.) } } 3) The Kevex is pretty old. Are we wasting money trying to get or keep } running something that is this old? I realize that we may need to replace } the detector, since it has been at room temp for about a year. Is there } anything else we should automatically consider replacing or upgrading as } part of the installation? } } 4) She also has been sending her samples out to do x-ray analysis of the } material to determine it's crystalline structure. The place she sends it } to has a special x-ray diffraction instrument. Could the x-ray } diffraction on the H-7100 be used to get equal results, or does this other } instrument have capabilities that the TEM doesn't. } } Thanks for any advice. } } Don
I responded to Eugenia's query on another listserver (MAS microprobe listserver), but since it is also discussed here, I will add a couple things.
I did some work on ammonium-bearing minerals (with ~ 3 - 5 wt%N), using BN and later synthetic buddingtonite (ammonium feldspar) as the N standard (see Microbeam Analysis 1994, New Orleans meeting). I avoided using any epoxy for the sample prep as a precaution against N artifacts from the mounting medium. The synthetic buddingtonite can be made ala Voncken et al., 1988 (Phys. Chem of Minerals, v. 15, p. 323).
We did "normal" carbon coating, using brass color as a guide to obtain 150-200A coating (standards and unknowns). I used a synthetic crystal, 2d 60A, and ran at 10kV, 25 nA probe current. I agree that a light C coat, done at same time/conditions as the standards is important. I don't recall a substantial problem with mobility at thnese operating conditions, although the N intensity yields were low and detection limits were on the order of 0.5wt% N. The minerals are feldpar and perhaps the ammonium ion stayed put better than it woud in zeolites. I agree though, it is a difficult analytical problem. But that makes it interesting.
Jim -- ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ James J. McGee (email: jmcgee-at-sc.edu) Department of Geological Sciences University of South Carolina Columbia, SC 29208
Tel: 803-777-6300 Fax: 803-777-6610
Carl Henderson 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 } -----------------------------------------------------------------------. } } Eugenia, } } You can avoid your concern with C coating by coating your standards } and unknowns at the same time. You should use the minimum coating } thickness--just enough to prevent beam charging. } } However, as stated below by Dr. Lachowski, ammonia volatility would } be a big problem, especially since you will need help from high beam } intensities and/or long counting times to get enough N counts to give } decent precision. The expected intensity measured with a 10kV beam } from a zeolite nominally of analcime composition (NaAlSi2O6.H2O) with } 5 wt% N would be about 2.5% that of the intensity from Si3N4. High N } intensity is difficult to achieve even with the best spectrometer and } crystal designs. And to add insult to injury, the addition of the C } coating absorbs even more of the N x-rays from the sample. The mass } absorption coefficient for N K-alpha emission with a C absorber is } huge (greater than 23,000). } } Plus, you should check for possible N peak shape differences between } Si3N4 (or other standards) and your ammonium-bearing zeolite. } } All in all, it is a very difficult analytical problem. } } Good luck! } } Carl } } At 4:24 PM +0000 2/3/00, Dr Eric Lachowski wrote: } } Eugenia, } } the microprobe is not the best tool for the analysis of } } ammonia in zeolites because ammonia is very volatile in the } } electron beam. If your zeolite is phase pure it would be } } far better to dissolve it and do a classical chemical } } analysis. } } } } regards, } } Eric } } On Thu, 3 Feb 2000 14:54:20 +0100 bengi {bengi-at-unimo.it} } } wrote: } } } } } } } } Dear listers, } } } I have started a research project on ammonium -bearing mineral (zeolites), } } } and now I'm trying to quantify N using electron microprobe analysis. My } } } problems of dealing with N are: } } } } } } a) The thickness of the coating (in the literateur = } } approximately 150 A°). } } } Does anyone have information on the model of coater to making these films? } } } Does anyone have experience with preparation of such } } } samples? } } } } } } b) Obtaining appropriate standards. (I have Si3N4, but it contains too much } } } N (=39.96%) towards my samples (5%). Does anyone known any good nitrogen } } } standards for such a sample ? } } } } } } Any other suggestions about quantitative nitrogen analysis would be greatly } } } appreciated !! } } } Thanks for your time. } } } } } } My best regards } } } } } } Eugenia } } } } } } } } } Eugenia Marchi } } } Università degli Studi di Modena } } } Dip. Scienze della Terra } } } P.le S.Eufemia n°.19 } } } 41100 Modena (Italy) } } } Tel. +39-059-417289 } } } Fax. +39-059-417399 } } } e-mail: bengi-at-unimo.it } } } } } } } } } } } } } ---------------------- } } Dr Eric Lachowski } } Department of Chemistry } } University of Aberdeen } } Meston Walk } } Old Aberdeen AB24 3UE } } Scotland } } +44 (0)1224 272934 fax 272921 } } e.lachowski-at-abdn.ac.uk } } ====================================== } Carl Henderson } Electron Microbeam Analysis Laboratory } University of Michigan } 2501 C.C. Little Bldg. } Ann Arbor, MI 48109-1063 USA } (734) 936-1550 FAX (734) 763-4690 } ======================================
Since the discussion on CM12 alignment shifted to several other problems, and eventually got some JEOL 2010 users involved (Larry Thomas), I thought of throwing in this alignment question regarding the OA in a 2010 with a LaB6 gun:
To keep the camera length at a consistent value when I am not using an internal standard, I record SA diffraction patterns with C2 fully defocused cw (this is by conventional jargon, although actually using the Brightness knob I am changing C3). This is supposed to make the illumination as parallel as possible and is easily reproducible. Then I focus the direct beam with the intermediate lens (diffraction focus). CBD and NBD are another problem.
The problem is to align the Objective Aperture under this condition. If the OA is centered around the direct beam in diffraction, when switching to image mode and increasing the illumination (Brightness) for imaging conditions, the aperture will be apparently out of center. This gives the impression that the voltage center changes between a fully defocused C2 and a moderately focused (still cw from crossover, converging the illumination on the sample) condenser. That shift is obvious in DIFF mode, the direct beam can be seen to shift as C2 is turned ccw from the fully cw position.
As was noted in the on-going discussion, the crossover planes move along the optic axis as the strength of C2 changes. The helicoidal path of the beam is straight down the optic axis of the lenses only piecewise, and at the level of the OA plane it stays centered only for a limited range of C2 settings--or convergence angles. In my case, the convergence changes considerably when I record images digitally in a 1kx1k CCD camera, which screams for brightness.
My two ways around this are to (1) recenter the OA in image mode, or (2) in DIFF mode, but keeping C2 at the setting I will use in MAG mode. Both methods are a pain, mostly when doing BF/DF work, and probably incorrect.
Is there an alignment procedure for the 2010 that will prevent this " OA shift " from DIFF to MAG modes?
Augusto Morrone Seagate Technology NRW-115 7801 Computer Ave S. (612) 844-5838 Fax: (612) 844-7301
Dear Paul, Why don't you use the Hitachi service from NSC? They are the best and the cheapest. At 12:12 PM 2/3/00 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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
Can anyone recommend an aqueous mounting medium that polymerises (or whatever) to form a permanent hard mount, and that does not autofluoresce? Thanks.
We had been using silver enhancement kits from Amersham and Nanoprobe. Of these, we found the Nanoprobe kit to be very difficult to work with given it's high viscosity. We also use a pre-embed protocol, and found that both silver enhancement kits caused collagen fibrils to denature (unravel). However, during our protocol the tissue is incubated for fairly long duration in the enhancement medium (15 minutes on ice, then warmed to 25 C in a water bath for an additional 5 minutes, all in the dark).
Our more recent experience is with the Nanoprobe gold enhancement kit. It has HUGE advantages over the silver system.
First, you can Osmicate the tissue. Second, the viscosity of the components is comparatively very low, easing the use of the product. It is not so sensitive to light, and it does not cause denaturation of collagen fibrils. We do see some variability in particulate size, but this is probably a problem associated with diffusion of the media through the tissue. We have found no disadvantages of the Gold enhancement v.s. Silver and now use the Gold method exclusively.
I hope this helps,
Doug
On Thu, 03 Feb 2000 12:44:23 -0500 Michael Plociniak {plocinia-at-aecom.yu.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 } -----------------------------------------------------------------------. } } } Dear Colleagues, } Has anyone used gold enhancement reagents from Nanoprobes, } Inc. (Stonybrook, NY - USA) as an alternative to silver } enhancement for enlarging 1 nm immunogold probes? } } Advertisements state that gold enhancement has lower } background and is compatible with osmium. Can anyone } verify this from personal experience? } } In addition to these advantages, I am hoping that milder pH } conditions will be less damaging to ultrastructure in } cultured neurons (using a preembedment protocol). } } Thank you, } Michael
---------------------- Douglas R. Keene Associate Investigator Shriners Hospital Research Facilities 3101 S.W. Sam Jackson Park Road Portland, Oregon 97201 phone: 503-221-3434 FAX: 503-412-6894 (9-5 PST) e-mail: DRK-at-shcc.org
-----Original Message----- } From: Ken Tiekotter {tiekotte-at-up.edu} ..One does not mixed and match objectives from different systems merely for the convenience of pricing.... ---------------------------------------------------------------------------- ---------------------------------------------------------------------------
Of course, I've heard this sort of statement many times before. But has anyone ever done any comparisons of mixed systems to see if there are certain brands that are particularly incompatible or particularly compatible?
I ask this both as a general question and because I use a Wild M7 for which I need a pair of high eyepoint oculars. I don't need the adjusting collar built into current models and haven't been able to find an older pair used. But before I buy a Nikon/Olympus/ etc. pair I'd love to know how different the complete system will be from what I've got now.
I know many people who have used third party photo eyepieces, in just the situation where you'd think quality would be of the utmost concern. Dealers are often not the best source of info since they tend to represent one brand and, even when they are completely honest, rarely have experienc ewith a wide range of brands (especially mixed up.
Dan Freidus freidus-at-wwnet.com
P.S. Of course, if anyone has a pair of Wild 10x high eyepoint oculars for sale, that would also solve my problem (I'd also consider other magnifications). (I'm also looking for accessory objective lenses and various other accessories. Please contact me off-list if you've got anything for sale or trade.)
At 09:20 AM 2/4/00 , you wrote: } } A client of ours wonders about a comparison between Zeiss and Olympus } } optics. The Zeiss 100X lense cost $10,000, and the Olympus about $4,000: } } does that mean the Olympus is inferior? That's what our client's } } colleagues imply. What measure of lense function is relevant? Does Olympus } } (or Zeiss) have a specification, like 'resolves particles to 0.1 micron' } } or something like that? Is there a measure of distortion? Best } } regards, Steve NiemelaVayTek, Inc.305 West Lowe AvenueFairfield, IA } } 52556email: sniemela-at-vaytek.com Phone: 515.472.2227Fax: 515.472.8131web: } } www.vaytek.comgeneral email: vaytek-at-vaytek.com } }
The NA is very important in achieving high resolution. The other factor is the NA of the condenser. An Abbe condenser is easily out gunned by the aplanatic condenser (NA=1.4). Olympus makes two PlanAPO 100X oil objectives. One has an NA of 1.3 while the other is 1.4. The 1.4 lens costs today new about $6000. The lower NA lens is about $4500. So this is probably the one you are comparing. I had a Zeiss PlanAPO and recall that it had an NA of 1.3. It was terrible. I replaced it with the Olympus 1.4 and got major improvements. Of the Zeiss objectives, their 63X PlanAPO is probably the best one they have made. The rest are so so. Some are easily beaten by Olympus flourites. Zeiss is more expensive because it is Zeiss, not necessarily because it is better. The cost of manufacturing in Germany is very high compared to Japan and other places that Olympus uses. The emerging problem I see is that Olympus is moving much of its scope production to China. I have purchased all of the Olympus items I need and that were made in Japan before being stuck with Chinese Olympus. It may reach a point where Zeiss is a better product despite the higher cost.
BTW, the 1.4NA lens at $6000 reflects the recent 10% across the board price increase by Olympus.
There are important differences between older and more recent TEMs, which go to the heart of this discussion.
On older dedicated HREM's (LaB6) imaging is done with the beam at or close to crossover on the specimen, and beam convergence is determined by the user using the C2 aperture. On newer TEM's such as the JEOL 2010 (FEG or not), you need to set C2 so crossover is pretty far from the specimen plane in order to get decent coherence for imaging. Whenever this is the case, the C2 aperture isn't incoherently filled, and some of the older theory won't apply (regardless of whether the machine is or isn't a FEG).
For example, the standard method of determining convergence in an image is by switching to diffraction under the same conditions used to image, and measuring the disc diameter. (see e.g. O'Keefe and Sanders, Acta Cryst A31, p.307). This assumes the beam being near or at crossover on the specimen aperture. It won't work if the beam is far from crossover on the specimen, because now the incident angle depends on position on the specimen. The disc diameters in diffraction only indicate what the total range of incident angles is over the entire illuminated area. But the area visible in the HREM image is much smaller than this, and over this latter area the range of illuminating angles is smaller. This is just another way of saying that the C2 aperture isn't incoherently filled. If it were, would be impossible to form a shadow image of the C2 aperture at the specimen plane, as one does by spreading the beam.
Think of the illumination on the specimen as a convolution of the source image with the circular C2 aperture: When the beam is at crossover, the aperture part is approximating a delta function - we see the source in the image. When the beam is spread very far, the illumination is a nice shadow image of the C2 aperture, with a little blurriness at the edge because of convolution with a source which is not quite point-like.
In the latter case, the angular spread locally is given by the source size (demagnification) in the diffraction pattern. This can be imaged by switching to diffraction and focusing with the intermediate lens to obtain sharp source images.
This doesn't depend on whether the machine is FEG or not, though the source will be very much smaller if it is. I have posted an example in which a tungsten filament is imaged sharply by defocusing with the intermediate lens with the illumination just slightly defocused. This should be a square pattern, so note that the optics introduce significant distortions in this extreme case.
++++++++++++++++++++++++++++++++++++++++++++++++++ Wharton Sinkler Argonne National Laboratory West P. O. Box 2528 Idaho Falls, ID 83403
} } } On Thu, 3 Feb 2000, Thomas, Larry wrote: } } } Laurie, } } } } Now I'm really confused. I used to think I understood this somewhat. } } } } 1. In the JEOL 2010F FEG (the "all bets are off" case in your item#7), } } changing the illumination spread by changing C2 profoundly moves the } } "diffraction plane" relative to the objective aperture. [There are actually } } two } } objective apertures at different heights in this microscope, but that's } } beside } } the point]. By diffraction plane, I mean the plane below the objective lens } } where the diffraction spots are in sharp focus. In other words (assuming } } fixed } } OL and condenser minilens excitations), in this microscope there is only one } } C2 } } value that allows the diffraction spot pattern and OL aperture to be in focus } } simultaneously. If you focus on the OL aperture (with the intermediate } } lens), } } the spot pattern can be focused at only this one C2 setting. At any other C2 } } setting, you can focus the diffraction pattern with the intermediate lens } } ("diffraction focus") but the OL aperture will then be defocused. Changing } } the } } condenser minilens excitation changes the diffraction plane (relative to the } } OL } } aperture) to a different C2 setting. This is actually quite a useful feature } } of } } the microscope, e.g., for getting very intense focused diffraction patterns, } } but } } a different story. } } } OK, the bets off case. In a FEG you have a somewhat coherent } range of incident directions. The "diffraction pattern" is therefore } something which is effected by the coherent aberrations of the microscope, } unlike the case with incoherent illumination. Consider the case with } focussed illumination, when the diffraction pattern shows an image of } the condensor aperture. In a FEG As a you can "focus" this (coherent) } image to a small spot by going out of focus in a true sense for the } diffraction pattern. This gives you a spot-like pattern, but you will also } see severe distortions at higher angles. With incoherent illumination you } cannot do this and going out of focus (in the diffraction pattern) will } give in general a blurred image of the condensor aperture - life is } simple. } What you need to do is focus "real diffraction" features, e.g. } Kikuchi lines, then not play with the diffraction focus at all. As you } change C2 the size of the spots will grow or shrink, this is fine. } } } 2. The reason for choosing to focus the diffraction spots rather than } } the K-lines is --to put it simply-- that's where the intensity is. In } } amplitude } } contrast imaging (conventional brightfield and darkfield imaging), the } } intensity } } contribution to the image is limited by the OL aperture. It's really } } important } } to have the defining aperture and diffraction spot pattern in focus } } simultaneously. In addition, if I aim to measure diffraction spot patterns } } there is little incentive to focus the K-lines instead of the spots. } } } Yes, you can always get a "spot" pattern with a FEG, and it may } be prettier. However, beware the distortions which make doing } measurements from it very dangerous. } } } 3. Textbooks such as Hirsch et al. often discuss the positional error } } in SA diffraction due to Cs. What readers often miss is the consequence of } } the } } fact that the error depends strongly on the diffraction angle. If } } measurements } } are confined to the low-index (low-angle) reflections, the selecting area can } } be } } reduced (to 200 nm or so at 200 kV) without big positional errors. Maybe } } that's } } an example of differences in experimental and theoretical viewpoints. } } } Hirsch et al is just a good reference to read, about this as well } as many other things! } } } 4. Reducing the condensor aperture size will give less illumination, } } but it won't give parallel illumination. It will change the size of the } } diffraction spots, but not their focus. } } } It makes it closer to parallel. Of course, with really parallel } illumination there is no intensity! } } } 5. I'm not quite sure why the illumination spread has such a strong } } effect on the apparent position of the diffraction plane relative to the OL } } aperture, but suspect it arises from the action of the strong } } condensor-objective in this microscope. I would also question that it has } } much } } to do with coherent/incoherent beams in a FEG or with prefield/postfield Cs } } effects. } } } Sorry, I don't think that is correct. I would suggested that } people look at the distortions with a standard sample and see how bad they } can be. } } } Larry } } } } } } Larry Thomas } } Pacific Northwest National Laboratory } } MSIN P8-16 } } P.O. Box 999 } } Richland, WA 99352 } } Phone: (509)372-0793 Fax: (509)376-6308 } } Email: mailto: Larry.Thomas-at-pnl.gov } } } } } } } } ---------- } } From: L. D. Marks } } Reply To: L-marks-at-nwu.edu } } Sent: Thursday, February 3, 2000 8:06 AM } } To: Radostin Danev } } Cc: MSA listserver } } Subject: Re: TEM:finding the parallel beam } } } } ------------------------------------------------------------------------ } } 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 } } -----------------------------------------------------------------------. } } } } } } Sorry, but I think we are getting a little confused here. } } } } 1) The diffraction plane is (by definition) the objective lens } } focal length below the sample - it is the focus of parallel } } directions coming from the sample. The focal length (and } } parameters such as Cs, Cc) change rapidly with the objective lens } } current so an eucentric position makes life simple but is not } } necessarily the best condition to use. (For HREM you generaly } } want to have the objective lens stronger and reduce Cs.) } } } } 2) In general C2 does not change the focal length at all. You } } can have some effect from coupling of the magnetic fields, but } } all C2 is supposed to do is change the convergence angle. } } } } 3) The objective aperature is (approximately) in the back-focal } } plane, given that it is finite and height adjustments are only } } done coarsely (for a specific value of the objective current } } only). } } } } 4) The selected area aperature is (approximately) in the same } } plane as the object for the same reasons as above. Remember that } } there are positional errors in SA diffraction due to Cs (see } } Hirsch et al for instance). } } } } 5) You should focus Kikuchi-lines, not go for the smallest spot, } } since these are real object in the diffraction pattern. What you } } will see is then representative of you incident beam, sometimes } } a Gaussian range of directions. } } } } 6) The simplest way to get more parallel illumination in general is } } to reduce the condensor aperture size. } } } } 7) All bets are off if you have a FEG. Rather than having incoherent } } illumination things like the Cs of the prefield (above the sample) } } and postfield (below) add coherently making it much more complicated. } } For instance, the illumination angle varies across the field of view. } } While this is also there with LaB6, it is a weak effect (except for } } certain classes of diffracted beams, e.g. those with a screw-axis). } } With } } a FEG you can get a "spot" pattern many ways, and most of these will } } have } } large aberrations (e.g. pincushion). } } } } ++++++++++++++++++++++++++++++++++++++++++++++++ } } Laurence Marks } } Department of Materials Science and Engineering } } Northwestern University } } fax: (847) 491-7820 } } mailto:l-marks-at-nwu.edu } } http://www.numis.nwu.edu } } ++++++++++++++++++++++++++++++++++++++++++++++++ } } } } } } } } ++++++++++++++++++++++++++++++++++++++++++++++++ } Laurence Marks } Department of Materials Science and Engineering } Northwestern University } fax: (847) 491-7820 } mailto:L-marks-at-nwu.edu } http://www.numis.nwu.edu } ++++++++++++++++++++++++++++++++++++++++++++++++ } }
We are looking to localize B-glucuronidase in the mouse cochlea. We'd like to stain for the enzyme and still be able to decalcify the cochleas and process them into Epon-Araldite for LM and TEM without losing the reaction product. Does anyone have any suggestions for staining or localization protocols for this enzyme? I'm not certain whether the tissue will be taken all the way to TEM, but I do know we'll be looking at LM (1 to 4 microns). I found a method by Hayashi and Hayashi ("Electron Microscopic Cytochemistry and Immunocytochemistry in Biomedicine", CRC Press, 1993) using hexazonium pararosanilin (red in LM and electron dense for EM) on 40 micron non-frozen sections (and then processed for EM) that sounds appropriate, but I'm concerned that it may not work en bloc (the cochlea is a twisty, windy beasty).
I posted this request a few months ago and received no assistance (just individuals also interested in similar protocols and a company promoting their antibody).
Thanks so much for any direction I can get,
Jaclynn M. Lett, Research Assistant jmlett-at-cid.wustl.edu
Fay and Carl Simons Center for Biology of Hearing and Deafness Central Institute for the Deaf 818 S. Euclid Ave. St. Louis, MO 63110
===} changing C3 (brightness) leads to a shift of the bright field (BF) beam in the back focal plane (BFP) causing the objective aperture to appear off centre after converging the beam?
If you have aligned the gun properly then I suspect that the (shift) wobble alignment has not been done properly. When this button is pressed the microscope goes over to diffraction and you should see a pair of discs or spots (which represent the two extremes in tilt when the beam is shifted to its own extremes). The beam tilts are adjusted to bring the two discs/spots into coincidence. This is done for both the X and Y shift coils separately. The X wobble button shifts the beam back and forth (X shift coils), but in the back focal plane the spot or disc should remain stationary. This is what you are trying to achieve with this alignment procedure. Afterwards this should give you a beam that expands about the same point in the back focal plane. You should find that the objective aperture remains centred in both diff and image modes after doing this alignment.
As for the crossovers, yes you are right, they do move up and down when changing the brightness. The back focal plane remains stationary (by definition as L.Marks said). Anything other than a parallel beam should give you a disc in the BFP (which represents the angular distribution of rays incident on the objective lens).
What did you mean by a voltage centre? My interpretation of this is that the source high tension is wobbled and any shift in the (previously focussed) image is minimised by tuning the beam tilts. This alignment would be used for energy filtered TEM (EFTEM) when doing elemental mapping on a Gatan Imaging Filter (GIF) for example.
******************************************************** Dr Jonathan Barnard
Analytical Materials Physics The Angstrom Laboratory, Uppsala University P O Box 534, SE-751 21 Uppsala, Sweden Phone: +46-(0)18-4716838 Fax: +46-(0)18-500131 http://www.angstrom.uu.se/analytical/home.html
The San Francisco Microscopical Society Announces its next regular meeting:
Wednesday, February 9, 2000
Randall Museum 199 Museum Way San Francisco, CA 94114-1429 (415) 554-9600
7:00 pm
Topic: Introduction to Phase Contrast Microscopy by Robert D. Griffin, President, SFMS And: Annual Business Meeting and Election of Officers
Further information, including an important Letter to the Members from President Griffin, is available at http://www.microdataware.com/sfms .
The San Francisco Microscopical Society (SFMS) is a small, informal group of light microscopists who meet on a monthly basis to discuss topics of common interest. All are invited and welcome, regardless of knowledge level or professional field.
Dear Steve, Because of the nomenclature and the way manufacturers determine their specifications, it is really difficult to compare optics on paper. When I find myself in these types of discussions, I find it more useful to compare optics of similar list price rather than specifications because generally speaking, in our industry, the more things cost, the more care is taken in their manufacture. Olympus has a new 100X objective with an NA of 1.65 that sells for $10k. That would be the optic I would compare to the $10k Zeiss.
Shane Collins Scientific Instruments
-----Original Message----- } From: Steve Niemela [mailto:sniemela-at-vaytek.com] Sent: Thursday, February 03, 2000 2:05 PM To: microscopy-at-sparc5.microscopy.com
To: {Microscopy-at-MSA.Microscopy.Com}
SECOND ANNOUNCEMENT: UBC LIVE-CELL COURSE: Please register by March!!
Hello all,
The faculty for the 2000 UBC 3D Live-cell Microscopy Course is now set and we welcome several new faces, including Stephan Hell, Andreas Kriete and Steve Potter. The whole list is below.
o Stephen Adams University of California-SD o Ping Chin Cheng SUNY, Buffalo o Elaine Humphrey Univ. of British Columbia o Stephan Hell Max Planck, Goettingen o Ted Inou Universal Imaging, PA o Iain Johnson Molecular Probes, OR o Larry Keenan Cell Robotics, NM o Ernst Keller Carl Zeiss, Thornwood, NY o Andreas Kriete University of Geissen o Felix Margadant University of Sydney o Tim Murphy University of British Columbia o Jim Pawley University of Wisconsin-Madison o Steve Potter California Institute of Tech o Jason Swedlow University of Aberdeen o Michael Weis Agriculture Canada
Basic info is below but you can get the entire brochure, including links to faculty home pages, at
Organized by Prof. James Pawley, (University of Wisconsin-Madison) (SEE SABBATICAL ADDRESS AT END OF MESSAGE)
in association with Dr. Elaine Humphrey UBC BioSciences Microscopy Facility: University of British Columbia Vancouver, BC, Canada
DATES
Applications must be received by March 1, 2000 Deposit due April 15, 2000 Registration 5:00 - 7:00 PM Sunday, June 18, 2000 First Lecture 7:30 PM Sunday, June 18, 2000 Live-cell Course ends, noon Thursday, June 29, 2000
APPLICATIONS DUE BY MARCH 1, 2000
More information at : http://www.cs.ubc.ca/spider/ladic/course/bulletin.html
or
REGARDING COURSE ORGANIZATION:
Prof. James B. Pawley, (on Sabbatical) Room LG 10, Madsen Building, F-09, University of Sydney, NSW, 2006 Australia
Dr. Elaine Humphrey, Biosciences EM Facility Biosciences Building Univ. of British Columbia 6270 University Blvd. Vancouver, BC, V6T-1Z4
THE PURPOSE OF THE COURSE
Modern methods of 3D light microscopy promise a revolutionary improvement in our ability to view living cells. To help convert this promise to reality for a wider selection of biological scientists, an intensive eleven-day residential course concentrating on all aspects of 3D Microscopy of Living Cells will be held at the University of British Columbia, in June of 2000. The course includes 4 days on 2D techniques, 6 days of 3D techniques and a summary presentation. It covers basic microscopy to the highest level confocal microscopy. (A half-day Pre-course is offered for any who may need to brush up on basic optics!).
Topics include: o Quantitative 2D light microscopy o How to keep your cells alive o 3D imaging in confocal microscopy o Widefield/deconvolution techniques o Two-photon excitation microscopy o Fluorescent and backscattered light signals o Dye design, characteristics and use o Pixelation: The Nyquist Criterion o Lasers and laser tweezers o Objectives and aberrations o Scanning-systems: AODs and mirrors o Optimal pinhole size/photon efficiency o Detectors: operation and performance o Video-rate confocal imaging o Measuring ion concentrations o Display and measurement of 3D data
Morning lecture/demonstrations lead to hands-on laboratory exercises each afternoon that will utilize most of the commercial instruments currently available for 3D microscopic imaging. Students will work in groups of 3 or 4 throughout the discussion and laboratory sessions, and may complete a live-cell 3D study on their own specimens. In the first four years, over 100 students from 22 countries have attended. Last year, 11 separate, 3D microscopical workstations were available for student use under the supervision of a 17-member international faculty. We expect to have even more workstations in 2000. Including manufacturers representatives, the teacher/ student ratio will be almost 2:1.
INTERNATIONAL FACULTY
o Stephen Adams University of California-SD o Ping Chin Cheng SUNY, Buffalo o Elaine Humphrey Univ. of British Columbia o Stephan Hell Max Planck, Goettingen o Ted Inou Universal Imaging, PA o Iain Johnson Molecular Probes, OR o Larry Keenan Cell Robotics, NM o Ernst Keller Carl Zeiss, Thornwood, NY o Andreas Kriete University of Geissen o Felix Margadant University of Sydney o Tim Murphy University of British Columbia o Jim Pawley University of Wisconsin-Madison o Steve Potter California Institute of Tech o Jason Swedlow University of Aberdeen o Michael Weis Agriculture Canada
TUITION
Course tuition is $2,150 US and includes lunches. On receipt of 50% deposit, students will receive preliminary group assignments and the textbook, Handbook of Biological Confocal Microscopy, (Plenum, 1995). The tuition fee includes the textbook, a binder of handouts, and tickets for the Opening Reception, the Manufacturer's Reception and the Beach Party. Accommodations and meals other than lunch are not included in the tuition fee. The Pre-course is $100 US.
APPLICATIONS
Applicants will complete a questionnaire to assess knowledge level and field of interest. Enrollment is limited to about 24 participants. Selection will be made on the basis of background and perceived need. Those without previous LM experience will be provided with basic texts on request to read before the course begins and are encouraged to take the Pre-course on the afternoon of June 18.
Application forms, and other course information from this and past years, can be downloaded from the WWW site at:
Dr. Elaine Humphrey, Biosciences EM Facility Biosciences Building Univ. of British Columbia 6270 University Blvd. Vancouver, BC, V6T-1Z4 Phone: 1-604.822-3354 FAX: 1-604.265-5315 Email: ech-at-unixg.ubc.ca.
Application deadlines:
Application forms are due by March 1, 2000! Deadlines are early to facilitate setting up groups. Successful applicants will be notified by April 1, and a deposit of 50% must be received by April 15, 2000 to reserve your position. In general, deposit refunds are only be possible if your position can be filled from the waiting list. The remainder of the fees is due at registration.
DATES
Applications must be received by March 1, 2000 Deposit due April 15 2000 Registration 5:00 - 7:00 PM Sunday, June 18, 2000 First Lecture 7:30 PM Sunday, June 18, 2000 Live-cell Course ends, noon Thursday, June 29, 2000
TEACHING PHILOSOPHY
As befits teaching in an area at the boundary of "what is now known," lecturers have been chosen based on their expertise as scientists working in the field rather than because they all agree. They are encouraged more to be provocative than to be prosaic. Students should expect discussion in areas where differences of opinion exist.
Prior to the course, students will be organized into groups and encouraged to communicate by email/phone, about the "Living-cell" group projects that they will pursue during the course and that will be presented to the class on the last day of the course. It has been found that group interactions make best use of students' prior experience and can be very effective in teaching the practical skills covered in a hands-on course of this type.
ARRANGEMENTS FOR LIVE SAMPLES
Students must contact Dr. Elaine Humphrey to make necessary arrangements for the transport and maintenance of cell lines etc. needed for their projects. Organisms linked in any way with human disease are not permitted because of safety considerations. Transport and customs arrangements for living specimens are entirely the responsibility of the student. Students also attending the 3D Image Processing Course, may be able to analyze, process and display some of the 3D data collected from their specimens
The workshop will cover 3D image processing for measurement and display. Enrollment is limited to those attending the 3D Microscopy course. Tuition : $850 US (lunches and snacks incl.)
WHO SHOULD ATTEND?
The course is designed for biologists working with multidimensional and possibly multicolor microscopical data sets. Getting the data is only half of the battle. Image data in 3, 4 or even 5 dimensions may be difficult to store let alone analyze or display. This course is to help students understand the hardware and software aspects of this problem and give them the techniques they need to make the best use of their data.
The course is designed for biologists who need to make measurements on 3D microscopical data sets and then display the results in an effective manner. The course will be taught on SGI, Macintosh and IBM-compatible computers. A wide variety of software designed for the 3D microscopy market will be described, demonstrated and available for use.
Workshop Organizers
o Andreas Kriete Giessen University, DE o Felix Margadant University of Sydney, Au
Faculty
o Ping Chin Cheng State U. of New York, Buffalo o Chris MacLean Vaytek Inc., IA
PLAN OF INSTRUCTION Classes will meet from 8:30-12:00 and 1:00-6:00 with lecture-demonstrations followed immediately by hands-on laboratory sessions using a variety of workstations. Students will "learn-by-doing" with two to a machine. Lab handouts will describe some specific exercises to be performed on "canned" data sets. Facilities and supervision will be available until 11:00 PM, for students to work on their own data. **************************************** Prof. James B. Pawley, (on Sabbatical) Ph. 61-2-9351-7548/2351 Room LG 10, Madsen Building, F-09, FAX 61-2-9351-7682 University of Sydney, NSW, 2006 Australia JBPAWLEY-at-FACSTAFF.WISC.EDU Come to the 5th 3D Live-cell Microscopy Course, June 19-29, 2000, UBC, Canada. Info and forms at: http://www.cs.ubc.ca/spider/ladic/course/bulletin.html "A scientist is not one who can answer questions but one who can question answers." Theodore Schick Jr., Skeptical Enquirer, 21-2:39
I recently inherited a used LKB Nova ultramicrotome from a medical facility in town. The previous owner could not locate the operating instructions and I am at a loss to figure out its proper operation. Does
anyone on the list have instructions for operation of this instrument?? I would be willing to reimburse anyone for a copy of the instruction manual.
Dr. Stanley A. Rice University of Tampa srice-at-alpha.utampa.edu (813) 253-3333 Ext. 3340
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I tried a Cy5 filtercube today on a upright flourescent microscope equiped with a Diagnostics Instrument Spot Camera. My tissue had been immunolabelled with Cy5 and I did not expect to see thru the microscope. So we took pictures with the Spot Camera and the immunolabeling seemed to work. The problem was that there was uneven illumination in the pictures, half moon of brightness and the rest of the field dark. The salesperson did check that the mercury bulb was aligned correctly. Also there was no ambient light from the room causing the shadow.
Anyone with an idea of what is causing this problem?
With emphasis on Tripod Polishing and Focused Ion Beam Specimen Preparation ....
..will be offered in conjunction with the Joint Annual Meeting of the Florida Society for Microscopy and the Florida American Vacuum Society in Orlando at the University of Central Florida March 15-17, 2000.
Instructors:
Ron Anderson, IBM Fred Stevie, Lucent Technologies Lucille Giannuzzi, UCF
Vendor Sponsors:
FEI Company Micro Optics South Bay Technology
For registration material please contact Lucille Giannuzzi at lag-at-mail.ucf.edu
******************************************************************* Lucille A. Giannuzzi, Ph.D.
Associate Professor, Dept. of Mechanical, Materials, and Aerospace Eng., University of Central Florida, PO Box 162450, 4000 Central Florida Blvd., Orlando, FL 32816-2450 USA phone (407) 823-5770 fax (407) 823-0208 email lag-at-mail.ucf.edu
Director, UCF/Cirent Materials Characterization Facility, 12443 Research Parkway, Suite 305 Orlando, FL 32826 phone (407) 275-4354,5,6 fax (407) 275-4321 -------------------------------------------------------------------- "Good judgement comes from experience.
Experience comes from making bad judgement."
Mark Twain ********************************************************************
The responses to this thread have been outstanding. Yet from my perspective the unasked and thus unanswerable question is: what is the purpose for the lens? A couple of the responses have brushed the issue, but it still hasn't been specifically stated. Assuming that resolution is the main issue, the concerns of flatness of field, NA, color correction, working distance, etc, all have to be factored into the evaluation of the different lenses. It should be possible to demo both (or any of a set of) lenses under the lab and experimental conditions of real life. Only then can one truly determine which lens is the "best" (or more correctly, the appropriate) lens. All of the theory and specifications in the world don't matter if the lens doesn't aid the microscopist in obtaining the data necessary to carry out the work. One thing that is unalterable is that even if one is considering a number of "infinity-corrected" lenses from different manufacturers, they may not be truly interchangeable. Make sure the lens works on the microscope body. Then make sure it provides the performance needed. That's the right lens.
Roger Moretz
On Thu, 3 Feb 2000 16:04:55 -0600, Steve Niemela 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 } -----------------------------------------------------------------------. } } } To: {Microscopy-at-MSA.Microscopy.Com} } Subject: A comparison of optics } Date: Thu, 3 Feb 2000 10:57:19 -0600 } MIME-Version: 1.0 } Content-Type: multipart/alternative; } boundary="----=_NextPart_000_001F_01BF6E35.70E8F2E0" } X-Priority: 3 } X-MSMail-Priority: Normal } X-Mailer: Microsoft Outlook Express 5.00.2615.200 } X-MimeOLE: Produced By Microsoft MimeOLE V5.00.2615.200 } } A client of ours wonders about a comparison between Zeiss and Olympus } optics. The Zeiss 100X lense cost $10,000, and the Olympus about $4,000: } does that mean the Olympus is inferior? That's what our client's } colleagues imply. What measure of lense function is relevant? Does Olympus } (or Zeiss) have a specification, like 'resolves particles to 0.1 micron' } or something like that? Is there a measure of distortion? Best } regards, Steve NiemelaVayTek, Inc.305 West Lowe AvenueFairfield, IA } 52556email: sniemela-at-vaytek.com Phone: 515.472.2227Fax: 515.472.8131web: } www.vaytek.comgeneral email: vaytek-at-vaytek.com } } }
Roger C Moretz, Ph.D.
_______________________________________________________ Get 100% FREE Internet Access powered by Excite Visit http://freeworld.excite.com
Lesley, How about CMC (Refractive Index=1.37). CMC10 (R.I.=1.39), CMCP-9 (R.I.=1.40) and CMCP-10 (R.I. 1.41)? This is a mixture of Polyvinyl alcohol (5%), phenol(25%), lactic acid (25%), and water (45%).
I believe you can get this from Polysciences Inc. This mountant causes problems with most stained specimens, but may work for you. Is glycerine jelly auto-fluorescening?
-Ken
-------------- Ken Tiekotter Dept. of Biology The University of Portland 5000N. Willamette Blvd. Portland, OR 97303
Dan, Mixing and matching objectives is one thing, while doing the same for oculars is probably not visually as big an issue. However, in highly corrected systems, compensating eyepieces are a critical consideration if one wishes to maintain image quality, i.e., plan apo objectives require compensating oculars to maintain the correction of the apochromatic objective. This is especially true for color reproduction in color photomicrograph. This is not as important in black and white photomicrography.
-Ken
---------------- Ken Tiekotter Dept. of Biology The University of Portland 5000 N Willamette Blvd. Portland, OR 97303
On Fri, 4 Feb 2000, Dan Freidus 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 } -----------------------------------------------------------------------. } } } -----Original Message----- } } From: Ken Tiekotter {tiekotte-at-up.edu} } ..One does not mixed and match objectives from different systems merely for } the convenience of pricing.... } ---------------------------------------------------------------------------- } --------------------------------------------------------------------------- } } Of course, I've heard this sort of statement many times before. But has } anyone ever done any comparisons of mixed systems to see if there are } certain brands that are particularly incompatible or particularly } compatible? } } I ask this both as a general question and because I use a Wild M7 for which } I need a pair of high eyepoint oculars. I don't need the adjusting collar } built into current models and haven't been able to find an older pair used. } But before I buy a Nikon/Olympus/ etc. pair I'd love to know how different } the complete system will be from what I've got now. } } I know many people who have used third party photo eyepieces, in just the } situation where you'd think quality would be of the utmost concern. Dealers } are often not the best source of info since they tend to represent one brand } and, even when they are completely honest, rarely have experienc ewith a } wide range of brands (especially mixed up. } } Dan Freidus } freidus-at-wwnet.com } } P.S. Of course, if anyone has a pair of Wild 10x high eyepoint oculars for } sale, that would also solve my problem (I'd also consider other } magnifications). (I'm also looking for accessory objective lenses and } various other accessories. Please contact me off-list if you've got anything } for sale or trade.) } } } } } }
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Reply to: RE: Aqueous mounting medium Dear Lesley,
You can make a very useful mounting medium from inexpensive ingredients. The main ingredient is poly(vinyl) alcohol which is soluble in water but will dry without shrinking too much. The water solubility means you can take previously mounted slides, rehydrate them and use them again.
Fading of fluorescent dyes can be retarded by including anti-fade compounds.
You can find the recipe at {http://www.hei.org/htm/moviol.htm} .
Details of anti-fade compounds can be found here (at the bottom of the page) {http://www.hei.org/htm/ifse.htm} . Ths site is getting a little outdated so if you feel there are details missing then please contact me with your questions.
Many thanks to Heinz Schwarz of Tuebingen for writing these protocols.
Regards,
Paul Webster
Paul Webster, Ph.D. Associate Scientist & Director Ahmanson Advanced Electron Microscopy & Imaging Center House Ear Institute 2100 West Third St. Los Angeles, CA 90057
by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id VAA13116 for dist-Microscopy; Sat, 5 Feb 2000 21:13:46 -0600 (CST) Received: from no_more_spam.com (sparc5.microscopy.com [206.69.208.10]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id VAA13113 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Sat, 5 Feb 2000 21:12:48 -0600 (CST) Received: from hme0.mailrouter01.sprint.ca (hme0.mailrouter01.sprint.ca [207.107.250.16]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id VAA13106 for {Microscopy-at-sparc5.Microscopy.Com} ; Sat, 5 Feb 2000 21:12:36 -0600 (CST) Received: from normandlaurier (spc-isp-mtl-58-8-589.sprint.ca [149.99.151.82]) by hme0.mailrouter01.sprint.ca (8.8.8/8.8.8) with SMTP id WAA18723; Sat, 5 Feb 2000 22:07:44 -0500 (EST) Message-ID: {000b01bf7046$84afd240$52976395-at-normandlaurier} "Dr. Gary Gaugler" {gary-at-gaugler.com} References: {v03007801b4bfabe09222-at-[129.78.149.225]} {4.2.2.20000204145256.00b2a820-at-pop.calweb.com}
I would call this a typical PARTIAL judgment. What you see is what you get. Thee only way, to the best of my experience, is to get both objectives, side by side if possible, and to do your own evaluation. I have honestly doubts concerning those values set on objectives. Things change and optic too and sometimes faster that we could expect. Norm ----- Original Message ----- } From: Dr. Gary Gaugler {gary-at-gaugler.com} To: MSA listserver {Microscopy-at-sparc5.Microscopy.Com} ; {sniemela-at-vaytek.com} Sent: Friday, February 04, 2000 5:02 PM
I am interested to know what the present state of the art is re: solid state electon emitters, total electron beam currents available and also beam current densities. If anyone could point out a recent review article, I would appreciate it. Thanks,
Don Marshall
Donald J. Marshall Relion Industries P.O. Box 12 Bedford, MA 01730 Ph: 781-275-4695 FAX: 781-271-0252 email dmrelion-at-world.std.com
Cathodoluminescence, mass spectroscopy, electron beam technology
"A weed is a flower out of place."
(Please note: Do not send email with attachments to this address. Instead, send it to donbarlen-at-aol.com. Thank you.)
In fact, if you're really interested in getting the BEST lens, try at least three examples of the ones you are interested in . All high NA lenses must meet some minimal standard but some are significantly better than that minimum and the only way to find one that is is to try THE lens you are going to buy - not just an example.
Bill Miller
At 10:04 PM 2/5/00 -0400, Norm wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Do not have difraction sofrware, but here are some URLs where you can download free simulation software I use for e-beam to solid interactions, hope it may be useful for you.
Casino is a freeware for monte carlo simulations: http://www.gme.usherb.ca/casino/
Monte carlo resources: http://dsa.dimes.tudelft.nl/pattern_definition/monte/monte.html
My FAVORITE monte carlo software: http://web.utk.edu/~srcutk/htm/simulati.htm
Valery Ray, MSEE, SEM engineer Applied Materials 001-208-8413744
Hi everybody While this has been an interesting discussion we seem to have got away from the original question.
This is the method I use - after of course ensuring normal alignments and adjustments have been carried out
Insert the objective aperture with a specimen in the beam. Focus the image of this aperture in Selected Area Diffraction using the first intermediate lens focus.
Focus the diffraction pattern (i.e. the smallest zero order spot) with the final condenser lens, this will give a parallel beam incident upon the specimen.
To test this, remove the objective aperture and insert the largest selected area aperture. If the beam is parallel moving this aperture will not move the diffraction spot.
Tilt - The purpose of this alignment is to ensure that when the beam is tilted it doesn't move on the specimen. This is double tilt correction and depends on using double deflector coils.
Shift - Double shift correction is meant to ensure that when the beam is shifted it doesn't tilt. The operation of this alignment is dependent upon the value of the first intermediate lens. The best value for this alignment is a parallel beam condition as described above. By the very nature of the beast it will be found to be impossible to perform this alignment correctly at some CBD conditions. This is meant to be a convergent beam mode after all not a parallel beam mode.
I understand that this is also referred to as tilt and shift purity.
Richard Hey
The views expressed herein are purely my own and do not reflect those of my employer
Richard Hey Technical Support Engineer Jeol (UK) Ltd Phone: (44) 1707 377117
Dear colleagues, I am now currently using TEM to observe a filamentous structure so-called hrp pilus produced by plant pathogen, Pseudomonas syringae which is grown on planta mimic medium agar plates. Even though I could easily find this structure through directly put the nickel-grid on the bacteria plate for a certain time,followed by the negative staining, I can't convince other people to believe it's the structure we are looking for as its size is around 7-10 nm, which is indistinguishable from that of bacterial flagellum. So we want to label this structure by using the specific antibody against its major structural protein followed by the 10 nm gold-conjugated second antibody. The difficulty I am encountering now is most pili are washed away after the whole immnuogoldlabeling process. Since I am a beginnner on TEM work, I don't know whether it's a very commonly encountered problem in immnogoldlabeling experiment or just a very special case happened to me, such as the pilus i'm interested in is too vulnerable to survive the exclusive wash, or I didn't master the whole immunogoldlabeling techinique. If it's a common problem, I hope someone could kindly provide me with your valuable experience on how to resolve this problem, how we can prevent them to be washed away. If it's due to my techinical problem, could you please send me a very detailed immunogoldlabeling protocol. Any kind of help will be highly appreciated have a nice week yours qiaoling jin jinq-at-msu.edu
Thank you for your responses. Please note that I do perform the TILT and SHIFT alignments (aka "pivot point alignment" in other instruments) on a regular basis, but this doesn't solve the problem.
Regarding: "...What did you mean by a voltage centre? My interpretation of this is that the source high tension is wobbled and any shift in the (previously focussed) image is minimised by tuning the beam tilts. This alignment would be used for energy filtered TEM (EFTEM) when doing elemental mapping on a Gatan Imaging Filter (GIF) for example".
This is a (standard) routine alignment for imaging as well. The voltage center alignment is done by first focusing the specimen at the optimum OL current (adjusting Z), then adjusting the Tilt (in image mode) and Shift (in diffraction mode), and finally adjusting the beam tilts to minimize the image shift at the center of the screen. The problem arises when the Brightness is changed, as is typically adjusted when optimizing the illumination on the sample to record an image (intense illumination) versus an SA diffraction pattern (fully overfocused condenser). Apparently this brightness change affects the voltage center as evidenced by a shift of the direct beam as that brightness change is effected in the SAD mode. This shift in the direct beam is equivalent to a beam tilt.
I regret to say that there are some important points which perhaps are being missed. Classic optics, i.e. most TEM textbooks and introductory optics texts are for one of two cases: a) Every point in the condensor aperture (CA) is incoherent (i.e is not related in phase in a statistical sense) with respect to any other point. b) Every point in the CA is coherent (i.e. has a fixed phase) with respect to any other point.
Approximation a) is used in most computer codes for HREM, approximation b) for STEM.
Alas, both a) and b) are only approximations! Current microscopes, particularly FEGs (but perhaps others as well) operate under conditions when neither of the two approximations are valid. While the analytic theory is well established (see the Chapter in Born and Wolff on partial coherence), it is not easy and there are no clean solutions that you can write down. (It can be solved numerically, but this would require big 4-D FFT's which are beyond most current computers.)
The simplest test is to fully focus the illumination. If the spot size is A nm, the coherence in the CA is about 1/A nm-1 (which you can convert to an angle as appropriate). If this is small relative to the CA radius you are working with case a) above. If it is about the same as the CA radius you have b). If it is 2-3 times less, neither approximation is correct!
The moral of the story: 1) If you have a), you can simulate an HREM image with confidence using any of the available programs. They are WRONG quantitatively otherwise. 2) If you have a), you can use simple "ray-diagram" optics, but not otherwise. 3) If you have b) the simple equations for STEM operation are valid, but not otherwise.
Sorry, but as microscopes improve the theory we need to understand them can change.
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University fax: (847) 491-7820 mailto:l-marks-at-nwu.edu http://www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
I am trying to get a handle on the scale of some old pictures. There are human RBC in theses pictures. Can anyone tell me the diameter of your average human RBC?
Thanks Mike
-- _________________________________________________ / Michael J. Herron / / U of MN,Medicine/Infectious Diseases / / herro001-at-maroon.tc.umn.edu / / http://128.101.243.213 / /_____________________________________________/
When a characteristic X-ray is given off from an atom, it carries 1 h-bar (Planck's constant divided by 2 pi) of angular momentum. The electronic transition that occurred for the X-ray to come off must conserve angular momentum of the system. Therefore, a transition from a 2s1/2 to a 1s1/2 state is forbidden by selection rules. You can't produce a Be X-ray. -Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Mike: According to Fawcett's CONCISE HISTOLOGY (page 32), "In the blood, erythrocytes are 8.5 micrometers in diameter, but in the dehydrated state, in dried blood smears, they measure 7.6 micrometers." --Kent
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ A. Kent Christensen, Professor Emeritus Department of Cell and Developmental Biology, Medical Science II Building University of Michigan Medical School, Ann Arbor, MI 48109-0616 Office: 2703A Medical Science II Building Tel (work) (734) 763-1287, Fax (work) (734) 763-1166 E-mail: akc-at-umich.edu http://www.umich.edu/~akc/ ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ --On Mon, Feb 7, 2000 9:20 AM -0600 Michael Herron {herro001-at-maroon.tc.umn.edu} wrote:
} All, } } I am trying to get a handle on the scale of some old pictures. There } are human RBC in theses pictures. Can anyone tell me the diameter of } your average human RBC? } } Thanks } Mike } } -- } _________________________________________________ } / Michael J. Herron / } / U of MN,Medicine/Infectious Diseases / } / herro001-at-maroon.tc.umn.edu / } / http://128.101.243.213 / } /_____________________________________________/
John, If they do, I hope that they will respond to the whole list because we have clients who have experienced similar problems, especially re: computers that are controlling VIS spectrometers collecting CL spectra. I know that there are in line RF filters available (e.g., Steward Co.) and there may be others but the final word is not in on how well these work yet.
Don Marshall
} From microprobe-at-microanalysis.org Mon Feb 7 11:43:28 2000 } } } From: John Hunt {hunt-at-ccmr.cornell.edu} } Subject: Computer lock-ups from static } To: microprobe-at-microanalysis.org } } } Dear Probe community, } It is winter in Central NY, and the room humidity is 33%. Static } electricity (I believe) has been causing frustrating computer lockups } requiring rebooting our microprobe workstation. } I am wondering if anyone has found the best solution for preventing these } problems. } } } } } John Hunt } Cornell Center for Materials Research } 607-255-0108 microprobe } -3789 SEM/TEM } -2365 fax }
Donald J. Marshall Relion Industries P.O. Box 12 Bedford, MA 01730 Ph: 781-275-4695 FAX: 781-271-0252 email dmrelion-at-world.std.com
Cathodoluminescence, mass spectroscopy, electron beam technology
"A weed is a flower out of place."
(Please note: Do not send email with attachments to this address. Instead, send it to donbarlen-at-aol.com. Thank you.)
I would like to tap the wealth of information out there.
We are experiencing a lot of problems with the tungsten filaments that we are currently using on our JEOL 5800 SEM. During long acquisitions (many hours) there is a lot of brightness drift and shifting, even when lengthy stabilization periods are given prior to the acquisitions. JEOL has checked out the SEM itself, and can't find the source for the drifting. I was thinking that it might be the filament. I was wondering if there are different types of tungsten filaments out there, and which is the most stable over long periods of time. I would like to try out all of them (considering trying to switch our entire system to a LaB6 filament is a bit much for now). Any recommendations or experiences you have would help us out a lot.
Thanks in advance!
Marisa Ahmad R&D Specialist Semiconductor Insights mahmad-at-semiconductor.com
weather report (for those interested): It's unusually sunny and warm for here in Ottawa.....only -10 degrees C (but it feels like -25 with the wind). It's such a nice day that I washed my car this morning since it's not really supposed to be brown - now all the doors and windows are frozen shut. {sigh}
In my experience, RBCs prepared for SEM by critical point drying or HMDS shrink even more and may be only 4 or 5 microns across.
Marie } } Mike: According to Fawcett's CONCISE HISTOLOGY (page 32), "In the blood, } erythrocytes are 8.5 micrometers in diameter, but in the dehydrated state, } in dried blood smears, they measure 7.6 micrometers." --Kent }
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology, U-2131 University of Connecticut Storrs, CT 06269-2131 Phone: 860-486-3588 Fax: 860-486-1936
It depends on the fixation/dehydration parameters. Somewhere in the nieghborhood of 4 microns, dependant on the angle of measurement and the angle of the sample with respect to the camera. } Date: Mon, 07 Feb 2000 09:20:44 -0600 } From: Michael Herron {herro001-at-maroon.tc.umn.edu} } X-Accept-Language: en } MIME-Version: 1.0 } To: Microscopy listserv {Microscopy-at-sparc5.Microscopy.Com} } Subject: Diameter of human RBC? } Content-Transfer-Encoding: 7bit } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Can anyone tell me what a Brandes dip is and how it is done? The only thing I know is that it can be used to detect virus particles from plant sap but I can't find any references on it.
Thank You!
Margaret
Margaret Dienelt
Plant Pathologist Electron Microscopy Lab
Floral and Nursery Plants Research Unit U.S. National Arboretum/Agricultural Research Service/USDA
B. 010A, Rm. 238, BARC-W 10300 Baltimore Avenue Beltsville MD. 20705 USA
Does anybody know any program capable of doing TEM simulation of dislocation network, e.g., misfit dislocations in plane view TEM? I know some programs can do one or two dislocations, but what I need is the simulation for a set of dislocations. Thanks a lot.
It has been very interesting reading about diffraction and electron beams but it is quite clear that we often give the instrument credit, "the instrument does it automatically", when credit is not due. Almost every "automatic" is a designers estimate or better still a calculation, it is a setting!
After 36 years with TEM I have to say that the only area that the instrument could set in the SA mode could be the approximate position of the first image plane. We normally move to the SA mode to free up the diffraction lens (1st intermediate) to enable it to be focussed on the diffraction (intermediate) aperture independent of the magnification system. We would then focus the image inside the aperture with the objective lens to ensure that the selected area IS the area in the diffraction pattern. Without this procedure electron rotation and misalignments could give you a diffraction pattern from an area not in the centre of the screen! Users are correct in that if they set the eucentric point or better still, adjust the specimen height to focus at the same objective lens current value, that done the first image plane will be at the same point within the diffraction (intermediate) lens.
So how could the Philips work? I would guess they set the diffraction lens at approximately the first image plane. Then as they do not use it within their lens scheme for the SA magnifications it is fixed at this focus, good enough I think?
A second point, parallel beams are very important if you are chasing the best image quality, biology or materials, most times we need high coherence. There are misunderstandings on how to obtain a parallel beam or high coherence, setting the final condenser underfocus is incorrect. The procedure for high coherence would be to use the smallest spot size you can tolerate (this probably means you must up the emission current). Once in this condition overfocus the final condenser (clockwise from crossover) the spot, what ever size it is now, becomes your virtual source. The further overfocus you go the greater the beam coherence and that is what we are after. You will deduce the smaller the condenser aperture the sharper the spot and the smaller the spot the greater the coherence for a given degree of overfocus.
Work with a design team and they expect everyone to overfocus, they expect everyone to use small condenser apertures and they expect everyone to use small spot sizes. They do not expect everyone to use too low an emission current (almost everyone does, we have talked before about filament life being the most important feature of many laboratories) because this makes the task too difficult! There must be people more knowledgeable than I who will cut out all the guesswork?
Steve Chapman Senior Consultant Protrain Electron microscopy courses world wide http://www.emcourses.com Tel & Fax 44 (0) 1280 814774
} I am trying to get a handle on the scale of some old pictures. There } are human RBC in theses pictures. Can anyone tell me the diameter of } your average human RBC?
When I was a lad...... in grad school, we were told it was a useful reference at 7um diameter.
Saturday, April 21, Sunday, April 22, 2000 Saturday, April 28, Sunday, April 29, 2000
An advanced course on polarized light microscopy which will cover the following topics: The nature of polarized light The origin and interpretation of interference colors Birefringence and crystal orientation The Indicatrix Compensation and variable compensators Interference figures and their interpretation
The workshop will consist of two consecutive weekends of lectures and hands on labs to cover the theoretical and practical aspects of polarized light microscopy.
The course instructors include;
Jan Hinsch of Leica, Inc.,
John Reffner of Trace Consulting,
N.Y.M.S. Instructor Donald O'Leary.
WHEN: April 22, 23, 28 & 29, 2000 from 10 A.M. to 4 P.M.
WHERE: Location To be Announced.
COST: $275 for N.Y.M.S. members, $295 for non-members (includes membership) Lunch and course materials are included. Checks made out to N.Y.M.S.
WHO: advanced course for those who have completed "The Use of the Microscope" or are experienced in microscopy and familiar with the theory of its use.
HOW: Register using the form below. Limited to the first 12 registrants.
FURTHER INFORMATION: Contact Donald O'Leary
eMail: mailto:donoleary-at-worldnet.att.net
(201) 797-8849 Voice Phone Number
PLEASE POST ------------------------------------------------------------------------------------------------------------
Return form to Donald O'Leary, 6 Chittenden Road, Fair Lawn, NJ 07410.
Registration Form
Polarized Light Microscopy, April 22, 23, 28 & 29, 2000
N.Y.M.S. Member _________________ ($275.00) Non-Member __________($295.00)
Name ___________________________________________________________________________
I'm just taking a guess. Is it possible, depending on the type of filters used in the cube, that infrared is passing through as flare if your first excitation filter is not blocking the IR? Or there may be a coating problem on your filter? We have an IR blocking filter right after the mercury lamp. The CCD cameras are extremely sensitive to IR and this has caused problems on our system. But in your situation I don't know why it would pass through. So it is probably something else.
Bob Morphology core U of Washington
On Fri, 4 Feb 2000, Patricia A. Glazebrook 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 tried a Cy5 filtercube today on a upright flourescent microscope equiped } with a Diagnostics Instrument Spot Camera. My tissue had been } immunolabelled with Cy5 and I did not expect to see thru the microscope. So } we took pictures with the Spot Camera and the immunolabeling seemed to } work. The problem was that there was uneven illumination in the pictures, } half moon of brightness and the rest of the field dark. The salesperson did } check that the mercury bulb was aligned correctly. Also there was no } ambient light from the room causing the shadow. } } Anyone with an idea of what is causing this problem? } } Thank-you, Pat Glazebrook } } } } }
One more thought to support Gary's observation about the importance of the NA of the condenser: The full equation for the limit of resolution is R = (1.22)(wavelength)/(NAobjective + NAcondenser). However, if you don't oil it to the back of the slide, you might as well use a lower NA condenser. Reminds me of the hematology scope built by an otherwise sensible company, meant to be used in ultra high resolution applications. The manufacturer was very good about providing a quality, high NA, oil immersion objective as well as a high NA condenser. The only problem was that you could never raise the condenser high enough to oil it to the back of the slide and, therefore, use it at the NA inscribed... only at about .95! What a waste of time and money!
Also, a reminder that the NAs engraved are the maximum working limit. Closing down an aperture iris or an iris in the back focal plane of the objective reduces the NA accordingly.
One last fact: the NA affects both resolution AND edge information, so, for the crispest, highest resolution images, go for high NA in both objective and condenser.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education}
At 03:02 PM 2/4/00 -0600, Dr. Gary Gaugler wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Marisa Ahmad: } I would like to tap the wealth of information out there. } } We are experiencing a lot of problems with the tungsten filaments that we } are currently using on our JEOL 5800 SEM. During long acquisitions (many } hours) there is a lot of brightness drift and shifting, even when lengthy } stabilization periods are given prior to the acquisitions. JEOL has } checked out the SEM itself, and can't find the source for the drifting. I } was thinking that it might be the filament. I was wondering if there are } different types of tungsten filaments out there, and which is the most } stable over long periods of time. I would like to try out all of them } (considering trying to switch our entire system to a LaB6 filament is a bit } much for now). Any recommendations or experiences you have would help us } out a lot. } Dear Marisa, Do you have control over the filament voltage? If so, set the voltage just a hair over that necessary to saturate the filament. If the voltage is too low, parts of the filament will not emit electrons which get through the wehnelt and into the beam; if too high, there may be significant evaporation of the filament; either may cause drifting and/or shifting. I have had very good luck with the stabilities of both Agar and Ebtec (now Energy Beam Sciences) filaments; not that others may not be as good, I haven't tried them. Ebtec makes several shapes of filament, and one may be best for long-term stability. I have no interest in Agar or Ebtec except as a satisfied customer. Yours, Bill Tivol
Have you considered a "local" remedy, like putting up a humidifier? I know that static can be an issue, but I have never heard of it being this bad before. In more humid environments, we often put a small enclosure around a light microscope then put out a dish of Drierite. Perhaps some simple plastic sheeting, like they sell for exterior storm windows/wind breaks, can be hung from the ceiling around your microprobe plus the humidifier will keep the humidity immediately around your microprobe more balanced.
Best of luck on a tough problem.
Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education}
At 12:06 PM 2/7/00 -0500, donald j marshall wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Is this a Spot Jr. or a SpotII? The IR filter in the Jr. cuts off at 650 nm, the SpotII puts it on a slider. Is it all the way out or [partially occluding the image? Bob Fern caught a lot of the possible issues, but is your lamp properly focussed and aligned? Are you picking up light through the eyepieces? We have a Nikon that will show a semi-circle of light in from a recessed ceiling light that projects to the scope at an angle.
Regards, Glen
Glen MacDonald Research Scientist Hearing Research Laboratories of the Virginia Merrill Bloedel Hearing Research Center Box 35-7923 University of Washington Seattle, WA 98195-7923 (206) 616-4156 glenmac-at-u.washington.edu
On Fri, 4 Feb 2000, Patricia A. Glazebrook 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 tried a Cy5 filtercube today on a upright flourescent microscope equiped } with a Diagnostics Instrument Spot Camera. My tissue had been } immunolabelled with Cy5 and I did not expect to see thru the microscope. So } we took pictures with the Spot Camera and the immunolabeling seemed to } work. The problem was that there was uneven illumination in the pictures, } half moon of brightness and the rest of the field dark. The salesperson did } check that the mercury bulb was aligned correctly. Also there was no } ambient light from the room causing the shadow. } } Anyone with an idea of what is causing this problem? } } Thank-you, Pat Glazebrook } } } } }
Out here in the dry West (here we have Summer humidity around the 33% value you mention, in Winter we go a lot further down) we have the same problem. There are a few methods to help:
1) increase humidity. Perhaps you can use a humidifier in your equipment room? 2) prevent charging. There are shoes available that prevent charging. 3) get rid of charging. There are several options here:
a) The easiest way and what I usually do if no other method is available, is to touch some metal part NOT of the PC but a chair or table before touching the PC. This requires a) that a suitable object is near the PC, b) that you're not afraid of "shocking" yourself, and c) that you don't move around a lot while working on the PC.
b) Purchase a grounding mat to place in front of the equipment. these mats are connected to the AC ground (use the same as the PC ground). If the mat is big enough so that everybody has to step on the mat first AND is wearing the conductive shoes, everything should be OK.
c) wear grounded wrist straps. These require of course that you put them on.
We use a combination of these techniques to protect our equipment both in the office and the production floor (believe me, it IS necessary here). As bad as it is for electronic equipment, though, it is just great for comfort levels outside. 90 degrees (F) feel just right for bicycling, climbing, rafting...
One company where you can buy this stuff is called "-at-once". Their web site is www.4atonce.com. I just happened to find their catalog first. There are many other companies out there that sell static control equipment. We have no financial interest in this company whatsoever.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
} ---------- } From: donald j marshall[SMTP:DMRELION-at-WORLD.STD.COM] } Sent: Monday, February 07, 2000 10:06:10 AM } To: hunt-at-ccmr.cornell.edu } Cc: Microscopy-at-sparc5.Microscopy.Com } Subject: Re: Computer lock-ups from static } Auto forwarded by a Rule } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
John, If they do, I hope that they will respond to the whole list because we have clients who have experienced similar problems, especially re: computers that are controlling VIS spectrometers collecting CL spectra. I know that there are in line RF filters available (e.g., Steward Co.) and there may be others but the final word is not in on how well these work yet.
Don Marshall
} From microprobe-at-microanalysis.org Mon Feb 7 11:43:28 2000 } } } From: John Hunt {hunt-at-ccmr.cornell.edu} } Subject: Computer lock-ups from static } To: microprobe-at-microanalysis.org } } } Dear Probe community, } It is winter in Central NY, and the room humidity is 33%. Static } electricity (I believe) has been causing frustrating computer lockups } requiring rebooting our microprobe workstation. } I am wondering if anyone has found the best solution for preventing these } problems. } } } } } John Hunt } Cornell Center for Materials Research } 607-255-0108 microprobe } -3789 SEM/TEM } -2365 fax }
Donald J. Marshall Relion Industries P.O. Box 12 Bedford, MA 01730 Ph: 781-275-4695 FAX: 781-271-0252 email dmrelion-at-world.std.com
Cathodoluminescence, mass spectroscopy, electron beam technology
"A weed is a flower out of place."
(Please note: Do not send email with attachments to this address. Instead, send it to donbarlen-at-aol.com. Thank you.)
I am planning to try EM immunolabelling with GFP antibodies. Does anyone out there know of the best source for GFP antibodies and had any luck with using them for TEM. I would like to try pre-embedding with a nanogold labelled secondary. Any suggestions would be appreciated. Thank you.
JoAnn Buchanan Molecular and Cellular Physiology Stanford University School of Medicine Stanford, CA 94305 650-723-5856
Just a quick comment for whatever it's worth. Your posting suggests that you are concerned about mechanical drifts of the filament -- the subject of a certain body of folklore in the microscopy community. It's been my experience that the principal drift in a tungsten filament is not the filament itself, but rather it is the mechanism supporting the filament which drifts.
The actual issue with drifting filaments is that mechanical stresses in the wire will gradually relieve under heat, causing the filament to distort slightly. This is a first-time-used issue. However, I believe most manufacturers include an annealing step in their process to remove these stresses before the filament is shipped to a customer (at least, that's what we used to do in another company when we manufactured our own filaments). In any case, the filament is operated at a very high temperature and it doesn't take long to anneal out these stresses under normal use, so long-term drift effects shouldn't be an issue.
My experience is that long-term mechanical drifts originate elsewhere in the gun mechanism. When you think about it, this makes sense. The filament itself has very little mass and quickly reaches its ultimate operating temperature distribution. However, the rest of the gun assembly is much more massive and, depending on various design considerations, may take hours to reach thermal equilibrium. Meanwhile, all of those intricate parts are expanding -- and many at different rates. Principal suspects, in my opinion, are lateral adjusting and clamping screws -- for example, a set of radial set screws is commonly used to center the ceramic base of the filament within a ring of some type. I used to work with a rather complex assembly which positioned the filament via lateral screws and sometimes had episodes of "filament drift". Subsequently, I have had nearly a decade of experience with a very simple design which supports the filament solely via axial pressure and find that "filament drift" isn't an issue. My conclusion is that a lot of what I used to call "filament drift" was actually mechanism drift. So if you are experiencing mechanical instabilities of the filament, I would suspect the way the filament is being supported more than the filament itself.
This opinion is based on my own experiences, however, and I will be very interested to hear if others have had different experience.
Fred Schamber RJ Lee Instruments Limited
Marisa Ahmad 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 would like to tap the wealth of information out there. } } We are experiencing a lot of problems with the tungsten filaments that we } are currently using on our JEOL 5800 SEM. During long acquisitions (many } hours) there is a lot of brightness drift and shifting, even when lengthy } stabilization periods are given prior to the acquisitions. JEOL has } checked out the SEM itself, and can't find the source for the drifting. I } was thinking that it might be the filament. I was wondering if there are } different types of tungsten filaments out there, and which is the most } stable over long periods of time. I would like to try out all of them } (considering trying to switch our entire system to a LaB6 filament is a bit } much for now). Any recommendations or experiences you have would help us } out a lot. } } Thanks in advance! } } Marisa Ahmad } R&D Specialist } Semiconductor Insights } mahmad-at-semiconductor.com } } weather report (for those interested): } It's unusually sunny and warm for here in Ottawa.....only -10 degrees C (but } it feels like -25 with the wind). It's such a nice day that I washed my } car this morning since it's not really supposed to be brown - now all the } doors and windows are frozen shut. {sigh}
We're interested in knowing a little bit more about the Hematology Slide Stainer RSG 61 commercialized by EMS. So if you have one can you please share your experience with us. Give the plus and also all the minus. If you have another equivalent brand in which you're fully satisfied of course say it. Naturally salesman can contact me (offline only please, so the whole community will not be annoided...)
Thank you for your help
Marc
PS hi Keith how is weather in Plymouth ? Here it's raining and we're singing...
------------------------------ SCHMUTZ Marc IGBMC 1 rue Laurent FRIES BP 163 F 67404 Illkirch Cedex FRANCE
Nasa has been working on micro-machined solid state electron emitters and Thomas George of that agency has published a design utilizing an array of them as the source for a "palm top" SEM in the August, 1998 issue of Nasa Tech Briefs. The proposed SEM relies upon the speculated use of an electon transparent membrane at the bottom of the lens column in this sealed, vacuum pumpless system. Resolution is anticipate in the micron range.
Nasa has also published designs for miniature x-ray tubes in an October 1995 issue of Nasa Tech Briefs.
As far as I know a solid state high energy electron gun exists only as a Star Wars project.
Nasa has been working on micro-machined solid state electron emitters and Thomas George of that agency has published a design utilizing an array of them as the source for a "palm top" SEM in the August, 1998 issue of Nasa Tech Briefs. The proposed SEM relies upon the speculated use of an electon transparent membrane at the bottom of the lens column in this sealed, vacuum pumpless system. Resolution is anticipate in the micron range.
Nasa has also published designs for miniature x-ray tubes in an October 1995 issue of Nasa Tech Briefs.
As far as I know a solid state high energy electron gun exists only as a Star Wars project.
Nasa has been working on micro-machined solid state electron emitters and Thomas George of that agency has published a design utilizing an array of them as the source for a "palm top" SEM in the August, 1998 issue of Nasa Tech Briefs. The proposed SEM relies upon the speculated use of an electon transparent membrane at the bottom of the lens column in this sealed, vacuum pumpless system. Resolution is anticipate in the micron range.
Nasa has also published designs for miniature x-ray tubes in an October 1995 issue of Nasa Tech Briefs.
As far as I know a solid state high energy electron gun exists only as a Star Wars project.
Just a friendly reminder about the upcoming American Chemical Society Course, "Applied Optical Microscopy". This course is not just for chemists!
If you need a refresher on a basic technique, need to know more about interpreting images gathered by a variety of contrast systems (including Hoffman Modulation Contrast and Polarized light), or are moving more into digital imaging, this is a great course for you!
Hands-on, 3 days of total immersion, basic principles through advanced techniques, quantitative polarized light.... and New Orleans, thrown in for good measure! $895 for ACS members; $995 for non-members (tuition and materials only).
Visit the MME website for details: {http://MME-Microscopy.com/education}
Best regards, Barbara Foster ACS Course Coordinator
Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ******************************************************
-----Original Message----- From: Michael Herron [SMTP:herro001-at-maroon.tc.umn.edu]
I am trying to get a handle on the scale of some old pictures. There are human RBC in theses pictures. Can anyone tell me the diameter of your average human RBC?
Mike-
I have recently subscribed to this list in the hopes of picking up some SEM pointers as I am beginning to dip my toes into the field. I certainly can't answer any SEM questions, but I CAN answer your question regarding the size of human RBCs. On a dried film, normal RBC's have a diameter of between 7.2-7.9 um. In certain disease states, they can deviate markedly from this. Microcytic RBC's can be well under 6 um, while macrocytes will have diameters greater than 9 um.
Good luck in your sizing!
{ {...} } Karen Hay, MS, MT(ASCP) Hematology Research, MC 2522 Loma Linda University Medical Center Loma Linda, CA 92354 Tel: (909) 558-4000 x45350 Fax: (909) 558-4189 Khay-at-ahs.llumc.edu {mailto:Khay-at-ahs.llumc.edu}
Michael, How about ~7.5 to 8.5um in diameter (depending on fixation/preparation) and ~2um in greatest thickness. -Ken
On Mon, 7 Feb 2000, Michael Herron 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 } -----------------------------------------------------------------------. } } } All, } } I am trying to get a handle on the scale of some old pictures. There } are human RBC in theses pictures. Can anyone tell me the diameter of } your average human RBC? } } Thanks } Mike } } -- } _________________________________________________ } / Michael J. Herron / } / U of MN,Medicine/Infectious Diseases / } / herro001-at-maroon.tc.umn.edu / } / http://128.101.243.213 / } /_____________________________________________/ } }
We have supplied a number of Cryostats & Microtomes to the automotive industry for sectioning paint finishes on steel body parts & plastic components e.g. bumper (fenders) door handles, door handle surrounds etc. Please view our Website to see our range of instruments.
If I can be of further assistance please come back to me.
Best Regards
Alan Bright
Bright Instrument Co.Ltd. St Margaret's Way Huntingdon PE18 6EB England
Tel No:+44 (0)1480 454528 Fax No:+44 (0)1480 456031 Email: AlanBright-at-brightinstruments.com Web Site: www.brightinstruments.com
-----Original Message----- } From: "csedax-at-alpha.arcride.edu.ar"-at-sparc5.Microscopy.Com [mailto:"csedax-at-alpha.arcride.edu.ar"-at-sparc5.Microscopy.Com] Sent: 08 February 2000 17:35 To: MICROSCOPY-at-sparc5.microscopy.com
Hi! My name is Jorgelina C. Altamirano. I'm an undegrated student of the licenciate in chemical sciences. I'm currently preparing my thesis, being its subjet: The morphological characterization and the chemical analysis of automotive paints with forensic purposes. I'm undergoing my work using SEM and EPMA techniques with a JEOL, JSM 35C and a microprobe EDAX PV9500. I've chosen some papers as references: - Beam et al, Analysis Protocol for discrimination for automotive paints by SEM-EDAX, Journal of Forensic Sciences, 35, 5, 1990 - Zieba-Palus et al, Mikrochim. Acta, 14, 1997.
I've taken same samples of automotive paints (such as Fiat, Peugeot, Renault, Ford, Chevrolet, Volkswagen) corresponding to the years 1994-99. I've found many difficulties while preparing them in: - separating the paint from the body work of the car; - cutting the specimen; - to stick specimen to the graphite specimen pedestal;
Though I've made the primary morphological and chemical analysis and I haven't observed great differences between the diferent automotive paints trades (while polyuretane paints and belayer metalizer paints already tested), but I'd found great differences between layers and layers of the paints specimen. As I'm not quite satisfied with the results obtained to be used for forensic purposes, I would greatly appreciate making contacts with specialist fo this subject, so as to receive any comments on this subject. Thank you in advance for your help. Jorgelina Altamirano CERIDE, Guemes 3450 (3000) Santa Fe, Argentina e-mail: csedax-at-arcride.edu.ar
Jorgelina SEM/EDS//EPMA is only one class of analytical methods that the Forensic community applies to paint analysis for comparison and association with original source. Although used routinely, probe methods are not used exclusively. The organic components in paints are generally more discriminating. Sample preparation usually entails some form of embedment and either microtomy or polishing in order to reveal internal structure. Removal of paint from an auto body is tricky, and requires practice.The manufacturers have designed their paint systems to prevent removal! I would be glad to provide you with additional resources.
Dennis
_________________________________________________________________________ Dennis C . Ward voice: 202-324-2982 FBI Laboratory fax: 202-324-4018 Microanalysis Laboratory email: DCWard-at-concentric.net
----- Original Message ----- } From: {"csedax-at-alpha.arcride.edu.ar"-at-sparc5.Microscopy.Com} To: {MICROSCOPY-at-sparc5.Microscopy.Com} Sent: Tuesday, February 08, 2000 9:35 AM
Jorgelina SEM/EDS//EPMA is only one class of analytical methods that the Forensic community applies to paint analysis for comparison and association with original source. Although used routinely, probe methods are not used exclusively. The organic components in paints are generally more discriminating. Sample preparation usually entails some form of embedment and either microtomy or polishing in order to reveal internal structure. Removal of paint from an auto body is tricky, and requires practice.The manufacturers have designed their paint systems to prevent removal! I would be glad to provide you with additional resources.
Dennis
_________________________________________________________________________ Dennis C . Ward voice: 202-324-2982 FBI Laboratory fax: 202-324-4018 Microanalysis Laboratory email: DCWard-at-concentric.net
----- Original Message ----- } From: "Bright, Alan" {ABright-at-brightinstruments.com} To: {"csedax-at-alpha.arcride.edu.ar"-at-sparc5.Microscopy.Com} ; {MICROSCOPY-at-sparc5.Microscopy.Com} Sent: Tuesday, February 08, 2000 12:59 AM
Jorgelina SEM/EDS//EPMA is only one class of analytical methods that the Forensic community applies to paint for characterization and association with original source. Although used routinely, probe methods are not used exclusively. The organic components in paints are generally more discriminating. Sample preparation usually entails some form of embedment and either microtomy or polishing in order to reveal layer structure. Removal of paint from an auto body is tricky, and requires practice.The manufacturers have designed their paint systems to prevent removal! I would be glad to provide you with additional resources.
Dennis
_________________________________________________________________________ Dennis C . Ward voice: 202-324-2982 FBI Laboratory fax: 202-324-4018 Microanalysis Laboratory email: DCWard-at-concentric.net
----- Original Message ----- } From: {"csedax-at-alpha.arcride.edu.ar"-at-sparc5.Microscopy.Com} To: {MICROSCOPY-at-sparc5.Microscopy.Com} Sent: Tuesday, February 08, 2000 9:35 AM
Dear all
I am planning to replace my Kr/Ar laser on a Zeiss 310 CSLM with two separate lasers: 488nm Ar ion and HeNe 543nm. Does anyone have experience of using a HeNe 543? I need this or something similar to excite Rhodamine for food structure applications. I'd appreciate any advice.
The reason I'm replacing the mixed gas laser is that has averaged 25% downtime over the past three years.
Mark
Mark Auty Dairy Products Research Centre Moorepark, Fermoy, Co. Cork, Ireland. mauty-at-moorepark.teagasc.ie tel 00353 2542447 fax 00353 2542340
Thank you for the many responses to my inquery about operation instructions for the LKB Nova ultramicrotome. For future reference, I have the instrument and the original instructions for the LKB Reichert ultramicrotome if anyone has acquired one of these without instructions. Thanks again for all of your responses.
On Mon, 7 Feb 2000 12:33:38 -0500 Marisa Ahmad {mahmad-at-semiconductor.com} wrote:
} We are experiencing a lot of problems with the tungsten filaments that } we are currently using on our JEOL 5800 SEM. During long acquisitions } (many hours) there is a lot of brightness drift and shifting, even when } lengthy stabilization periods are given prior to the acquisitions. } JEOL has checked out the SEM itself, and can't find the source for the } drifting
Marisa,
You can easily enough to see if it is filament drift. First, start the filament alignment program to ensure alignment. Then with the beam on, wait a few minutes for the filament to drift, then switch off the autobeam function, and manually check the alignment using the 2 filament alignment knobs. If you can make the image brighter, then there is filament drift. With my experiences with the 5800 though, I suspect this is not the case.
W. L. (Buddy) Steffens, Ph.D Dept. of Pathology University of Georgia
On Mon, 7 Feb 2000 16:35:56 -0500 (EST) William Tivol {tivol-at-wadsworth.org} 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 Marisa, Do you have control over the filament voltage? If so, } set the voltage just a hair over that necessary to saturate the } filament. If the voltage is too low, parts of the filament will not } emit electrons which get through the wehnelt and into the beam; if too } high, there may be significant evaporation of the filament; either may } cause drifting and/or shifting. I have had very good luck with the } stabilities of both Agar and Ebtec (now Energy Beam Sciences) } filaments; not that others may not be as good, I haven't tried them. } Ebtec makes several shapes of filament, and one may be best for } long-term stability. I have no interest in Agar or Ebtec except as a } satisfied customer. Yours, } Bill Tivol
Marisa,
This is a good point that Bill makes...if you are slightly undersaturated, there will be fluctuations in the rate of electron emission from the filament. The 5800 automatically aligns and saturates the filament when the Autobeam mode is selected...and on our 5800, saturation in this mode is a bit low. If you turn off the autobeam switch and use the filament emission knob to slightly increase the emission current, this might solve the problem at a cost of only a small decrease in filament life.
W. L. (Buddy) Steffens, Ph.D Dept. of Pathology University of Georgia
John Hunt wrote: } } Dear Probe community, } It is winter in Central NY, and the room humidity is 33%. Static } electricity (I believe) has been causing frustrating computer lockups } requiring rebooting our microprobe workstation. } I am wondering if anyone has found the best solution for preventing these } problems. } John we had this same problem for many years while we were using the old RT-11 based Tracor Northern and Kevex x-ray analyzers. The problem was especially bad with the TN-5500. This was due to the keyboard with built in scroll knob. The motion of turning that knob on the keyboard was perfect for generating a little static. (This is pre- GUI and mouse days, except for the Mac users!) Because of the low humidity conditions and severe static accumulation, in winter months we were forced to set the entire keyboard on a grounded pad. We also attached an antistatic strip to the front of the keyboard. Because of the nature of the job, and the movement to and from the computer and the microscope controls, we could not wear the wrist straps. But with the conductive grounding strip across the entire front of the keyboard we found that we could very easily get into the habit of grounding our wrist or thumb on the strip before touching the rest of the computer or keyboard. Then as we used the keyboard we would make some effort to touch the grounding strip from time to time. This virtually eliminated the problem. By the way I believe it was such a problem that Tracor engineered a static discharge switch into the keyboard connection.
Brad Huggins BPAmoco, Naperville, Microscopy and Microanalysis
At 9:20 AM -0600 2/7/0, Michael Herron wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Confocal Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
Hi! My name is Jorgelina C. Altamirano. I'm an undegrated student of the licenciate in chemical sciences. I'm currently preparing my thesis, being its subjet: The morphological characterization and the chemical analysis of automotive paints with forensic purposes. I'm undergoing my work using SEM and EPMA techniques with a JEOL, JSM 35C and a microprobe EDAX PV9500. I've chosen some papers as references: - Beam et al, Analysis Protocol for discrimination for automotive paints by SEM-EDAX, Journal of Forensic Sciences, 35, 5, 1990 - Zieba-Palus et al, Mikrochim. Acta, 14, 1997.
I've taken same samples of automotive paints (such as Fiat, Peugeot, Renault, Ford, Chevrolet, Volkswagen) corresponding to the years 1994-99. I've found many difficulties while preparing them in: - separating the paint from the body work of the car; - cutting the specimen; - to stick specimen to the graphite specimen pedestal;
Though I've made the primary morphological and chemical analysis and I haven't observed great differences between the diferent automotive paints trades (while polyuretane paints and belayer metalizer paints already tested), but I'd found great differences between layers and layers of the paints specimen. As I'm not quite satisfied with the results obtained to be used for forensic purposes, I would greatly appreciate making contacts with specialist fo this subject, so as to receive any comments on this subject. Thank you in advance for your help. Jorgelina Altamirano CERIDE, Guemes 3450 (3000) Santa Fe, Argentina e-mail: csedax-at-arcride.edu.ar
Dear Scott, I've seen a Be Ka x-ray peak in a Link advertisement and we have generated one in the Quartz XOne applications lab with a really good light element deteector. You will find the Be Ka1 peak listed at 0.110 keV in most x-ray tables. At 10:52 AM 2/7/00 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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
we are looking at desmosomes in mouse epidermis. My question: From looking at sections we got the impression that demosomes are rather uniform, round knob-like structures. As we did not do any serial sections, we are not sure if this is true. Does anybody know of a publication dealing with this?
Thanks for your help,
Christoph
Christoph Bauer Ph.D. University of Chicago Molecular Genentics and Cell Biology 5841 S. Maryland Ave/MC 1028 Chicago, Il 60637
I received the following fax from Ms. Irene Piscopo, EM Consultant, regarding electron diffraction procedures for the CM-12 or CM-120 TEM's from FEI/Philips. I'll copy the information for those interested.
ELECTRON DIFFRACTION PROCEDURES
There are three methods for doing electron diffraction in the CM-12 involving two different techniques: Method I is an aperture limiting method while Methods II and III are probe limiting. In all cases, the sample must be in the eucentric position and focused.
METHOD I: (area } 1 µm) SAD
1. Select the SA mode (the diffraction aperture and image are focused in the same plane). 2. Insert and center the SA aperture around the area of interest.* 3. Select D. 4. Remove the objective aperture. 5. Overfocus the second condenser to sharpen the pattern (i.e. parallel illumination conditions). 6. Focus the pattern with the focus control which is now changing the diffraction lens). 7. Change the size of the pattern (i.e. camera length) by changing the magnification.
Size of the SA aperture * Size of area selected = ---------------------------------------------------------------------------- ------------ Objective lens magnification in D aperture plane (~ 30x**)
** Actual magnification is focal length dependent.
METHOD II: (areas down to 40 nm with w filament; 20 nm with LaB6)
1. Locate an area of interest. 2. Remove the objective aperture. 3. Using Condensers 1 and 2, bring the beam down to a size slightly smaller than the area of the area from which the diffraction pattern is to be obtained. 4. Go to D and select the desired CL with the magnification control. 5. Focus the pattern with the focus control. Note: While it's sometimes difficult to determine exact focus, it's often easier to insert an objective aperture and focus the edge of the aperture to determine diffraction focus. 6. The size of the discs within the pattern can be decreased by decreasing the size of the condenser aperture. Be sure the aperture is well-centered.
METHOD III: NANOPROBE and/or STEM (to 2 nm)
For diffraction and/or chemical information in the TEM mode from regions of the sample less than 40 nm in diameter requires operating the instrument in either the NANOPROBE or STEM modes.
NANOPROBE:
1. Depress NANOPROBE. 2. Follow instructions in Method II.
STEM:
1. Obtain a focused STEM image using the smallest C2 aperture. 2. Stop the scan. 3. Lower the main screen (CBED pattern) 4. Slightly larger probes and more parallel beam conditions can be obtained using UUD and focusing with INTENSITY (C2 lens).
NOTES ON ELECTRON DIFFRACTION
1. The sample must always be eucentric and focused. 2. In METHOD I, the area from which the diffraction pattern is obtained is determined by the selected area - (diffraction) aperture, when in SA mode. To sharpen the ED pattern, one overfocuses the second condenser (C2) 3. In METHODS II and III, the area from which the diffraction pattern is selected depends on the size of the beam. The size of the beam can be altered by changing condenser lenses C1 and C2 (i.e. spot size and intensity. To sharpen the ED pattern , one reduces the size of the C2 aperture. 4. Spherical aberration limits METHOD I to ~1 µm. 5. In METHODS II and III, spherical aberration of the objective lens is not the limiting factor, the minimum probe size is. 6. For identifying an ED pattern, at least three rings are required. 7. Be sure the sample and the standard are in the eucentric position. Changes in the objective current will cause variations in CL.
Bruce F. Ingber, Biologist USDA-ARS, SRRC 1100 Robert E. Lee Blvd. New Orleans, LA 70124
I'm wrong. I guess I should have looked up at the X-ray periodic table above my head and saw the 0.108 keV value for Be. The reason that I was wrong was that I did not consider the solid state aspects. I am still right about not having a 2s1/2 to a 1s1/2 transition and that an X-ray carries away 1h-bar of angular momentum. What I didn't do was think about the band structure of a solid. If you have the solid, the 2p bands of Be most be spilling into or be the conduction band for the electrons. That must be the source for the electronic transition. I guess I should have engaged brain before typing. My most humble apologies. -Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies." --
} -----Original Message----- } From: microprobe [mailto:chris.salter-at-materials.oxford.ac.uk] } Sent: Tuesday, February 08, 2000 7:10 AM } To: Walck. Scott D. } Subject: Re: Be X-ray peaks -I don't think so } } } Dear Walck, } } You may not, in theory, be able to produce Be X-rays, but we } can detect them on our JEOL JXA 8800, they correspond to the position } in the tables for Be Ka!, and the target is pure Be. } } On Mon, 7 Feb 2000 10:52:29 -0500 "Walck. Scott D." {walck-at-ppg.com} } wrote: } } -----Original Message----- } From: Ekstrom, Harry [mailto:harry.ekstrom-at-honeywell.com] } Sent: Monday, February 07, 2000 7:40 PM } To: 'Walck. Scott D.' } Subject: RE: Be X-ray peaks -I don't think so } } } I don't agree Scott. An L or M line for any element would } not be located in } the area } of Be Ka. And the peak I normally get there with an ultra thin window } is a well defined peak with great resolution. } Please re evaluate your theory. } } Harry Ekstrom } } } } -------------------------------------------------------------- } ---------- } } 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 } } } } -------------------------------------------------------------- } ---------. } } } } } } When a characteristic X-ray is given off from an atom, it } carries 1 h-bar } } (Planck's constant divided by 2 pi) of angular momentum. } The electronic } } transition that occurred for the X-ray to come off must } conserve angular } } momentum of the system. Therefore, a transition from a } 2s1/2 to a 1s1/2 } } state is forbidden by selection rules. You can't produce a } Be X-ray. } } -Scott } } } } Scott D. Walck, Ph.D. } } PPG Industries, Inc. } } Glass Technology Center } } Guys Run Rd. (packages) } } P. O. Box 11472 (letters) } } Pittsburgh, PA 15238-0472 } } } } Walck-at-PPG.com } } } } (412) 820-8651 (office) } } (412) 820-8161 (fax) } } } } } } } } ------------------- } microprobe } chris.salter-at-materials.ox.ac.uk } } * This e-mail message was sent with Execmail V5.0.x * }
It has been very interesting reading about diffraction and electron beams but it is quite clear that we often give the instrument credit, "the instrument does it automatically", when credit is not due. Almost every "automatic" is a designers estimate or better still a calculation, it is a setting!
After 36 years with TEM I have to say that the only area that the instrument could set in the SA mode could be the approximate position of the first image plane. We normally move to the SA mode to free up the diffraction lens (1st intermediate) to enable it to be focussed on thefirst image plane and the diffraction (intermediate) aperture, independent of the magnification system. We would then focus the image inside the aperture with the objective lens to ensure that the selected area IS the area in the diffraction pattern. Without this procedure electron rotation and misalignments could give you a diffraction pattern from an area not in the centre of the screen! Users are correct in that if they set the eucentric point or better still, adjust the specimen height to focus at the same objective lens current value, that done the first image plane will be at the same point within the diffraction (intermediate) lens.
So how could the Philips work? I would guess they set the diffraction lens at approximately the first image plane. Then as they do not use it within their lens scheme for the SA magnifications it is fixed at this focus, good enough I think?
A second point, parallel beams are very important if you are chasing the best image quality, biology or materials, most times we need high coherence. There are misunderstandings on how to obtain a parallel beam or high coherence, setting the final condenser underfocus is incorrect. The procedure for high coherence would be to use the smallest spot size you can tolerate (this probably means you must up the emission current). Once in this condition overfocus the final condenser (clockwise from crossover) the spot, what ever size it is now, becomes your virtual source. The further overfocus you go the greater the beam coherence . You will deduce the smaller the condenser aperture the sharper the spot and the smaller the spot the greater the coherence for a given degree of overfocus.
Work with a design team and they expect everyone to overfocus, they expect everyone to use small condenser apertures and they expect everyone to use small spot sizes. They do not expect everyone to use too low an emission current (almost everyone does, we have talked before about filament life being the most important feature of many laboratories) because this makes the task too difficult!
Steve Chapman Senior Consultant Protrain Electron microscopy courses world wide http://www.emcourses.com Tel & Fax 44 (0) 1280 814774
} I am planning to replace my Kr/Ar laser on a Zeiss 310 CSLM with two } separate lasers: 488nm Ar ion and HeNe 543nm. Does anyone have } experience of using a HeNe 543? I need this or something similar to } excite Rhodamine for food structure applications. I'd appreciate any } advice.
} The reason I'm replacing the mixed gas laser is that has } averaged 25% downtime over the past three years.
} Mark
} Mark Auty } Dairy Products Research Centre } Moorepark, Fermoy, Co. Cork, Ireland. } mauty-at-moorepark.teagasc.ie } tel 00353 2542447 } fax 00353 2542340
I have been using the Argon ion (488nm) and HeNe 543nm combination on different Zeiss LSM310's and 410's for the past 9 years. The HeNe 543nm is fairly low power, but it has been perfectly adequate for scanning the fluorochromes I have needed to see including TRITC, Texas Red, Cy3, propidium iodide, and Sirius Red. I have never had a problem with the laser going down either, but one would expect that with a HeNe laser.
Slàinte,
Matthew J. Schibler Ph.D. UCLA Brain Research Institute 1524A Gonda (Goldschmied) Center for Neuroscience and Genetics Los Angeles, CA 90095-1761
Hi All, Does anyone have experience with the Pelco UVC2 or UVC1 Cryo Chamber for UV curing of resins (especially LR White) or with something similar? We're contemplating buying something like that for UV polymerization of LR White at -20 C. Thanks for your help, Kristen Kristen A. Lennon Cell, Molecular & Developmental Biology Group Department of Botany & Plant Sciences University of California Riverside, CA 92521 kalen-at-citrus.ucr.edu
Scott, I'm not sure what brought this up, but my reaction is "thems fightin' words". I do not currently have a wavelength reflecting crystal to measure that low in the periodic table, but I did on another instrument. I distinctly
recall seeing a pretty hefty peak at the Be K-alpha position when I focused the beam down on a piece of pure Be metal. I guess rules are made to be broken. Are your calculations correct? Or am I misunderstanding something here?
Jim -- ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ James J. McGee (email: jmcgee-at-sc.edu) Department of Geological Sciences University of South Carolina Columbia, SC 29208
Tel: 803-777-6300 Fax: 803-777
"Walck. Scott 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 } -----------------------------------------------------------------------. } } When a characteristic X-ray is given off from an atom, it carries 1 h-bar } (Planck's constant divided by 2 pi) of angular momentum. The electronic } transition that occurred for the X-ray to come off must conserve angular } momentum of the system. Therefore, a transition from a 2s1/2 to a 1s1/2 } state is forbidden by selection rules. You can't produce a Be X-ray. } -Scott } } Scott D. Walck, Ph.D. } PPG Industries, Inc. } Glass Technology Center } Guys Run Rd. (packages) } P. O. Box 11472 (letters) } Pittsburgh, PA 15238-0472 } } Walck-at-PPG.com } } (412) 820-8651 (office) } (412) 820-8161 (fax)
Dear Listers, I have responses to two of today's postings, but they are quite unrelated. 1. I did not see the original question about Be x-rays, only Scott Walk's reply, so I don't know what info was being sought. We used a Be-containing mineral to evaluate microprobes before buying. I won't embarrass those who failed, but only one manufacturer succeeded. The reason the others failed is probably that in compounds the x-ray energy is subject to largish chemical shifts relative to pure Be because there is no shielding by outer electrons and you need to search on either side of the tabulated value of about 0.1keV. So there IS a Be x-ray and a reasonable WDS spectrometer should detect it.
2. Filament drift. Fred Schamber's reply is right most of the time, but I did once have a batch of 6 JEOL-type filaments made by an EM supply house where 2 of them drifted until the tips were right at the edge of the hole and stayed there even when the filament was cooled down again. This was the only time in about 15 years of using JEOL microscopes.
Eric ---------------------- Dr Eric Lachowski Department of Chemistry University of Aberdeen Meston Walk Old Aberdeen AB24 3UE Scotland +44 (0)1224 272934 fax 272921 e.lachowski-at-abdn.ac.uk
Hi List I am using live blood staining light microscopy, is there anyone who has tried or is using this technique. I would like to swap notes. Dean Armytage PhD Hillstream Health Centre Australia
I'm getting more curious as I read on here. Speaking strictly energy now and leaving out wavelength, the characteristic energy of Be Ka is about .11 KeV like Mary stated. I've had an occasional peak show up there before and thought it may have been Be. Now I understand that it may have been "noise" all along. Would "noise" be a pretty defined and relatively well resolved peak at that energy level using an ultra thin window? Depending on the low energy cutoff setting, one might truly have Be and never detect it. Would lowering the KV reduce the noise artifact?
I guess I'll get out my Be planchet and try a few things.
Harry
-----Original Message----- } From: Jim McGee [mailto:mcgee-at-geol.sc.edu] Sent: Tuesday, February 08, 2000 3:09 PM To: 'Microscopy'
Scott, I'm not sure what brought this up, but my reaction is "thems fightin' words". I do not currently have a wavelength reflecting crystal to measure that low in the periodic table, but I did on another instrument. I distinctly
recall seeing a pretty hefty peak at the Be K-alpha position when I focused the beam down on a piece of pure Be metal. I guess rules are made to be broken. Are your calculations correct? Or am I misunderstanding something here?
Jim -- ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ James J. McGee (email: jmcgee-at-sc.edu) Department of Geological Sciences University of South Carolina Columbia, SC 29208
Tel: 803-777-6300 Fax: 803-777
"Walck. Scott 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 } -----------------------------------------------------------------------. } } When a characteristic X-ray is given off from an atom, it carries 1 h-bar } (Planck's constant divided by 2 pi) of angular momentum. The electronic } transition that occurred for the X-ray to come off must conserve angular } momentum of the system. Therefore, a transition from a 2s1/2 to a 1s1/2 } state is forbidden by selection rules. You can't produce a Be X-ray. } -Scott } } Scott D. Walck, Ph.D. } PPG Industries, Inc. } Glass Technology Center } Guys Run Rd. (packages) } P. O. Box 11472 (letters) } Pittsburgh, PA 15238-0472 } } Walck-at-PPG.com } } (412) 820-8651 (office) } (412) 820-8161 (fax)
My personal experience with silver enhancement dates back to when Janssen Life Sciences first introduced the IntenSE M silver enhancement kit in late 80's (this product was later taken over by Amersham). At the time, I felt the IntenSE M was easy to use, but its fast reaction made it hard to control the particle growth. Just like you, the desire for a better results has kept me searching and testing whenever a new protocol or product became available.
Danscher's method gives wonderful intensification, but its low pH sometimes damages the ultrastructure when used for the pre-embedding immunogold labeling. Burry's method and the Nanoprobes HQ silver made progress with pH, however they inherited Danscher's high viscosity and light sensitivity, which limit their penetration in pre-embedding silver enhancement and render it more difficult to handle from a practical standpoint. I have also tested the Nanoprobes GoldEnhance kit for pre-embedding immunogold labeling recently. At the LM level the results were decent. I have not spent a lot of time evaluating its performance at the EM-level. So far, the particle size homogeneity is not as good as what I've found with Danscher's method. But I will refrain from further judgment until more work has been done.
In my opinion, the first breakthrough in intensifying gold particles since Danscher's method was realized when Aurion, a Dutch company, first introduced the R-gent SE-EM silver enhancement kit last June. As you wished for in your post, it has a near-neutral pH, low viscosity, and it is light insensitive. Moreover, it gives a great enhancement efficiency and even particle size and shape. Background remains minimal even after two hours enhancement on the bench. Morphology preservation after enhancement is superior to any other enhancement reagent I have used. I have been testing this kit profusely for both post- and pre-embedding immunogold labeling on various types of samples (including cell cultures and vibratome sections), with many different primary and secondary antibodies, and am very pleased with its performance. I use 0.5% OsO4 for 20 min and have never had any problem with silver disappearing. If you like, I would be very happy to e-mail some images to you so you can evaluate them yourself.
Hong ================= Hong Yi Emory University Department of Neurology 6215 Woodruff Memorial Research Building 1639 Pierce Dr. Atlanta, GA 30341 Phone: (404) 727-8692 Fax: (404) 727-3157 Email: hyi-at-emory.edu
On Thu, 3 Feb 2000, Michael Plociniak 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 Colleagues, } Has anyone used gold enhancement reagents from Nanoprobes, Inc. } (Stonybrook, NY - USA) as an alternative to silver enhancement for } enlarging 1 nm immunogold probes? } } Advertisements state that gold enhancement has lower background and is } compatible with osmium. Can anyone verify this from personal experience? } } In addition to these advantages, I am hoping that milder pH conditions will } be less damaging to ultrastructure in cultured neurons (using a } preembedment protocol). } } Thank you, } Michael }
Here is the way I found yesterday for finding the diffraction plane. By diffraction plane I mean the plane at which diffraction pattern is formed (crossover, Fourier of the object wave) not the theoretical BFP of the objective.
First perform standard alignment for brightfield mode. Then go to 10 000X. There are several ways to perform:
1. Fixed illumination and specimen position (i.e. OL current)
a) Adjust the desired brightness with C2. Adjust the desired OL current and specimen position. b) Insert OL aperture and move it so that the edge of the aperture to cross through the middle of the screen. If the aperture is exactly at the diff. plane then you'll not be able to see its edge - just the whole beam will fade uniformly ... so you have the diff. plane. c) If you see aperture edge then it is not at diff. plane. Now two ways: - change the aperture z-position (not recommended ... but if it is very far ..) - If your microscope has condenser minilens you can try tweaking both C2 and CML in order to keep the intensity same and just make the aperture edge disappear (i.e. the whole screen uniformly darkened).
2. Fixed specimen (i.e. OL current).
a) Focus the specimen b) same as 1b c) Change C2 excitement in order to make the edge disappear
3. Fixed illumination
a) Set desired C2 excitement b) same as 1b c) Change the objective lens current until the aperture edge disappears. Then move specimen in z-direction until it is focused.
In all these methods after adjusting the aperture at diff. plane you can try tweaking C2 or OL ... you will see that the aperture is not at diff. plane anymore, The edge will appear on one or the other side (i.e. mirrored) depending on the direction of the lens current change. Another thing - after adjusting the aperture at diff. plane you can check if beam shift tilts the beam (by tilting here I mean - if the specimen is illuminated with plane wave then shifting the beam should not tilt the wave front). Shift the beam and if the brightness changes (when the aperture edge partially covers the zero diff. spot) then the beam is tilting. I wish there was a IL1 tweaking knob allowing for change of the IL1 current while in brightfield mode. This wold make the things much more easier.
About the methods for making the illumination at the specimen parallel. They will work only if: - The OL aperture is positioned by the manufacturer exactly at the theoretical BFP (I think that in most of the TEMs it is not) - The OL current should be set to zero deviation (i.e. at the value for which the BFP position was calculated) - The intermediate lens system should be properly calibrated by the manufacturer so that if both of the above conditions are satisfied and the specimen is at front focal plane of the OL then it to be in focus. In other words the theoretical BFP of the objective to coincide with the theoretical front focal plane of the IL1.
I don't think parallel illumination is so very important ... using spherical wave will give the same results but just the diff. planes will be shifted.
All of the above is just my opinion. I could be mistaking somewhere ...
Best wishes,
Rado
--------------------------------------------------------------------- Radostin Danev Laboratory of Ultrastructure Research National Institute for Physiological Sciences Myodaiji-cho, Okazaki 444-8585, JAPAN e-mail: rado-at-nips.ac.jp ---------------------------------------------------------------------
----- Original Message ----- } From: "Dennis Ward" {DCWard-at-concentric.net} } } Jorgelina } SEM/EDS//EPMA is only one class of analytical methods that the Forensic } community applies to paint analysis for comparison and association with } original source. Although used routinely, probe methods are not used } exclusively. The organic components in paints are generally more } discriminating. } Sample preparation usually entails some form of embedment and either } microtomy or polishing in order to reveal internal structure. } Removal of paint from an auto body is tricky, and requires practice.The } manufacturers have designed their paint systems to prevent removal! } I would be glad to provide you with additional resources.
I have never do this but this is what i would try first.
One thing I would try on would be removing the metal from the paint. A weak elecrolite solution and a low DC voltage connected to reverse electoplate the metal away sould get rid of almost all of it and the last bit could be removed with acid or evaporated as the cathode in a vacum chamber.
Once you get the metal down to a bunch of small islands you might be able to let it rust free of the paint.
I have no idea what this would do to the paint but most paint films I have delt with are pretty tough.
Gordon
Gordon Couger gcouger-at-couger.com
Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
we are massively working on analytical and structural TEM characterization of semiconductors, mainly InGaAs/GaAs, quantum well, wires and dots.
Specimen preparation is of course a crucial point of our work.
We have just switched to PIPS (we used to make our samples using the Gatan Duo Mill), and we are getting controversial results about the damage introduced by the milling procedure.
We are perfectly aware of all the differences between the two instruments, especially the absence of cooling stage and the higher beam current.
Has anyone performed a systematic and careful analysis to try to evaluate the milling damage on similar materials systems. We are willing to start a systematic work to assess this issue, but would like to know if anyone has already done anything on this topic. Also, any collaboration will be more than welcome.
in our lab we have prepared a number of GaAs, AlGaAs, InGaAs devices. Mechanical thinning is by the tripod wedge technique, but typically we end up with more damage than we find for Si samples so there is a need to leave samples thicker and use more extensive ion milling for final thinning.
We also have a PIPS and I have found its capabilities essential for achieving good thin samples in these materials. I found more success by using low angles and low KeV for long times. Typically I would be using 5-6 degrees and 4-4.5 KeV initially, with 3-4 degrees and 3-3.5 KeV for the final ion polishing. On samples 2-3 microns thick, this would amount to 2-3 hours total milling time. I did not observe any artefacts introduced with milling under these conditions, and the junction structures and inherent defects were clearly observed.
The conditions above should work with any ion miller capable of low angle milling, but our experience is only with the PIPS. If you use the wedge technique for mechanical thinning, I would recommend that you use Mo grids to mount your specimens as the extended milling times at low angles will really chew up a Cu grid. Also, Mo grids are considerably thinner, allowing angles as low as 3-4 degrees on the grid hole side for a sample positioned in the middle of a 1 mm grid hole.
If you would like, I can send you some typical images of devices we have prepared, including images showing the removal of the mechanical polishing damage.
Philip L. Flaitz IBM Analytical Services, Hopewell Junction, NY http://www.chips.ibm.com/services/asg Ph.......(914) 892-3094, FAX -2003 flaitz-at-us.ibm.com
Massimo Catalano {massimo.catalano-at-ime.le.cnr.it} on 02/09/2000 03:43:03 AM
To: Microscopy-at-sparc5.Microscopy.Com cc:
Dear Listservers,
we are massively working on analytical and structural TEM characterization of semiconductors, mainly InGaAs/GaAs, quantum well, wires and dots.
Specimen preparation is of course a crucial point of our work.
We have just switched to PIPS (we used to make our samples using the Gatan Duo Mill), and we are getting controversial results about the damage introduced by the milling procedure.
We are perfectly aware of all the differences between the two instruments, especially the absence of cooling stage and the higher beam current.
Has anyone performed a systematic and careful analysis to try to evaluate the milling damage on similar materials systems. We are willing to start a systematic work to assess this issue, but would like to know if anyone has already done anything on this topic. Also, any collaboration will be more than welcome.
Hello ..... we have an old ETEC Omniscan SEM, and we are searching for ETEC company or any people that can supply manuals ....
any help is welcome
best regards
=================================================== Fernando D. Balducci Laboratorio de Microscopia Electrnica Facultad de Ingeniera - Bioingenieria Universidad Nacional de Entre Rios Argentina e-mail: microsc-at-fi.uner.edu.ar tel: 54 43 975100 fax: 54 43 975077
I am working with archival specimens of formalin-fixed paraffin embedded tissues and I am getting tremendous autofluorescence. I looked at tissue sections after the sections have been de-waxed using either FITC or rhodamine filter sets and red cells and connective tissues are brightly fluorescent. Is there a way of suppressing the autofluorescence and still retain reactivity of tissue to antibodies? I would appreciate any comments or suggestions.
******************************************************* Corazon D. Bucana, Ph.D. Department of Cancer Biology U.T. M.D. Anderson Cancer Center 1515 Holcombe Blvd. Box 173 Houston, Texas 77030 Phone: (713) 792-8106 FAX: (713) 792-8747 Email:bucana-at-audumla.mdacc.tmc.edu FAX: (713) 792-8747
I don't have a UTW detector here, so I'll leave this to the EDS experts. But, on the JEOL 8800/8900 I used to run we had a thin window detector that could see B (not Be) if the low end noise peak (which is huge) was properly discriminated. Get out your C planchett while you are getting the Be one, and see if you get the noise peak at Be. Far better to use wavelength than EDS down at that end.
Jim McGee -- ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ James J. McGee (email: jmcgee-at-sc.edu) Department of Geological Sciences University of South Carolina Columbia, SC 29208
Tel: 803-777-6300 Fax: 803-777-6610
"Ekstrom, Harry" 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'm getting more curious as I read on here. Speaking strictly energy now } and leaving out wavelength, the characteristic energy of Be Ka is about .11 } KeV like Mary stated. I've had an occasional peak show up there before and } thought it may have been Be. Now I understand that it may have been "noise" } all along. Would "noise" be a pretty defined and relatively well resolved } peak at that energy level using an ultra thin window? Depending on the low } energy cutoff setting, one might truly have Be and never detect it. Would } lowering the KV reduce the noise artifact? } } I guess I'll get out my Be planchet and try a few things. } } Harry } } -----Original Message----- } } From: Jim McGee [mailto:mcgee-at-geol.sc.edu] } Sent: Tuesday, February 08, 2000 3:09 PM } To: 'Microscopy' } Subject: Re: Be X-ray peaks -I don't think so } } ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } Scott, } I'm not sure what brought this up, but my reaction is "thems fightin' } words". I do not currently have a wavelength reflecting crystal to measure } that low in the periodic table, but I did on another instrument. I } distinctly } } recall seeing a pretty hefty peak at the Be K-alpha position when I focused } the beam down on a piece of pure Be metal. I guess rules are made to be } broken. Are your calculations correct? Or am I misunderstanding something } here? } } Jim } -- } ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ } James J. McGee (email: jmcgee-at-sc.edu) } Department of Geological Sciences } University of South Carolina } Columbia, SC 29208 } } Tel: 803-777-6300 Fax: 803-777 } } "Walck. Scott 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 } } -----------------------------------------------------------------------. } } } } When a characteristic X-ray is given off from an atom, it carries 1 h-bar } } (Planck's constant divided by 2 pi) of angular momentum. The electronic } } transition that occurred for the X-ray to come off must conserve angular } } momentum of the system. Therefore, a transition from a 2s1/2 to a 1s1/2 } } state is forbidden by selection rules. You can't produce a Be X-ray. } } -Scott } } } } Scott D. Walck, Ph.D. } } PPG Industries, Inc. } } Glass Technology Center } } Guys Run Rd. (packages) } } P. O. Box 11472 (letters) } } Pittsburgh, PA 15238-0472 } } } } Walck-at-PPG.com } } } } (412) 820-8651 (office) } } (412) 820-8161 (fax)
-- ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ James J. McGee (email: jmcgee-at-sc.edu) Department of Geological Sciences University of South Carolina Columbia, SC 29208
I am looking for TEM images that show cross-sectional views of skeletal muscle sarcomeres during the *contracted state*. In particular, I am interested in seeing the spacing of the thin filaments when they **overlap each other** (i.e., when those from one side of the sarcomere overlap those from the other side of the sarcomere in the contracted state). In addition, I am interested in seeing the distribution of the thin filaments as they pass through the M line. It would be greatly appreciated if anyone could tell me if they know of any research papers or review articles that contain such TEM images.
Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 75 N. Eagleville Rd. U-125 Storrs, CT 06269 Knecht-at-uconnvm.uconn.edu 860-486-2200 860-486-4331 (fax) home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html ************************************************************
Christoph: I don't have any references at hand, but there is a whole literature on desmosomes. A search on Medline (or PubMed on the internet) should provide you with a good list of EM related publications on desmosomes--probably more than you ever wanted to know! As an aside, I think there may be some books that cover this as well, but since it is not an area of personal expertise or current research interest, I am afraid they have all gone from my memory banks.
Roger
On Tue, 08 Feb 2000 09:56:35 -0800, Christoph Bauer 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 at desmosomes in mouse epidermis. My question: From } looking at sections we got the impression that demosomes are rather } uniform, round knob-like structures. As we did not do any serial } sections, we are not sure if this is true. Does anybody know of a } publication dealing with this? } } Thanks for your help, } } Christoph } } } } Christoph Bauer Ph.D. } University of Chicago } Molecular Genentics and Cell Biology } 5841 S. Maryland Ave/MC 1028 } Chicago, Il 60637 }
Roger C Moretz, Ph.D.
_______________________________________________________ Get 100% FREE Internet Access powered by Excite Visit http://freeworld.excite.com
Christoph, I have examined alot of mouse and pig skin at the TEM level and the desmosomes and hemidesmosomes appear rather typical, that is, small, long bridge-like structures between the cells.
Mary Gail Engle Electron Microscopy & Imaging Facility University of Kentucky
At 09:56 AM 2/8/00 -0800, Christoph Bauer wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
The Department of Microscopy and Microanalysis at Abbott Laboratories has two summer internships available for 8-12 weeks this summer. One position is in Biological Microscopy, and one is in Materials Analysis and Microscopy. We're looking for students who are considering a career in microscopy, especially students interested in the pharmaceutical industry.
Abbott Laboratories is a diversified healthcare company that produces pharmaceuticals, diagnostic devices, and hospital products. The Microscopy department analyzes samples related to all these functions. We use polarized light, fluorescence, TEM, SEM, EDXS, confocal microscopy, and flow cytometry to solve problems related to products, as well as to provide support for pharmaceutical Discovery and Development basic research and drug safety.
Housing and a stipend are provided for the summer. We're located near Lake Michigan and the Wisconsin state border, about an hour's drive or train-ride north of Chicago. There's lots to see and do in the area, and there are planned activities with other interns, as well. The Abbott Summer Internship Program has been nationally recognized as one of the best in the country.
If you're interested, please send a resume to:
Jane A. Fagerland, Ph.D. Abbott Laboratories D-45M/AP31 200 Abbott Park Rd. Abbott Park IL 60064-6202
You may also fax a resume to me at (847) 938-5027 or e-mail it to me at jane.a.fagerland-at-abbott.com.
If you'd like further information, I can be reached by telephone at (847) 935-0104, or by e-mail. I prefer e-mail, but will be more than happy to talk to discuss our projects and laboratory by telephone.
There is an easy way to determine the milling damage between the two machines and the amount of ion milling damage (subjectively); compare them with samples prepared using the small angle cleavage technique. Look at John McCaffrey's paper on using the small angle cleavage technique in Ultramicrotomy 38, (149) 1991. He shows the difference between high angle milling, low angle milling and the small angle cleavage technique. Of course, the small angle cleavage technique showed no ion milling damage. It is perfect for these types of materials. If you don't need a site specific technique, it is the way to go for semiconductors.
You should also look at his paper in the MRS TEM Sample Prep series III (vol 254) that also shows a comparison for a SiGe/Si layer structure. We have a detailed pictorial outline with tips and tricks on how to do it in the MRS TEM Sample Prep series IV (vol 480).
I know that you invested a lot in your ion mill, but you can do these samples very cheaply. SouthBay Technology sells a Microcleave kit that gets you started with the technique.
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies." --
} -----Original Message----- } From: Massimo Catalano [mailto:massimo.catalano-at-ime.le.cnr.it] } Sent: Wednesday, February 09, 2000 3:43 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: PIPS and milling damage } } } -------------------------------------------------------------- } ---------- } 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 Listservers, } } we are massively working on analytical and structural TEM } characterization } of semiconductors, mainly InGaAs/GaAs, quantum well, wires and dots. } } Specimen preparation is of course a crucial point of our work. } } We have just switched to PIPS (we used to make our samples } using the Gatan } Duo Mill), and we are getting controversial results about the damage } introduced by the milling procedure. } } We are perfectly aware of all the differences between the two } instruments, } especially the absence of cooling stage and the higher beam current. } } Has anyone performed a systematic and careful analysis to try } to evaluate } the milling damage on similar materials systems. We are } willing to start a } systematic work to assess this issue, but would like to know } if anyone has } already done anything on this topic. Also, any collaboration } will be more } than welcome. } } Thanks. } } Massimo } } } } } Dr. Massimo Catalano } CNR-IME } Campus Universitario } Via Arnesano } 73100 Lecce - ITALY } tel: + 39 0832 322362 } fax: + 39 0832 325299 } email: massimo.catalano-at-ime.le.cnr.it } http://www.ime.le.cnr.it } http://www.ime.le.cnr.it/sime/sime.htm } } }
It has been brought to my attention that no dates were attached to this course reminder:
March 10-12, 2000 Hyatt Regency New Orleans, LA.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education}
At 07:57 PM 2/7/00 -0500, Barbara Foster wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Suggest you contact Chuck Garber at Structure Probe: www.2spi.com I'm sure that he has some sort of derivative of his tacky dots which would be helpful
At 08:48 PM 8/27/70 +0000, Linda Chicoine wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I know this is off topic however a change like this could make this type of forum impossible.
} Date: Tue, 8 Feb 2000 08:13:53 -0700 } X-Sender: liperez-at-cnmailsvr.nmsu.edu } Mime-Version: 1.0 } To: all-at-biology.nmsu.edu } From: liperez-at-NMSU.Edu (LSPSaldana) } Subject: Congress to allow email charges } Sender: owner-all-at-biology.nmsu.edu } Precedence: bulk } X-Keywords: } X-UID: 43 } Status: O } } } } } Congress to allow email charges } } } } } } Please pass this on to all you know since many of us use e-mail for } } } business and to keep up with friends and family, I thought you'd like } } } to know the following. } } } } } } Please jump on it right away and forward this to others. } } } } } } CNN has reported that within the next two weeks Congress is going to } } } vote on allowing telephone companies to CHARGE A TOLL FEE for } } } internet access. } } } Translation: Every time we send long distance e-mail we will receive a } } } long distance charge. This will get costly. Please visit the following web } } } site and file a complaint. Complain to your Congressperson. We can't } } } allow this to pass. The following address will allow you to send an } } } e-mail on this subject DIRECTLY to your Congressperson. } } } } } } http://www.house.gov/writerep {http://www.house.gov/writerep} } } } } } } Pass this on to your friends. It is urgent! I hope all of you will pass } } } this on to all your friends and family. We should ALL have } } } an interest in this one. } } } } } } WAIT, THERE'S MORE. IN ADDITION, The last few months have revealed an } } } alarming trend in the Government of the United States attempting to } } } quietly push through legislation that will affect your use of the } } Internet. } } } Under } } } proposed legislation the U.S. Postal Service will be attempting to bill } } } email users out of "alternate postage fees". Bill 602P will permit the } } } Federal Govt. to charge a 5 cent surcharge on every email delivered, by } } } billing Internet Service Providers at source. The consumer would then be } } } billed in turn by the ISP. Washington D.C. lawyer Richard Stepp is } } } working without pay to prevent this legislation from becoming law. The } } U.S. } } } Postal Service is claiming that lost revenue due to the proliferation of } } } email } } } is costing nearly $230,000,000 in revenue per year. You may have noticed } } } their recent ad campaign "There is nothing like a letter". } } } Since the average citizen received about 10 pieces of email per day in } } } 1998, } } } the cost to the typical individual would be an additional 50 cents per } } } day, or over $180 dollars Per year, above and beyond there regular } } Internet } } } costs. } } } Note that this would be money paid directly to the U.S. Postal Service } } } for a service they do not even provide. The whole point of the Internet is } } } democracy and non-interference. If the federal government is permitted } } } to tamper with our liberties by adding a surcharge to email, who knows } } Where } } } it will end. You are already paying an exorbitant price for snail mail } } } because of bureaucratic inefficiency. It currently takes up to 6 days for } } a } } } letter to be delivered from New York to Buffalo. If The U.S. Postal } } Service } } } is } } } allowed to tinker with email; it will mark the end of the "free" } } } Internet in the United States. } } } One congressman, Tony Schnell has even suggested a "twenty to forty } } } dollar per month surcharge on all internet service" above and beyond the } } } government's proposed email Charges. Note that most of the major } } } newspapers have ignored the story, the only exception being the } } } Washingtonian } } } which called the idea of email surcharge "a useful concept who's time has } } } come" } } } (March 6, 1999) Editorial. } } } } } } Don't sit by and watch your freedoms erode away! Send this e-mail to } } } EVERYONE on your list, and tell all your friends and relatives to write } } } to their congressman and say "No!" to Bill 602P. } } } } } } } } } It will only take a few moments of your time, and could very well be } } } instrumental in killing a bill we don't want. } } } } } } PASS THIS ON TO EVERYONE YOU KNOW WHO USES EMAIL REMEMBER THESE ARE TWO } } } SEPARATE ISSUES THAT EFFECT ALL OF US ONLINE. LET YOU VOICE BE HEARD } } } NOW, NOT AFTER } } } } } } } } } } } } } } } } } } } } }
Dear Donald, the Institute of Applied Physics (http://www.sandy.ru/science/science/appl.new/frames.html) constructs, produces and uses solid state high energy electron gun for the High-power electronics and plasma physics researches. May be they will help you.
Best regards, Dr. S.A.Gusev
******************************************** * Institute for Physics of Microstrutures * * Russian Academy of Science * * ( IPM RAS ) * * * * Niznii Novgorod, GSP-105 * * 603600 * * RUSSIA * * * ********************************************
Hello, We have got a JEM3010 Transmission electron microscopy.I have got a one problem about water cooling system.I dont know,Which kind of water to used?I think so, We should use pure water for water cooling?Besides My city water is not clean.What do you think about this problem? Thanks for your interest
********************************** ********************************** ** Research.Asst.ERDEM YASAR ** ** University of Kirikkale ** ** Department of Physics ** ** TEM LABORATORY ** ** 71450 KIRIKKALE/TURKEY ** ** erdem.yasar-at-physics.org ** ** yasar-at-science.ankara.edu.tr ** ** yasar-at-turkuaz.kku.edu.tr ** ********************************** **********************************
I am looking for TEM images that show cross-sectional views of skeletal muscle sarcomeres during the *contracted state*. In particular, I am interested in seeing the spacing of the thin filaments when they **overlap each other** (i.e., when those from one side of the sarcomere overlap those from the other side of the sarcomere in the contracted state). In addition, I am interested in seeing the distribution of the thin filaments as they pass through the M line. It would be greatly appreciated if anyone could tell me if they know of any research papers or review articles that contain such TEM images.
Thank you!
Izak Paul Biological Sciences Mount Royal College
The following papers are classics and would make good starting point. Huxley, H.E. & Hanson, J. (1954). Nature, 173, 973-976. Huxley, A.F. & Niedergerke, R.M. (1954). Nature, 173, 971. Huxley, H.E. (1957). Biophys. & Biochem. Cytol. 3, 631-648.
Thin filaments 'pass through' the Z-line. Try John Squire and Pradeep Luther (3D reconstruction of fish Z-disc) and Mike Reedy (insect) or Bellinda Bullard.
Matt Walker School of Biomedical Sciences Leeds University, UK
I'm sorry to bring this topic up yet again, but I have just come across the specification for a flat-bed scanner which looks as if it would be suitable for scanning e.m. cut-film. It is the Epson Perfection 1200 Photo which includes a 5x4inch film adapter.
The UK web site address is: http://www.epson.co.uk/sohoprod/imaging/scanner/perf1200/perf1200photo/spec.htm
and the specification is: A4 Colour flatbed with 5"x4" film image scanner Single pass scanning Optical resolution: 1200 x 2400 (30600 pixels/line) Maximum output resolution of 9600x9600 dpi 36 bits per pixel in colour (24 bit output) 12 bits per pixel in black (8 bit output) Optical Density 3.2D USB (Type B)
Does anyone have any experience of this machine because it is about a fifth of the price of the Umax flatbed film scanners? My only reservation is that it is a SOHO product rather than professional.
Thanks in advance
Malcolm
Malcolm Haswell Electron Microscopy School of Sciences Fleming Building University of Sunderland SUNDERLAND SR1 3SD Tyne and Wear UK
Tel (0191) 515 2872 e-mail: malcolm.haswell-at-sunderland.ac.uk
This is a recurring post about nothing more than a hoax.
Search for internet hoaxes and so forth and you will find this one and many other interesting yet equally invalid assertions.
gary g.
At 10:58 PM 2/9/00 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
The statement that the smallest spot size gives the best coherence can be a misleading. The full statement should be the spot size with the smallest angular distribution should be used. Anyone who has tried to do off-axis holography in nanoprobe mode will testify to this. The spatial coherence envelope, roughly the distance over which the beam is considered 'coherent', is the Fourier transform of the angular distribution of intensity at the source (Van Cittert Vernike theorem).
Born & Wolf (section 10.4.2) "Hence if the linear dimensions of the source and the distance between P1 and P2 (points in the imaging plane) are small compared to the distance of these points from the source, the degree of coherence, |mu_12| is equal to the absolute value of the normalized Fourier transform of the intensity function of the source"
The reason holography and high resolution work is done with te most parallel beam possible becomes clear, the angular distribution of a converged (or diverged beam) is wide enough to reduce the spatial coherence envelope. In off-axis holography this is even more stringent since two interfering points (reference wave & object wave) can be hundreds of nanometers, microns even, apart. Elliptical illumination is used to preserve coherence in the one important direction (minimum angular width) and converged in the other to improve the intensity.
******************************************************** Dr Jonathan Barnard
Analytical Materials Physics The Angstrom Laboratory, Uppsala University P O Box 534, SE-751 21 Uppsala, Sweden Phone: +46-(0)18-4716838 Fax: +46-(0)18-500131 http://www.angstrom.uu.se/analytical/home.html
I read the following just the other day, from the US Postal Service
http://www.usps.gov/news/press/99/99045new.htm
FOR IMMEDIATE RELEASE May 21, 1999 Release No. 45
E-MAIL RUMOR COMPLETELY UNTRUE
WASHINGTON â A completely false rumor concerning the U.S. Postal Service is being circulated on Internet e-mail. As a matter of fact, the Postal Service has learned that a similar hoax occurred recently in Canada concerning Canada Post.
The e-mail message claims that a "Congressman Schnell" has introduced "Bill 602P" to allow the federal government to impose a 5-cent surcharge on each e-mail message delivered over the Internet. The money would be collected by Internet Service Providers and then turned over to the Postal Service.
No such proposed legislation exists. In fact, no "Congressman Schnell" exists.
The U.S. Postal Service has no authority to surcharge e-mail messages sent over the Internet, nor would it support such legislation.
-30-
Evex Analytical X-ray Analyzers and Digital Imaging Systems 857 StateRoad Princeton, NJ 08540 609-252-9192 T 609-252-9091 F www.evex.com info-at-evex.com
In a message dated 2/10/00 12:26:56 AM Eastern Standard Time, chbennet-at-nmsu.edu writes:
{ { Subj: FYI: Congress to allow email charges Date: 2/10/00 12:26:56 AM Eastern Standard Time From: chbennet-at-nmsu.edu (Christina bennett) To: microscopy-at-sparc5.microscopy.com
------------------------------------------------------------------------ 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 know this is off topic however a change like this could make this type of forum impossible.
} Date: Tue, 8 Feb 2000 08:13:53 -0700 } X-Sender: liperez-at-cnmailsvr.nmsu.edu } Mime-Version: 1.0 } To: all-at-biology.nmsu.edu } From: liperez-at-NMSU.Edu (LSPSaldana) } Subject: Congress to allow email charges } Sender: owner-all-at-biology.nmsu.edu } Precedence: bulk } X-Keywords: } X-UID: 43 } Status: O } } } } } Congress to allow email charges } } } } } } Please pass this on to all you know since many of us use e-mail for } } } business and to keep up with friends and family, I thought you'd like } } } to know the following. } } } } } } Please jump on it right away and forward this to others. } } } } } } CNN has reported that within the next two weeks Congress is going to } } } vote on allowing telephone companies to CHARGE A TOLL FEE for } } } internet access. } } } Translation: Every time we send long distance e-mail we will receive a } } } long distance charge. This will get costly. Please visit the following web } } } site and file a complaint. Complain to your Congressperson. We can't } } } allow this to pass. The following address will allow you to send an } } } e-mail on this subject DIRECTLY to your Congressperson. } } } } } } http://www.house.gov/writerep {http://www.house.gov/writerep} } } } } } } Pass this on to your friends. It is urgent! I hope all of you will pass } } } this on to all your friends and family. We should ALL have } } } an interest in this one. } } } } } } WAIT, THERE'S MORE. IN ADDITION, The last few months have revealed an } } } alarming trend in the Government of the United States attempting to } } } quietly push through legislation that will affect your use of the } } Internet. } } } Under } } } proposed legislation the U.S. Postal Service will be attempting to bill } } } email users out of "alternate postage fees". Bill 602P will permit the } } } Federal Govt. to charge a 5 cent surcharge on every email delivered, by } } } billing Internet Service Providers at source. The consumer would then be } } } billed in turn by the ISP. Washington D.C. lawyer Richard Stepp is } } } working without pay to prevent this legislation from becoming law. The } } U.S. } } } Postal Service is claiming that lost revenue due to the proliferation of } } } email } } } is costing nearly $230,000,000 in revenue per year. You may have noticed } } } their recent ad campaign "There is nothing like a letter". } } } Since the average citizen received about 10 pieces of email per day in } } } 1998, } } } the cost to the typical individual would be an additional 50 cents per } } } day, or over $180 dollars Per year, above and beyond there regular } } Internet } } } costs. } } } Note that this would be money paid directly to the U.S. Postal Service } } } for a service they do not even provide. The whole point of the Internet is } } } democracy and non-interference. If the federal government is permitted } } } to tamper with our liberties by adding a surcharge to email, who knows } } Where } } } it will end. You are already paying an exorbitant price for snail mail } } } because of bureaucratic inefficiency. It currently takes up to 6 days for } } a } } } letter to be delivered from New York to Buffalo. If The U.S. Postal } } Service } } } is } } } allowed to tinker with email; it will mark the end of the "free" } } } Internet in the United States. } } } One congressman, Tony Schnell has even suggested a "twenty to forty } } } dollar per month surcharge on all internet service" above and beyond the } } } government's proposed email Charges. Note that most of the major } } } newspapers have ignored the story, the only exception being the } } } Washingtonian } } } which called the idea of email surcharge "a useful concept who's time has } } } come" } } } (March 6, 1999) Editorial. } } } } } } Don't sit by and watch your freedoms erode away! Send this e-mail to } } } EVERYONE on your list, and tell all your friends and relatives to write } } } to their congressman and say "No!" to Bill 602P. } } } } } } } } } It will only take a few moments of your time, and could very well be } } } instrumental in killing a bill we don't want. } } } } } } PASS THIS ON TO EVERYONE YOU KNOW WHO USES EMAIL REMEMBER THESE ARE TWO } } } SEPARATE ISSUES THAT EFFECT ALL OF US ONLINE. LET YOU VOICE BE HEARD } } } NOW, NOT AFTER } } } } } } } } } } } } } } } } } } } } }
----------------------- Headers -------------------------------- Return-Path: {Microscopy-request-at-sparc5.Microscopy.Com} Received: from rly-yd01.mx.aol.com (rly-yd01.mail.aol.com [172.18.150.1]) by air-yd01.mail.aol.com (v67_b1.24) with ESMTP; Thu, 10 Feb 2000 00:26:56 -0500 Received: from ultra5.microscopy.com ([206.69.208.10]) by rly-yd01.mx.aol.com (v67_b1.24) with ESMTP; Thu, 10 Feb 2000 00:26:44 -0500 Received: (from daemon-at-localhost) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id XAA23514 for dist-Microscopy; Wed, 9 Feb 2000 23:10:53 -0600 (CST) Received: from no_more_spam.com (sparc5.microscopy.com [206.69.208.10]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id XAA23511 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Wed, 9 Feb 2000 23:09:56 -0600 (CST) Received: from bubba.NMSU.Edu (bubba.NMSU.Edu [128.123.3.39]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id XAA23504 for {microscopy-at-sparc5.Microscopy.Com} ; Wed, 9 Feb 2000 23:09:44 -0600 (CST) Received: from dns1.NMSU.Edu (dns1.NMSU.Edu [128.123.3.5]) by bubba.NMSU.Edu (8.9.3/8.9.0) with ESMTP id WAA15989 for {microscopy-at-sparc5.Microscopy.Com} ; Wed, 9 Feb 2000 22:04:53 -0700 (MST) Received: from nestor.NMSU.Edu (nestor.NMSU.Edu [128.123.34.146]) by dns1.NMSU.Edu (8.9.3/8.9.0) with ESMTP id WAA29784 for {microscopy-at-sparc5.Microscopy.Com} ; Wed, 9 Feb 2000 22:04:54 -0700 (MST) Received: from [128.123.62.47] (analog-ts3-11.NMSU.Edu [128.123.62.47]) by nestor.NMSU.Edu (8.9.3/8.9.0) with ESMTP id WAA266830 for {microscopy-at-sparc5.Microscopy.Com} ; Wed, 9 Feb 2000 22:04:53 -0700 X-Sender: chbennet-at-cnmailsvr.nmsu.edu Message-Id: {v0310280db4c7f4c978e0-at-[128.123.62.47]} Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii" Date: Wed, 9 Feb 2000 21:58:18 -0700 To: microscopy-at-sparc5.microscopy.com From: Christina bennett {chbennet-at-nmsu.edu} Subject: FYI: Congress to allow email charges Errors-to: Microscopy-request-at-sparc5.Microscopy.Com
Is this yet another hoax eating time & bandwidth by being posted without confirmation (like the last 3-4 times I saw this message in the pas year or two), or is it real?
I could find no reference to it at the FCC site.... Those people who are REALLY in charge of telecommunication regulations....
Woody
New format, more pix: http://www.geocities.com/capecanaveral/3722
Weelll... For most systems, if the low end noise is not inhibited (most are), the noise generated, apparent x-ray intensity, tends to increase with lower ev. This would not produce a gaussian-like peak, but a monotonic rise in intensity with lower ev. OTOH, pulse processor circuitry *could* be designed/setup to produce such an effect. Seeing Be x-rays from 100% element is difficult to impossible with most EDS systems. If compounded, the odds get worse.
In the early 1980's Philips believed that their research labs had successfully developed a new electron source suitable for electron microscopes. Indeed there was a time when they were planning to sell microscopes with the new sources. I never heard what went wrong. There were rumors that the lifetime was not good enough.
Anyone who wants details can find them in their library: "An Efficient Silicon Cold Cathode for High Current Densities" Van Gorkom and Hoeberechts Philips Journal of Research 39 (1984) 51-60
Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvannia 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
This hoax is designed, like many of the others, to eat up bandwidth and get people involved in a general uproar.
Hopefully, no one panicked here! It may be worth the time to bookmark this hoax site, or a similar one, so that when we get messages such as this in our email, we can check their validity, before contributing to its spread. No insult intended towards anyone who falls victim to these emails, as I'm sure most of us have at one time or another.
Sincerely,
David Bell Scientist Electron Microscopy Lab Millipore Corporation 80 Ashby Road Bedford, MA 01730 (781) 533-2108
This is one of those internet hoaxes. It is not true, do not send it to your friends, do not write your congressperson. Mining Co has a great web page that debunks these hoaxes and myths {http://urbanlegends.miningco.com/culture/urbanlegends/library/blhoax.htm?pid=27 33&cob=home} and is a good place to check before sending on any email that urges you to send it to everyone you know. This message is a combination of two old and popular hoaxes, see {http://urbanlegends.miningco.com/culture/urbanlegends/library/blemtax2.htm} and {http://urbanlegends.about.com/culture/urbanlegends/library/weekly/aa012099.htm} Hopefully this will die here. Scott
} } } } Congress to allow email charges } } } } } } } } Please pass this on to all you know since many of us use e-mail for } } } } business and to keep up with friends and family, I thought you'd like } } } } to know the following. } } } } } } } } Please jump on it right away and forward this to others. } } } } } } } } CNN has reported that within the next two weeks Congress is going to } } } } vote on allowing telephone companies to CHARGE A TOLL FEE for } } } } internet access. } } } } Translation: Every time we send long distance e-mail we will receive a } } } } long distance charge. This will get costly. Please visit the following web } } } } site and file a complaint. Complain to your Congressperson. We can't } } } } allow this to pass. The following address will allow you to send an } } } } e-mail on this subject DIRECTLY to your Congressperson. } } } } } } } } http://www.house.gov/writerep {http://www.house.gov/writerep}
..sniped... } } } } } } } } PASS THIS ON TO EVERYONE YOU KNOW WHO USES EMAIL REMEMBER THESE ARE TWO } } } } SEPARATE ISSUES THAT EFFECT ALL OF US ONLINE. LET YOU VOICE BE HEARD } } } } NOW, NOT AFTER
-------------------note: new mailing address---------------------- Scott Wight e-mail: scott.wight-at-nist.gov NIST 222/A113 W voice: 301-975-3949 100 Bureau Dr STOP 8371 | fax: 301-417-1321 Gaithersburg, MD 20899-8371 \|/ disclaimer: Any opinion expressed is my own and does not represent those of my employer.
A technical position is available in the Neurobiology of Aging program at the Mount Sinai School of Medicine in New York. We are seeking an experienced Electron Microscopist with a BS/BA or MS in Biology/Life Sciences. Applicants should have excellent communication and organizational skills, an understanding of basic laboratory procedures, and the ability to manage a large and varied workload. The successful candidate will participate in ultrastructural studies focusing on the effects of: estrogen and aging on hippocampal circuitry which is implicated in learning and memory, quantitative excitatory amino acid receptor (NMDA and AMPA) distribution within the central nervous system of transgenic models, as well as manipulated primate and rodent models. Qualifications include at least 2 years of experience in routine transmission electron microscopy procedures, ultramicrotomy, immunogold labelling, specimen preparation, digital photography, and routine maintenance of equipment. Experience with immunofluorescence and confocal microscopy is an asset.
We offer a salary commensurate with experience and excellent benefits. For consideration, please mail/email your resume to:
Bill Janssen Neurobiology of Aging Box 867, EB9-02 Mount Sinai School of Medicine, One Gustave L. Levy Place, New York, NY 10029-6574
We are an equal opportunity employer fostering diversity in the workplace. Bill Janssen Neurobiology of Aging Laboratories Mount Sinai School of Medicine, Box 1639 One Gustave L. Levy Place New York, NY 10029
I think the best solution would be to use a water recirculatory system. Fill it with clean water and it should stay clean for a long time. We use a 'Neslab'
Hope this helps
Alan Walker.
********************************************* Alan Walker Dept of Electronic and Electrical Engineering University of Sheffield Mappin Street Sheffield S1 3JD United Kingdom
this is of course a recurring theme in image analysis. There really is no easy answer to this. Counting particles is easier as you can perhaps separate the particles and arrive at the correct count, but if you don't know what is occluded, you cannot measure it. However, if you can use some information that is available to make some guesses or extrapolations what the shape is, it can be done. A simple example: If you look at a heap of coins, they may overlap. It is nevertheless possible to measure the individual coins because I know that they are all round. So I can take the "protruding" part of a coin and simply fit a circle and that should give me the size pretty accurately.
Are ice crystals like snowflakes? Snowflakes, if I remember correctly, have a six-fold symmetry. Can't you just measure one "branch" of a snowflake (the one that you can see), and multiply that by 6? I am not an expert on ice crystals (although I love to ski), so this may be oversimplified. You may have to think about all the information you have about ice crystals and can then perhaps make some guesses about the occluded part.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
} ---------- } From: Rosemary Walsh[SMTP:RW9-at-PSU.EDU] } Sent: Tuesday, February 08, 2000 3:24:35 PM } To: microscopy-at-sparc5.Microscopy.Com } Subject: Size determination of overlapping particles } Auto forwarded by a Rule } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Listers, I have a student interested in sizing ice crystals which overlap. Any suggestions? Rosemary
We have a tedious task of liquid nitrogen transfer and fill-up on our instruments such as SEMs, TEM and for cryo-ultramicrotomes. Is there any compact unit on market which would condense nitrogen from air into liquid form? I know that Liquid oxygen would be a problem. TIA;
I just read the two excellent replies from Scott and Phillip and fully agree with them. Small angle cleavage technique (SACT) and the tripod polisher are two very useful TEM prep techniques. In my experience, SACT is easier to learn than tripod polishing but tripodding is applicable to more materials and yields larger thin areas than does SACT.
As Phillip mentions, low energy milling in the final step of the process is extremely important for reducing artifacts. I would finish using the lowest possible energy that still yields effective milling.
Another approach is to use reactive ion beam etching (RIBE) or chemically assisted ion beam etching (CAIBE). In both these techniques a reactive gas (usually Iodine) is used. In RIBE, iodine is actually ionized and is used as the milling gas. In CAIBE, iodine gas is introduced into the milling chamber directly adjacent to the sample during argon ion milling. Both these techniques have been shown to eliminate the ion beam damage for binary type III-V compound semiconductors containing InP. Chew and Cullis (Ultramicroscopy 23/1987/175-198) also report improvements of ion milled surfaces for ternary type III-V semiconductors containing InGaAs (materials you mentioned) using RIBE.
I believe that Gatan offers an optional CAIBE attachment for the PIPS. This may be the easiest thing to try first. Remember that iodine is very corrosive and can damage ion milling parts if not used properly. Check with Gatan for their recommendations.
Hope this helps,
Eric W.
Eric S. Windsor Physical Scientist NIST 100 Bureau Dr. Stop 8371 Gaithersburg, MD 20899-8371 (301) 975-3930 Fax: (301) 417-1321 Eric.Windsor-at-NIST.Gov
On Feb 9 -at- 09:43 Massimo Catalano wrote:
Dear Listservers, we are massively working on analytical and structural TEM characterization of semiconductors, mainly InGaAs/GaAs, quantum well, wires and dots. Specimen preparation is of course a crucial point of our work. We have just switched to PIPS (we used to make our samples using the Gatan Duo Mill), and we are getting controversial results about the damage introduced by the milling procedure. We are perfectly aware of all the differences between the two instruments, especially the absence of cooling stage and the higher beam current. Has anyone performed a systematic and careful analysis to try to evaluate the milling damage on similar materials systems. We are willing to start a systematic work to assess this issue, but would like to know if anyone has already done anything on this topic. Also, any collaboration will be more than welcome. Thanks. Massimo
Eric S. Windsor Physical Scientist NIST 100 Bureau Dr. Stop 8371 Gaithersburg, MD 20899-8371 (301) 975-3930 Fax: (301) 417-1321 Eric.Windsor-at-NIST.Gov
I have found the basic description on the US/Canada website so I assume its available there too - see address below: http://www.epson.com/cam_scan/scanners/perfection1200/index.html
The price inclusive of transparency adapter appears to be about 200 UK pounds over here. There seems to be three models: 1200s (standard print scanner - SCSI interface) 1200u (standard print scanner - USB interface) 1200photo (standard print scanner + 5x4inch transparency adapter - USB interface) It's the last of these that is of interest.
Malcolm Haswell ------------------------------------------ Maureen Petersen wrote: } } Malcolm: } } Will you divulge what this gem costs? Do you know if it is available in the } US? } } Thank you, Maureen Petersen } Dept Plant Pathology } University of Florida } Gainensville, FL 32611 } } I'm sorry to bring this topic up yet again, but I have just come across } the specification for a flat-bed scanner which looks as if it would be } suitable for scanning e.m. cut-film. It is the Epson Perfection 1200 } Photo which includes a 5x4inch film adapter. } } The UK web site address is: } http://www.epson.co.uk/sohoprod/imaging/scanner/perf1200/perf1200photo/spec. } htm } } and the specification is: } A4 Colour flatbed with 5"x4" film image scanner Single pass scanning } Optical resolution: 1200 x 2400 (30600 pixels/line) } Maximum output resolution of 9600x9600 dpi } 36 bits per pixel in colour (24 bit output) } 12 bits per pixel in black (8 bit output) } Optical Density 3.2D } USB (Type B) } } Does anyone have any experience of this machine because it is about a } fifth of the price of the Umax flatbed film scanners? My only } reservation is that it is a SOHO product rather than professional. } } Thanks in advance } } Malcolm } } Malcolm Haswell } Electron Microscopy } School of Sciences } Fleming Building } University of Sunderland } SUNDERLAND SR1 3SD } Tyne and Wear } UK } } Tel (0191) 515 2872 } e-mail: malcolm.haswell-at-sunderland.ac.uk
I am considering the purchase of a new digital camera. Some of the new cameras like the Spot-RT and Magnafire have the option of running as three shot color or one shot gray-scale acquisition. If someone wants to do dual channel fluorescence like FITC/Rhod or annexin/PI, we can do that as either acquiring each image with a separate filter cube and combining in software, or designing a dual filter cube and acquiring in color (or I guess acquiring separate color images iwth the single cubes and combining??). What is people real world experience with the relative merits of these two approaches. It seems that most of what I read is done by separate filter cubes combined in software, but there also seems to be a push toward fast cooled color cameras. The advantage I can see to the color approach would be using a color chip camera you get multiple channels simultaneously. However, do you have to expose significantly longer to acheive this compared to multiple gray scale acquisitions? I can't see an obvious advantage to the three shot color approach because I suspect the light throughput is lower and the filters sets I suspect are expensive to add on if you already have the single fluor filters. The only real advantage I see to three shot acquisition is the ability to easily image stained/histology transmitted light slides and not for fluorescence. Any thoughts appreciated. Thanks- Dave
Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 75 N. Eagleville Rd. U-125 Storrs, CT 06269 Knecht-at-uconnvm.uconn.edu 860-486-2200 860-486-4331 (fax) home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html ************************************************************
This nonsense resurfaces every few months. There has never been any truth to it. (Of course, there's no assurance that Congress will not act so foolishly in the future.)
Kal
On Wed, 9 Feb 2000, Christina bennett 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 know this is off topic however a change like this could make this type of } forum impossible. } } } } } } } Date: Tue, 8 Feb 2000 08:13:53 -0700 } } X-Sender: liperez-at-cnmailsvr.nmsu.edu } } Mime-Version: 1.0 } } To: all-at-biology.nmsu.edu } } From: liperez-at-NMSU.Edu (LSPSaldana) } } Subject: Congress to allow email charges } } Sender: owner-all-at-biology.nmsu.edu } } Precedence: bulk } } X-Keywords: } } X-UID: 43 } } Status: O } } } } } } } } Congress to allow email charges } } } } } } } } Please pass this on to all you know since many of us use e-mail for } } } } business and to keep up with friends and family, I thought you'd like } } } } to know the following. } } } } } } } } Please jump on it right away and forward this to others. } } } } } } } } CNN has reported that within the next two weeks Congress is going to } } } } vote on allowing telephone companies to CHARGE A TOLL FEE for } } } } internet access. } } } } Translation: Every time we send long distance e-mail we will receive a } } } } long distance charge. This will get costly. Please visit the following web } } } } site and file a complaint. Complain to your Congressperson. We can't } } } } allow this to pass. The following address will allow you to send an } } } } e-mail on this subject DIRECTLY to your Congressperson. } } } } } } } } http://www.house.gov/writerep {http://www.house.gov/writerep} } } } } } } } } Pass this on to your friends. It is urgent! I hope all of you will pass } } } } this on to all your friends and family. We should ALL have } } } } an interest in this one. } } } } } } } } WAIT, THERE'S MORE. IN ADDITION, The last few months have revealed an } } } } alarming trend in the Government of the United States attempting to } } } } quietly push through legislation that will affect your use of the } } } Internet. } } } } Under } } } } proposed legislation the U.S. Postal Service will be attempting to bill } } } } email users out of "alternate postage fees". Bill 602P will permit the } } } } Federal Govt. to charge a 5 cent surcharge on every email delivered, by } } } } billing Internet Service Providers at source. The consumer would then be } } } } billed in turn by the ISP. Washington D.C. lawyer Richard Stepp is } } } } working without pay to prevent this legislation from becoming law. The } } } U.S. } } } } Postal Service is claiming that lost revenue due to the proliferation of } } } } email } } } } is costing nearly $230,000,000 in revenue per year. You may have noticed } } } } their recent ad campaign "There is nothing like a letter". } } } } Since the average citizen received about 10 pieces of email per day in } } } } 1998, } } } } the cost to the typical individual would be an additional 50 cents per } } } } day, or over $180 dollars Per year, above and beyond there regular } } } Internet } } } } costs. } } } } Note that this would be money paid directly to the U.S. Postal Service } } } } for a service they do not even provide. The whole point of the Internet is } } } } democracy and non-interference. If the federal government is permitted } } } } to tamper with our liberties by adding a surcharge to email, who knows } } } Where } } } } it will end. You are already paying an exorbitant price for snail mail } } } } because of bureaucratic inefficiency. It currently takes up to 6 days for } } } a } } } } letter to be delivered from New York to Buffalo. If The U.S. Postal } } } Service } } } } is } } } } allowed to tinker with email; it will mark the end of the "free" } } } } Internet in the United States. } } } } One congressman, Tony Schnell has even suggested a "twenty to forty } } } } dollar per month surcharge on all internet service" above and beyond the } } } } government's proposed email Charges. Note that most of the major } } } } newspapers have ignored the story, the only exception being the } } } } Washingtonian } } } } which called the idea of email surcharge "a useful concept who's time has } } } } come" } } } } (March 6, 1999) Editorial. } } } } } } } } Don't sit by and watch your freedoms erode away! Send this e-mail to } } } } EVERYONE on your list, and tell all your friends and relatives to write } } } } to their congressman and say "No!" to Bill 602P. } } } } } } } } } } } } It will only take a few moments of your time, and could very well be } } } } instrumental in killing a bill we don't want. } } } } } } } } PASS THIS ON TO EVERYONE YOU KNOW WHO USES EMAIL REMEMBER THESE ARE TWO } } } } SEPARATE ISSUES THAT EFFECT ALL OF US ONLINE. LET YOU VOICE BE HEARD } } } } NOW, NOT AFTER } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } }
Someone at a remote location within our company has decided to purchase an RG Lee SEM. I am looking for information and opinions from people who have one of these microscopes. I'm interested in any service issues, reliability issues and usability concerns that anyone may have. The model they are interested in is the PSM 75LS. Also, if anyone has the URL for their website, that would be appreciated, as well.
Thanks in advance,
David Bell Scientist Electron Microscopy Lab Millipore Corporation 80 Ashby Road Bedford, MA 01730 (781) 533-2108
A poster does not mean anything. I have seen them with the Lithium-Ka energy listed, although quantum physics does forbid this X-ray line. But I can assure you the Be-K line does exist.
On modern EDX systems Be can be detected, provided you have either a windowless detector or a detector with a modern polymer type window (SUTW, SATW, Norvar, or whatever the EDX supplier calls it. My apologies if I break any trade-marks here...). On most demonstration setups your EDS specialist will be pleased to show you a Be peak. But probably from a pure Be sample only. The absorption coefficient (MAC) of Be in basically any matrix is so huge, and the X-ray fluorescence yield so small, that in any compound with less than 50 atomic percent Be you will not detect a visible Be peak. For this reason most EDX installations "in the field" are not setup to detect Be-K radiation.
WDS has a better P/B ratio, so if you have the proper multilayer crystal you can get results. But peak shifts and peak shape alterations will make quantification extremely difficult, not to mention the fact that for most MACs of Be-K in any matrix we only have a ball-park figure.
Other techniques, like PEELS, are much more suitable for Li and Be analysis. Best regards,
Hans Dijkstra
Disclaimer: This is my opinion, and not necessarily the one from EDAX Inc. ------------------------------------------------------------- EDAX Europe www.edax.com Ringbaan Noord 103 Tel.: +31-13-5364000 P.O. Box 4144 Fax.: +31-13-5356279 5004 JC Tilburg E-mail: hans.dijkstra-at-edax.nl the Netherlands -------------------------------------------------------------
} -----Original Message----- } From: Walck. Scott D. [mailto:walck-at-ppg.com] } Sent: Tuesday, February 08, 2000 6:57 PM } To: 'microprobe'; 'harry.ekstrom-at-honeywell.com' } Cc: 'Microscopy' } Subject: RE: Be X-ray peaks -I don't think so } } } ------------------------------------------------------------------------ } 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'm wrong. I guess I should have looked up at the X-ray periodic table } above my head and saw the 0.108 keV value for Be. The reason that I was } wrong was that I did not consider the solid state aspects. I am still right } about not having a 2s1/2 to a 1s1/2 transition and that an X-ray carries } away 1h-bar of angular momentum. What I didn't do was think about the band } structure of a solid. If you have the solid, the 2p bands of Be most be } spilling into or be the conduction band for the electrons. That must be the } source for the electronic transition. I guess I should have engaged brain } before typing. } My most humble apologies. } -Scott } } Scott D. Walck, Ph.D. } PPG Industries, Inc. } Glass Technology Center } Guys Run Rd. (packages) } P. O. Box 11472 (letters) } Pittsburgh, PA 15238-0472 } } Walck-at-PPG.com } } (412) 820-8651 (office) } (412) 820-8161 (fax) } } } "The opinions expressed are those of Scott D. Walck and not of PPG } Industries, Inc. nor of any PPG-associated companies." } -- }
The first meeting of the millenium for NESM (New England Society for Microscopy) will be held on March 8, 2000 at Polaroid Corporation in Wayland, MA. Registration will begin at 5:00pm with tours of the (new) facility following at 5:30pm. Attendees will also be asked to fill outa questionnaire (optional) on "Professional Imaging" at this time.
A buffet supper will be served from 6-7pm followed by three scientific presentations. The speakers are : Gabriel Rojano, Staff Reliability Engineer-MIA-Com (Tyco Electronics Company), Paul Bain from the Harvard Medical Library and Dr. Hjalmar Brismer from the Karolinska Institute in Stockhold, Sweden.
Advance registration is encouraged (by March 3rd). The registration fee for members is $5.00 and $20.00 for non-members (this includes a current one-year membership in the society). Interested parties should contact: Peggy Sherwood, Corresponding Secretary (MESnesm-at-aol.com).
Different subject title in case other posting got filtered out.
} Date: Thu, 10 Feb 2000 08:08:10 -0800 } To: Microscopy-at-MSA.Microscopy.Com } From: "Dr. Gary Gaugler" {gary-at-gaugler.com} } Subject: urban myths and legends - Congress to allow e-mail charges } } Here are two URLs showing that this is just another hoax; } } } http://www.urbanmyths.com/email_internettax602p.html } } } } http://www.ulrc.com.au/html/report.asp?CaseFile=ULRR0027&Page=1&View=Request } } gary g.
The insidious nature of this hoax/scam/... is that not only does the original hoax get circulated, using up bandwidth, but then the replies of the people who identified the hoax, many with the full hoax message attached, also get circulated. And then the triple play (baseball and spring training is near) is when people like me comment on the aforesaid. Please, please don't reply to this note (which should have no attachments!) and make it a home run!
Don Marshall
Donald J. Marshall Relion Industries P.O. Box 12 Bedford, MA 01730 Ph: 781-275-4695 FAX: 781-271-0252 email dmrelion-at-world.std.com
Cathodoluminescence, mass spectroscopy, electron beam technology
"A weed is a flower out of place."
(Please note: Do not send email with attachments to this address. Instead, send it to donbarlen-at-aol.com. Thank you.)
This is not true. It is an internet hoax. Here's the info from an urban legends website: --------------------------- Claim: Congress will soon be voting on whether or not your phone company will be allowed to charge you long-distance rates for accessing the Internet.
Status: False.
Example: [Collected on the Internet, 1999]
Please forward this to everyone you can...
There is a new bill in the US Congress that will affect ALL INTERNET USERS. CNN stated that the Government would in two weeks time decide whether to allow or not allow a Charge to YOUR phone bill equal to a long distance call each time you access the Internet.
This affects us all! We cannot allow this to happen! Please visit the following URL and fill out the necessary form to let your Congressman know how you feel! The address is http://www.house.gov/writerep/. Write your representative!
Synopsis: Neither the FCC nor Congress is considering -- much less voting on -- legislation that would impose (or allow phone companies to impose) per-minute access fees on Internet users. Recent decisions by the FCC in this area have dealt only with the issue of how phone companies reimburse each other for handling calls placed to Internet service providers, not with the prospect of allowing phone companies to charge their customers long-distance rates for Internet use.
Origins: As soon as we get hold of something we really like at a reasonable price, we start worrying that it will be banned, taxed, or made too expensive for us to afford. The Internet is no exception, as our old friend -- the capitalized, exclamation pointed, "send this to everyone you know" anonymous e-mail message -- is here to tell you.
Way back when in 1987, the Federal Communications Commission did consider imposing a surcharge for transmitting data over the public telephone network, but they ultimately rejected the idea (thanks in part to the more than 10,000 letters of complaint they received). Unable to believe our good fortune (the government wasn't going to make us pay through the nose for dialing up all those neat computer bulletin boards we'd discovered), we couldn't leave well enough alone, and in 1991 a flood of urgent messages warning us that the FCC was again considering a proposal they'd rejected three years earlier hit e-mail systems all over the country (and the nascently popular Internet). Like the ubiquitous Craig Shergold message, the "modem tax" warning would long outlive the validity of the information it conveyed.
Fast forward to 1998. On-line services, the Internet, the World Wide Web, e-mail, and chat rooms are more popular than ever, a daily part of many people's lives. Somebody -- the government, the phone companies, Bill Gates, the Grinch -- must be on his way to spoil the party. Sure enough, we're now being told the phone companies and the government are in cahoots to ruin our good time.
First of all, a little background. Most of us still have to dial up over a modem and connect to an Internet Service Provider (ISP) to access the Internet. If your ISP is in your local dialing area, you probably don't pay anything at all for the call, no matter how long you stay connected. This means you get to tie up a phone line with your modem for hours and hours on end, every day, at no charge beyond the price of basic phone service. And the party at the other end of the line -- your ISP -- isn't paying anything extra, either. It's easy to see that somebody has a chance to reap some windfall profits here. If the phone companies were allowed to charge you a per-minute fee for accessing the Internet (or the government were allowed to tax your use of the Internet), their coffers would soon overflow with cash.
Scary thought, isn't it? All the phone companies need, we hear, is to get the FCC to reclassify and/or regulate ISPs, and then the phone companies can charge gobs of extra money for handling Internet traffic. And Congress is just about to vote on that very issue, we're told.
In fact, there is no such proposal before Congress, and there never has been.
The only real issue before the FCC concerning Internet usage (and the genesis of this latest round of scaremongering) is the subject of "reciprocal compensation." In short, reciprocal compensation means that when you place a local call to someone who is serviced by a different phone company, your phone company has to compensate his phone company for completing the call. (On the other hand, when you place a long-distance call, the long distance carrier who handles the traffic has to pay access charges to your phone company for originating the call.) But if the "person" you're calling is an ISP, should your phone company have to compensate the ISP's phone company?
The subject of recpirocal compensation has been a hot issue of late because new local phone companies have been springing up all over the place. The bigger phone companies, figuring that they would have many more customers than their newer (and smaller) competitors, negotiated reciprocal compensation agreements with the new phone companies. Every time one of these little phone companies' customers placed a call to a destination outside his local service that ended up on the bigger phone company's network, the big phone company would get to collect money from the little phone company. Not a bad scheme, the big phone companies thought.
Ah, but some of the little guys had a neat trick up their sleeves. They started offering their services to Internet service providers -- ISPs with banks and banks of modems that received thousands and thousands of calls every day, but never made any outgoing calls. All the reciprocal compensation started flowing one direction, from the big phone companies to the little phone companies, which wasn't what the big guys had in mind at all. "Foul," they cried. "Internet traffic flows all over the world," they noted. "Internet traffic is therefore interstate in nature and should be classified as long-distance, so the little guys should be paying us for originating the calls," they insisted. "We're not paying," they sputtered.
Enter the FCC to resolve the dispute, which they did (for now) on 25 February 1999 by ruling that phone companies are bound by whatever reciprocal compensation agreements they've negotiated with each other, whether they think they're fair or not. That was the only issue before the FCC. But most of us are already struggling with a glut of information, and we don't have the time to familiarize ourselves with details like interconnection agreements and reciprocal compensation, so when we hear reports with buzzwords like "Internet," "FCC," "access fees," and "long distance," we assume the worst, even though the real story is something quite different. And even if we make the effort to understand the whole story, we find all too often that we're reading information that has been misreported by others -- often the mainstream media -- who didn't make enough of an effort to understand the whole story themselves. If we can't even depend upon the people whose business it is to supply us with accurate information to get the facts straight, what more can we do? (See, for example, the misleading headline on the CNN article referenced in the "Additional information" section below.)
A few important points related to the recent FCC decision:
Didn't the FCC rule that Internet connections are long-distance calls?
Sort of. The FCC declared that "Internet traffic is jurisdictionally mixed and appears to be largely interstate in nature," which is the technical definition of "long distance." But that doesn't mean -- as is often misreported -- that Internet users will be paying long distance rates for dial-up connections, since the Internet has been, and still is, exempt from interstate access charges. The FCC did nothing to abolish that exemption.
Won't the phone companies just pass the cost of carrying Internet traffic to customers by raising their phone rates?
There is no guarantee that phone rates won't go up in the future, of course. However, since most states require phone companies to charge a flat rate for unlimited local usage, you won't have to pay per-minute charges for accessing the Internet (as long as your ISP has a dial-up number within your local service area).
What if the FCC changes their mind?
The possibility exists that the FCC might someday decide that additional fees can be imposed for Internet access. But as Bill Kennard, the chairman of the FCC, has stated on more than one occasion: "I want to say this as clearly as I can . . . as long as I'm chairman of the Federal Communications Commission this agency will not regulate the Internet. It's not going to happen. The FCC has no intention of making computer users pay long distance fees for dial-up access to the Internet, as people now pay when they make long-distance telephone calls. These rumors get on the Internet that the big bad FCC is going to impose all this regulation on the Internet. Now I know this painfully because every so often when one of these rumors flares up I get, literally, about 600 e-mail messages a day by people who are telling me to keep my hands off the Internet." [Note: this is not a direct quote from Kennard; it is pieced together from multiple statements of his.]
Additional information:
No Consumer Per-Minute Charges to Access ISPs (FCC)
Users, Advertisers Await FCC Decision on Internet Charges (CNN)
Update: In April 1999, a Canadian version of this message was unleashed on the Internet. Unlike its American counterpart, this version is not mere misinformation based on a flawed understanding of actual events or legislation -- it is an outright hoax:
Please read the following carefully if you intend to stay online and continue using email:
The last few months have revealed an alarming trend in the Government of Canada attempting to quietly push through legislation that will affect your use of the Internet. Under proposed legislation Canada Post will be attempting to bilk email users out of "alternate postage fees".
Bill 602P will permit the Federal Govt to charge a 5 cent surcharge on every email delivered, by billing Internet Service Providers at source. The consumer would then be billed in turn by the ISP. Toronto lawyer Richard Stepp QC is working without pay to prevent this legislation from becoming law.
The Canada Post Corporation is claiming that lost revenue due to the proliferation of email is costing nearly $23,000,000 in revenue per year. You may have noticed Canada Post's recent ad campaign "There is nothing like a letter". Since the average citizen received about 10 pieces of email per day in 1998, the cost to the typical individual would be an additional 50 cents per day, or over $180 dollars per year, above and beyond their regular Internet costs. Note that this would be money paid directly to Canada Post for a service they do not even provide. The whole point of the Internet is democracy and non-interference. If the Canadian Government is permitted to tamper with our liberties by adding a surcharge to email, who knows where it will end. You are already paying an exhorbitant price for snail mail because of beaurocratic inefficiency. It currently takes up to 6 days for a letter to be delivered from Mississauga to Scarborough. If Canada Post Corporation is allowed to tinker with email, it will mark the end of the "free" Internet in Canada. One back-bencher, Liberal Tony Schnell (NB) has even suggested a "twenty to forty dollar per month surcharge on all Internet service" above and beyond the government's proposed email charges. Note that most of the major newspapers have ignored the story, the only exception being the Toronto Star that called the idea of email surcharge "a useful concept who's time has come" (March 6th 1999 Editorial) Don't sit by and watch your freedoms erode away! Send this email to all Canadians on your list and tell your friends and relatives to write to their MP and say "No!" to Bill 602P.
Kate Turner Assistant to Richard Stepp QC Berger, Stepp and Gorman Barristers at Law 216 Bay Street Toronto, ON MlL 3C6
Yes, Americans are not alone in their belief that when the need for the primary service provided by a governmental agency diminishes or disappears, that agency will come up with draconian schemes to perpetuate its existence at taxpayer expense rather than modernizing or closing up shop. In this case, however, the message is simply too riddled with errors to be anything but a hoax:
There is no "Bill 602P" currently before the Canadian parliament. The designations of Canadian parliamentary bills take the form of the letter 'C' or 'S' followed by a number (depending upon whether they originated in the House of Commons or the Senate). Besides having the wrong prefix, this purported bill is assigned a number far too high to be one currently being considered in parliament, as you can see on the list of Canadian government bills.
Despite the claim in the message, nothing about this alleged bill is to be found. Also, there was no editorial about this on the 6 March 1999 OpEd page of Toronto Star. (Maybe "major papers have ignored the story" because it's a work of fiction?)
There is no Richard Stepp QC; law firm by the name of Berger, Stepp and Gorman; or 216 Bay Street in Toronto.
A list of Canadian Members of Parliament contains no MP by the name of Tony Schnell.
Of course, it didn't take long before the same thing started circulating in an Americanized version:
Dear Internet Subscriber:
Please read the following carefully if you intend to stay online and continue using email: The last few months have revealed an alarming trend in the Government of the United States attempting to quietly push through legislation that will affect your use of the Internet. Under proposed legislation the U.S. Postal Service will be attempting to bilk email users out of "alternate postage fees".
Bill 602P will permit the Federal Govt to charge a 5 cent surcharge on every email delivered, by billing Internet Service Providers at source. The consumer would then be billed in turn by the ISP. Washington D.C. lawyer Richard Stepp is working without pay to prevent this legislation from becoming law.
The U.S. Postal Service is claiming that lost revenue due to the proliferation of email is costing nearly $230,000,000 in revenue per year. You may have noticed their recent ad campaign "There is nothing like a letter". Since the average citizen received about 10 pieces of email per day in 1998, the cost to the typical individual would be an additional 50 cents per day, or over $180 dollars per year, above and beyond their regular Internet costs. Note that this would be money paid directly to the U.S. Postal Service for a service they do not even provide. The whole point of the Internet is democracy and non-interference. If the federal government is permitted to tamper with our liberties by adding a surcharge to email, who knows where it will end. You are already paying an exorbitant price for snail mail because of bureacratic efficiency. It currently takes up to 6 days for a letter to be delivered from New York to Buffalo. If the U.S. Postal Service is allowed to tinker with email, it will mark the end of the "free" Internet in the United States. One congressman, Tony Schnell (r) has even suggested a "twenty to forty dollar per month surcharge on all Internet service" above and beyond the government's proposed email charges. Note that most of the major newspapers have ignored the story, the only exception being the Washingtonian which called the idea of email surcharge "a useful concept who's time has come" (March 6th 1999 Editorial) Don't sit by and watch your freedoms erode away!
Send this email to all Americans on your list and tell your friends and relatives to write to their congressman and say "No!" to Bill 602P.
Kate Turner Assistant to Richard Stepp Berger, Stepp and Gorman Attorneys at Law 216 Concorde Street, Vienna, Va.
Does anyone know where I can find a troubleshooting manual/guide for paraffin sectioning? I am having trouble with the tissue "smearing" and with it looking chopped-up. Is this a vibration issue? I welcome any comments from those of you with experience in this area!
} Date: Thu, 10 Feb 2000 09:03:00 -0800 } To: microscopy-at-sparc5.microscopy.com } From: Laurie Wallin {lwallin-at-ucsd.edu} } Subject: paraffin sectioning troubleshooting guide } Cc: } Bcc: } X-Attachments: } } Does anyone know where I can find a troubleshooting manual/guide for } paraffin sectioning? I am having trouble with the tissue "smearing" and } with it looking chopped-up. Is this a vibration issue? I welcome any } comments from those of you with experience in this area! } } Thanks in advance. } } Sincerely.
Laurie Wallin UCSD Department of Anesthesiology 9500 Gilman Drive, 0629, La Jolla, CA 92093 (858) 822-3271
There has been some very significant work done by Dr. Arpad Barna et al in Hungary. Some of the papers he has presented include:
Ion Energy Effect on Surface Amorphisation of Semiconductor Crystals/A. Barna, et al
Amorphisation and Surface Morphology Development at Low Energy Ion Milling/A. Barna, B. Pecz.
Analysis of the Development of Large Surface Topography During Ion Etching/A. Barna; P. Barna; et al
Model Considerations of Ion Beam Thinning for Preparing TEM Samples/A.Barna; et al
Possibility of Surface Polishing by Ion Beam Thinning/ A. Barna
Ion Beam Thinning on the Basis of Topographic Kinetics/A. Barna
Low Angle and Low Energy Ion Beam Etching for TEM Sample Preparation/A. Barna
TEM Sample Preparation by Ion Milling / Amorphization/A. Barna, B. Pecz, M. Menyhard
I also have the following paper that may be of interest:
Preparation of InGaAs/GaAs Multilayered Materials for TEM by One Side Non-Rotation Ion Beam Thinning/J.Y. Yao; G.L. Dunlop
I do not have the complete references available in front of me, but we do have copies of all of these papers in our technical library. I would be pleased to mail copies to you if that would be of interest. We also have a list of over 250 papers dealing with various aspects of sample preparation (much of it TEM sample preparation) which may be of interest. If you would like to review that list, I can send it over to you in MS Excel format and you can select any other papers that would be of interest.
I hope this helps.
DISCLAIMER: South Bay Technology, Inc. markets the IV3 Ion Mill in both high and low energy (down to 200ev) versions which is based on the work of Dr. Barna. We also produce the XLA 2000 computer controlled Low Angle Ion Mill so we have a vested interest in promoting their use.
David Henriks Vice President 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
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by Massimo Catalano } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Listservers,
we are massively working on analytical and structural TEM characterization of semiconductors, mainly InGaAs/GaAs, quantum well, wires and dots.
Specimen preparation is of course a crucial point of our work.
We have just switched to PIPS (we used to make our samples using the Gatan Duo Mill), and we are getting controversial results about the damage introduced by the milling procedure.
We are perfectly aware of all the differences between the two instruments, especially the absence of cooling stage and the higher beam current.
Has anyone performed a systematic and careful analysis to try to evaluate the milling damage on similar materials systems. We are willing to start a systematic work to assess this issue, but would like to know if anyone has already done anything on this topic. Also, any collaboration will be more than welcome.
Well, this urban legend has finally made it to the microscopy listserver! And that is just what it is--a legend, a myth, a fable. As a stamp collector and one who keeps abreast of all of this kind of stuff, this urban legend has been debunked in both Linn's Stamp News and Stamp Collector.
All of us have important issues to consider. Please, put this one to rest.
Roger Moretz
On Wed, 9 Feb 2000 21:58:18 -0700, Christina bennett 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 know this is off topic however a change like this could make this type of } forum impossible. } } } } } } } Date: Tue, 8 Feb 2000 08:13:53 -0700 } } X-Sender: liperez-at-cnmailsvr.nmsu.edu } } Mime-Version: 1.0 } } To: all-at-biology.nmsu.edu } } From: liperez-at-NMSU.Edu (LSPSaldana) } } Subject: Congress to allow email charges } } Sender: owner-all-at-biology.nmsu.edu } } Precedence: bulk } } X-Keywords: } } X-UID: 43 } } Status: O } } } } } } } } Congress to allow email charges } } } } } } } } Please pass this on to all you know since many of us use e-mail for } } } } business and to keep up with friends and family, I thought you'd like } } } } to know the following. } } } } } } } } Please jump on it right away and forward this to others. } } } } } } } } CNN has reported that within the next two weeks Congress is going to } } } } vote on allowing telephone companies to CHARGE A TOLL FEE for } } } } internet access. } } } } Translation: Every time we send long distance e-mail we will receive a } } } } long distance charge. This will get costly. Please visit the following web } } } } site and file a complaint. Complain to your Congressperson. We can't } } } } allow this to pass. The following address will allow you to send an } } } } e-mail on this subject DIRECTLY to your Congressperson. } } } } } } } } http://www.house.gov/writerep {http://www.house.gov/writerep} } } } } } } } } Pass this on to your friends. It is urgent! I hope all of you will pass } } } } this on to all your friends and family. We should ALL have } } } } an interest in this one. } } } } } } } } WAIT, THERE'S MORE. IN ADDITION, The last few months have revealed an } } } } alarming trend in the Government of the United States attempting to } } } } quietly push through legislation that will affect your use of the } } } Internet. } } } } Under } } } } proposed legislation the U.S. Postal Service will be attempting to bill } } } } email users out of "alternate postage fees". Bill 602P will permit the } } } } Federal Govt. to charge a 5 cent surcharge on every email delivered, by } } } } billing Internet Service Providers at source. The consumer would then be } } } } billed in turn by the ISP. Washington D.C. lawyer Richard Stepp is } } } } working without pay to prevent this legislation from becoming law. The } } } U.S. } } } } Postal Service is claiming that lost revenue due to the proliferation of } } } } email } } } } is costing nearly $230,000,000 in revenue per year. You may have noticed } } } } their recent ad campaign "There is nothing like a letter". } } } } Since the average citizen received about 10 pieces of email per day in } } } } 1998, } } } } the cost to the typical individual would be an additional 50 cents per } } } } day, or over $180 dollars Per year, above and beyond there regular } } } Internet } } } } costs. } } } } Note that this would be money paid directly to the U.S. Postal Service } } } } for a service they do not even provide. The whole point of the Internet is } } } } democracy and non-interference. If the federal government is permitted } } } } to tamper with our liberties by adding a surcharge to email, who knows } } } Where } } } } it will end. You are already paying an exorbitant price for snail mail } } } } because of bureaucratic inefficiency. It currently takes up to 6 days for } } } a } } } } letter to be delivered from New York to Buffalo. If The U.S. Postal } } } Service } } } } is } } } } allowed to tinker with email; it will mark the end of the "free" } } } } Internet in the United States. } } } } One congressman, Tony Schnell has even suggested a "twenty to forty } } } } dollar per month surcharge on all internet service" above and beyond the } } } } government's proposed email Charges. Note that most of the major } } } } newspapers have ignored the story, the only exception being the } } } } Washingtonian } } } } which called the idea of email surcharge "a useful concept who's time has } } } } come" } } } } (March 6, 1999) Editorial. } } } } } } } } Don't sit by and watch your freedoms erode away! Send this e-mail to } } } } EVERYONE on your list, and tell all your friends and relatives to write } } } } to their congressman and say "No!" to Bill 602P. } } } } } } } } } } } } It will only take a few moments of your time, and could very well be } } } } instrumental in killing a bill we don't want. } } } } } } } } PASS THIS ON TO EVERYONE YOU KNOW WHO USES EMAIL REMEMBER THESE ARE TWO } } } } SEPARATE ISSUES THAT EFFECT ALL OF US ONLINE. LET YOU VOICE BE HEARD } } } } NOW, NOT AFTER } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } }
Roger C Moretz, Ph.D.
_______________________________________________________ Get 100% FREE Internet Access powered by Excite Visit http://freeworld.excite.com
Does anyone know if GFP is quenched after OsO4 postfixation? Hank Adams Integrated Microscopy Core Molecular and Cell Biology Baylor College of Medicine Houston, TX 77030
I'm working with a InSb sample prepared with tripod polishing. Since the layer is quite thick, more than 3 micron plus a 2 micron buffer layer, I am experimenting some difficult to prepare a good sample for cross section observation. The final polishing is done in a Gatan ion milling set using a low angle. The problem with my samples is how to get a uniform thickness from the top of the sample to the substrate. Usually the top of the sample is milled much faster than near to the substrate. I can't mechanically polish the sample too much, since the sample is somewhat fragile (brittle). Even in old papers people report this kind of problem when preparing InSb samples for TEM using ion milling. Does someone else suffered from this kind of problem: the top part of a sample being milled away? Oh yes, when I polish the sample I try to make wedge perpendicular to the sample surface, so the thickness of the sample is the same from the InSb layer to the substrate after the mechanical polishing. There is some correlation between the ion milling angle/energy with this effect? The Gatan epoxy I use to glue a Si piece on top of the sample should have some influence (charging)? Thanks.
Kazuo
o-------------------------------------------------------o | Carlos Kazuo Inoki | | Department of Physics - University at Albany | | 1400 Washington Ave.- Albany - NY - 12222 | o-------------------------------------------------------o
I think that I know understand the reason why you get Be X-ray and why there are energy shifts when it is in different compounds. My next question then is do the elements between B and F have measurable energy shifts depending on what material they are in? The key word here is measurable. The Ka1 and Ka2 peaks are due to transitions form the 2p1/2 and 2p3/2 (LII and LIII) to the 1s1/2 state. But these would be valence/conductance bands for the pure elements. (Assuming you collect your X-rays from solid N2, O2, or F2) The band structure would be different for different materials. For example, do you see any shifts between B2O3? and BN or C(diamond) and C(graphite) or TiC? Any microprobers following this thread?
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
} -----Original Message----- } From: Hans Dijkstra [mailto:hans.dijkstra-at-edax.nl] } Sent: Thursday, February 10, 2000 11:01 AM } To: Walck. Scott D.; 'microprobe' } Cc: 'Microscopy' } Subject: RE: Be X-ray peaks -I don't think so } } } -------------------------------------------------------------- } ---------- } 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 Scott, } } A poster does not mean anything. I have seen them with the } Lithium-Ka energy } listed, although quantum physics does forbid this X-ray line. } But I can } assure you the Be-K line does exist. } } On modern EDX systems Be can be detected, provided you have either a } windowless detector or a detector with a modern polymer type } window (SUTW, } SATW, Norvar, or whatever the EDX supplier calls it. My } apologies if I break } any trade-marks here...). } On most demonstration setups your EDS specialist will be } pleased to show you } a Be peak. But probably from a pure Be sample only. The absorption } coefficient (MAC) of Be in basically any matrix is so huge, } and the X-ray } fluorescence yield so small, that in any compound with less } than 50 atomic } percent Be you will not detect a visible Be peak. For this } reason most EDX } installations "in the field" are not setup to detect Be-K radiation. } } WDS has a better P/B ratio, so if you have the proper } multilayer crystal you } can get results. But peak shifts and peak shape alterations will make } quantification extremely difficult, not to mention the fact } that for most } MACs of Be-K in any matrix we only have a ball-park figure. } } Other techniques, like PEELS, are much more suitable for Li } and Be analysis. } Best regards, } } Hans Dijkstra } } Disclaimer: This is my opinion, and not necessarily the one } from EDAX Inc. } ------------------------------------------------------------- } EDAX Europe www.edax.com } Ringbaan Noord 103 Tel.: +31-13-5364000 } P.O. Box 4144 Fax.: +31-13-5356279 } 5004 JC Tilburg E-mail: hans.dijkstra-at-edax.nl } the Netherlands } ------------------------------------------------------------- } } } } -----Original Message----- } } From: Walck. Scott D. [mailto:walck-at-ppg.com] } } Sent: Tuesday, February 08, 2000 6:57 PM } } To: 'microprobe'; 'harry.ekstrom-at-honeywell.com' } } Cc: 'Microscopy' } } Subject: RE: Be X-ray peaks -I don't think so } } } } } } } -------------------------------------------------------------- } ---------- } } 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'm wrong. I guess I should have looked up at the X-ray } periodic table } } above my head and saw the 0.108 keV value for Be. The } reason that I was } } wrong was that I did not consider the solid state aspects. } I am still } right } } about not having a 2s1/2 to a 1s1/2 transition and that an } X-ray carries } } away 1h-bar of angular momentum. What I didn't do was } think about the } band } } structure of a solid. If you have the solid, the 2p bands } of Be most be } } spilling into or be the conduction band for the electrons. } That must be } the } } source for the electronic transition. I guess I should } have engaged brain } } before typing. } } My most humble apologies. } } -Scott } } } } Scott D. Walck, Ph.D. } } PPG Industries, Inc. } } Glass Technology Center } } Guys Run Rd. (packages) } } P. O. Box 11472 (letters) } } Pittsburgh, PA 15238-0472 } } } } Walck-at-PPG.com } } } } (412) 820-8651 (office) } } (412) 820-8161 (fax) } } } } } } "The opinions expressed are those of Scott D. Walck and not of PPG } } Industries, Inc. nor of any PPG-associated companies." } } -- } } } }
I did InSb and AlInSb by the small angle cleavage technique when I was at Wright Patterson AFB. I assume since you are using the Tripod Technique that you require site-specific results. If not, try it. It works really well on this material.
I am getting to sound like a broken record on this topic, aren't I? See my post yesterday.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies." --
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
} -----Original Message----- } From: Carlos Kazuo Inoki [mailto:kazuo-at-csc.albany.edu] } Sent: Thursday, February 10, 2000 1:05 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: TEM: sample preparation for InSb } } } -------------------------------------------------------------- } ---------- } 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'm working with a InSb sample prepared with tripod polishing. } Since the layer is quite thick, more than 3 micron plus a 2 } micron buffer } layer, I am experimenting some difficult to prepare a good sample for } cross section observation. The final polishing is done in a Gatan ion } milling set using a low angle. The problem with my samples is } how to get a } uniform thickness from the top of the sample to the } substrate. Usually the } top of the sample is milled much faster than near to the substrate. I } can't mechanically polish the sample too much, since the sample is } somewhat fragile (brittle). Even in old papers people report } this kind of } problem when preparing InSb samples for TEM using ion milling. Does } someone else suffered from this kind of problem: the top part of a } sample being milled away? Oh yes, when I polish the sample I } try to make } wedge perpendicular to the sample surface, so the thickness } of the sample } is the same from the InSb layer to the substrate after the mechanical } polishing. There is some correlation between the ion milling } angle/energy } with this effect? The Gatan epoxy I use to glue a Si piece on } top of the } sample should have some influence (charging)? Thanks. } } Kazuo } } o-------------------------------------------------------o } | Carlos Kazuo Inoki | } | Department of Physics - University at Albany | } | 1400 Washington Ave.- Albany - NY - 12222 | } o-------------------------------------------------------o } }
I hate to add to the bandwidth clutter but these are very old hoaxes.
Damian
} Is this yet another hoax eating time & bandwidth by being posted without } confirmation (like the last 3-4 times I saw this message in the pas year or } two), or is it real?
Yes, after OsO4 GFP is quenched, moreover it is quenched significantly (three times or more) even with 0.05% of glutaraldehyde (10 min.
Dr. Alexander A. Mironov Jr., MD, PhD Unit of Morphology Dept. of Cell Biology and Oncology Consorzio Mario Negri Sud Via Nazionale, S.Maria Imbaro (Ch) 66030 Italy
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Does anyone know if GFP is quenched after OsO4 postfixation? } Hank Adams } Integrated Microscopy Core } Molecular and Cell Biology } Baylor College of Medicine } Houston, TX 77030 } } }
Thanks to all of you who have shown an interest in our internships. I'd like to provide a few more details and answer some questions I've received so far about the summer internships at Abbott.
First: Are the internships open to international students? Technically, yes. However, interns are employees of Abbott Laboratories during their time here. Thus, the logistics of processing the necessary paperwork for non-US citizens for a 12-week employment period would not be practical. So, in truth, the answer has to be that, except in highly unusual circumstances, we will limit applications to students in the United States.
Second: Are the internships intended for college or high school students? The internships are for college students working towards a Bachelor's degree or higher. Students in the Associate degree programs at Madison Area Technical College and San Joaquin Delta College are eligible, but priority will be given to students who already have a Bachelor's degree.
Third: How to apply? Go to the Abbott Laboratories website at abbott.com and follow the links: Careers, Entry Level Positions, Summer Internship Program. There you will find an electronic resume form that can be submitted from your computer. THE DEADLINE FOR SUBMISSION IS MARCH 1.
Along with submitting the electronic resume, please send me either an e-mailed or faxed copy of your resume, or a hardcopy via snail mail. The electronic resume ends up in Corporate Staffing, where your skills will be matched with the internship openings at Abbott. If I have a copy of your resume, I can make sure that you are considered for positions in the microscopy department.
If there are any other questions, please feel free to contact me.
Thanks,
Jane A. Fagerland, Ph.D. Dept. Microscopy and Microanalysis D-45M/AP31 200 Abbott Park Rd. Abbott Park IL 60064-6202 voice: (847) 935-0104 fax: (847) 938-5027 e-mail: jane.a.fagerland-at-abbott.com
There was a paper by Cowen, Haven and Burnstock 1985, using Pontamine Sky Blue couterstain to reduce autofluorescence.
Also a paper by Kittleburger, Davis, and Stephans in Acta Histochem 89, using Eriochrome Black T.
Bob Morphology Core U of Washington
On Wed, 9 Feb 2000, Corazon Bucana wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I am working with archival specimens of formalin-fixed paraffin embedded } tissues and I am getting tremendous autofluorescence. I looked at tissue } sections after the sections have been de-waxed using either FITC or } rhodamine filter sets and red cells and connective tissues are brightly } fluorescent. Is there a way of suppressing the autofluorescence and still } retain reactivity of tissue to antibodies? I would appreciate any comments } or suggestions. } } ******************************************************* } Corazon D. Bucana, Ph.D. } Department of Cancer Biology } U.T. M.D. Anderson Cancer Center } 1515 Holcombe Blvd. Box 173 } Houston, Texas 77030 } Phone: (713) 792-8106 } FAX: (713) 792-8747 } Email:bucana-at-audumla.mdacc.tmc.edu } FAX: (713) 792-8747 } } }
At 8:51 AM +0100 4/2/00, Alexander Mironov Jr. wrote: } I have tried GoldEnhance for preembedding protocol to label intracellular } structures. It is marvelous. Buy it and use it - you will not be } unsatisfied. I have no any interest in Nanoprobes, I just like these dense } gold particles. } Practically all they claim is true: } - light insensitive } - no self-nucleation } - do not react with buffer ions } - is not dissolved by uranil and osmium (I have treat for 1 h)
I have been using BioCell's silver enhancement kit for years and particularly like it for the light insensitivity - one can monitor the reaction under the light microscope. Is it possible to do this with the GoldEnhance as well?
GoldEnhance sounds perfect - has anyone had any problems with it?
Diana
Diana van Driel Department of Clinical Ophthalmology Sydney University GPO Box 4337 Sydney NSW AUSTRALIA 2001
Reply to: Re:FYI: Congress to allow email charges -Hoax? I too had my suspicions. It looks very much like the "Internet Access Tax" letter from a few years ago. See: {http://urbanlegends.miningco.com/culture/urbanlegends/library/weekly/aa010798.htm} for details.
There are many sites documenting this sort of stuff. {http://www.hoaxkill.com/} is an example .
Disclaimer: I have no affiliation to this site.
Regards, Paul Webster
Paul Webster, Ph.D. Associate Scientist & Director Ahmanson Advanced Electron Microscopy & Imaging Center House Ear Institute 2100 West Third St. Los Angeles, CA 90057
} ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } Woody, } } I hate to add to the bandwidth clutter but these are very old hoaxes. } } Damian } } } } Is this yet another hoax eating time & bandwidth by being posted without } } confirmation (like the last 3-4 times I saw this message in the pas year or } } two), or is it real? } } Damian's response is indeed correct -- see posted responses by "David_Bell-at-millipore.com", "EvexAnalyt-at-aol.com", Scott Wight, Kalman Rubin, Dr. Gary Gaugler, donald j. marsh, Roger Moretz, Laurie Wallin and Damian. Being new to the MSA listserver, I'm very relieved that such notices (err, BANDWIDTH-OCCUPYING, ANNOYING HOAXES) are determined to be false by very astute readers. I fell for this, and, upon reading other people's notices, I am upset that someone would suggest spreading such hoaxes around like the classic junk chain mail activities done at school (or even college). Such notices clog up the space needed for important issues regarding microscopy. Anyway .. I'm responding merely to alert other GULLIBLE people (like myself) not to believe such notices, as well as be thankful that this has been a recurring event in the MSA listserver. Nelson Conti
_______________________________________________________ Get 100% FREE Internet Access powered by Excite Visit http://freeworld.excite.com
IMHO, the main problem with GoldEnhance is some inhomogenity in particle size but for my purposes it is irrelevant. Also the kit works good in my hands at pH 7.4 but does not at pH 6.5 (as claimed Nanoprobes).
Sincerely, Dr. Alexander A. Mironov Jr., MD, PhD Unit of Morphology Dept. of Cell Biology and Oncology Consorzio Mario Negri Sud Via Nazionale, S.Maria Imbaro (Ch) 66030 Italy
Dear readers, could anyone give me some hints or tips how I can separate a formvarfilm (0.5% in chloroform) from the glassslide ? Breathing or freezing doesn't help at all. It works better with non-washed slides, but then I get an uneven and dirty surface of the film. I appreciate every kind of support. Best regards
Guido Luond Institute of Oral Microbiology and General Immunology Univ. of Zurich Plattenstr. 11 CH-8028 Zurich, Switzerland E-mail: lueoend-at-zzmk.unizh.ch
Carlos, sounds like you may be having a problem with differential milling rates. Try milling over a restricted angle; you can use oscillation (if your ion mill is set up to do this) or make C-shaped Ta plate shields which you fix on the sample holding plates of your ion mill, with the sample at the centre of the 'C'. If you put your sample in the plates such that the interface is parallel to the opening of the shield, the average direction of the ion beam will be perpendicular to the interface. This will prevent the ion beam milling parallel to the glue line and stop the layers disappearing before the substrate. If you stick to angles below 10 degrees you will get better results, and of course use liquid nitrogen cooling and/or iodine. My apologies if this isn't very clear - it's hard to describe everything with just words! I can send you a picture of the sample holder I use if you like.
Best regards,
Richard Beanland
} ------------------------------------------------------------------------ } 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'm working with a InSb sample prepared with tripod polishing. } Since the layer is quite thick, more than 3 micron plus a 2 micron buffer } layer, I am experimenting some difficult to prepare a good sample for } cross section observation. The final polishing is done in a Gatan ion } milling set using a low angle. The problem with my samples is how to get a } uniform thickness from the top of the sample to the substrate. Usually the } top of the sample is milled much faster than near to the substrate. I } can't mechanically polish the sample too much, since the sample is } somewhat fragile (brittle). Even in old papers people report this kind of } problem when preparing InSb samples for TEM using ion milling. Does } someone else suffered from this kind of problem: the top part of a } sample being milled away? Oh yes, when I polish the sample I try to make } wedge perpendicular to the sample surface, so the thickness of the sample } is the same from the InSb layer to the substrate after the mechanical } polishing. There is some correlation between the ion milling angle/energy } with this effect? The Gatan epoxy I use to glue a Si piece on top of the } sample should have some influence (charging)? Thanks. } } Kazuo } } o-------------------------------------------------------o } | Carlos Kazuo Inoki | } | Department of Physics - University at Albany | } | 1400 Washington Ave.- Albany - NY - 12222 | } o-------------------------------------------------------o }
============================================================== Richard Beanland Caswell Technology, Caswell, Towcester, Northants NN12 8EQ
e-mail richard.beanland-at-gecm.com Tel. +44 1327 356363 Fax. +44 1327 356398 ============================================================== "The information contained in this message is legally privileged and confidential information intended for the eyes of the individual or entity named above. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution or copying of this message is strictly prohibited. If you have received this message in error, please notify us immediately by telephone. Caswell Technology is the trading name of Marconi Caswell Limited. Registered in London No. 3694360 Registered Office: One Bruton Street London W1X 8AQ. Holding Company: Marconi plc."
Is your end goal hi res imaging or ratioing? If the latter, I saw an interesting technology at Cell Bio which might be of help: the PARISS Spectral imaging system from LightForm. I saw very early versions of this technology some years ago at PITTCON but it has really matured and offers an interesting alternative. PARISS can acquire 240 spectra simultaneously, full field or in a region of interest. It really crashes through the old barriers imposed by filter cubes. Also, despite the radical differences in intensity, it can acquire the transmitted light image simultaneously with the fluorescence image and present the combined results with perfect registration. The system retrofits to existing microscopes and is moderately priced, considering all the accessories (2 cameras, spectrometer, motorized stage, software). If you are interested, visit their website at lightforminc.com or contact Jeremy Lerner (908-281-9098).
Caveat: MME has no financial interest in this product.
Hope this is helpful.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education}
At 10:23 AM 2/10/00 -0500, David Knecht wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Sorry, I think a small correction/clarification is requires. This is ONLY for an incoherently filled condensor aperture, i.e. a large beam with C2 fully focussed. This approximation is not true with the newer microscopes!
On Thu, 10 Feb 2000, Jonathan Barnard wrote:
} Born & Wolf (section 10.4.2) } "Hence if the linear dimensions of the source and the distance between P1 } and P2 (points in the imaging plane) are small compared to the distance of } these points from the source, the degree of coherence, |mu_12| is equal to } the absolute value of the normalized Fourier transform of the intensity } function of the source" }
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University fax: (847) 491-7820 mailto:l-marks-at-nwu.edu http://www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
WDS spectra of Boron, Carbon, Nitrogen and Oxygen in many compounds have been extensively recorded by Bastin and Heijligers in the 1980's and early 1990's. They report that peak shifts and peak shape alterations are clearly present in Boron compounds. In the case of TiB crystals it was even found that the crystal orientation influences the peak shape and position, i.e. if you rotate the sample 90 degrees you get a different peak shape. For Carbon the peak shapes are practically unaffected by the bonding with other elements.
Please check "Quantitative Electron Probe Microanalysis of Boron in Binary Borides" by Bastin and Heijligers, University of Technology Eindhoven, the Netherlands, 1997, ISBN 90-3860-898-5.
In EDX the peak shifts are undetectable. Modern EDX systems give a FWHM of around 60-65 eV for these very light elements, so a peak shift of a few eV is basically undetectable. For many users this makes EDX the prefered technique to quantify Boron compounds, since with WDS you need to record either the full peak (tedious) or use fixed area-peak-fraction correction methods. In EDX we can simple ignore peak shape and peak position changes.
Best regards,
Hans Dijkstra
Disclaimer: This is my opinion, and not necessarily the one from EDAX Inc. ------------------------------------------------------------- EDAX Europe www.edax.com Ringbaan Noord 103 Tel.: +31-13-5364000 P.O. Box 4144 Fax.: +31-13-5356279 5004 JC Tilburg E-mail: hans.dijkstra-at-edax.nl the Netherlands -------------------------------------------------------------
} -----Original Message----- } From: Walck. Scott D. [mailto:walck-at-ppg.com] } Sent: Thursday, February 10, 2000 7:51 PM } To: 'Microscopy' } Subject: RE: Be X-ray peaks, solid state physics, how about 2nd row } elements } } } ------------------------------------------------------------------------ } 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 think that I know understand the reason why you get Be X-ray and why } there are energy shifts when it is in different compounds. My next question } then is do the elements between B and F have measurable energy shifts } depending on what material they are in? The key word here is measurable. } The Ka1 and Ka2 peaks are due to transitions form the 2p1/2 and 2p3/2 (LII } and LIII) to the 1s1/2 state. But these would be valence/conductance bands } for the pure elements. (Assuming you collect your X-rays from solid N2, O2, } or F2) The band structure would be different for different materials. For } example, do you see any shifts between B2O3? and BN or C(diamond) and } C(graphite) or TiC? Any microprobers following this thread? } } -Scott } } Scott D. Walck, Ph.D. } PPG Industries, Inc. } Glass Technology Center } Guys Run Rd. (packages) } P. O. Box 11472 (letters) } Pittsburgh, PA 15238-0472 } } Walck-at-PPG.com } } (412) 820-8651 (office) } (412) 820-8161 (fax) }
first of all I would like to thank of the people who spent part of their time trying to share their knowledge with me, and also to the manufacturers who of course replied my messages.
All messages contained very important suggestions, that I need to study and evaluate, and I will contact personally the people that offered to share their previous experience and/or to collaborate.
Our work on InGaAs/GaAs quantum wells, single and stacked quantum wires, single and stacked quantum dots, consists of conventional TEM, high resolution transmission electron microscopy and high spatial resolution analytical electron microscopy, that we perform in close collaboration with Arizona State University, using a FEG/STEM machine equipped with EELS and EDX. One of the goals of the research is to evaluate compositional fluctuations in the well and in the dots at nanoscale.
As everybody knows, the requirements from a TEM sample for imaging are completely different from the requirements necessary to perform high spatial resolution analytical work, were effects such as amorphisazion, surface contamination are critical.
We have a pretty well estabilished specimen preparation technique, but before publishing results into the scientific community, we want to be sure that what we observe is what is in the sample (isn't this the main concern of every microscopist?).
That's why all suggestions, opinions and critics are strongly appreciated.
I don't wash the slides, just wipe the dry slide with a Kim-Wipe a few times. Works like a charm, whatever the weather! (Very grey and nasty here on "the island" today, by the way).
Tamara Howard CSHL
On Fri, 11 Feb 2000, GUIDO L[ISO-8859-1] üöND 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 readers, } could anyone give me some hints or tips how I can separate a formvarfilm } (0.5% in chloroform) from the glassslide ? Breathing or freezing doesn't } help at all. It works better with non-washed slides, but then I get an } uneven and dirty surface of the film. } I appreciate every kind of support. } Best regards } } Guido Luond } Institute of Oral Microbiology and } General Immunology } Univ. of Zurich } Plattenstr. 11 } CH-8028 Zurich, Switzerland } E-mail: lueoend-at-zzmk.unizh.ch } } }
There is a product called "Victawet", which is available here in the U.S. from various EM suppliers. It comes in a small vial that lasts forever and is an off-white, waxy or soapy substance. We used to take our slides and clean them very well with lint-free papers, then place them in a rack inside a vacuum evaporator, with the side intended for the Formvar film facing a tungsten basket connected to the electrodes. Then we place a piece of Victawet about the size of a grain of rice in the basket, pump down the system to high vacuum, and slowly heat the tungsten basket until it glows dull red. (If you increase the current too quickly the little piece will jump out of the basket.) The slides will begin to look fogged, as if someone had breathed moisture onto them. You can stop at that point---you only need a little.
Store these coated slides until needed, then take them as required and polish them very carefully with lint-free paper. They should feel slippery. Use these cleaned slides to dip into the Formvar solution and the film should separate easily.
Please note that the type of slide and the humidity levels in the workplace also have a large effect. I have worked in places where films would separate from any slide, every day, with no Victawet coating. I have also worked in places where we could only get Formvar films on occasion (who knows why?), and then we made a bunch of them at once.
Hope this helps.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 (573) 882-8304 (573) 884-5414 (fax) email: tindallr-at-missouri.edu {mailto:tindallr-at-missouri.edu} http://biotech.missouri.edu/emc {mailto:tindallr-at-missouri.edu}
Dear readers,
could anyone give me some hints or tips how I can separate a formvarfilm
(0.5% in chloroform) from the glassslide ? Breathing or freezing doesn't
help at all. It works better with non-washed slides, but then I get an
uneven and dirty surface of the film.
I appreciate every kind of support.
Best regards
Guido Luond
Institute of Oral Microbiology and
General Immunology
Univ. of Zurich
Plattenstr. 11
CH-8028 Zurich, Switzerland
E-mail: lueoend-at-zzmk.unizh.ch
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 (573) 882-8304 (573) 884-5414 (fax) email: tindallr-at-missouri.edu {mailto:tindallr-at-missouri.edu} http://biotech.missouri.edu/emc {mailto:tindallr-at-missouri.edu}
Hasn't anyone realized that the "hoax" email is a success even as a failure. Everyone is responding so much about the email being a hoax that we're using up band width and cluttering email accounts. We all know it's a hoax..., let's leave it at that.....
Regards,
Mike Santana, Jr. Fab25 Product Engineering Advanced Micro Devices Desk - (512) 602-4172 Pager - (512) 622-2494 email - mike.santanajr-at-amd.com
On Thursday, February 10, 2000 10:27 PM, Paul Webster [SMTP:pwebster-at-hei.org] 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 } -----------------------------------------------------------------------. } } } Reply to: Re:FYI: Congress to allow email charges -Hoax? } I too had my suspicions. It looks very much like the "Internet Access Tax" letter from a few years ago. See: {http://urbanlegends.miningco.com/culture/urbanlegends/library/weekly/aa 010798.htm} for details. } } There are many sites documenting this sort of stuff. {http://www.hoaxkill.com/} is an example . } } Disclaimer: I have no affiliation to this site. } } Regards, } Paul Webster } } Paul Webster, Ph.D. } Associate Scientist & Director } Ahmanson Advanced Electron Microscopy & Imaging Center } House Ear Institute } 2100 West Third St. } Los Angeles, CA 90057 } } Phone: (213) 273-8026 } Fax: (213) 413-6739 } e-mail: pwebster-at-hei.org } http://www.hei.org/htm/aemi.htm }
I would like to contact users of Bal-Tec RES 010 ion thinner to comment on performance of this equipment. We have found over the years frequent stability problems of the guns, which are not solved only with cleaning. We are at the moment checking any possible source of unstability. We would like advise from anyone who has had similar problems with this apparatus, to know if we are missing something and what would be the best course of action.
Thanks in advance
Jess Ricote ------------------------------------------------------------- Dr. Jess Ricote Departamento de Materiales Ferroelctricos Instituto de Ciencia de Materiales de Madrid Consejo Superior de Investigaciones Cientificas (CSIC) Cantoblanco 28049 Madrid SPAIN
first of all I would like to thank of the people who spent part of their time trying to share their knowledge with me, and also to the manufacturers who of course replied my messages.
All messages contained very important suggestions, that I need to study and evaluate, and I will contact personally the people that offered to share their previous experience and/or to collaborate.
Our work on InGaAs/GaAs quantum wells, single and stacked quantum wires, single and stacked quantum dots, consists of conventional TEM, high resolution transmission electron microscopy and high spatial resolution analytical electron microscopy, that we perform in close collaboration with Arizona State University, using a FEG/STEM machine equipped with EELS and EDX. One of the goals of the research is to evaluate compositional fluctuations in the well and in the dots at nanoscale.
As everybody knows, the requirements from a TEM sample for imaging are completely different from the requirements necessary to perform high spatial resolution analytical work, were effects such as amorphisazion, surface contamination are critical.
We have a pretty well estabilished specimen preparation technique, but before publishing results into the scientific community, we want to be sure that what we observe is what is in the sample (isn't this the main concern of every microscopist?).
That's why all suggestions, opinions and critics are strongly appreciated.
Ah. Bit of a problem, that. I didn't mention how far away the shield is from the sample - about 3 or 4 mm - and that for low angles I use a wedge-shaped shield to deflect ions away from the specimen, rather than back on to it. Did you really try it that quickly? I only sent the e-mail a few hours ago! I did expect anyone who was going to have a go to ask me for a picture of my holder. As to cleaning your sample, I think you can only ion mill it some more, with amended shields...
Richard
} Dear Richard, } } I did as you suggested in my sample preparation. However, the Ta was } sputtered on the surface of the } specimen. Could you suggest me how to avoid or how to clean the Ta on the } specimen surface. } } Chengge Jiao } H.H.Wills Physics Lab } University of Bristol, U.K } c.g.jiao-at-bristol.ac.uk
============================================================== Richard Beanland Caswell Technology, Caswell, Towcester, Northants NN12 8EQ
e-mail richard.beanland-at-gecm.com Tel. +44 1327 356363 Fax. +44 1327 356398 ============================================================== "The information contained in this message is legally privileged and confidential information intended for the eyes of the individual or entity named above. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution or copying of this message is strictly prohibited. If you have received this message in error, please notify us immediately by telephone. Caswell Technology is the trading name of Marconi Caswell Limited. Registered in London No. 3694360 Registered Office: One Bruton Street London W1X 8AQ. Holding Company: Marconi plc."
One of the benefits from the small angle cleavage technique on the III-V compounds is that often a "step-like" sample is produced. When this happens, you have a perfect sample for both High resolution and analytical work. In the very thin steps, you can do HREM and EELS. In the thicker steps, you can do EDS analysis. -Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
} -----Original Message----- } From: Massimo Catalano [mailto:massimo.catalano-at-ime.le.cnr.it] } Sent: Friday, February 11, 2000 10:23 AM } To: Microscopy-at-sparc5.Microscopy.Com } Subject: PIPS and milling damage: Second message } } } -------------------------------------------------------------- } ---------- } 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 Listservers: } } first of all I would like to thank of the people who spent } part of their } time trying to share their knowledge with me, and also to the } manufacturers } who of course replied my messages. } } All messages contained very important suggestions, that I } need to study and } evaluate, and I will contact personally the people that } offered to share } their previous experience and/or to collaborate. } } Our work on InGaAs/GaAs quantum wells, single and stacked } quantum wires, } single and stacked quantum dots, consists of conventional TEM, high } resolution transmission electron microscopy and high spatial } resolution } analytical electron microscopy, that we perform in close } collaboration with } Arizona State University, using a FEG/STEM machine equipped } with EELS and } EDX. One of the goals of the research is to evaluate compositional } fluctuations in the well and in the dots at nanoscale. } } As everybody knows, the requirements from a TEM sample for } imaging are } completely different from the requirements necessary to perform high } spatial resolution analytical work, were effects such as } amorphisazion, } surface contamination are critical. } } We have a pretty well estabilished specimen preparation } technique, but } before publishing results into the scientific community, we } want to be sure } that what we observe is what is in the sample (isn't this the } main concern } of every microscopist?). } } That's why all suggestions, opinions and critics are strongly } appreciated. } } Best regards } } Massimo } } } Dr. Massimo Catalano } CNR-IME } Campus Universitario } Via Arnesano } 73100 Lecce - ITALY } tel: + 39 0832 322362 } fax: + 39 0832 325299 } email: massimo.catalano-at-ime.le.cnr.it } http://www.ime.le.cnr.it } http://www.ime.le.cnr.it/sime/sime.htm } } }
I've always had good luck getting the film off by polishing the slide first with Ross Optical Lens Tissue. My major professor taught me this, and his explaination was that it has silicone in it. Once you have cast the film onto the slide and allowed it to dry, cut through the film at the edges and bottom of the slide by running a different slide along them. Then breathe on the film and lower it into the water dish.
Good luck, Heather Owen
p.s. I have no affiliation with the company that makes, or any of the EM supply houses that sell Ross lens paper.
Heather A. Owen, Director Electron Microscope Laboratory Department of Biological Sciences University of Wisconsin - Milwaukee (414)229-6816
Dear Scott, In the Microbeam Analysis 1985, G.F. Bastin and H.J.M. Heijligers have the first article, entitled: "Quantitative Electtron-Probe Microanalysis of Very Light Elements" (page 1). They have several articles in that and succeeding years in which they try to analyse every element and compound in the B to F range material that they can get to sit still under the EPMA beam. They did a very valuable set of studies that is worth looking into if you want to do any very light element analysis. They cover peak shifts, peak shape changes, even crystallographic direction sensitivities. At 01:51 PM 2/10/00 -0500, you wrote:
} } I think that I know understand the reason why you get Be X-ray and why } there are energy shifts when it is in different compounds. My next question } then is do the elements between B and F have measurable energy shifts } depending on what material they are in? The key word here is measurable. } The Ka1 and Ka2 peaks are due to transitions form the 2p1/2 and 2p3/2 (LII } and LIII) to the 1s1/2 state. But these would be valence/conductance bands } for the pure elements. (Assuming you collect your X-rays from solid N2, O2, } or F2) The band structure would be different for different materials. For } example, do you see any shifts between B2O3? and BN or C(diamond) and } C(graphite) or TiC? Any microprobers following this thread? } } -Scott 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
Hello Casey/Marty I cannot answer for the first question as we use Diotome knives and I have no experience with the other.
But the answer to the second question is "Yes." At least for us it is. We are a facility used by the Faculty of Medicine and Science so many and varied samples come through here for electron, light, and confocal microscopy. We have been working up some protocols for difficult specimens such as nematodes, drosophila larvae and plant material. The power supply allows control so that you don't have to do any other processing to the specimens than toss them in the fixative. Since the fix and post fix enter the specimens at low power ( {200 watts) without damaging the specimens (full power is usually about 800 watts) we can fix in about 45 minutes for something which would normally be in 24 hours. And we don't have to punch holes in the specimen.
We have just been playing with the introduction of fluorescent dyes for light microscopy and confocal in C. elegans, without the use of agents to distrupt the membranes to get the dye in. So far we have got the timings for beautiful staining of the tail region and partial staining of the thicker sections. We still have to get it right but the initial experiments are very positive.
No-one seems to know why the microwaves work, but the hypothesis is that at low power, the cell membranes are "jiggled about" to let the fix/dye in without distrupting them.
We are a very busy lab, and our experiments are constantly getting interupted so it is taking longer than we'd like to pin down a finished protocol.
I would be interested in receiving protocols from labs who have already got it to work for their specimens. We are finding that every specimen is different and the protcols have to be tweaked for each one. Elaine
} } A colleague would like your responses to these questions. I believe that I } say something on the first question about a year ago and apologize for } asking again.
} } Here's what I/we need to know. } } } } 1) Are Microstar diamond knives equal to Diatome. We have an offer that } } will save us about $1200 on three knives ( one 3mm ultra with 45 degree } } angle, one 3mm ultra with 35 degree angle, one 10 mm histo knife). } } } } } } 2) Is the power controller feature on the Pelco Microwave Tissue processor } } (model #3451) worth it? (costs about $1300 additional compared to the } } Pelco 3450 without the power controller). I plan to do immunolocalization } } work, but am not sure this power controller feature is necessary or worth } } the cost. } } } } Casey
Marty Reed
Dr. Elaine Humphrey Biosciences Electron Microscopy Facility 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
Dear Guido, I have learned long ago not to use films cast on slides. It is easier and much cleaner to spread your films on the surface of water in a trough. You can vary the thickness by the amount of drops you use and chose the area you want to place your grids. This method gave me beautiful and sturdy films. Regards, Maria
Maria Fazio-Zanakis Bioimaging and Molecular Histology Aventis Pharmaceuticals 1-908-231-3357 Fax: 1-908-231-3962 Email: Maria.Fazio-Zanakis-at-Aventis.com
-----Original Message----- } From: GUIDO LND [mailto:lueoend-at-zzmk.unizh.ch] Sent: Friday, February 11, 2000 4:31 AM To: EM-Tips
Dear readers, could anyone give me some hints or tips how I can separate a formvarfilm (0.5% in chloroform) from the glassslide ? Breathing or freezing doesn't help at all. It works better with non-washed slides, but then I get an uneven and dirty surface of the film. I appreciate every kind of support. Best regards
Guido Luond Institute of Oral Microbiology and General Immunology Univ. of Zurich Plattenstr. 11 CH-8028 Zurich, Switzerland E-mail: lueoend-at-zzmk.unizh.ch
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Reply to: RE: TEM Formvar Film Cast on Glass Hi Guido,
Here is my recipe which has worked on four continents at altitudes up to 2,000m and in all weathers (humid to dry, cold to hot). Plus, it does not use those very unscientific body fluids I often see being used on this list!
Dissolve the formvar in chloroform as a concentration higher than will make the correct film thickness (film thickness will vary with the speed the glass is drawn out of the solution as well as with formvar concentration).
Take a glass slide (it is important to use precleaned 25x75 mm slides, VWR Scientific, Cat number 48312-002) and dip it in absolute ethanol. Immediately dry it with Ross lens tissue. If you wash it any other way or dry it with anything other than the Ross lens tissue, the film will not come off the glass.
Coat the slide immediately and let the film dry. Score the edges of the film so that it will easily detach from the glass and float it onto the water surface. Evaluate the film thickness and adjust the formvar concentration by adding chloroform to the solution.
I know there are many versions of this method that work. However, this is the only method I have found that is completely oblivious to the weather (still sunny in LA) and has been reproduced by many people. in my experience, the formvar dissolved in choroform does not seem to have the short shelf life I read about recently. I have used the same solution for years, topping it up with chloroform to keep it at a concentration that will produce thin films.
Regards,
Paul Webster
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm
GUIDO LND 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 readers, } could anyone give me some hints or tips how I can separate a formvarfilm } (0.5% in chloroform) from the glassslide ? Breathing or freezing doesn't } help at all. It works better with non-washed slides, but then I get an } uneven and dirty surface of the film. } I appreciate every kind of support. } Best regards } } Guido Luond } Institute of Oral Microbiology and } General Immunology } Univ. of Zurich } Plattenstr. 11 } CH-8028 Zurich, Switzerland } E-mail: lueoend-at-zzmk.unizh.ch } } } } } RFC822 header } ----------------------------------- } } Received: from ultra5.microscopy.com [206.69.208.10] by mailhouse.hei.org } with ESMTP } (SMTPD32-5.05) id A182210B0080; Fri, 11 Feb 2000 10:14:26 -0800 } Received: (from daemon-at-localhost) } by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id DAA27990 } for dist-Microscopy; Fri, 11 Feb 2000 03:36:30 -0600 (CST) } Received: from no_more_spam.com (sparc5.microscopy.com [206.69.208.10]) } by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id DAA27987 } for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Fri, 11 Feb 2000 } 03:35:32 -0600 (CST) } Received: from alpha.zzmk.unizh.ch (zzmkmail.unizh.ch [130.60.67.4]) } by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id DAA27980 } for {Microscopy-at-sparc5.microscopy.com} ; Fri, 11 Feb 2000 03:35:21 -0600 (CST) } Received: from [192.168.7.192] by alpha.zzmk.unizh.ch } (8.8.8/1.1.22.3/03Jan00-0358PM) } id KAA0000026325; Fri, 11 Feb 2000 10:30:33 +0100 (MET) } User-Agent: Microsoft Outlook Express Macintosh Edition - 5.0 (1513) } Date: Fri, 11 Feb 2000 10:31:14 +0100 } Subject: TEM Formvar Film Cast on Glass } From: GUIDO L=?ISO-8859-1?B?/PY=?=ND {lueoend-at-zzmk.unizh.ch} } To: EM-Tips {Microscopy-at-sparc5.microscopy.com} } Message-ID: {B4C99572.725%lueoend-at-zzmk.unizh.ch} } Mime-version: 1.0 } Content-type: text/plain; charset="US-ASCII" } Content-transfer-encoding: 7bit } Errors-to: Microscopy-request-at-sparc5.Microscopy.Com } X-RCPT-TO: {pwebster-at-hei.org} } X-UIDL: 242867993 } Status: U } Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm
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hi all-
just got a request to view surface structure of the inside of a 12" metal pipe. its too big for my chamber and they'd prefer a non-destructive technique. will replication of the surface with acetate/acetone and SEM work. i've never tried this before so any pointers would be welcome...
thx! brian ---------------------------------------------- Brian McIntyre Electron Microscopy Lab- River Campus Univ of Rochester Rochester, NY 14627 716-275-3058/4875
Laurie, Frieda Carson's book about histologic technique is the "gold standard for histotechs right now. It is available at Amazon.com and Barnes and Noble's website, and can be special ordered at the major bookstores, as it is currently in print. (Unfortunately I am at home and do not have the actual title in front of me) Wanda Shotsberger ----- Original Message ----- } From: Laurie Wallin {lwallin-at-ucsd.edu} To: {microscopy-at-sparc5.Microscopy.Com} Sent: Thursday, February 10, 2000 11:03 AM
I was asked a couple of questions about fluorescent microscopy and I didn't have a clue. Are all the dyes used with UV or are there other wavelengths that can fluoresce them? Are the dyes used dangerous/hazardous? Are they transparent to visible? Are they expensive? Can you get them in quantity?
-Clueless in Pittsburgh
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
I knew a very good, reputable company in the Southern California region that serviced ETEC SEMs. The name was Scan Service in California, Earl Weltmer 714 area code.
- Jeff
-----Original Message----- } From: LAB. DE MICROSCOPIA ELECTRONICA - FI - UNER [mailto:microsc-at-fi.uner.edu.ar] Sent: Wednesday, February 09, 2000 5:10 AM To: microscopy-at-sparc5.microscopy.com
Hello ..... we have an old ETEC Omniscan SEM, and we are searching for ETEC company or any people that can supply manuals ....
any help is welcome
best regards
=================================================== Fernando D. Balducci Laboratorio de Microscopia Electrnica Facultad de Ingeniera - Bioingenieria Universidad Nacional de Entre Rios Argentina e-mail: microsc-at-fi.uner.edu.ar tel: 54 43 975100 fax: 54 43 975077
In regards to the diamond knives, both companies make an excellent product and both provide good service. Because of price, the last 6 knives I've bought have been from Microstar (two well-used but dull Diatome knives have been traded during these transactions) . All six knives have been more than suitable for the intended purpose. I have no commercial interest in either company.
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A colleague would like your responses to these questions. I believe that I say something on the first question about a year ago and apologize for asking again.
} Here's what I/we need to know. } } 1) Are Microstar diamond knives equal to Diatome. We have an offer that } will save us about $1200 on three knives ( one 3mm ultra with 45 degree } angle, one 3mm ultra with 35 degree angle, one 10 mm histo knife). } } } 2) Is the power controller feature on the Pelco Microwave Tissue processor } (model #3451) worth it? (costs about $1300 additional compared to the } Pelco 3450 without the power controller). I plan to do immunolocalization } work, but am not sure this power controller feature is necessary or worth } the cost. } } Casey } } } Thanks for your time } Marty Reed Equipment Technician Biology Department Humboldt State University Arcata CA 95521 707-826-3234 707-826-3201 FAX mmr7001-at-axe.humboldt.edu
by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id QAA00380 for dist-Microscopy; Fri, 11 Feb 2000 16:46:21 -0600 (CST) Received: from no_more_spam.com (sparc5.microscopy.com [206.69.208.10]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id QAA00372 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Fri, 11 Feb 2000 16:45:23 -0600 (CST) Received: from renko.ucs.ed.ac.uk (renko.ucs.ed.ac.uk [129.215.13.3]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id QAA00361 for {microscopy-at-sparc5.microscopy.com} ; Fri, 11 Feb 2000 16:45:04 -0600 (CST) Received: from inveresk (dialup-76.publab.ed.ac.uk [129.215.38.76]) by renko.ucs.ed.ac.uk (8.8.7/8.8.7) with SMTP id WAA11995; Fri, 11 Feb 2000 22:40:11 GMT Message-Id: {200002112240.WAA11995-at-renko.ucs.ed.ac.uk}
Date sent: Fri, 11 Feb 2000 14:01:04 -0500 To: Microscopy-at-sparc5.Microscopy.Com } From: Brian McIntyre {mcintyre-at-optics.rochester.edu}
Depends on the detail needed, but acetate replication should work for almost all situations. Of course, everything you see will be inside out... Often, selecting inverse video helps one perceive thing properly. If the pipe is dirty or corroded, you will likely pick up some deposit with the replica. This can be distracting, but can also be helpful if x-ray analysis is required... Woody White McDermott Technology My place: http://www.geocities.com/capecanaveral/3722
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hi all-
just got a request to view surface structure of the inside of a 12" metal pipe. its too big for my chamber and they'd prefer a non-destructive technique. will replication of the surface with acetate/acetone and SEM work. i've never tried this before so any pointers would be welcome...
thx! brian ---------------------------------------------- Brian McIntyre Electron Microscopy Lab- River Campus Univ of Rochester Rochester, NY 14627 716-275-3058/4875
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If any one has a PGT IMIX X-ray Analysis/Imaging system 4-40 (SUN Sparc) that is in storage, or not in use, I would be interested in acquiring its Image Signal Processor ISP module (Part# MO328D-00).
Stop destroying your pipe. With our large chamber SEM we can analyse your pipe up to a length of 35 inches and a diameter of 28 inches. Maximum weight of the sample should not exceed 500 pounds.
Because of the positionable electron gun it is easy to observe the inside surface.
Best regards Martin Klein
Disclaimer: VisiTec is the manufacturer of large chamber SEMs. And we like large samples.
----- Original Message ----- } From: Brian McIntyre {mcintyre-at-optics.rochester.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Friday, February 11, 2000 8:01 PM
I am in desperate need of a control unit for a LKB ultratome III, since mine is beyond repair. I would appreciate if anyone coud sell me an unused unit from old apparatus, even if it does not work, I might be able to repair mine with the pieces. Please reply to the e-mail below
Thanks
Dr. A.P. Alves de Matos Dental Medical School Lisbon University apmatos-at-ip.pt
ELECTRON MICROSCOPY MATERIALS SPECIALIST UNL Center for Materials Research and Analysis
Analyze/characterize materials using electron microscopy, materials preparation and computer instrumentation. Supervise/train students, faculty and visiting researchers utilizing these methods. Bachelor's with major in physical science, engineering or related field plus three years experience in the operation, repair or design of electron microscopes or other scientific instrumentation. Master's preferred. Must have excellent computer and interpersonal/communication skills. Knowledge of x-ray diffraction, TEM/SEM principles and sample preparation experience preferred. Excellent benefits. Submit cover letter, resume and names, addresses and telephone numbers of three professional references to Professor David J. Sellmyer, 112 Brace Lab, Lincoln, NE 68588-0113. Review of resumes will begin March 1. Position will remain open until a suitable candidate is found. UNL is committed to AA/EEO and ADA/504. If you require accommodation, please call (402) 472-8762.
Try a different brand of slide. Corning is the only brand we use in our EM Class. There must be different kinds of surfactants used on them. Some other brands of slides just don't release the formvar. We usually wipe them off thoroughly with a Kimwipe only. Dip the slide into the formvar and let dry. But don't dry for too long. Try using a diamond tipped (or carbide) pen to score a "rectangle" on the slide. Slowly insert the slide at about a 30 degree angle into the water. Wait until you see the formvar start to release before you continue inserting into the water. This method usually works in our lab and for students.
Dear Linda, I have a DuPont Sorvall MT2B or a Porter Blum MT1 I would be interested in selling if the price is right. If you are interested, let me know via my email address: tiekotte-at-up.edu. Cheers! Ken ----------- Ken Tiekotter Dept. of Biology The University of Portland 5000 N. Willamette Blvd. Portland, OR 97303
On Mon, 27 Aug 1956, Linda Chicoine wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Does anyone in the Connecticut area have an old microtome they would } like to sell? I would be interested in anything from an LKB to RMC to } Reichert/ and Leica. Please contact me by email. Thanks. Linda } Chicoine } } }
} To: MICROSCOPY BB {Microscopy-at-sparc5.microscopy.com} } Subject: SEM pipe replication } Date: Sat, 12 Feb 2000 10:44:32 -0500 } From: "Garber, Charles A." {cgarber-at-2spi.com} } X-Mailer: E-Mail Connection v3.1a } Content-Length: 3106
} } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } Brian McIntyre wrote: } ========================================= } just got a request to view surface structure of the inside of a 12" metal } pipe. its too big for my chamber and they'd prefer a non-destructive } technique. will replication of the surface with acetate/acetone and SEM } work. i've never tried this before so any pointers would be welcome... } ========================================== } You did not indicate how far into the pipe you want to do your looking, but } cellulose acetate is certainly one approach. The biggest problem you are } going to have is to balance the need to do this all "nondestructively" vs. } the need to see what might be underneath material covering the top surface
} (after all, it is the metal substrate that you really want to see). In } other words, if the acetone should be a solvent for the contaminating } material, (or if it lifts off metal oxide or other aspects of a scale) you } will not be doing this very non-destructively. One technique is to make } repetitive replicas on the same area, each additional one taking off more of } the contaminating material, in order to get to the bare metal surface. But } this might not be considered, by a court of law, to be nondestructive. And } if you are under an order to look at it nondestructively, this could result } in a problem with the court. } } When we have been faced with this dilemma, in order to first document what } was there originally, we like to use our SPI "Wet Replica" kit, it is a } silicone based system, and is described on our website. Certain dental } replication systems such as "Sil-Flow" might work as well, but in general, } for this kind of work, we believe our own kit is better. The silicone resin } in general won't remove surface contamination or scale. The drawback of this } technique is that after about 800X in the SEM you start to see "structure" } from the replication system itself. A "plus" however is that you can make a } "replica of the replica", doing your SEM viewing on a positive, which does } look just like the original inside surface. } } Once you have documented the state of the inside surface that way, you } should be able to get permission to try the cellulose acetate replication } method. There is additional information on the SPI Supplies website relative } to cellulose acetate replication methods. } } Disclaimer: SPI Supplies offers both cellulose acetate replicating tapes } and sheets as well as the silicone based SPI "Wet Replica Kit" for use in } electron microscopy applications. } } Chuck } } ============================================ } } Charles A. Garber, Ph. D. Ph: 1-610-436-5400 } President 1-800-2424-SPI } SPI SUPPLIES FAX: 1-610-436-5755 } PO BOX 656 e-mail:cgarber-at-2spi.com } West Chester, PA 19381-0656 USA } Cust.Service: spi2spi-at-2spi.com } } Look for us! } ######################## } WWW: http://www.spi.cc } ######################## } ============================================ } } } }
Hi, I made an embedding of the endodermis of an iris root with LR-Wihte and would like to make ultrathin sections and stain them. My problem is how to stain the sections and how to receive permantly stained sections. Is there anyone who has experiences with the staining of suberin in the 3rd endodermis? Please inform me about the handling. Thank you so much Dagmar Preusse
I run a 1830 Amray S.E.M. and work with failed aircraft components. From time to time I am asked to replicate a surface using the acetate tape & acetone method. The coating I use is a combination of Au and Pd. My problem is that when I have to stay in one particular area of the replica, the beam leaves a nice little square embedded into the surface of the replica. I try to rapidly focus in full field instead of partial field, and use between 6 - 10Kv. Does anyone have any advise for me on how to avoid the embedded square?
It is the total current that you put into the specimen that is causing the material to erode.
If you use a smaller spot size, smaller final aperture or smaller emission current you should be better off? I do not know how good your SEM is at low kV but if it was me I would use 2kV on this type of specimen; it will be very sensitive even though it is coated.
To run happily at 2kV you need a filament to cathode distance that will give you at least 30uA with your normal bias settings. If you do a good deal of work on this type of specimen I would be inclined to lift the anode by 5mm also. This will make the gun more efficient and make the job easier.
Good luck
Steve Chapman Protrain for Electron Microscopy Courses World Wide E-mail - protrain-at-emcourses.com Web Site - www.emcourses.com
I have been trying to develop a styrene replica fluid so I can use polorized light.So far the styrene clumps ups in little islands insted of making a nice film. I am using acetone as a solvent
Has anyone had any luck with this?
Gordon
Gordon Couger gcouger-at-couger.com
Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
} A colleague would like your responses to these questions. I believe that I
} say something on the first question about a year ago and apologize for
} asking again.
}
}
} } Here's what I/we need to know.
} }
} } 1) Are Microstar diamond knives equal to Diatome. We have an offer that
} } will save us about $1200 on three knives ( one 3mm ultra with 45 degree
} } angle, one 3mm ultra with 35 degree angle, one 10 mm histo knife).
} }
} }
} } 2) Is the power controller feature on the Pelco Microwave Tissue processor
} } (model #3451) worth it? (costs about $1300 additional compared to the
} } Pelco 3450 without the power controller). I plan to do immunolocalization
} } work, but am not sure this power controller feature is necessary or worth
} } the cost.
} }
} } Casey
} }
} }
} }
} Thanks for your time
} }
} Marty Reed
} Equipment Technician
} Biology Department
} Humboldt State University
} Arcata CA 95521
} 707-826-3234
} 707-826-3201 FAX
} mmr7001-at-axe.humboldt.edu
}
}
{/fontfamily} Cheers, Mark **************************************************** Mark A. Sanders University of Minnesota Program Director Twin Cities Campus Imaging Center St. Paul, MN 55108 23-35 Snyder Hall ph: 612-624-3454 1475 Gortner Ave. fax: 612-624-1799 http://biosci.umn.edu/imagingcenter President: Minnesota Microscopy Society http://resolution.umn.edu/MMS/
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------- Start of forwarded message -------
Rules, FAQ Docs - Microscopy ListServer To: NewSub-at-sparc5.Microscopy.Com } From: "Nestor J. Zaluzec" {zaluzec-at-sparc5.Microscopy.Com}
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To: NewSub-at-MSA.Microscopy.Com } From: "Nestor J. Zaluzec" {zaluzec-at-microscopy.com}
Here is a final reminder and IMPORTANT CORRECTION for the Call for Papers for Microscopy and Microanalysis 2000.
The title for symposium #05 in the Call for Papers was listed incorrectly. The correct title should be "Phase Transitions" (If you check the symposium titles on the electronic data submission forms it is listed correctly). X-ray microanalysis papers should be directed to symposia 25 and 26.
The deadline for papers is Tuesday February 15, which is almost upon us.
Stuart McKernan (Program Chair)
__________________ Stuart McKernan stuartm-at-tc.umn.edu Director Office:(612) 626-7594 IT Characterization Facility, University of Minnesota Desk:(612) 624-6009 100 Union Street S. E., Minneapolis, MN 55455 NEW-} Fax:(612) 625-5368
} X-Sender: mcintyre-at-optics.rochester.edu } Date: Fri, 11 Feb 2000 14:01:04 -0500 } To: Microscopy-at-sparc5.Microscopy.Com } From: Brian McIntyre {mcintyre-at-optics.rochester.edu} } Subject: metal pipe- inside surface } Errors-to: Microscopy-request-at-sparc5.Microscopy.Com } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
The best way to make replicas is to use dental casting compound.
It has the ability to replicate very fine detail and accepts even damp surfaces.
We used "Kerr Extrude Wash" (two part polyvinylsiloxane impression material) to replicate (damp) tuna fish skin.
Once cured, we made epoxy positive casts from the negative casts, then mounted, gold coated etc. for regular SEM.
Your local dental supply house will have this.
Kerr USA is at 28200 Wick Road, Romulus MI 48174-2600
Dr. Mel Dickson, Deputy Director, The Electron Microscope Unit, Adjunct Associate Professor, School of Microbiology & Immunology The University of New South Wales Sydney, Australia. Phone +612 9385 6383 Fax +612 9385 6400
True optical resolution 1600x3200. Dynamic range 3.3 Although it is not from a scientific point of view the authors claim that this is the best scanner for the price they've ever seen.
Good luck,
Rado
Disclaimer: I'm not related to Epson company in any way.
--------------------------------------------------------------------- Radostin Danev Laboratory of Ultrastructure Research National Institute for Physiological Sciences Myodaiji-cho, Okazaki 444-8585, JAPAN e-mail: rado-at-nips.ac.jp --------------------------------------------------------------------- ----- Original Message ----- } From: Malcolm Haswell {malcolm.haswell-at-sunderland.ac.uk} To: Microscopy (MSA) {microscopy-at-sparc5.microscopy.com} Sent: Thursday, February 10, 2000 8:26 PM
On Sat, 12 Feb 2000, Gordon Couger wrote:
} I have been trying to develop a styrene replica } fluid so I can use polarized light.So far the } styrene clumps up in little islands instead of } making a nice film. I am using acetone as } a solvent
Polystyrene is only partially soluble in acetone, so the low MW goes into solution while the high MW forms clumps or whatever. BUTANONE (MEK) should work fine, but you'll have to give it perhaps 4 times the drying time you'd need with acetone. For samples that are sensitive to polar solvents such as these, you can also use toluene, but that takes an even longer drying time.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
} I run a 1830 Amray S.E.M. and work with failed aircraft components. From } time to time I am asked to replicate a surface using the acetate tape & } acetone method. The coating I use is a combination of Au and Pd. My } problem is that when I have to stay in one particular area of the replica, } the beam leaves a nice little square embedded into the surface of the } replica. I try to rapidly focus in full field instead of partial field, and } use between 6 - 10Kv. Does anyone have any advise for me on how to avoid } the embedded square?
Polystyrene MIGHT be a better material to use than cellulose acetate, which tends to chemically self-destruct under electron beams: in fact, when there is acetate remaining on replicas for TEM I sometimes burn it off SLOWLY with the electron beam (too fast and you leave carbonaceous stuff). The aromatic nature of PS also allows it to take up more energy.
You could replicate with PS, as follows:
Make a solution of PS in butanone (MEK) in a glass Petri dish. Allow to evaporate slowly (if you can fume it off from a closed dish in an oven at about 50^C, that's better). This film can then either be cast onto the component with butanone, or you can melt-form it above the glass transition of PS, say about 150^C. Don't use PS windows directly from envelopes, because the films contain a lot of built-in-strain.
I'm not sure of it's performance in SEM, because I've used it for TEM replication of acetone-sensitive specimens.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
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So your companyÕs SEM operator just retired. You have used light microscopes for years and they have asked you to run the SEM also. So now what do you do? The SEM manuals are long gone and do you really want to crack open that 800 page SEM/x-ray microanalysis textbook?
DonÕt worry, consider taking Electron Microscopy and Microanalysis a one-day short course to be held on Sunday, March 12, at PITTCON 2000 in New Orleans. The course is an introduction to SEM, TEM and x-ray microanalysis and is designed for analytical chemists and others who need to use these techniques for industrial problem-solving and product research and development.
For further information and online registration visit www.pittcon.org (Course #2029) or contact me offline.
Mark S. Germani, Ph.D. MicroMaterials Research, Inc. 136 Shore Drive Suite 200 Burr Ridge, IL 60521
by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id IAA06667 for dist-Microscopy; Mon, 14 Feb 2000 08:52:52 -0600 (CST) Received: from no_more_spam.com (sparc5.microscopy.com [206.69.208.10]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id IAA06661 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Mon, 14 Feb 2000 08:51:54 -0600 (CST) Received: from mercury.uwe.ac.uk (mercury.uwe.ac.uk [164.11.132.23]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id IAA06651 for {microscopy-at-sparc5.Microscopy.Com} ; Mon, 14 Feb 2000 08:51:37 -0600 (CST) Received: from fas655.uwe.ac.uk ([164.11.205.145]) by mercury.uwe.ac.uk (2.0.4/SMS 2.0.4-devel) with SMTP id OAA04217; Mon, 14 Feb 2000 14:46:35 GMT
Electron Optical Services Ltd. make a replacement control box. Their address is given below.
Dave
52 Higher Road, Urmston, Manchester, M41 9AP England UK
Dave On Fri, 11 Feb 2000 19:12:34 -0600 "A.P. Alves de Matos" {apmatos-at-ip.pt} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I am in desperate need of a control unit for a LKB ultratome III, since } mine is beyond repair. } I would appreciate if anyone coud sell me an unused unit from old } apparatus, even if it does not work, I might } be able to repair mine with the pieces. } Please reply to the e-mail below } } Thanks } } Dr. A.P. Alves de Matos } Dental Medical School } Lisbon University } apmatos-at-ip.pt } } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
Dear Microscopy Listserver members, can someone provide me: - a detailed protocol for embedding Chlamydomonas into the Lowicryl Resin K4M (fixation media, desiccation solutions, times and concentrations that are suitable for Chlamydomonas) - a fixation protocol for cryosectioning of Chlamydomonas
Thank you very much, Yvonne Nemcova
-- ***************************************** Mgr. Yvonne Nemcova Department of Botany, Charles University, Prague Benatska 2. 128 01 Praha 2, Czech Republic tel: 00-420-2-21953127, fax: 00-420-2-21953125 e-mail address: ynemcova-at-natur.cuni.cz *****************************************
Thank you for the information on the MSA/MAS format that you sent me some weeks ago.
The MSA/MAS spectra file format is now implemented in the EELS and XAS database, therefore the spectra can be downloaded in this format. Please let me know if you have any other problems. Any more suggestions ?
We are having an Edwards Vacuum Evaporator model E12E2 with a control unit EVM052 in excess. This instrument has not been used for several year and it may need some maintanance work. If you are interested please contact Tea Meulia (meulia.1-at-osu.edu) or Pat Ashbaugh (ashbaugh.11-at-osu.edu).
Tea Meulia
Tea Meulia Research Scientist and Head Molecular and Cellular Imaging Center OSU/OARDC 1680 Madison Ave. Wooster OH 44691
Thanks to all that responded. I have a lot better handle on how to do it. I will report back my results.
Thanks to all for providing a great source of infomation.
Gordon
Gordon Couger gcouger-at-couger.com
Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00 ----- Original Message ----- } From: "Robert H. Olley" {r.h.olley-at-reading.ac.uk} } On Sat, 12 Feb 2000, Gordon Couger wrote: } } } I have been trying to develop a styrene replica } } fluid so I can use polarized light.So far the } } styrene clumps up in little islands instead of } } making a nice film. I am using acetone as } } a solvent } } Polystyrene is only partially soluble in acetone, so the low MW goes into } solution while the high MW forms clumps or whatever. BUTANONE (MEK) } should work fine, but you'll have to give it perhaps 4 times the drying } time you'd need with acetone. For samples that are sensitive to polar } solvents such as these, you can also use toluene, but that takes an even } longer drying time. }
Epson has a new ink jet printer out-Stylus pro 5000-that is expensive but they claim it is much better than the much cheaper models many of us know and love. Would anyone know whether this is true of just hype-hard to tell from the published specs...? Thanks...Tom Reese
Dear Gordon, Try xylene. It works quite nicely as a mounting medium of a differenting refractive index from that of say, canada balsam or Permount. Email me if you have questions. Good luck! Cheers, Ken
------------- Ken Tiekotter Dept. of Biology The Univerisity of Portland 5000 N. Willamette Blvd. Portland, OR 97303
On Sat, 12 Feb 2000, Gordon Couger 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 trying to develop a styrene replica } fluid so I can use polorized light.So far the } styrene clumps ups in little islands insted of } making a nice film. I am using acetone as } a solvent } } Has anyone had any luck with this? } } Gordon } } Gordon Couger gcouger-at-couger.com } } Stillwater, OK www.couger.com/gcouger } 405 624-2855 GMT -6:00 } } } }
You wrote "I am working with archival specimens of formalin-fixed paraffin embedded tissues and I am getting tremendous autofluorescence. I looked at tissue sections after the sections have been de-waxed using either FITC or rhodamine filter sets and red cells and connective tissues are brightly fluorescent. Is there a way of suppressing the autofluorescence and still retain reactivity of tissue to antibodies? I would appreciate any comments or suggestions."
It is an unfortunate fact that most fixatives cause high autofluorescence making the studies with conventional fluorescence microscope hard. The disturbance of autofluorescence can be avoided by using time-resolved fluorescence microscope and special labels (See the following articles: Visl M, Sevus L, Kuusisto A, Harju R, and Soini E: Time- resolved fluorescence microscopy: Elimination of autofluorescence in tissue specimens for Image cytometry with fluorescent labelled probes. Micron and Microscopica Acta 21; 3, (1990).
Sevus L, Kuusisto A, Visl M, Hemmil I., and Soini E: Lanthanide chelates as labels in in situ hybridization and immunohistochemistry, Micron and Microscopica Acta 21; 3, (1990).
Sevus L, Visl M, Syrjnen S, Sandberg M, Kuusisto A, Harju R, Salo J, Hemmil I, Kojola H, and Soini E: Time-Resolved Fluorescence Imaging of Europium Chelate Label in Immunohistochemistry and In Situ Hybridization, Cytometry 13; 329, (1992).
Sevus L, Kuusisto A, Visl M, Hemmil I., Kojola H., Roomans G. M. and Soini E.: Use of Fluorescent Europium Chelates as Labels in Microscopy Allows Glutaraldefyde Fixation and Permanent Mounting and Leads to Reduced Autofluorescence and Good Long-Time Stability, Microscopy Research and Technique 27; 00, (1994).
I saw the Epson web spec and it looks quite impressive - especially as they seem to be quoting speeds for real picture printing, not 5% coverage or text.
My only concern is that Epson now market two printers with "10 years light fastness" (the Epson Stylus photo 870 - A4 and 1270 A3) so you have to wonder which is better for e.m. prints.
Malcolm Haswell Electron Microscopy School of Sciences Fleming Building University of Sunderland SUNDERLAND SR1 3SD Tyne and Wear UK
Tel (0191) 515 2872 e-mail: malcolm.haswell-at-sunderland.ac.uk
Tom Reese 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 } -----------------------------------------------------------------------. } } Epson has a new ink jet printer out-Stylus pro 5000-that is expensive } but they claim it is much better than the much cheaper models many of } us know and love. Would anyone know whether this is true of just } hype-hard to tell from the published specs...? Thanks...Tom Reese
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Dear Carol,
We were looking up to 70 mm deep inside cylinder bores and engine blocks several times during the last months with our large-chamber SEM. Roughness was typically very small (less than 10 µm), but for small diameter tubes the positioning system needs large travels, tilts, and rotary axes to inspect every interesting area.
Disclaimer: Visitec manufactures the large-chamber SEM.
Best regards Peter ___________________________ Dr. Peter Marienhoff VisiTec Microtechnik GmbH Karl-Marx-Str. 14 D-23936 Grevesmuehlen Germany
"Orth99-at-aol.com"-at-sparc5.Microscopy.Com wrote: } How often does this community need to look at the inside of metal pipes or } pipes in general? } } And, what is the typical surface roughness you are dealing with, i.e. would } an instrument with 100 um to 200 um of Z range be able to handle it? } } And, what is the typical x,y range, i.e. would an instrument with 2 mm image } field be acceptable? } } Just wondering. Thanks for any input you are willing to share. } } } Carol Rabke, Ph.D. } Surface Metrology Marketing & Applications Consultant } 716-425-0976 }
The Epson 5000 is a printer/proofer using 6 color inkjet technology. The 6 color technology expands the available color gamut significantly. It is available both with or without a Fiery RIP. The Fiery RIP is used for Postscript processing of complex files from Quark, Pagemaker, Illustrator,etc. Without the Fiery box, an Epson software RIP is included. Print quality is excellent although without the Fiery RIP, printing times can be long,especially for 11x17 or larger prints. For more information see: http://prographics.epson.com/products/stypro5000/index.html
Canon also has an excellent printer, the BJC-8500 that also uses 6 color technology. The BJC-8500 also handles up to 13x19 paper with excellent print quality. Canon utilizes a technology that optimizes ink drying and water resistance. For more information see: http://www.bjc8500.com/index2.html
Neither of these printers are speed demons in terms of page throughput, but quality is excellent. I will be happy to answer any questions anyone may have regarding output devices.
George Laing National Graphic Supply
-----Original Message----- } From: Tom Reese [mailto:treese-at-mbl.edu] Sent: Monday, February 14, 2000 6:57 PM To: Microscopy-at-sparc5.microscopy.com
Epson has a new ink jet printer out-Stylus pro 5000-that is expensive but they claim it is much better than the much cheaper models many of us know and love. Would anyone know whether this is true of just hype-hard to tell from the published specs...? Thanks...Tom Reese
} -----Original Message----- } From: Wright, Cheryl W [mailto:CWright-at-Sikorsky.com] } Sent: Saturday, February 12, 2000 7:25 AM } To: 'Microscopy-at-MSA.Microscopy.Com' } Subject: Photography of Coated Replicas } } } -------------------------------------------------------------- } ---------- } 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/MicroscopyListser} ver/FAQ.html } } } -------------------------------------------------------------- } ---------. } } } I run a 1830 Amray S.E.M. and work with failed aircraft } components. From } time to time I am asked to replicate a surface using the } acetate tape & } acetone method. The coating I use is a combination of Au and Pd. My } problem is that when I have to stay in one particular area of } the replica, } the beam leaves a nice little square embedded into the surface of the } replica. I try to rapidly focus in full field instead of } partial field, and } use between 6 - 10Kv. Does anyone have any advise for me on } how to avoid } the embedded square? } } Thanks, Cheryl }
I am a technical recruiter. Currently, I am working on a search for one of my Northeast clients for a Senior Metallographic Technician:
Will perform metallographic evaluation of coatings including work with SEM (Scanning Electron Microscopy) and X-ray. Will be responsible for the metallurgy laboratory, including organization of work, and operation and calibration of equipment. Will write technical and engineering reports. Will prepare projects for the metallurgy laboratory. Will maintain communication with customers.
AAS/BS plus fours years experience in a metallography laboratory.
Resumes should be sent by email, mail or fax. For best results, please send emails as attached Word.docs - I have Word 97.
Pearl Martin Image Associates Inc. 5254 Merrick Road Massapequa, NY 11758 Phone (516)798-3993 Fax (516)797-8703 Email: pearl-at-jobspot.com
With regard to diamond knives, I think it depends on what you're cutting and what quality of section you require.
We cut steel and cross sections of coatings (some very hard and brittle) on steel, and the only knife that approaches satisfactory result is Diatome 35 degree, sharpened to extra fine edge. We traded our big old Microstar knife for a 2nd Diatome, because it couldn't make good, electron transparent sections.
On the other hand, if your material cuts as well with a glass knife as with a diamond but you don't want to bother making your own glass knives, then go ahead and save some dollars.
(In my view, the same reasoning applies to the brand of ultramicrotome to use. My company is very frugal, and there is always strong pressure to save a dollar here and there, but we found that only Reichert/Leica can consistently produce good samples in our demanding application; and the higher initial cost has been more than offset by savings in time and effort, and by the quality of results. (I.e., often the ability to just get any kind of a result).)
Valdemar Furdanowicz, Ph.D. Research Labs - G165 Bethlehem Steel Bethlehem PA 18016
No interest in Microstar, Diatome, Leica; and this not reflect an opinion or position of my employer....etc.
-----Original Message----- From: Chuck Butterick [mailto:cbutte-at-ameripol.com] Sent: Friday, February 11, 2000 5:11 PM To: Microscopy-at-sparc5.Microscopy.Com; mmr7001-at-humboldt.edu Subject: Re: questions for MSA members
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In regards to the diamond knives, both companies make an excellent product and both provide good service. Because of price, the last 6 knives I've bought have been from Microstar (two well-used but dull Diatome knives have been traded during these transactions) . All six knives have been more than suitable for the intended purpose. I have no commercial interest in either company.
Chuck Butterick Engineered Carbons, Inc.
______________________________ Reply Separator _________________________________ Subject: questions for MSA members Author: Marty Reed {mmr7001-at-humboldt.edu} at INTERNET-MAIL Date: 1/4/80 8:23 AM
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A colleague would like your responses to these questions. I believe that I say something on the first question about a year ago and apologize for asking again.
} Here's what I/we need to know. } } 1) Are Microstar diamond knives equal to Diatome. We have an offer that } will save us about $1200 on three knives ( one 3mm ultra with 45 degree } angle, one 3mm ultra with 35 degree angle, one 10 mm histo knife). } } } 2) Is the power controller feature on the Pelco Microwave Tissue processor } (model #3451) worth it? (costs about $1300 additional compared to the } Pelco 3450 without the power controller). I plan to do immunolocalization } work, but am not sure this power controller feature is necessary or worth } the cost. } } Casey } } } Thanks for your time } Marty Reed Equipment Technician Biology Department Humboldt State University Arcata CA 95521 707-826-3234 707-826-3201 FAX mmr7001-at-axe.humboldt.edu
We are considering the purchase of a separate deconvolution system to augment our imaging lab's confocal system Any users or vendors with information about such systems, please contact me offline.
thanks in advance
steve
Dr. Steven Barlow, Associate Director EM Facility/Biology Department San Diego State University 5500 Campanile Drive San Diego CA 92182-4614 phone: (619)594-4523 fax: (619) 594-5676 email: sbarlow-at-sunstroke.sdsu.edu website: http://www.sci.sdsu.edu/emfacility/
Chairman, Educational Outreach subcommittee promoting access to microscopes Microscopy Society of America http://www.msa.microscopy.com/
We are in the market for a new oven for curing blocks.... Any suggestions? This lab uses as a standard Eponate 12 as the epoxy resin of choice.... The oven we currently have after being fixed several times does not see to keep a constant temp.....
One of the members of this listserver group just asked me privately about the fact that he had noticed that the reading of the thermocouple gauge on his vacuum evaporator changed markedly when he jiggled the wires running from the gauge tube to the gauge control unit. This is a matter that is discussed in some detail on page 87 of my book 'Vacuum Methods in Electron Microscopy' (see http://www.2spi.com/catalog/books/book48.html for a description)
Actually, this is a rather common problem with thermocouple gauges which all people using them should be aware of. It arises because the sensing element in a thermocouple gauge is an arrangement of very fine wires that form one or more thermocouple junctions. The output of these gauges is thus the electrical potential produced by these junctions that is of the order of only 10 or 15 millivolts DC. As a consequence, the signal detected by the gauge controller can be strongly influenced by minor changes the resistance of the electrical lines connecting the gauge to the controller. Typically, unplugging one of the connectors in this line and reconnecting it can change the resistance of the line sufficiently to cause a significant change in the reading produced by the controller.
For this reason, considerable care must be exercised in installing and maintaining thermocouple gauges if acceptable readings are to be obtained. As a minimum, the same cables used in calibrating a gauge must subsequently be used when it is put into operation, and the pins and sockets of the plugs involved in the circuit must be kept clean and firmly connected.
Because of the low DC voltages that must be detected, the meters used in the gauge circuit must be rather delicate and sensitive. We have encountered situations when a static electric charge on the plastic window of a meter affected the movement of the needle inside the meter enough to prevent an accurate response. Coating the meter window with an anti-static solution cured this situation.
As discussed in my book, Thermocouple gauges are rugged, inexpensive, and very convenient for use in many applications, but they must be handled with care if they are to give acceptable service.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
A graduate student is getting ready for a rather large scale preparation of rat brains for TEM using vascular perfusion. Luckily (since I'm a botanist) her major professor has had experience with the procedure. He doesn't know for sure whether he used EM or Biological grade glutaraldehyde previously, but considering the large volume that will be required, and the cost difference, I'm hoping that the Biological grade will suffice.
I would appreciate advice from those of you that have experience with this method of fixation.
Thank you in advance for any information.
Heather Owen
Heather A. Owen, Director Electron Microscope Laboratory Department of Biological Sciences University of Wisconsin - Milwaukee (414)229-6816
I have attempted to purchase an epifluorescence nose from 'Carl Zeiss' to fit the old "Universal" model scopes, but was dismayed to find out that it is no longer available. If anyone has one that is not being used and wishes to sell it, I would be very interested.
Thanks
JOSE ULLAN SERRANO, Ph.D., M.D. ÑÑÑÑÑÑÑÑÑÑÑÑÑÑÑÑ E-mail: jullan-at-mixcoac.upmx.mx
Departamento de ANATOMIA Escuela de MEDICINA Universidad Panamericana c/ Donatello, 59 Col. Insurgentes-Mixcoac 03920 MEXICO, D.F. ÑÑÑÑÑÑÑÑÑÑÑÑÑÑÑ http://www.mixcoac.upmx.mx Telef. 55 98 33 02 / 55 63 13 43 FAX: 56 11 35 85
Thermocouple gauges are more like gross indicators of vacuum. As a resistive bridge, they are driven by constant current sources and the bridge or sensor output is fed to a differential amplifier. This amplifier has a gain and zero adjust potentiometer. If either of these are dirty or defective, readings will be off. But if a thermocouple sensor is exhibiting erratic readings, and it uses the standard vacuum tube socket connector, I'd replace the detector right off the bat. They do eventually fail. A $25 throw away isn't worth the hassle.
Most instruments, like SEMs, use the TC gauge to indicate that the mechanical pump is working. Once the turbo pump spins up to } 80% RPM, the TC gauge is rather useless. Better readings are from cold cathode gauges....which tend not to fire very regularly. Hence, the optimum readings are from the ion pump current.
At 01:11 PM 2/15/00 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We perfuse 4% formaldehyde in 1x PBS only, than prepared the brain's slices .3-.4 mm thick and fix them in a mixture 3-4% formaldehyde 2% GA (all - EM grade) in 1x PBS. If you would like to add GA into perfusion solution - it is only 0.1% - not a big deal. From another hand, I don't think Biological grade GA will affects fixation quality. But, who knows, every time it is a "game": if fixation is good - no problem, if - no, we will start to think what's going on and "biological" grade may be a "case"... To avoid such troubles I always used fresh GA and formaldehyde from non-opened ampoules.
Good luck, Sergey
} Date: Tue, 15 Feb 2000 18:02:57 -0600 (CST) } From: Heather A Owen {owenha-at-csd.uwm.edu} } Subject: TEM - glutaraldehyde for perfusion } To: MSA Listserver {Microscopy-at-sparc5.Microscopy.Com} } } ------------------------------------------------------------------------ } 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
As for my knowledge (mostly from physic's course at 7th grade Russian school) the thermocouple gauge has two "thermocouples": the probe itself and reference thermocouple inside of gauge (separate "references" for each type of probe, I believe). All together they generate enough response to be easily measured. I agree that the major problem in your case is a bad electrical contact in the connectors or even broken wire (it is well possible, because some material uses for thermocouples are very fragile). In my point of view, thermocouple gauges is very reliably and most accurate devices and they do not need any special care.
I am using BARNANT-100 Thermocouple Gauge and REX-P96 Thermocouple Controller with type "T" thermocouple sensors (copper/constantan, I believe, but not sure) for many years without any problem. The sensor goes into the vacuum chamber through regular 8-pin feed-thought. I was using same as thermocouple material for screws to connect thermocouple's wires to the feed-though wiring. For some reason, soldering does not work. I took that screws from disassembled thermocouple connectors (looks like small plugs) most suppliers sells for thermocouple probes (a dollar each). That connectors should correspond to the thermocouple probe you are using. Usually those connectors are color-coded. For instance connectors for my type "T" probe was coded in blue. BARNANT-100 Thermocouple Gauge and REX-P96 Controller performs self-calibration test every time you shut it ON or disconnect/connect probe. During that calibration they "compensate" some electrical abnormalities in the circuit like difference in the probe length or as in my case the current deviation on the enter and end of the feed-through. In theory, the "break" in thermocouple wiring on feed-through should affect the gauge accuracy. In practice, I did not find a difference between "solid" and with-feed-though probe up to 0.1oC accuracy (which is sufficient for my needs). REX-P96 Thermocouple Controller is just amazing. You may program complicated profiles for cooling/heating cycles and it holds temperature pretty well. It has special auto-calibration function to adjust heat rate depending from parameters of your system. In practice it mean that your sample will not overheated because of heater inertia. I am using that Controller in my high-resolution shadowing device to control temperature from -150 to +50oC.
I have no interest in described products (only happy user) mostly because I even don't know the names of real manufacturers. I remember it was distributed by Cole-Parmer.
Sergey
} Date: Tue, 15 Feb 2000 15:11:33 -0400 } From: Wil Bigelow {bigelow-at-engin.umich.edu} } Subject: Thermocouple gauges } X-Sender: bigelow-at-srvr5.engin.umich.edu } To: Microscopy Listserver {microscopy-at-sparc5.Microscopy.Com} } } ------------------------------------------------------------------------ } 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
MEK worked great. I broke up the clear plastic from a CDROM case and made a thick solution by adding MEK. cleaned a slide and spread a drop of the solution on the slide and put a celery leaf on it I covered the whole thing with saran warp and gently pressed another slide on it to get good contact. Then removed the slide and Saran wrap.
After drying about 30 minutes I pealed the leaf off and took a look under a Nomarski phase scope. It was the best image I have seen on the scope. The Nomarski colors were extremely strong and I could get three bands of color in some features/artifacts. The effect of the styrene on the polarized light were not as strong as I had hoped for but they do increase the contrast and Nomarski colors. It will make lovely photos. Now if my color CCD camera would just get here. Monochrome just won't do it justice.
I has been a long time since I looked at a leaf but the stoma showed up just like the books show and I remember. Individual cells were very clearly visible.
The film is extremely fragile and probably needs a little plastizer added and probably go to a go to a multi coat system if the film needs to be pealed from a surface and mounted. with the first coats being thin with little or no plastizier and the following coats being thicker and carrying more plastisizer.
My thanks to all the helped on this. I will probably try other solvents but MEK does not set off my asthma like xylene and toluene do. So I will probably put up with the slower drying times.
I was very impressed with the detail that was replicated.
Thanks Gordon
Gordon Couger gcouger-at-couger.com
Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
} From: Robert H. Olley [mailto:r.h.olley-at-reading.ac.uk] } On Sat, 12 Feb 2000, Gordon Couger wrote: } } } I have been trying to develop a styrene replica } } fluid so I can use polarized light.So far the } } styrene clumps up in little islands instead of } } making a nice film. I am using acetone as } } a solvent } } Polystyrene is only partially soluble in acetone, so the low MW goes into } solution while the high MW forms clumps or whatever. BUTANONE (MEK) } should work fine, but you'll have to give it perhaps 4 times the drying } time you'd need with acetone. For samples that are sensitive to polar } solvents such as these, you can also use toluene, but that takes an even } longer drying time.
Dear Josi, Try contacting John Oren at Vermont Optechs. I cannot tell you immediately what his email or web site address is but his mail address is: P.O. Box 69, Charlotte, VT 05445: Tel. (802) 425-2040; FAX. (802) 425-2074. Good luck! -Ken
On Tue, 15 Feb 2000, Jos[ISO-8859-1] é Ullán Serrano 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 Colleagues: } } I have attempted to purchase an epifluorescence nose from 'Carl Zeiss' } to fit the old "Universal" model scopes, but was dismayed to find out that } it is no longer available. } If anyone has one that is not being used and wishes to sell it, I would be } very interested. } } Thanks } } JOSE ULLAN SERRANO, Ph.D., M.D. } } E-mail: jullan-at-mixcoac.upmx.mx } } Departamento de ANATOMIA } Escuela de MEDICINA } Universidad Panamericana } c/ Donatello, 59 } Col. Insurgentes-Mixcoac } 03920 MEXICO, D.F. } } http://www.mixcoac.upmx.mx } Telef. 55 98 33 02 / 55 63 13 43 } FAX: 56 11 35 85 } }
I have heard recently that Diatome invent some coating of their diamond knives that significantly improve the quality of cryosections (they appear more smooth, less compressed, practically without folds). Is it true or just usual story to increase sales?
Sincerely, Dr. Alexander A. Mironov Jr., MD, PhD Unit of Morphology Dept. of Cell Biology and Oncology Consorzio Mario Negri Sud Via Nazionale, S.Maria Imbaro (Ch) 66030 Italy
Speaking of thermocouples, I need to replace (probably) the T1 and T2 thermocouples in our OLD Denton evaporator. Where, other than Denton, can I look for these? I also need service work done on our old Ultracut (the model before the "E" series). I refuse to work with Leica. Is there anyone in the "real" midwest area?
I've also seen this type of behavior caused by a "cold" solder joint on one or more of the wires in the guage connector socket. Re-flowing the joints did wonders for stability.
============================================================ Bede Willenbring Phone: (651)236-5470 H.B. Fuller Company FAX: (651)236-5430 Research Chemist E-mail: Bede.Willenbring-at-HBFuller.com P.O. Box 64683 St. Paul, MN 55164-0683
} } } Wil Bigelow {bigelow-at-engin.umich.edu} 02/15 1:11 PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
One of the members of this listserver group just asked me privately about the fact that he had noticed that the reading of the thermocouple gauge on his vacuum evaporator changed markedly when he jiggled the wires running from the gauge tube to the gauge control unit. This is a matter that is discussed in some detail on page 87 of my book 'Vacuum Methods in Electron Microscopy' (see http://www.2spi.com/catalog/books/book48.html for a description)
Actually, this is a rather common problem with thermocouple gauges which all people using them should be aware of. It arises because the sensing element in a thermocouple gauge is an arrangement of very fine wires that form one or more thermocouple junctions. The output of these gauges is thus the electrical potential produced by these junctions that is of the order of only 10 or 15 millivolts DC. As a consequence, the signal detected by the gauge controller can be strongly influenced by minor changes the resistance of the electrical lines connecting the gauge to the controller. Typically, unplugging one of the connectors in this line and reconnecting it can change the resistance of the line sufficiently to cause a significant change in the reading produced by the controller.
For this reason, considerable care must be exercised in installing and maintaining thermocouple gauges if acceptable readings are to be obtained. As a minimum, the same cables used in calibrating a gauge must subsequently be used when it is put into operation, and the pins and sockets of the plugs involved in the circuit must be kept clean and firmly connected.
Because of the low DC voltages that must be detected, the meters used in the gauge circuit must be rather delicate and sensitive. We have encountered situations when a static electric charge on the plastic window of a meter affected the movement of the needle inside the meter enough to prevent an accurate response. Coating the meter window with an anti-static solution cured this situation.
As discussed in my book, Thermocouple gauges are rugged, inexpensive, and very convenient for use in many applications, but they must be handled with care if they are to give acceptable service.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
} Heather A Owen wrote: } } } Hello everyone, } } } } A graduate student is getting ready for a rather large scale preparation } } of rat brains for TEM using vascular perfusion. Luckily (since I'm a } } botanist) her major professor has had experience with the procedure. He } } doesn't know for sure whether he used EM or Biological grade } } glutaraldehyde previously, but considering the large volume that will be } } required, and the cost difference, I'm hoping that the Biological grade } } will suffice. } } } } Heather, } } It's been a while but, back when I did such things, I had bouts of } non-uniform fixation when using biological grade glutaraldehyde solutions } even in plant tissues. However, the solutions had been kept much longer than } I'd now use and there are other preparation technique changes that confound } the determination of causality. In the cases where I've perfused small } animals, I've used EM grade GA since, as expensive as it is, it's cheaper } than repeating the experiment. } } However, I did a full systemic perfusion. One random thought ( I haven't } tried it): What's the vascular volume of a rat brain? If you just need just } the brain, I'd think a little creative clamping (really small hemostats?) in } the thorax should let you reduce the volume needed to make even a large } experiment affordable with "the good stuff". } } cheers and good luck, } John }
Heather:
I agree with John on both counts. We clamp the aorta (and anything eles that gets in the way) high in the abdomen to avoid perfusing organs we are not interested in. Saves time and money. Also we tilt the animal once perfusion is underway so that the head is lower.
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 have had a lot of years' experience doing vascular perfusion of the CNS of rodents (rat and mouse). We have used the EM Sciences Biological Grade glutaraldehyde, 50%, as our primary source for perfusion without problems related to fixation. However, we have not used straight glutaraldehyde, but found that 2% paraformaldehyde + 1% glutaraldehyde in 0.1M cacodylate buffer at pH 7.0 to 7.2 gave consistent results with excellent preservation of structures, morphology and cytoplasmic and nuclear components. We have used cacodylate primarily because we use uranyl acetate stain either in the dehydration/processing protocol and/or on the thin sections. Use of phosphate buffers invariably results in precipitate formation, even when used only on sections. Another reason for the paraform/glut/cacodylate combination is that we have studied the leakage of tracers (e.g. HRP, microperoxidase) across the blood brain barrier, and had to be able to move into Tris buffer for DAB reactions, and phosphate buffer gave consistently poorer results.
Hope this helps.
Roger Moretz Dept. of Toxicology BI Pharmaceuticals, Inc.
I have no personal or financial interest in EM Sciences--just a satisfied user.
On Tue, 15 Feb 2000 18:02:57 -0600 (CST), Heather A Owen 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 everyone, } } A graduate student is getting ready for a rather large scale preparation } of rat brains for TEM using vascular perfusion. Luckily (since I'm a } botanist) her major professor has had experience with the procedure. He } doesn't know for sure whether he used EM or Biological grade } glutaraldehyde previously, but considering the large volume that will be } required, and the cost difference, I'm hoping that the Biological grade } will suffice. } } I would appreciate advice from those of you that have experience with this } method of fixation. } } Thank you in advance for any information. } } Heather Owen } } Heather A. Owen, Director } Electron Microscope Laboratory } Department of Biological Sciences } University of Wisconsin - Milwaukee } (414)229-6816 } } }
Roger C Moretz, Ph.D.
_______________________________________________________ Get 100% FREE Internet Access powered by Excite Visit http://freeworld.excite.com
} ------------------------------------------------------------------------ } 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 everyone, } } A graduate student is getting ready for a rather large scale preparation } of rat brains for TEM using vascular perfusion. Luckily (since I'm a } botanist) her major professor has had experience with the procedure. He } doesn't know for sure whether he used EM or Biological grade } glutaraldehyde previously, but considering the large volume that will be } required, and the cost difference, I'm hoping that the Biological grade } will suffice. } } I would appreciate advice from those of you that have experience with this } method of fixation. } } Thank you in advance for any information. } } Heather Owen } } Heather A. Owen, Director } Electron Microscope Laboratory } Department of Biological Sciences } University of Wisconsin - Milwaukee } (414)229-6816
Heather,
It's been a while but, back when I did such things, I had bouts of non-uniform fixation when using biological grade glutaraldehyde solutions even in plant tissues. However, the solutions had been kept much longer than I'd now use and there are other preparation technique changes that confound the determination of causality. In the cases where I've perfused small animals, I've used EM grade GA since, as expensive as it is, it's cheaper than repeating the experiment.
However, I did a full systemic perfusion. One random thought ( I haven't tried it): What's the vascular volume of a rat brain? If you just need just the brain, I'd think a little creative clamping (really small hemostats?) in the thorax should let you reduce the volume needed to make even a large experiment affordable with "the good stuff".
cheers and good luck, John
John Heckman MSM Department Michigan State University
We use Biological grade to perfuse rats (whole body, we haven't tried clamping bits off, though that might be a good idea since we're interested in just the head). After the perfusion is finished, we remove the interesting bit, which is huge by EM standards, and fix some more in Karnowski's. Then we go on to osmium, and continue processing. We get acceptable EM pictures at high magnification, even though our blocks are so large and processing times are therefore extended. So I can recommend Biological grade glutaraldehyde. Hope this helps.
Lesley Weston.
On Tue, 15 Feb 2000, Heather A Owen 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 everyone, } } A graduate student is getting ready for a rather large scale preparation } of rat brains for TEM using vascular perfusion. Luckily (since I'm a } botanist) her major professor has had experience with the procedure. He } doesn't know for sure whether he used EM or Biological grade } glutaraldehyde previously, but considering the large volume that will be } required, and the cost difference, I'm hoping that the Biological grade } will suffice. } } I would appreciate advice from those of you that have experience with this } method of fixation. } } Thank you in advance for any information. } } Heather Owen } } Heather A. Owen, Director } Electron Microscope Laboratory } Department of Biological Sciences } University of Wisconsin - Milwaukee } (414)229-6816 } } } }
} From: RLevenson-at-cri-inc.com Subject: RE: autofluorescence quenching: spectral imaging can help
You asked about strategies for reducing autofluorescence.
In addition to various strategies which can include using near-IR dyes like Cy5.5 and exciting far in the red, you can also treat tissues with sodium borohydride or toluidine blue (these latter observations I pass on from others--have not tried them myself).
I have recently had very promising results using spectral imaging to discriminate autofluorescence from desired fluorescence. [Caution: I am describing a product sold by my company]. CRI makes a liquid crystal tunable filter (LCTF) that can be used to collect a stack of images at different wavelengths. This yields a spectrum at every pixel of an image. (A product with similar capabilities is sold by Applied Spectral Imaging, but uses a different technology.) CRI's new LCTF, the FluoroSpec, uses special optics to combine polarization states; along with other improvements, it has more than twice the light throughput compared to previous models.
The sample was formalin-fixed brain probed with a Cy2-labeled antibody to a low-abundance protein. By eye it was very difficult to distinguish the Cy2 signal from the bright green to green-orange autofluorescence. Using a spectral data set (15 images from 510 nm to 650 nm at 10 nm intervals) and linear combination algorithms for pixel-unmixing, it was possible to completely separate the Cy2 and autofluorescence images.
I would be glad to share these images with any interested parties.
Richard
Richard Levenson, MD Technical Director, Biomedical Systems Cambridge Research & Instrumentation (CRI), Inc. 80 Ashford St. Boston, MA 02134
I just purchased one for printing images, x-ray maps, etc. obtained from my new JEOL 8900 microprobe. Previous to this I was utilizing an Epson Photo 700 printer with my older Cameca MBX probe. I was very pleased with the 700 and for a few hundred dollars sure was worth it! Not a fast printer but gave high quality photo-type prints. After reading the 5000 specs I decided to get one for the new probe. In a "nutshell" here are my observations:
I purchased just the 5000 printer (not the Fiery RIP server that interfaces to it for network printing). Cost: around $3000. for the 5000 only. The 5000 is a much more robust printer than the 700 type Epson models. It can print up to 13"X19" formats. It is faster than the 700 series and the ink cartridges and paper capacity are much greater.
To realistically compare the printers I ran a speed and print quality comparison between my two models. I printed a full 8 1/2" X 11" full color bleed consisting of 3 x-ray maps, a BSE image and an EDS spectrum, along with some text. Printing on the same PC, under the same conditions the print quality is about the same. (A little disappointing here). However the print speed was a full 4 times faster on the 5000 as compared to the 700. Actual print time with the 5000 was about 2.5 min. at 1440 DPI. I found an excellent web site which compares various Epson Photo printers, including the 5000 and 700 series, and demonstrates print quality issues quite vividly:
http://www.tssphoto.com/sp/dg/news/dot_comp.html
All in all I believe the 5000 is a much higher caliber printer than the 700 series, but unfortunately the print quality is about the same.
One final note: the paper makes ALL the difference in the world with this class of printers. Email me if you need info and suggestions on paper issues...
Chuck Burilla United Technologies Research Center 400 Silver Lane East Hartford, CT 06108
(860) 610-7388 burilact-at-utrc.utc.com
-----Original Message----- } From: Malcolm Haswell [mailto:malcolm.haswell-at-sunderland.ac.uk] Sent: Tuesday, February 15, 2000 9:03 AM To: Microscopy (MSA)
I saw the Epson web spec and it looks quite impressive - especially as they seem to be quoting speeds for real picture printing, not 5% coverage or text.
My only concern is that Epson now market two printers with "10 years light fastness" (the Epson Stylus photo 870 - A4 and 1270 A3) so you have to wonder which is better for e.m. prints.
Malcolm Haswell Electron Microscopy School of Sciences Fleming Building University of Sunderland SUNDERLAND SR1 3SD Tyne and Wear UK
Tel (0191) 515 2872 e-mail: malcolm.haswell-at-sunderland.ac.uk
Tom Reese 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 } -----------------------------------------------------------------------. } } Epson has a new ink jet printer out-Stylus pro 5000-that is expensive } but they claim it is much better than the much cheaper models many of } us know and love. Would anyone know whether this is true of just } hype-hard to tell from the published specs...? Thanks...Tom Reese
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Hello,
Does anyone know the name of a supplier of wall posters suitable for an EM lab. I am thinking of cut-out diagrams of TEM and SEM, history of EM (I think JEOL once had one like that), specimen preparation etc. Regards,
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm
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My apologies to the list, but: could Gordon Cougar email me? I get a fatal error when trying to email you: The following addresses had permanent fatal errors ----- } {gcouger-at-rfdata.net} [...] } 550 {gcouger-at-rfdata.net} ... User unknown
Thanks!
Phil
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Paul, My favorite has always been the DIATOME calendars. Next are the Gatan calanders. Dennis Kunkel has great original SEMs of diatomes etc, but I don't know if he has any posters. David Sharpe has the old standby insect SEMs. The students at SanJoaquin Delta College have put together some good calendars also.
} Date: 16 Feb 00 15:54:53 -0800 } From: Paul Webster {pwebster-at-mailhouse.hei.org} } Subject: General:Wall Posters } To: MSA listserver submission {Microscopy-at-sparc5.microscopy.com} } X-Priority: 3 } MIME-Version: 1.0 } Reply-To: Paul Webster {pwebster-at-mailhouse.hei.org} } X-MIME-Autoconverted: from quoted-printable to 8bit by } ultra5.microscopy.com id RAA14282 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Very best regards, Steve D'Angelo Equipment Resurrection 650-738-0351
Does anyone out there have direct experience of any benefit(s) gained by the use of Santovac 5 in the diffusion pumps of a JEOL 840, instead of the recommended Lion S?
And did they have to fiddle with either the heating elements or heater supply voltages in order to acheive the benefits?
I know that Santovac is well thought of, but it's expensive, and my experience of Lion S in an older JEOL (JXA-5A) has been very positive. I think there's something in there about optimising the pump design to a particular fluid.
As an economical alternative, mentioned in Wil Bigelow's book, has anyone tried NEOVAC SY, marketed by Varian, in an 840?
Does anyone know what chemical family Lion S belongs to?
Perhaps someone from JEOL, or even from the Lion company might care to reply.
cheers
Ritchie
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Hello, I'm cleaning out some things in the laboratory and we have a Mikros Vacuum Evaporator Manual C-20, 1964 here. We no longer have this evaporator, so we no longer need the manual. If anyone would have need of it, please email me personally and we'll send it out to you.
I'm amazed at the quality of this older manual - actual blue prints included!
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ 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
There was a fairly extensive discussion about Dp oils on the listserver a while ago. That should be in the archives somewhere.
} } Does anyone out there have direct experience of any benefit(s) gained } by the use of Santovac 5 in the diffusion pumps of a JEOL 840, } instead of the recommended Lion S? }
"Yes". We have been using Santovac 5 in our early 6400 machine for about 5 months now. The diffusion pumps appear to be the same as on an 840 and the 6400 had Lion S in it beforehand.
} And did they have to fiddle with either the heating elements or } heater supply voltages in order to acheive the benefits? }
No. The vacuum seems to be as good as ever without doing any of this. It all looks very clean.
} I know that Santovac is well thought of, but it's expensive, and my } experience of Lion S in an older JEOL (JXA-5A) has been very } positive. I think there's something in there about optimising the } pump design to a particular fluid. } } As an economical alternative, mentioned in Wil Bigelow's book, has } anyone tried NEOVAC SY, marketed by Varian, in an 840? } } Does anyone know what chemical family Lion S belongs to?
That should all be in the archives somewhere, or perhaps on certain web sites. Jim Darley and/or Will Bigelow had the answers.
} } Perhaps someone from JEOL, or even from the Lion company might care } to reply. }
We were told that the Santovac 5 is more tolerant of the occasional gulp of air.....so presumably it should last longer.
Cheers
Arthur Day, Electron Microscope Unit Phone: 61-2-9717-3457 Ansto Materials Division Fax: 61-2-9543-7179 PMB 1, Menai (Sydney), NSW, 2234 Email: ard-at-ansto.gov.au Australia www: http://www.ansto.gov.au/
I am using Photoshop 5.5 on a G3 Macintosh and can no longer get a preview of the image when the file is selected in FILE } OPEN. What gives?
In previous versions (say 4.0 and earlier) when the file was selected, I got a thumbnail preview of the image prior to opening it. This is very convenient and I miss this capability.
Thanks,
John
#################################################################### John J. Bozzola, Ph.D., Director Micro-Imaging and Analysis Center 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 ####################################################################
Santovac is a great oil to use in Jeol instruments. Backstreaming is less and it will withstand a lot more abuse than Lion oil if the vac system has a glitch. Just put a little less charge in than the normal Lion oil. Regards Steve Parkins
WOW, thanks soooo much for all the responses to my question. I have alot of techniques to try so that should keep me busy for awhile. This is a very informative site and I appreciate the help!!
The Department of Biological Sciences at the University of Iowa must find a home for a Philips 300 Transmission Electron Microscope by June 2000. It is in excellent condition and has been under service contract for nearly 30 years. We are remodeling our building and will no longer maintain support for this instrument. Free to anyone willing to come and take it away (plus Haskris chiller, carbon evaporator, film dessicator, etc.).
Please contact: Dean Abel Biological Sciences 138BB University of Iowa Iowa City IA 52242 USA 319-335-1070 (fax 319-335-1069) dean-abel-at-uiowa.edu
We are currently using Cool Prep in our Coolwell cooler system. This is a liquid cleaner & slime preventer that helps keep mineral deposits to a minimum in the cooling system. It also helps prevent corrosion in the water passages without harming hoses, gaskets or seals. Since Coolwell is no longer in business does anyone know of a replacement product for Cool Prep.
Can we simply switch to ethylene glycol? Robert Champaign Raytheon Electronic Systems Labs - Texas Region r-champaign-at-raytheon.com 2501 W.University, MS 8011 McKinney Texas, 75070 972-952-3165
Hi Y'all: We have an JEOL 845 SEM (TMP pumped) and we have been having problems with the TMP REV light staying on. Because of the microscopes' timing circuit, the scope will shut off after 20 minutes. The shutting off/timing issue can be overridden by keeping the key turned clockwise: the microscope will eventually go to the correct vacuum, but the darn TMP light never goes out. JEOL told us of a trick of depressing the standby button on the TMP controller, sometimes remedies this situation, but it didn't work in our case. We have been in contact with JEOL and so far we/they haven't figured out the problem. The TMP pump and the rough pump both seem OK: the TMP 80 % relay kicks in at 80% RPM as it's supposed to and we changed the rough pump oil, just to be sure). I have a few questions to ask of you 845 owners: 1) Have you seen this type of problem, and if so, how did you remedy it? 2) Does anyone out there have wiring diagrams, manuals for the TMP 845 (not the DP, we already have that), if so can we get a copy? JEOL doesn't seem to have anything.
Your help will be greatly appreciated. Mike Coviello Lab Manager UT Arlington Arlington, TX 817 272-5496
We have an old Photomicroscope from Zeiss. It's grey in color and about 20 years old, but works pretty good. Until now we have an old 1 inch video camera on top. We want to change this video camera to a state of art CCD-camera with 1/2 inch CCD-chip. To enlarge the field of view, we want to use a video adapter with a magnification 0.5x. Unfortunately several oprtical corrections are made within this adapter. Such an adapter is not offered by Zeiss. Does anybody out there knows a company that sells such a special adapter or can build one?
We have an old Photomicroscope from Zeiss. It's grey in color and about 20 years old, but works pretty good. Until now we have an old 1 inch video camera on top. We want to change this video camera to a state of art CCD-camera with 1/2 inch CCD-chip. To enlarge the field of view, we want to use a video adapter with a magnification 0.5x. Unfortunately several oprtical corrections are made within this adapter. Such an adapter is not offered by Zeiss. Does anybody out there knows a company that sells such a special adapter or can build one?
FORWARDED TO LISTSERVER, PLEASE RESPOND TO THE ADDRESS SHOWN BELOW:
{randy_anderson-at-ameritech.net}
I am a member of MSA and I work for the University of Chicago in the section of Nephrology. I was wondering if you would know of any universities or other types of institutions that would offer summer courses for the beginner and advance areas of immunofluorescence microscopy. Any help that you can give me in matter would greatly be appreciated.
Thank you
Randy Anderson
#################################################################### John J. Bozzola, Ph.D., Director Micro-Imaging and Analysis Center 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 ####################################################################
The JEOL Schematics are somewhat ambiguous as the logic from the turbo pump activates several relays. The relays are NOT connected in the manner that one would think, i.e. when the turbo pump speed is 80% the relay is activated and signals the vacuum logic that all is well.
Earl Weltmer
Michael Coviello 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 Y'all: } We have an JEOL 845 SEM (TMP pumped) and we have been having problems with the } TMP REV light staying on. Because of the microscopes' timing circuit, the } scope will shut off after 20 minutes. The shutting off/timing issue can be } overridden by keeping the key turned clockwise: the microscope will } eventually go to the correct vacuum, but the darn TMP light never goes out. } JEOL told us of a trick of depressing the standby button on the TMP } controller, sometimes remedies this situation, but it didn't work in our } case. We have been in contact with JEOL and so far we/they haven't figured } out the problem. The TMP pump and the rough pump both seem OK: the TMP 80 % } relay kicks in at 80% RPM as it's supposed to and we changed the rough pump } oil, just to be sure). I have a few questions to ask of you 845 owners: } 1) Have you seen this type of problem, and if so, how did you remedy it? } 2) Does anyone out there have wiring diagrams, manuals for the TMP 845 (not } the DP, we already have that), if so can we get a copy? JEOL doesn't seem to } have anything. } } Your help will be greatly appreciated. } Mike Coviello } Lab Manager } UT Arlington } Arlington, TX } 817 272-5496
ELECTRON MICROSCOPY TECHNICIAN A full time position for a senior research technician is available at the University of Chicago in the Department of Molecular Genetics and Cell Biology. The lab of Elaine Fuchs Ph.D. is interested in the cytoskeleton, in cell-cell adhesion and in the cell biology of skin development. The successful candidate will perform routine transmission electron microcopy, including specimen acquisition, tissue processing, ultramicrotomy, operation of a Philips CM120 electron microscope and darkroom procedures/digital imaging. In addition he/she will be trained in on-section immunolabeling using Lowicryl K4M. Applicant Qualifications: Minimum requirements are a Bachelors degree and 2 years of relevant experience. If you are motivated to work in an excellent scientific environment with state of the art instrumentation please contact:
Christoph Bauer Ph.D. University of Chicago, Department of Molecular Genetics and Cell Biology 5841 S. Maryland Ave/MC 1028 Chicago, Il 60637
I'm reinitializing the server. You may see some old and/or duplicate messages. I will try to restore those that I think did not get through. If you tried to post a message in the last day or so and haven't seen it please wait ~ 12 hours after that time if you do not see it appearby then , you should presume it was lost and repost your original question/comment.
} } Ms. Lalini Pillayi Would like to take both a TEM and SEM short } } course. She is located at the at the US ARmy Dental Research Great } } Lakes Lab. } } Her applications will be both biological and biological } } combined with material. If you offer such a course please contact her } } directly at } } lalini70-at-hotmail.com } } } } Thanks
Gregory W. Erdos, Ph.D. Ph. 352-392-1295 Assistant Director, Biotechnology Program PO Box 110580 Fax: 352-846-0251 University of Florida Gainesville, FL 32611
The JEOL Schematics are somewhat ambiguous as the logic from the turbo pump activates several relays. The relays are NOT connected in the manner that one would think, i.e. when the turbo pump speed is 80% the relay is activated and signals the vacuum logic that all is well.
Earl Weltmer
Michael Coviello 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 Y'all: } We have an JEOL 845 SEM (TMP pumped) and we have been having problems } with the } TMP REV light staying on. Because of the microscopes' timing circuit, the } scope will shut off after 20 minutes. The shutting off/timing issue can be } overridden by keeping the key turned clockwise: the microscope will } eventually go to the correct vacuum, but the darn TMP light never goes out. } JEOL told us of a trick of depressing the standby button on the TMP } controller, sometimes remedies this situation, but it didn't work in our } case. We have been in contact with JEOL and so far we/they haven't figured } out the problem. The TMP pump and the rough pump both seem OK: the TMP 80 % } relay kicks in at 80% RPM as it's supposed to and we changed the rough pump } oil, just to be sure). I have a few questions to ask of you 845 owners: } 1) Have you seen this type of problem, and if so, how did you remedy it? } 2) Does anyone out there have wiring diagrams, manuals for the TMP 845 (not } the DP, we already have that), if so can we get a copy? JEOL doesn't seem to } have anything. } } Your help will be greatly appreciated. } Mike Coviello } Lab Manager } UT Arlington } Arlington, TX } 817 272-5496
We would like to resurrect 2 older Kevex SiLi detectors for scintillation counting, and have lost or misplaced the manuals for the preamp output connectors, ie. voltages and pinouts. The model numbers of the preamps are 2002 and 2003.
If anyone has this info, could you please fax or email?
Thanks in advance
Fred Pearson
******************************************************** Fred Pearson Brockhouse Institute for Materials Research McMaster University 1280 Main St. West Hamilton, Ontario Canada L8S 4M1
ELECTRON MICROSCOPY TECHNICIAN A full time position for a senior research technician is available at the University of Chicago in the Department of Molecular Genetics and Cell Biology. The lab of Elaine Fuchs Ph.D. is interested in the cytoskeleton, in cell-cell adhesion and in the cell biology of skin development. The successful candidate will perform routine transmission electron microcopy, including specimen acquisition, tissue processing, ultramicrotomy, operation of a Philips CM120 electron microscope and darkroom procedures/digital imaging. In addition he/she will be trained in on-section immunolabeling using Lowicryl K4M. Applicant Qualifications: Minimum requirements are a Bachelors degree and 2 years of relevant experience. If you are motivated to work in an excellent scientific environment with state of the art instrumentation please contact:
Christoph Bauer Ph.D. University of Chicago, Department of Molecular Genetics and Cell Biology 5841 S. Maryland Ave/MC 1028 Chicago, Il 60637
Hi Y'all: We have an JEOL 845 SEM (TMP pumped) and we have been having problems with the TMP REV light staying on. Because of the microscopes' timing circuit, the scope will shut off after 20 minutes. The shutting off/timing issue can be overridden by keeping the key turned clockwise: the microscope will eventually go to the correct vacuum, but the darn TMP light never goes out. JEOL told us of a trick of depressing the standby button on the TMP controller, sometimes remedies this situation, but it didn't work in our case. We have been in contact with JEOL and so far we/they haven't figured out the problem. The TMP pump and the rough pump both seem OK: the TMP 80 % relay kicks in at 80% RPM as it's supposed to and we changed the rough pump oil, just to be sure). I have a few questions to ask of you 845 owners: 1) Have you seen this type of problem, and if so, how did you remedy it? 2) Does anyone out there have wiring diagrams, manuals for the TMP 845 (not the DP, we already have that), if so can we get a copy? JEOL doesn't seem to have anything.
Your help will be greatly appreciated. Mike Coviello Lab Manager UT Arlington Arlington, TX 817 272-5496
Heather A Owen wrote: } } Hello everyone, } } A graduate student is getting ready for a rather large scale preparation } of rat brains for TEM using vascular perfusion. Luckily (since I'm a } botanist) her major professor has had experience with the procedure. He } doesn't know for sure whether he used EM or Biological grade } glutaraldehyde previously, but considering the large volume that will be } required, and the cost difference, I'm hoping that the Biological grade } will suffice. } } I would appreciate advice from those of you that have experience with this } method of fixation. } } Thank you in advance for any information. } } Heather Owen } } Heather A. Owen, Director } Electron Microscope Laboratory } Department of Biological Sciences } University of Wisconsin - Milwaukee } (414)229-6816
Heather,
It's been a while but, back when I did such things, I had bouts of non-uniform fixation when using biological grade glutaraldehyde solutions even in plant tissues. However, the solutions had been kept much longer than I'd now use and there are other preparation technique changes that confound the determination of causality. In the cases where I've perfused small animals, I've used EM grade GA since, as expensive as it is, it's cheaper than repeating the experiment.
However, I did a full systemic perfusion. One random thought ( I haven't tried it): What's the vascular volume of a rat brain? If you just need just the brain, I'd think a little creative clamping (really small hemostats?) in the thorax should let you reduce the volume needed to make even a large experiment affordable with "the good stuff".
cheers and good luck, John
John Heckman MSM Department Michigan State University
Santovac is a great oil to use in Jeol instruments. Backstreaming is less and it will withstand a lot more abuse than Lion oil if the vac system has a glitch. Just put a little less charge in than the normal Lion oil. Regards Steve Parkins
We use Biological grade to perfuse rats (whole body, we haven't tried clamping bits off, though that might be a good idea since we're interested in just the head). After the perfusion is finished, we remove the interesting bit, which is huge by EM standards, and fix some more in Karnowski's. Then we go on to osmium, and continue processing. We get acceptable EM pictures at high magnification, even though our blocks are so large and processing times are therefore extended. So I can recommend Biological grade glutaraldehyde. Hope this helps.
We have an old Photomicroscope from Zeiss. It's grey in color and about 20 years old, but works pretty good. Until now we have an old 1 inch video camera on top. We want to change this video camera to a state of art CCD-camera with 1/2 inch CCD-chip. To enlarge the field of view, we want to use a video adapter with a magnification 0.5x. Unfortunately several oprtical corrections are made within this adapter. Such an adapter is not offered by Zeiss. Does anybody out there knows a company that sells such a special adapter or can build one?
Does anyone have the geometry settings for a Kevex Quantum detector on a JEOL 2000FX? I am using DTSA and need the values. I would like to have the sample to detector distance and the azimuthal angle. I am using +45 for the azimuthal angle and 90 for the detector angle. Are these correct? I know that the takeoff angle is 70. -Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
} From: RLevenson-at-cri-inc.com Subject: RE: autofluorescence quenching: spectral imaging can help
You asked about strategies for reducing autofluorescence.
In addition to various strategies which can include using near-IR dyes like Cy5.5 and exciting far in the red, you can also treat tissues with sodium borohydride or toluidine blue (these latter observations I pass on from others--have not tried them myself).
I have recently had very promising results using spectral imaging to discriminate autofluorescence from desired fluorescence. [Caution: I am describing a product sold by my company]. CRI makes a liquid crystal tunable filter (LCTF) that can be used to collect a stack of images at different wavelengths. This yields a spectrum at every pixel of an image. (A product with similar capabilities is sold by Applied Spectral Imaging, but uses a different technology.) CRI's new LCTF, the FluoroSpec, uses special optics to combine polarization states; along with other improvements, it has more than twice the light throughput compared to previous models.
The sample was formalin-fixed brain probed with a Cy2-labeled antibody to a low-abundance protein. By eye it was very difficult to distinguish the Cy2 signal from the bright green to green-orange autofluorescence. Using a spectral data set (15 images from 510 nm to 650 nm at 10 nm intervals) and linear combination algorithms for pixel-unmixing, it was possible to completely separate the Cy2 and autofluorescence images.
I would be glad to share these images with any interested parties.
Richard
Richard Levenson, MD Technical Director, Biomedical Systems Cambridge Research & Instrumentation (CRI), Inc. 80 Ashford St. Boston, MA 02134
Hi all. I have just joined this list and am posting without the usual "lurking time" due to time constraints - please forgive if recently covered. I am also a novice in all microscopy techniques, especially fluorescence.
I am trying to detect GFP-containing cells in liver tissue and having some problems.
1. I can't determine the spectra for the filter blocks I am using (on a Nikon Eclipse TE300 with TE-FM Epifluorescence attachment). The blocks are marked "UV", "B-2" (blue from source, green in field) and "G" (green from source, red in field). I have e-mailed Nikon and to be fair they've only had a few days but I'm under some pressure. No help from the manual. I have been using B-2 for GFP.
2. I get quite a lot of background fluorescence even with frozen sections (fixed in neutral buffered formalin). Can anyone suggest a way to reduce this? (eg any extra filters?)
3. I have read conflicting opinions on fixation. Most previous work has used thick sections (50 microns cut eg with Vibratome, ? to avoid having to embed tissue blocks). GFP is interfered with by acetone (and probably other organic solvents) but I would have thought that after fixation with formalin it should be stabilized and resistant to the xylene/alcohol used in paraffin embedding. I have seen fluorescence retained after this treatment but perhaps it can be improved.
4. For those with liver fluoro experience - on "UV" setting I see bright fluorescence which photobleaches. I believe I am looking at retinoids in stellate cells, although the bleaching is incomplete and a bit slower than I have seen before. Can anyone confirm this?
Apologies for long post. Any help much appreciated.
David Lockwood University of Qld Dept of Surgery PA Hospital Brisbane Australia
Hello Robert, Preparation of the water in water chillers is a strange world of its own and you may get many responses to your question. The majority of my experience has been with the Haskris systems but the chemistry may be the same. In about 30 years of looking after many chillers, my formula for success goes like this: 1 - Use deionized or distilled water. The water will leach enough products from the hoses, microscope lenses, pumps and so forth to reach a equilibrium and then be "ideally suited" for your system. 2 - Don't use any fungicide or other bug killer because it will slime up the works. 3 - Put 4 or 5 drops of oil on top of the water reservoir. This will form a barrier by creating a monolayer on top of the water and keep things from growing in the water. I have used this proceedure in tropical climates and in the north and in both dry and humid environments and it has always worked very well. I can almost gurantee you that ethyline glycol will gum up the works.
Best wishes and good luck.
Alex Greene SCIENTIFIC INSTRUMENTATION SERVICES, INC. PMB-499, 1807 West Slaughter Lane, Number 200 Austin, Texas 78748-6200 Phone:512-282-5507 FAX 512/280-0702
QUALITY ELECTRON MICROSCOPE REPAIRS
-----Original Message----- } From: Robert Champaign {r-champaign-at-raytheon.com} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
I'm using a CM30 FEG with GIF PEELS atached to it. I'm getting confused about focussing an energy filtered image.
GATAN proposes to focus the image on the TV camere at a loss of 100eV.Apparently it will be focused more or less OK at a higher loss say 741eV. DM uses the voltage offset which changes the HT of the microscope. So as far as I understand the focus should be OK for every energy loss(no refocussing should be needed at all) . Although in practice it is not. The trick with focussing at 100eV works better than not focussing at all but it is not satisfactory for high spatial resolution.
In the Phd of M.Schenner (TU Wien "The quantification problem in EELS Microanalysis") I found a callibration procedure to find a defocus to correct for chromatic abberation as a function of the energy loss you are interested in. But he seems to use the drift tube in the GIF.
The question now: How should I focus my ESI images and what is the explanation behind it?
Thanks in advance,
Jo
************************************************************* * Jo Verbeeck * * University of Antwerp * * Dept. EMAT (Electron Microscopy for Materials Research) * * e-mail: joverbee-at-ruca.ua.ac.be * * tel: +32(0)3 218 02 49 * * fax: +32(0)3 218 02 57 * *************************************************************
Dear All, We are considering trying to label aspects of innervation using a quinacrine fluorescence technique. So far the preliminary trials have not gone well. Could anyone give advice on this, especially on a source for a reliable fluorescent quinacrine dye?
Thanks in advance
Alex Black Department of Anatomy National University of Ireland, Galway
We have specialists in this area who can offer customized, on-site courses, using your equipment and speaking specifically to your application. If you are interested in further details, or in how U. Chi can sponsor such a course, please contact me directly.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education}
At 04:59 PM 2/21/00 -0600, John J. Bozzola wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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I perfused dozens of rat brains over the years. I would recomend using the purified glutaraldehyde. However, you can use a slightly less concentrated solution since the fixation is so much more efficient tham immersion fixation. We primarily fixed for ICC but only the choice of fix would vary not the method. For normal ultrastructure, I would try 2-3% Glut or half-strength Karnovsky's in a phosphate buffer system with added NaCl and Mg at physiological concentrations and see which fixation I prefered by examining the desired tissues prior to fixing critical animals.
We would anesthetize the animal, open the chest cavity and clamp off the descending aorta, thus limiting the perfusion to the upper half of the animal. The cannula would then be inserted into a small slit in the left ventricle, positioned up into the aorta, and held with a hemostat. Perfusion lines should have been filled with normal saline or ringers so that as soon as the cannula is in place, the flow can be started to wash out the blood. A small slit is made in the right atrium to permit release of venous blood, etc. As soon as the return is relatively pale, switch to your fixative and run the desired amount through. The condition of the liver is a reasonable criteria for the efficiency of the perfusion. It should be very pale yellow as the blood is removed and should stiffen up within a few minutes. In fact the entire upper part of the animal should become quite stiff as the tissues are fixed. Figure on a couple of hundred mls. of fix per animal.
A couple of suggestions are to use the initial ringer wash and fixatives at room temperature or slightly above so as not to cause vessels to contract. Do any further washes and post fix the portions of interest at 4oC. Also make sure you have a consistant but gental perfusion rate so as not to break delicate capillaries. Usually this can be accomplished just by raising the vessels containing the buffer and fix above the animal and let gravity do the rest. Good luck, Debby
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
--------------------------------------
Heather A Owen wrote: } } Hello everyone, } } A graduate student is getting ready for a rather large scale preparation } of rat brains for TEM using vascular perfusion. Luckily (since I'm a } botanist) her major professor has had experience with the procedure. He } doesn't know for sure whether he used EM or Biological grade } glutaraldehyde previously, but considering the large volume that will be } required, and the cost difference, I'm hoping that the Biological grade } will suffice. } } I would appreciate advice from those of you that have experience with this } method of fixation. } } Thank you in advance for any information. } } Heather Owen } } Heather A. Owen, Director } Electron Microscope Laboratory } Department of Biological Sciences } University of Wisconsin - Milwaukee } (414)229-6816
Heather,
It's been a while but, back when I did such things, I had bouts of non-uniform fixation when using biological grade glutaraldehyde solutions even in plant tissues. However, the solutions had been kept much longer than I'd now use and there are other preparation technique changes that confound the determination of causality. In the cases where I've perfused small animals, I've used EM grade GA since, as expensive as it is, it's cheaper than repeating the experiment.
However, I did a full systemic perfusion. One random thought ( I haven't tried it): What's the vascular volume of a rat brain? If you just need just the brain, I'd think a little creative clamping (really small hemostats?) in the thorax should let you reduce the volume needed to make even a large experiment affordable with "the good stuff".
cheers and good luck, John
John Heckman MSM Department Michigan State University
I have a Beseler 45M Enlarger and Kodak Royal Print Processor for sale. Both are in working condition. We are going digital! Equipment is located in Easton, PA.
Well the DNS (Domain Name Server) failed and was probably pretty sick over most of last week. Alot of mail probably just ended up in a black hole somewhere on the Net. Some of you may have gotten messages, but I see that most of the mail I sent out during my trip DownUnder to the ACEM-16 meeting in Canberra never made it to it's destination.
The DNS "looks" fixed now but I'm not confident enough to proclaim all as back to normal yet. Let's see if this message actually gets through to everyone this time.
Sorry gang....
Nestor Your Friendly (Frustrated) Neighborhood SysOp
I have 2 queries pertaining to the use of the lipid colourant Sudan Black B, I hope someone can be of some help:
1. I have been able to stain oil traces in the glands of citrus fruit BUT the epoxy resin Procure/Araldite also stains quite strongly. Is this typical of epoxy resins and would Spurrs show the same problem?
- Should I reduce the staining time from 30 mins? - Adjust the temperature from 60 degrees C for staining? - Rinse my sections for longer with 70% ethanol after staining?
2. I have used osmium tetroxide to fix lipids in the citrus rind tissues, with mixed results. I have also seen some success in oil fixation by applying Sudan Black B to the tissue prior to chemical fixation. I assume it binds and reduces the oil solubility. I am going to test this by treating tissue with SBB prior and post chemical fix, and comparing.
- SBB is made up in 70% ethanol. As a postfix, would it be most logical to introduce it at the 70% step in my ethanol dehydration series? (10 to 100%)
Department of Horticulture, Viticulture and Oenology The University of Adelaide Plant Research Centre Waite Campus, PMB 1 Glen Osmond, SA 5064 AUSTRALIA
has anyone out there any experiences with immuno-labeling of osmificated and Spurr embedded samples. We have tried out etching, oxidizing sections with hydrogen peroxide (10% for 15 min) or sodium methylat (1% for 15 min) without any significant increase of label intensity (Ab used is polyclonal from rabbit, protein A purified)
It made it her ok. I thought I was getting pretty good service all week. But I know what evils hide in name servers. Not nearly to the extent you do:)
As always you efferots are welcome. You provide the pipe for one of the best resorces on the net.
Gordon
G. C,. Couger 624 Cheyenne Stillwater, OK 74075-1411 405 624 2855 between 3:00 pm to 1:00 am CST 405 742 2758 cell ----- Original Message ----- } From: "Nestor J. Zaluzec" {zaluzec-at-sparc5.microscopy.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, February 22, 2000 7:20 PM
} } Dear All: } } I was wondering if anyone out there is aware of a company } that sells pinhole grids. I am looking for grid(2D) of } pinholes that are approximately 15-20 microns in diameter } with a center to center separation of about 150-200 microns. } } I would greatly appreciate any information that you could } provide regarding the commercial availability of similar } pinhole grids. } } TIA, } Fatima Merchant } } } {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} } } Fatima Merchant, Ph.D. } Senior Research Engineer } Perceptive Scientific Instruments, Inc. } 2525 South Shore Blvd., Suite 100 } League City, Texas 77573 } } Telephone: (281) 334-3027 Ext: 230 } Toll Free: (800) 288-3027 Ext: 230 } Facsimile: (281) 538-2222 } Email: merchant-at-persci.com } } {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} } }
-----Original Message----- } From: Valdes Erica R Dr. SBCCOM Sent: Tuesday, February 22, 2000 1:26 PM To: 'microscopy-at-sparc5.Microscopy.Com'
We got a sputter-coater system (VG Microtech SC500) that we would like to convert into a glow discharge. Does anybody have done it already? We got electronic schemes to do the High Voltage unit control, but we miss the mecanical design to fit into our vacuum chamber (of the SC500). Does anybody know how to do? We know that an acessory (for the Microtech) have been developped some years ago (GD350A) but it is not anymore available. What are the modification made to use this chamber? Jean-Jacques LACAPERE INSERM U 410 Facult de Mdecine X. Bichat Paris FRANCE
I'm using a CM30 FEG with GIF PEELS atached to it. I'm getting confused about focussing an energy filtered image.
GATAN proposes to focus the image on the TV camere at a loss of 100eV.Apparently it will be focused more or less OK at a higher loss say 741eV. DM uses the voltage offset which changes the HT of the microscope. So as far as I understand the focus should be OK for every energy loss(no refocussing should be needed at all) . Although in practice it is not. The trick with focussing at 100eV works better than not focussing at all but it is not satisfactory for high spatial resolution.
In the Phd of M.Schenner (TU Wien "The quantification problem in EELS Microanalysis") I found a callibration procedure to find a defocus to correct for chromatic abberation as a function of the energy loss you are interested in. But he seems to use the drift tube in the GIF.
The question now: How should I focus my ESI images and what is the explanation behind it?
Thanks in advance,
Jo
************************************************************* * Jo Verbeeck * * University of Antwerp * * Dept. EMAT (Electron Microscopy for Materials Research) * * e-mail: joverbee-at-ruca.ua.ac.be * * tel: +32(0)3 218 02 49 * * fax: +32(0)3 218 02 57 * *************************************************************
Material fixed like yours works only rarely. You might try etching with saturated sodium metaperiodate for one hour followed by water washes before incubating with the primary. That is assuming that your antigen is not periodate sensitive.
At 07:26 AM 02/23/2000 +0100, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Gregory W. Erdos, Ph.D. Ph. 352-392-1295 Assistant Director, Biotechnology Program PO Box 110580 Fax: 352-846-0251 University of Florida Gainesville, FL 32611
} } I was wondering if anyone out there is aware of a company } } that sells pinhole grids. I am looking for grid(2D) of } } pinholes that are approximately 15-20 microns in diameter } } with a center to center separation of about 150-200 microns. } } } } I would greatly appreciate any information that you could } } provide regarding the commercial availability of similar } } pinhole grids. } }
Dear Fatima, I asked about the same question and got a reply from Janko Otto (otto-at-quantifoil.com). He wrote me:
"I read a discussion about making holey films . We had solving the problem using semiconductor lithographic techniques.
We have been producing QUANTIFOIL, a perforated foil with pre-definable hole size shape and arrangement. The variation among the hole sizes is small: the standard deviation for their distribution is well below 0.1 µm within a batch; the reproducibility for the average hole size between batches is 0.2 µm. The shapes of the holes in the foils produced so far are circular and square. Until now, their arrangements have been orthogonal.
Foils with any desired hole size in the micrometer range, and with any shape and arrangement can be fabricated. Any of these three parameter can also be varied in a defined manner within one foil.
Until now, we produce three different types of QUANTIFOIL:
Type Hole Repetition distancee um um R1.2/1.3 ~1.2 2.5 (circular holes) R2/2 ~2 4 (circular holes) S7/2 ~7 9 (square holes)
QUANTIFOIL is generally delivered as a carbon foil; it can also be obtained strengthened with plastic film.
If you interested in these perforated foil, I can send you some samples."
He may be able to produce what you want. I have not yet tried his grids out, but I intend to when I get my specimens ready. I am not affiliated with Quantifoil, just a potential user. Good luck. Yours, Bill Tivol
************************************************************* * Jo Verbeeck * * University of Antwerp * * Dept. EMAT (Electron Microscopy for Materials Research) * * e-mail: joverbee-at-ruca.ua.ac.be * * tel: +32(0)3 218 02 49 * * fax: +32(0)3 218 02 57 * *************************************************************
---------- Forwarded message ----------
} We are currently using Cool Prep in our Coolwell cooler system. This is a } liquid cleaner & slime preventer that helps keep mineral deposits to a } minimum in the cooling system. It also helps prevent corrosion in the } water passages without harming hoses, gaskets or seals. Since Coolwell is } no longer in business does anyone know of a replacement product for Cool } Prep. } } Can we simply switch to ethylene glycol?
Dear Robert, The suggestion of using distilled water and letting the system come to equilibrium might work in a truly closed system composed of only one material, but if you need to add water, or if you have several metals in the system--as we do--this will be unsatisfactory in the long run. We have found that Aqua-Treet 42 works very well in our Haskris system. This product is made by Aqua Laboratories, Inc., PO Box 645, 8 Indus- trial Way, Amesbury MA, (978) 388-3989. I have no relationship to this company except as a satisfied user of their product. Yours, Bill Tivol
I am running two copies of DTSA, one is an older version and one is the newer one that will run on a power Mac. I am also fairly new to DTSA. The old version does not have the MSA format capability for reading and writing files, but the newer one does. However, the new one will read two versions of MSA format, 1.0 and 1.1 but will only save in 1.1. Now I'm confused. I asked Nestor where I could get the latest MSA description of the MSA format and he sent me what I already have, EMMFF.total from of the EMMPDL/MAS-Lib files. I know that Nestor was on that committee. So what gives with MSA version 1.1 in the latest version of DTSA? Why don't they have the 1.0 option? The keywords are not even the same.
-Confused in Pittsburgh.
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Why not use a liquid that has NO water in it. That solved my problem in vibratory polishing of dissimilar metals in an alumina slurry. Ethylene glycol or propylene glycol, (less toxic), are possibilities.
Dear Bernie,
That will work unless there is some microorganism which will
grow in ethylene glycol. One could also just buy commercial anti-freeze
and get some additional corrosion protection. I don't think that there
would be any problem with the circulation pump, since the viscosity
of glycol is similar to that of water. Keeping the fluid in the system for
some decades has different requirements from using it over a period
Look for a "Write MSA 1.0" plug-in in the "extras Folder" of your DTSA download and move it to the "Plug-ins" folder at the same level as DTSA.
At 11:42 AM -0500 2/23/00, Walck. Scott D. wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
====================================== Carl Henderson Electron Microbeam Analysis Laboratory University of Michigan 2501 C.C. Little Bldg. Ann Arbor, MI 48109-1063 USA (734) 936-1550 FAX (734) 763-4690 ======================================
Greetings, I wonder if anyone could explain the following observation to me. I have been sputter coating with platinum and viewing the sample with a FESEM (Hitatchi 4700, below lens type). In a certain type of sample, the image appears darker than the background where there is sample. Now these samples are perhaps a bit unusual. They are very thin, starting off life as 1.75 um methacrylate sections of a plant root adhered to a glass coverslip, followed by extraction in hot acidic peroxide to remove all organic material except crystalline cellulose. The cellulosic cell walls left behind on the coverslip are nominally 100 nm thick and perhaps flattened further by the processing. The cell walls are present in the sample as either cross sections or as small regions laying in the plane of the coverslip. These regions can be anywhere from the size of a cell (10 x 40 or so microns) down to small portions thereof. All of this remaining cell wall is darker than the surrounding background. I am putting on a thin coat of Pt, somewhere around 2 nm, and imaging at 2 kV (I have seen the same thing at 5 kV). It really puzzles me that my sample is darker than background. Any explanations???
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Hi, Well I guess we never run out of new challenges when it comes to specimen prep. This one concerns a product that is made of a cereal bound together with a sugar syrup. The investigators want to see how the syrup behaves as to structural integrety over a period of time with and without the addition of various stabilizing components. Information would be collected at the light microscopic level. We first thought of cryosectioning followed by examination using LM. However, the product is filled with air pockets and literally disintegrates when microtomed. It is easily cut into pieces at temperatures above freezing as long as the pieces are no thinner than3-4 mmAccempts to infiltrate it with OCT compoound prior to freezing were not satisfactory. New thought is to use a low temperature resin to infiltrate the sample followed by UV or chemical polymerization. butThe resulting blocks could then be microtomed at room temperature and imaged. It would be desirable to cut sections of 4-8 µm in thickness which is too thick for an ultramicrotome with glass or diamond knives. Also UV polymerization would probably limit us to very small sample size which may be a problem. Any suggestions would be appreciated as to resins to use which have low viscosity, could be used to infiltrate pieces of ~0.5-1cm sq, polymerize at low temperatures (+4 to 0oC range) using UV or chemical polymerization, and are preferably soft enough to section with a metal microtome blade. Any other suggestions of preparations techniques to try would be very appreciated.
Thanks in advance for your input. Debby
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
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Dear listservers, I need to determine the value of a Philips 410 that has been on service contract until it was decommissioned last year. I need this information for an insurance claim. Is there a company or dealer of used em equipment that can help me? Thanks Hank Adams Integrated Microscopy Core Molecular and Cellular Biology Baylor College of Medicine Houston, Tx
Sorry if this is a second post, but we just got an e-mail saying the first was rejected as spam, so here goes:
Hi:
I want to be able to "Glow Discharge" carbon coated copper grids for the purpose of making them hydrophilic and negatively charged, so that I can spread aqueous suspensions on them. I have a Denton Vacuum Desk ll sputter coater with a "etch" mode. The question I have for my fellow list servers is: will "etching" the grids do the same thing as "Glow Discharging" them? Many Thanks, Tim
Timothy Schneider, Director of Electron Microscopy Department of Pathology Thomas Jefferson University Room 229 Jeff Hall 1020 Locust St. Philadelphia, Pa. 19107 215-503-4798 Timothy.Schneider-at-Mail.TJU.EDU
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We are interested in localizing arsenic in biological tissues (toxicology research), but have been unable to come up with a technique that will give us the cellular localization information we're looking for (sub-cellular would be even better).
We have tried TEM with energy dispersive x-ray microanalysis and our concentrations are apparently too low, so we are unable to detect arsenic.
A few years ago we looked into using the autometallographic techniques that have been published by Dr. Gorm Dansher & his colleagues. We were told that the photographic emulsions required were no longer available.
In a brainstorming session yesterday several techniques were bantered about, but in truth we are not familiar enough with the techniques to know if they would suit our needs. Does EELS have anything to offer, or perhaps wavelength dispersive spectoscopy (WDS)? How about some of the new FT-IR instruments? Are there any other techniques that might be able to localize arsenic and/or indicate its state of oxidation?
Vendors are welcome to reply.
Yours, Doug Cromey .................................................................... : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : Research Specialist, Principal University of Arizona : : (office: AHSC 4212A) P.O. Box 245044 : : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: doug-cromey-at-ns.arizona.edu): :...................................................................: http://www.pharmacy.arizona.edu/exp_path.html Home of: "Microscopy and Imaging Resources on the WWW"
Go to http://www.cstl.nist.gov/div837/837.02/dtsa.html and click on "supporting files" page then download "preferences and plugins" you will find 2 versions of the MSA write 1.0 (one is Kevex specific). It would make life much easier on us microscopists if all manufacturers adopted the MSA format. Good luck, Scott
} I am running two copies of DTSA, one is an older version and one is the } newer one that will run on a power Mac. I am also fairly new to DTSA. The } old version does not have the MSA format capability for reading and writing } files, but the newer one does. However, the new one will read two versions } of MSA format, 1.0 and 1.1 but will only save in 1.1. Now I'm confused. I } asked Nestor where I could get the latest MSA description of the MSA format } and he sent me what I already have, EMMFF.total from of the EMMPDL/MAS-Lib } files. I know that Nestor was on that committee. So what gives with MSA } version 1.1 in the latest version of DTSA? Why don't they have the 1.0 } option? The keywords are not even the same. } } -Confused in Pittsburgh. } } Scott D. Walck, Ph.D. } PPG Industries, Inc. } Glass Technology Center } Guys Run Rd. (packages) } P. O. Box 11472 (letters) } Pittsburgh, PA 15238-0472 } Walck-at-PPG.com } (412) 820-8651 (office) } (412) 820-8161 (fax)
For the weather watchers: Mostly sunny, 60 F, snow is melted away.
-------------------note: new mailing address---------------------- Scott Wight e-mail: scott.wight-at-nist.gov NIST 222/A113 W voice: 301-975-3949 100 Bureau Dr STOP 8371 | fax: 301-417-1321 Gaithersburg, MD 20899-8371 \|/ disclaimer: Any opinion expressed is my own and does not represent those of my employer.
Dear all, I am doing the image analysis of SEM micrographs. The micrographs contain both dense (light) and air cell (dark) areas. But two areas are not discrete. I'm having a difficulty of quantifying those separated fractions. I wonder if anyone has any ideas of how to solve the problem. Looking forward for your suggestions. Regards, Supanee
Hello all, I need to know how to draw an ellipse to measure at Image Tool software. Thanks a lot. Rejane Pimentel Rejane Magalhes Pimentel Galindo Functional Plant Morphology Universidade Federal Rural de Pernambuco ggalindo-at-elogica.com.br Rua Maria Carolina, 417/804 51020-220 Boa Viagem
I thought I'd try this again in case it got lost in space:
Hi-
We are going to be processing rat eyes (from perfused animals) for embedding in epoxy and I was wondering if anyone has any suggestions for processing. We are interested in the retina, optic nerve, cornea and uveal tract.
Ahh! That is always the rub, trying to get the areas of interest to stand out as a distinct range of gray scales.
If it is just a question of noise pixels in what is otherwise a distinct region, then you might try filtering (smoothing) your image before thresholding. Or you might try changing your image acquisition protocol to get less noisy images.
You might try a different signal to find something that is less subject to overlaps. Usually I resort to backscattered electron imaging since the dependence on atomic number of that signal typically results in distinct gray levels.
If that won't do it, you might be able to perform some sort of image processing to separate the regions. There are techniques available, but there seems to be as much art as science to them. I would refer you to John Russ' "Image Processing Handbook" or a similar text, because there is not a short answer to that question.
Warren Straszheim
At 03:40 PM 2/23/2000 -0500, you wrote: } Dear all, } I am doing the image analysis of SEM micrographs. The } micrographs contain both dense (light) and air cell (dark) areas. But two } areas are not discrete. I'm having a difficulty of quantifying those } separated fractions. I wonder if anyone has any ideas of how to } solve the problem. } Looking forward for your suggestions. } Regards, } Supanee
} From: {"wft03-at-health.state.ny.us"-at-sparc5.microscopy.com} To} Why not use a liquid that has NO water in it. That solved my problem in } vibratory polishing of dissimilar metals in an alumina slurry. Ethylene } glycol or propylene glycol, (less toxic), are possibilities. } } } Dear Bernie, } } That will work unless there is some microorganism which will } } grow in ethylene glycol. One could also just buy commercial anti-freeze } } and get some additional corrosion protection. I don't think that there } } would be any problem with the circulation pump, since the viscosity } } of glycol is similar to that of water. Keeping the fluid in the system for } } some decades has different requirements from using it over a period } } of hours, though.
Glycols won't have near the heat capacity of water. I doubt you will find a bug that will live in ethylene glycol but you can probably find many that can live in propylene glycol.
Any cooling system needs to have the coolant changed periodically. Following the manufactures recommendation is the safe way to go.
Gordon
Gordon Couger gcouger-at-couger.com
Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
This is something that we can probably help you with. We do similar jobs for several other companies. If you could fax a drawing with the material, OD, Thickness, number of holes, etc. to 1-802-878-8074 we can get you a quote.
Thanks,
John Arnott Chairman --
LADD RESEARCH 131 Dorset Lane Williston, VT 05495
TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net web site http://www.ladd.cc
--
LADD RESEARCH 131 Dorset Lane Williston, VT 05495
TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net web site http://www.ladd.cc
We are considering to purchase a cryotransfer system for our 2010F TEM. There are two major suppliers: Gatan and Oxford. We are not sure which one works better since this is the first time dealing with this technique.
Could you please send me your comments and experiences on these two vendors? Our samples are mostly soft tissues: Polymers, surfactants, cell membranes, and proteins. Your help is highly appreciated.
Maoxu Qian **************************** * Maoxu Qian, Ph.D. * * Dept of MSE, box 352120 * * University of Washington * * Seattle, WA 98195 * * mxq-at-u.washington.edu * * (206)616-3973(phone) * * (206)543-3100(fax) * ****************************
This course will cover all aspects of pre-thinning and focus on final thinning via Tripod Polishing. Due to the limited class size and the extensive hands-on opportunities, this course is well suited to novices as well as advanced Tripodders. Attendees will also learn the latest techniques available in ion milling, plasma cleaning and ion beam sputter deposition and etching for TEM and SEM samples.
The workshop will be held in San Clemente, CA on May 26-27, 2000. Please call for more information or visit our web site ( www.southbaytech.com ) for registration information.
David Henriks Vice President 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
*************************************************************************** } } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
} } } Hi, There, } } We are considering to purchase a cryotransfer system for our 2010F TEM. } There are two major suppliers: Gatan and Oxford. We are not sure which one } works better since this is the first time dealing with this technique. } Could you please send me your comments and experiences on these two } vendors? Our samples are mostly soft tissues: Polymers, surfactants, cell } membranes, and proteins. Your help is highly appreciated. } } Maoxu Qian } **************************** } * Maoxu Qian, Ph.D. * } * Dept of MSE, box 352120 * } * University of Washington * } * Seattle, WA 98195 * } * mxq-at-u.washington.edu * } * (206)616-3973(phone) * } * (206)543-3100(fax) * } **************************** } } } } }
A related question from India. We have been using DM (demineralised) water in our JEOL 2000 EX II cooled by a Neslab chiller for the last ten years. This requires change every 6 months approximately. What additives can be used to prevent algae (? it is green and slimy and I am not a biologist) growth? Something obtainable locally (chemical name rather than trade names) will be most useful to me. Thanks very much.
-----Original Message----- } From: William Tivol [SMTP:tivol-at-wadsworth.org] Sent: Thursday, February 24, 2000 10:25 AM To: Robert Champaign Cc: microscopy-at-sparc5.microscopy.com
} We are currently using Cool Prep in our Coolwell cooler system. This is a } liquid cleaner & slime preventer that helps keep mineral deposits to a } minimum in the cooling system. It also helps prevent corrosion in the } water passages without harming hoses, gaskets or seals. Since Coolwell is } no longer in business does anyone know of a replacement product for Cool } Prep. } } Can we simply switch to ethylene glycol?
Dear Robert, The suggestion of using distilled water and letting the system come to equilibrium might work in a truly closed system composed of only one material, but if you need to add water, or if you have several metals in the system--as we do--this will be unsatisfactory in the long run. We have found that Aqua-Treet 42 works very well in our Haskris system. This product is made by Aqua Laboratories, Inc., PO Box 645, 8 Indus- trial Way, Amesbury MA, (978) 388-3989. I have no relationship to this company except as a satisfied user of their product. Yours, Bill Tivol
Dear Colleagues Does anybody have experience on the Allied MultiPrep Polishing System, especially in terms of getting a good TEM sample? How well do the Allied's Precision Cross-Section Tool and TEM Wedge Polisher work on this semi automatic multiprep polisher? Also, how does their TEM Wedge Polisher compare with the South Bay Tripod Polisher? TIA Anita ******************************************* Dr. Anita Garg NASA Glenn Research Center at Lewis Field Advanced Metallics Branch Mail Stop 49-3 21000 Brookpark Road Cleveland, OH 44135 Phone : (216) 433-8908 Fax : (216) 977-7132 E-mail : Anita.Garg-at-grc.nasa.gov *******************************************
Fatima Merchant, I used to work for a company called Buckbee Mears. They have been making tight tolerance electroformed Ni pinholes for many, many years. They are located in St. Paul, MN and their phone number is 651-228-6400. Good luck. Dan May
} Hi- } } We are going to be processing rat eyes (from perfused animals) for } embedding in epoxy and I was wondering if anyone has any suggestions for } processing. We are interested in the retina, optic nerve, cornea and uveal } tract. } } Thank you very much, } Sandy Perkins
**************************** Hi Sandra,
Eyes are a real pain. There are so many layers of different cell types, and each seems to infiltrate differently. Having worked on a fir number of eyes, I woauld make the following suggestions: Process the cornea separately. It is almost pure collagen. It will not osmicate all that noticeably, so don't be surprised when it doesn't turn dark brown or black. Give it long infiltration times, in vaccuum if possible.
If you don't need the optic nerve attached to the eye, dissect it free and process it. Give it slighlty longer than usual dehydration and infiltration steps to be sure you'e got the myelin taken care of.
As for the rest: remove the lens and the vitreous so that you have good acess to the retina. Be careful not to disrupt the retina when you remove the vitreous. Once the lens is out, you can usually get the vitreous out by gently blotting against a Kimwipe. You may want to split the orb into hemispheres. Just be sure not to take any sections from the cut edge, since you will disturb the structure there when you cut. Again, you'll nees longer dehyration and infiltration times, since the sclera is really tough. I use 30 minutes in each step of graded acetone, propylene oc=xide then a graded infiltraion with PO:resin (Epon analog) 2:1 for 30 min, 1:1 overnight, 1:2 2 hours, pure resin overnight, on a rotator.
I'm sure there ore others out there, with their own "pet protocol" for eyes. Its all a bit of magic and luck, isn't it?
Good luck, Lee
Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Confocal Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
Not sure this went out earlier...I've inherited an Olympus BH-2 RFCA upright that I'd like to equip with WI lenses for live cell fluorescence work. This is a 160mm tube length scope, and Olympus is now only manufacturing the 60X WI fluor objective (I think/hope). Any leads to sources of compatible WI objectives in the 10-60X range much appreciated. Please reply offline. Thanks.
Mike
Michael Ferrari Department of Biological Sciences 629 Science Engineering (SCEN) University of Arkansas Fayetteville, Arkansas 72701 Ph: 501-575-6372 Fax: 501-575-4010
We are interested in selling our Perkin-Elmer Model 1010M microdensitometer at a very reasonable price. It was purchased as a reconditioned model from Perkin-Elmer eight years ago and is in very good condition. Technical manual is included.
If interested, please contact:
Amy Kendall Department of Molecular Biology Vanderbilt University Box 1820, Station B Nashville TN 37232 (615) 322-2012 amy-at-helix.molbio.vanderbilt.edu
R. Divakar wrote, "A related question from India. We have been using DM (demineralised) water in our JEOL 2000 EX II cooled by a Neslab chiller for the last ten years. This requires change every 6 months approximately. What additives can be used to prevent algae (? it is green and slimy and I am not a biologist) growth? Something obtainable locally (chemical name rather than trade names) will be most useful to me."
I quote here recommendations from the Haskris Co. (we have several Haskris chillers):
1. If algae is present, flush the system with one of two alternatives: add 1 cup of chlorine bleach or one pint of hydrogen peroxide per 15 gallons of water. Circulate 20-30 minutes. Drain the system and refill with clean water.
2. To prevent regrowth of algae, one of the following additives are recommended: a. 10% solution of lab grade (no additives) ethylene glycol. b. Dichlorophene. This is an insoluble white powder which is a fungicide/bacteriacide. It should be sprinkled evenly across the entire surface of the water in the chiller's tank. c. 8 parts per million chlorine.
---------------------------------------------------------------------- Russell E. Cook, Ph.D. Electron Microscopy Center for Materials Research Argonne National Laboratory Materials Science Division, Building 212 9700 South Cass Avenue Argonne, IL 60439-4838 (630)252-7194 FAX: (630)252-4289 recook-at-anl.gov
Dear Tobias, The most logical explanation for this phenomenon is that the carbon-based cell material is of a lower average atomic number, even with the thin Pt coating, than the glass coverslip (SiO2) it is located on, also with the Pt coat. The secondary electron yield is directly proportional to the average atomic number, if topographic effects are not in evidence, which they won't be on such a thin sample. At 11:39 AM 2/23/00 -0600, you wrote: } Greetings, } I wonder if anyone could explain the following observation } to me. I have been sputter coating with platinum and viewing the } sample with a FESEM (Hitatchi 4700, below lens type). In a certain } type of sample, the image appears darker than the background where } there is sample. Now these samples are perhaps a bit unusual. They } are very thin, starting off life as 1.75 um methacrylate sections of } a plant root adhered to a glass coverslip, followed by extraction in } hot acidic peroxide to remove all organic material except crystalline } cellulose. The cellulosic cell walls left behind on the coverslip are } nominally 100 nm thick and perhaps flattened further by the } processing. The cell walls are present in the sample as either cross } sections or as small regions laying in the plane of the coverslip. } These regions can be anywhere from the size of a cell (10 x 40 or so } microns) down to small portions thereof. All of this remaining cell } wall is darker than the surrounding background. I am putting on a } thin coat of Pt, somewhere around 2 nm, and imaging at 2 kV (I have } seen the same thing at 5 kV). It really puzzles me that my sample is } darker than background. Any explanations??? } } THanks, } Tobias
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
You have a basic problem - free lipids interfere with the polymerization of epoxies. Since you are having mixed results with osmium not all lipids may be fixing adequately. If possible, try keeping the specimens in osmium overnight, on a rotator. (After the first hour of exposure to osmium, change to fresh osmium). I have done this very successfully with lipid rich structures. Hope it works.
} ------------------------------------------------------------------------ } 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 Microscopist, } } has anyone out there any experiences with immuno-labeling of osmificated and } Spurr embedded samples. We have tried out etching, oxidizing sections with } hydrogen peroxide (10% for 15 min) or sodium methylat (1% for 15 min) } without any significant increase of label intensity (Ab used is polyclonal } from rabbit, protein A purified) } } Any suggestions are welcome. Manfred } } } Hello,
You may have a problem which you do not recognize. Spurr's really has no place in immunocytochemistry. It is highly crosslinked. As you might know, epoxies not only crosslink, they also chemically combine with tissue proteins making them unavailable. Also, a highly stable, highly crosslinked embeddding medium will cut a shiny, very smooth surface on sections. The surface relief of such a section is extremely low, and, of course, the more surface relief, the more chance for the antigen to be exposed to markers. You might also have a paucity of antigen which compounds all problems.
If you want to stay with epoxies, I would suggest switching to a soft Epon-Araldite for starters. If that does not work, then my suggestion is to go to LR Gold. A block of LR Gold does not "cut", but cleaves, causes something like a triple increase in surface relief thus giving more antigens the chance to be exposed.
If I am not making myself clear, please e-mail me. I have just gone through 2yrs of frustration with stuff like this, but this week the paper is going out!!!!
Bye, Hildy Crowley University of Denver Denver, CO
P.S. I am not making all this up. I can reference everything, but do not have the time to do that in this format
Before adding glycol to your Haskris water chiller check with the manufacture of the microscope. If you use glycol to cool a Hitachi microscope you will damage the plastic water lines and they will all need to be replaced after a few years. I have seen this happen a few times now after people have used this advice from the list server. Please check with your service engineer first.
To those users of Hitachi S2500/S570, glycol is used to cool the objective lense but only on these 2 instruments.
I am a service engineer for Hitachi in Canada and the above opinion is my opinion and not that of Hitachi.
} I quote here recommendations from the Haskris Co. (we have several Haskris } chillers): } } 1. If algae is present, flush the system with one of two alternatives: add } 1 cup of chlorine bleach or one pint of hydrogen peroxide per 15 gallons of } water. Circulate 20-30 minutes. Drain the system and refill with clean } water. } } 2. To prevent regrowth of algae, one of the following additives are } recommended: } a. 10% solution of lab grade (no additives) ethylene glycol. } b. Dichlorophene. This is an insoluble white powder which is a } fungicide/bacteriacide. It should be sprinkled evenly across the entire } surface of the water in the chiller's tank. } c. 8 parts per million chlorine. } } ---------------------------------------------------------------------- } Russell E. Cook, Ph.D. } Electron Microscopy Center for Materials Research } Argonne National Laboratory } Materials Science Division, Building 212 } 9700 South Cass Avenue } Argonne, IL 60439-4838 } (630)252-7194 } FAX: (630)252-4289 } recook-at-anl.gov } }
I've been using dichlorophene in our Haskris chillers following the incident related below. If you want to buy some, dichlorophene is listed in several chemical company catalogs as:
2,2'-Methylenebis(4-chlorophenol)
Considering how I was planning to use it, I bought technical grade (90%), since it was substantially less expensive than higher grades and have not experienced any problems.
A complete replacement of our chiller water (along with the water in the scope and lines) was required about a year ago when the water pump in our TEM Haskris had to be replaced. Our sevice engineer flushed the system using the concentrated hydrogen peroxide as mentioned above, and a truly amazing amount of material spewed forth, so much so that small connectors to the power supply clogged up a few weeks down the line, causing our HV to shut down. Cleaning out the connectors and flushing the system again with distilled water took care of the problem.
At the time, we were cautioned (by Haskris, I think) to use distilled water and not deionized water in the chillers. Does anyone know why?
Heather Owen
Heather A. Owen, Director Electron Microscope Laboratory Department of Biological Sciences University of Wisconsin - Milwaukee (414)229-6816
What is the real answer? When we had our Hitachi S-3200N installed two years ago, the service engineer used an ethylene glycol/water solution in the Haskris chiller. And it was pretty strong, too. Something like 1:3 glycol:water.
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====================================== Carl Henderson Electron Microbeam Analysis Laboratory University of Michigan 2501 C.C. Little Bldg. Ann Arbor, MI 48109-1063 USA (734) 936-1550 FAX (734) 763-4690 ======================================
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If you can puncture the eye, or preferably take off the anterior chamber, so that the chemicals can get in, there is absolutely no problem. Standard embedding, using longish times, works well. For instance, 3 hrs in osmium, 1 hr in UAc, 20 mins in each of the alcohols/acetone, 1 hr in resin/acetone (dilute), overnight infiltration in resin/acetone (more concentrated), 1 hr in warm resin, embed. If you need to have the eye unpunctured, it may be difficult. I've never embedded a discrete eye, but I do know that it is really difficult to get even isolated sclera well embedded, as it seems to act as a very effective barrier to resin. The eye is a well designed ball that doesn't want to deflate, but that also means nothing much will get in.
Contact me if you want further info. I've embedded more eyes than I care to remember, including some rats eyes.
Diana
Diana van Driel Department of Clinical Ophthalmology Sydney University GPO Box 4337 Sydney NSW AUSTRALIA 2001
You are correct Carl you can use glycol to cool your sem(all Hitachi sems) as only the diffusion pump is being cooled and Hitachi uses a vinyl hose that isn't effected by the Glycol. For those that have Hitachi TEM's the water lines for the lenses and diffusion pumps are the smaller black appearing lines, these will be adversely effected by the glycol.
Sorry for the confusion I will be more specific in the future. So in the sem's that use the clear reinforced vinyl hoses only, glycol is okay. In Hitachi TEM which use a small black plastic hose for the lenses and diffusion pumps, do not use glycol.
} OK, now I am concerned.
} What is the real answer? When we had our Hitachi S-3200N installed } two years ago, the service engineer used an ethylene glycol/water } solution in the Haskris chiller. And it was pretty strong, too. } Something like 1:3 glycol:water.
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I am interested in doing some PC based image analysis of fibers and fiber properties on crimping. Is there anyone doing IA of crimp angle/amplitude or crimps per inch? Is there commercial IA software that will do these types of measurements?
Any help would be appreciated. Thanks.
Duane Krueger 7635 Harold Avenue Golden Valley, Minnesota 55427
} Glycols won't have near the heat capacity of water. I doubt } you will find a bug that will live in ethylene glycol but you can } probably find many that can live in propylene glycol. }
Dear Gordon, I've been trying to look up the heat capacity of glycols in my Handbook of Chemistry and Physics. It doesn't have the data I want, but it does have the Cp for water (~4 j/gK), butane (~0.006 j/molK = ~10^-4 j/gK, if I'm reading my cal's and Cal's right), and 1-propanol (~0.01 j/molK with the same caviat). } From these, I'd guess Cp for ethylene glycol is ~0.014 j/molK, which is, indeed, much less than that of water. Propylene glycol's Cp should be similar. Yours, Bill Tivol
} A related question from India. We have been using DM (demineralised) water in our JEOL 2000 EX II cooled by a Neslab chiller for the last ten years. This requires change every 6 months approximately. What additives can be used to prevent algae (? it is green and slimy and I am not a biologist) growth? Something obtainable locally (chemical name rather than trade names) will be most useful to me. Thanks very much.
Dear Divakar, Russell's suggestion of dichlorophene is good--it's also what we use. To prevent growth in the tubing, we added some CuSO4--being careful to keep the pH slightly above 7.5. We have copper tubing in the system, so the Cu++ should not be a problem, and it will inhibit algal growth. At that pH, CuSO4 is not too soluble, so go slowly (if at all). Yours, Bill Tivol
} A complete replacement of our chiller water (along with the water in the } scope and lines) was required about a year ago when the water pump in our } TEM Haskris had to be replaced. Our sevice engineer flushed the system } using the concentrated hydrogen peroxide as mentioned above, and a truly } amazing amount of material spewed forth, so much so that small connectors } to the power supply clogged up a few weeks down the line, causing our HV } to shut down. Cleaning out the connectors and flushing the system again } with distilled water took care of the problem. } }
We added inline filters--the ones with the fiber cartridges--to trap all the particulate material in the lines, and over a period of several months, we probably got as much material as you got all at once. I highly recommend adding filters.
At the time, we were cautioned (by Haskris, I think) to use distilled
} water and not deionized water in the chillers. Does anyone know why? }
Hard to say. Deionized water can contain material leached out of the resin used, and maybe that is bad for the system, while volatiles (mostly organics, I'd guess) from distillation may not be. Yours, Bill Tivol
A student wishes to take a number of serial sections through a bee brain mounted in glycerine. The total distance is around 500µm and she would like to include three reference points which enable the sections to be correctly orientated throughout the brain. Embedding three small rods seems to be the most likely option but the material would need to be stable and able to be cut by a vibratome. Keeping the rods parallel would also be beneficial.
If anyone has tried this sort of thing or has any suggestions as to what we could use for the markers I'd appreciate hearing your bright ideas.
Regards
Andrew McNaughton
______________________________________________________________________________ Andrew McNaughton South Campus Electron Microscope Unit C/-Department of Anatomy and Structural Biology School of Medical Sciences University of Otago PO Box 913, Dunedin NEW ZEALAND
We are using sputter coater in "etch" mode for this purpose. It is doing the job flawlessly. I don't know about your coater but I'll give you the settings we are using here. The coater has maximum operating voltage of 1400V. The grids are put on glass plate (so they do not make contact with the lower electrode) and then the plate is put on the lower electrode. The current we use is 5 mA for 20 seconds (the current depends strongly on the pressure and the voltage). The pressure is about 20 Pa.
Good luck !
Rado
--------------------------------------------------------------------- Radostin Danev Laboratory of Ultrastructure Research National Institute for Physiological Sciences Myodaiji-cho, Okazaki 444-8585, JAPAN e-mail: rado-at-nips.ac.jp --------------------------------------------------------------------- ----- Original Message ----- } From: Timothy Schneider {Timothy.Schneider-at-mail.tju.edu} To: {microscopy-at-sparc5.microscopy.com} Sent: Thursday, February 24, 2000 4:29 AM
Dear Microscopy users,
I just want to inform you about a new web site (http://www.nanofactory.com) with some useful information on scanning probe microscopy (SPM)
There are, for example: - Weekly update of SPM research articles - Suggestions of some SPM literature - Links to many SPM research groups - Links to almost all SPM companies
There are also some interesting mpeg-movies with TEM-STM measurements of two tunneling tips jumping into each other.
Best regards, Hakan Olin -- Dr Hakan Olin Physics and Engineering Physics Chalmers University of Technology SE-412 96 Goteborg Sweden tel. +46-31-772 3338 fax +46-31-772 3367 mobile +46-70-30 88 99 0 e-mail hakan.olin-at-fy.chalmers.se home page http://fy.chalmers.se/~f4aolin
An effective way to prevent your cooling system from growing green or blue-green algae is to ensure that it is totally light-tight. Algae require light for photosynthesis. The Philips engineers who installed our CM120 Biotwin recommended we connect up the chiller with the black-lined vinyl hose used in drinks vending machines, where microorganism growth in the water lines is obviously undesirable. The product we used is trademarked "Aquavend", is opaque white outside and black inside. It appears to have worked so far, though the ethylene glycol we use would also discourage algae. Yours Chris Jeffree
Date sent: Thu, 24 Feb 2000 18:08:15 -0500 } From: William Tivol {tivol-at-wadsworth.org} To: microscopy-at-sparc5.microscopy.com
Dear Debby, Some ten yaers ago we were doing research on sugar syrup in cooperation with a sugar company. They were interested in the stucture of mixtures of sugar syrups. W e had some succes using freeze-fracture and looking at the replicas in the TEM. Of course these were not mixtures of cereals with sugar syrup and the specimens were allowed to be a lot smaller but this could be a suggestion. Succes ! Wim Jacob Ctr.of Electron Microscopy University of Antwerp Belgium
Debby Sherman 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, } Well I guess we never run out of new challenges when it comes to specimen prep. This one concerns a product that is made of a cereal bound together with a sugar syrup. The investigators want to see how the syrup behaves as to structural integrety over a period of time with and without the addition of various stabilizing components. Information would be collected at the light microscopic level. } We first thought of cryosectioning followed by examination using LM. However, the product is filled with air pockets and literally disintegrates when microtomed. It is easily cut into pieces at temperatures above freezing as long as the pieces are no thinner than3-4 mmAccempts to infiltrate it with OCT compoound prior to freezing were not satisfactory. } New thought is to use a low temperature resin to infiltrate the sample followed by UV or chemical polymerization. butThe resulting blocks could then be microtomed at room temperature and imaged. It would be desirable to cut sections of 4-8 µm in thickness which is too thick for an ultramicrotome with glass or diamond knives. Also UV polymerization would probably limit us to very small sample size which may be a problem. } Any suggestions would be appreciated as to resins to use which have low viscosity, could be used to infiltrate pieces of ~0.5-1cm sq, polymerize at low temperatures (+4 to 0oC range) using UV or chemical polymerization, and are preferably soft enough to section with a metal microtome blade. Any other suggestions of preparations techniques to try would be very appreciated. } } Thanks in advance for your input. } Debby } } Debby Sherman, Manager Phone: 765-494-6666 } Microscopy Center in Agriculture FAX: 765-494-5896 } Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu } Purdue University } 1057 Whistler Building } West Lafayette, IN 47907-1057
Dear Doug, Why don't you try imaging SIMS? I would not surprised if such instrument(s) are present at the University of Arizona. Of course this would restrict the resolution, I gess, to 0.1 - 0.5 micrometer, with the newer instruments. Regards Wim Jacob Ctr. of Electron Microscopy University of Antwerp Belgium
Doug Cromey 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 } -----------------------------------------------------------------------. } } Colleagues, } } We are interested in localizing arsenic in biological tissues (toxicology } research), but have been unable to come up with a technique that will give } us the cellular localization information we're looking for (sub-cellular } would be even better). } } We have tried TEM with energy dispersive x-ray microanalysis and our } concentrations are apparently too low, so we are unable to detect arsenic. } } A few years ago we looked into using the autometallographic techniques that } have been published by Dr. Gorm Dansher & his colleagues. We were told } that the photographic emulsions required were no longer available. } } In a brainstorming session yesterday several techniques were bantered } about, but in truth we are not familiar enough with the techniques to know } if they would suit our needs. Does EELS have anything to offer, or perhaps } wavelength dispersive spectoscopy (WDS)? How about some of the new FT-IR } instruments? Are there any other techniques that might be able to localize } arsenic and/or indicate its state of oxidation? } } Vendors are welcome to reply. } } Yours, } Doug Cromey } .................................................................... } : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : } : Research Specialist, Principal University of Arizona : } : (office: AHSC 4212A) P.O. Box 245044 : } : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : } : (FAX: 520-626-2097) (email: doug-cromey-at-ns.arizona.edu): } :...................................................................: } http://www.pharmacy.arizona.edu/exp_path.html } Home of: "Microscopy and Imaging Resources on the WWW"
Please visit my new homepage at http://syli.homepage.com at your convinience! It contains not only my current research work, but also LINKS to useful journal informations, instruction for authors, on-line EELS database, periodic table, physical constants, online TEM course and etc.
I am looking forward to your invaluable suggestions! Yours sincerely,
Shu-You Li
************************************************** Shu-You Li, Dr. Institut fuer Physikalische Chemie Johannes Guttenberg Universitaet Jakob-Welder-Weg 11 D-55099 Mainz, Germany
Becky, I do not know if it is okay in the S4700 to use glycol. If it is working as in why change?
X-From_: r-holdford-at-ti.com Thu Feb 24 19:53:24 2000
X-From_: Patrick.Kilcran-at-hii-hitachi.com Thu Feb 24 23:55:28 2000 } From: "Kilcran, Patrick-HIIEX" {Patrick.Kilcran-at-hii-hitachi.com} To: dhoyle-at-istar.ca Cc: "Lima, Rick-HIIEX" {Rick.Lima-at-hii-hitachi.com}
Dear Researcher:
The University of Florida College of Medicine Electron Microscopy Facility is hosting a two-day workshop on immunogold technique. The workshop will include two identical sessions on April 10/11 and 13/14. Dr. Jan Leunissen from the Aurion Immunogold Reagent & Accessories, an internationally known expert in the field, will be the instructor for the workshop. Attached is the program information. If you are interested in attending this workshop, please contact Dr. Verlander or Ms. Hong Yi at the addresses provided below. Due to practical considerations, the workshop will be limited to a maximum of 20 participants for each session. Therefore, we encourage you to register as soon as possible. If you are not able to attend, we would appreciate if you could pass the message onto someone else who might be interested in learning about immunogold techniques. Thank you very much.
Dr. Jill W. Verlander Reed (workshop faculty) Director, University of Florida College of Medicine Electron Microscopy Facility Phone: (352) 846-0820 Fax: (352) 846-3299 verlaj-at-medicine.ufl.edu
Hong Yi (technical coordinator) Supervisor, Emory Neurology Microscopy Core Laboratory Phone: (404) 727-8692 Fax: (404) 727-3157 Email: hyi-at-emory.edu
University of Florida, College of Medicine, Electron Microscopy Facility Aurion Immunogold Reagents & Accessories / Electron Microscopy Sciences
Present Immunogold Workshop
April 10-14, 2000 Gainesville, FL
WORKSHOP HOST Dr. Jill Verlander Reed Director, Electron Microscopy Facility University of Florida College of Medicine Phone: (352) 846-0820 Fax: (352) 846-3299 verlaj-at-medicine.ufl.edu
WORKSHOP INSTRUCTORS
Dr. Jan L.M. Leunissen is an internationally known scientist in the field of ultrastructural localization. He received his Ph.D in molecular cell biology at the University of Utrecht in the Netherlands. Dr. Leunissen is the inventor of ultra small colloidal gold probes and also the founder and president of the company Aurion, which specializes in immunogold reagents and custom immunogold labeling. He has been invited to many international microscopy conferences and workshops and is especially experienced in providing hands-on training. His previous workshops in Europe, Asia, and the US have been fully attended and very well received.
Dr. Jill W. Verlander is the Director of the Electron Microscopy Facility for the University of Florida College of Medicine. Dr. Verlander has 15 years of experience in ultrastructural research in kidney and is known internationally for her work examining structure-function relationships in renal transport epithelia. Her work has been published in such journals as the Journal of Clinical Investigation, the American Journal of Physiology, Journal of the American Society of Nephrology, and Kidney International. These studies have been accomplished using light microscopic immunohistochemistry and in situ hybridization, scanning and transmission electron microscopy, morphometric analyses, and immunogold and immunoperoxidase cytochemistry at the ultrastructural level.
GENERAL INFORMATION Objectives To provide researchers the opportunity to learn the theory and practice of immunogold labeling To permit participants to process their own samples using these techniques under expert guidance To promote technology exchange and research collaboration
Rooms will be reserved in the above hotel under the name "UF EM workshop". Shuttle buses are available between the hotel and UF campus
Enrollment Note Two sessions (A; April 10-11, B: April 13-14) will be hosted at UF. Each session will be limited to a maximum of 20 participants. The workshops will provide samples to those who prefer not to bring their own. Each participant will be provided with a sample of gold conjugate of their choice at the end of the workshop.
MAIN CURRICULUM The properties of gold particles and their protein conjugates. Theories underlying immunogold labeling protocols. Silver enhancement of gold particles Imunogold labeling on a variety of sample preparations for LM. Immunogold labeling for EM a. Pre-embedding immunogold labeling using ultra_small gold conjugates and silver enhancement. b. Post-embedding immunogold labeling using ultra small and conventional colloidal gold conjugates.
CONTACT PERSON AND TECHNICAL COORDINATOR Hong Yi Phone: (404) 727-8692 Fax: (404) 727-3157 Email: hyi-at-emory.edu
Jill Verlander Reed, D.V.M. Associate Scientist Office of the Dean University of Florida College of Medicine P.O. Box 100215 Gainesville, FL 32610 verlaj-at-medicine.ufl.edu Phone: (352) 846-0820 Fax: (352) 846-3299
Molecular Imaging is updating our contact information.
If you are interested in scanning probe microscopy, would you please take a moment and update your information at http://www.molec.com/info/form.html
Thank you
George
_______________________ George Sibbald, President Molecular Imaging: Technology Leader In Advanced Bio-AFM Systems 9830 South 51 Street, Ste 124A Phoenix AZ 85044 (480) 753-4311 www.molec.com
In fact, we have an S-4700 and the service technician told us that there are tons of problems with using glycol in the chillers. We have a Haskris chiller. Apparently, the glycol volatilizes some of the hose components.
} } Looking to find and to determine what is the best printer (photographic and } archive quality) } for TEM digital images. } } Currently considering Kodak 8670PS, Codonics, or Fuji Printers } Would be interested in comments from people using photoraphic qualtiy } printers } for TEM applications } thoughts, "next time's" etc. } } Thank You } } Gregory Argentieri } Novartis Pharmacuticals Corp } gregory.argentieri-at-pharma.novartis.com } } } }
Debby Sherman Talk to Mark Cavaleri at 3M. He made a polished cross sectgion of M%M candies and preserved tyhe sugr layer
Sam Purdy
} ---------- } From: Debby Sherman } Sent: February 2000 1:17 PM } To: message to: MSA list } Subject: Food Science Prep. } } ------------------------------------------------------------------------ } 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, } Well I guess we never run out of new challenges when it comes to } specimen prep. This one concerns a product that is made of a cereal bound } together with a sugar syrup. The investigators want to see how the syrup } behaves as to structural integrety over a period of time with and without } the addition of various stabilizing components. Information would be } collected at the light microscopic level. } We first thought of cryosectioning followed by examination using LM. } However, the product is filled with air pockets and literally } disintegrates when microtomed. It is easily cut into pieces at } temperatures above freezing as long as the pieces are no thinner than3-4 } mmAccempts to infiltrate it with OCT compoound prior to freezing were not } satisfactory. } New thought is to use a low temperature resin to infiltrate the sample } followed by UV or chemical polymerization. butThe resulting blocks could } then be microtomed at room temperature and imaged. It would be desirable } to cut sections of 4-8 µm in thickness which is too thick for an } ultramicrotome with glass or diamond knives. Also UV polymerization would } probably limit us to very small sample size which may be a problem. } Any suggestions would be appreciated as to resins to use which have low } viscosity, could be used to infiltrate pieces of ~0.5-1cm sq, polymerize } at low temperatures (+4 to 0oC range) using UV or chemical polymerization, } and are preferably soft enough to section with a metal microtome blade. } Any other suggestions of preparations techniques to try would be very } appreciated. } } Thanks in advance for your input. } Debby } } Debby Sherman, Manager Phone: 765-494-6666 } Microscopy Center in Agriculture FAX: 765-494-5896 } Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu } Purdue University } 1057 Whistler Building } West Lafayette, IN 47907-1057 } }
I received this today. If anyone could help, please reply to the sender.
Thanks.
ML
} Date: Fri, 25 Feb 2000 17:14:04 -0600 } From: Howard Hsu {hhsu-at-kumc.edu} } Organization: KU Medical Center } X-Accept-Language: en } MIME-Version: 1.0 } To: wong-at-msg.ucsf.edu } Subject: sem } } I am a researcher at the University of Kansas Med. Center and would like } study a biological sample for the presence of mineral using SEM } approach. If you can provide external service on cost or collaborative } basis, please let me know. Otherwise, I would greatly appreciate if you } would let me know any academic or industrial sources that provide this } type of service. Thank you. Howard Hsu, Ph.D. } Mei Lie Wong Electron Microscope Laboratory Department of Biochemistry HHMI-UCSF Ph. 415-476-4441 Fax 415-476-1902 http://util.ucsf.edu/agard/wong/index.html email wong-at-msg.ucsf.edu
We are a hospital based research lab. starting to use the electron microscope to identify or confirm pathogens that cause gastrointestinal disturbances. We would like to know if someone out there could recommend appropriate specific EM protocols (negative staining, processing of the gastrointestinal contents, etc....) to visualize these pathogens (ranging from viruses to bacteria and protozoans). We have a Jeol 1010 EM and perform the routine EM preparations designed to examine tissues.
In the interest of bring quality healthcare to our people, we will be very much obliged,
Mei Lie Wong wrote: ================================================== I received this today. If anyone could help, please reply to the sender.
Thanks.
ML
} Date: Fri, 25 Feb 2000 17:14:04 -0600 } From: Howard Hsu {hhsu-at-kumc.edu} } Organization: KU Medical Center } X-Accept-Language: en } MIME-Version: 1.0 } To: wong-at-msg.ucsf.edu } Subject: sem } } I am a researcher at the University of Kansas Med. Center and would like } study a biological sample for the presence of mineral using SEM } approach. If you can provide external service on cost or collaborative } basis, please let me know. Otherwise, I would greatly appreciate if you } would let me know any academic or industrial sources that provide this } type of service. Thank you. Howard Hsu, Ph.D. } Mei Lie Wong Electron Microscope Laboratory Department of Biochemistry HHMI-UCSF Ph. 415-476-4441 Fax 415-476-1902 http://util.ucsf.edu/agard/wong/index.html email wong-at-msg.ucsf.edu ============================================================ Although you have specifically asked about SEM, I am aware only of a TEM approach for this kind of analysis. We at Structure Probe, Inc. do perform this kind of work for clients on a commercial basis. You can see an example of the final sample that is analyzed by TEM on our website at page URL http://www.2spi.com/catalog/instruments/etchers4.html
The method is pretty much described so anyone really could do with, provided they did have access by a plasma etcher such as the SPI Plasma Prep II. If you wanted to do it yourself and were having difficulties, let us know, perhaps we could give you assistance. We have found that the removal of the organics is usually quite important in order to remove the source of the unwanted Bremsstrahlung radiation.
We have heard from persons trying to do this by SEM/EDS, where by the section is deposited on a polished beryllium mount, and then the organics etched away but they seem to have not been successful.
Disclaimer: SPI Supplies manufactures the plasma etching equipment needed to practice this procedure and our Structure Probe® analytical services part of our business performs this kind of sample preparation and analysis for clients. We realize there might be other methods for doing this, but we have favored the approach that works for us.
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.spi.cc ######################## ============================================
We have some experience in using micro-PIXE (proton induced X-ray emission) and the so-called nuclear microprobe in toxicological research. It is more sensitive than EDS and WDS, is very easy to quantify (standardless technique). In principle you can have detection limits of the order of 1 ppm in point analysis mode. Specimens can be of any thickness and their actual thickness is assessed by the simultaneous use of the BS technique (proton backscatering spectrometry). What in my opinion makes it really attractive for this purpose is its very good scanning capability. Scan sizes are from ca. 2.5 mm x 2.5 mm down to the beam resolution, which is of the order of few micrometer (can be down to 1 micrometer or even less, but in most of our uses few micrometers is good enough). The sample preparation is similar as for EDS, using cryotechniques to avoid elemental losses and redistribution. We have so far done studies on Ni, Mo, Cd. The detection limits for As are better than for Mo and Cd. Of course, this technique cannot indicate the state of oxidation, you can really treat it as a "more sensitive EDS".
If you want more information, I can refer you to our earlier work and perhaps you might be interested in contacting me.
Best regards
xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx Dr Wojciech J. Przybylowicz Materials Research Group National Accelerator Centre PO Box 72 Faure 7131 South Africa E-mail: przybylowicz-at-nac.ac.za Fax: +27-21-8433543 Phone: +27-21-8433820 http://www.nac.ac.za/IBA/ http://www.nac.ac.za/materials/ xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx
We have some experience in using micro-PIXE (proton induced X-ray emission) and the so-called nuclear microprobe in toxicological research. It is more sensitive than EDS and WDS, is very easy to quantify. In principle you can have detection limits of the order of 1 ppm in point analysis mode. Specimens can be of any thickness and their actual thickness is assessed by the simultaneous use of the BS technique (proton backscatering spectrometry). What in my opinion makes it really attractive for this purpose is its very good scanning capability. Scan sizes are from ca. 2.5 mm x 2.5 mm down to the beam resolution, which is of the order of few micrometer (can be down to 1 micrometer or even less, but in most of our uses few micrometers is good enough). The sample preparation is similar as for EDS, using cryotechniques to avoid elemental losses and redistribution. We have so far done studies on Ni, Mo, Cd. The detection limits for As are better than for Mo and Cd. Of course, this technique cannot indicate the state of oxidation, you can really treat it as a "more sensitive EDS".
If you want more information, I can refer you to our earlier work and perhaps you might be interested in contacting me.
Best regards
xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx Dr Wojciech J. Przybylowicz Materials Research Group National Accelerator Centre PO Box 72 Faure 7131 South Africa E-mail: przybylowicz-at-nac.ac.za Fax: +27-21-8433543 Phone: +27-21-8433820 http://www.nac.ac.za/IBA/ http://www.nac.ac.za/materials/ xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx
An information about a new principle in vibration damping to all microscopy users, who have problems with particularly low frequency vibrations from natural environment.
I invite you kindly to visit our home page: http://www.hwlbioanalytic.com
We show there active vibration isolation systems for various applications with damping frequencies from 0,6 Hz to 100/200 Hz and load capacities (depending from system) from max. 90 kg up to 330/660 kg and max. 1.500 kg. Thank you for your time.
We use many of the techniques and the atlas from the following excellent book. "EM in Diagnostic Virology" by Doan and Anderson,1987, Cambridge University Press, ISBN 0 521 24311 4.
-- Tracy Gales Fox Chase Cancer Center Electron Microscopy Facility 7701 Burholme Ave. Philadelphia PA 19111 Tel. 215-728-2484 Fax 215-728-2412
A friend recently retired and gave me a TEM diffraction plate reader. This was made by Ernest F. Fullam, Inc. in the early 1970's. It is still in the wooden shipping crate, is in unused condition and looks complete except there is no manual. It is a basic no-frills manual version. It has a high quality Starrett vernier scale and could be used for measurement of any electron micrograph, or other type of transparency, or it may be useful for teaching the history and theory of diffraction pattern measurement.
I would be willing to send it to someone willing to pay the shipping costs.
Thanks, Louie
Louie Kerr Research and Education Support Coordinator Marine Biological Laboratory 7 MBL Street Woods Hole, MA 02543 508-289-7273 508-540-6902 (FAX) 508-292-0289 (Cell phone)
The University of Kentucky invites applications for a Research Associate in the area of electron microscopy. The Electron Microscopy Facility houses two SEMs, two TEMs one AFM and extensive sample preparation equipment that are used by undergraduate and graduate students in support of research and education. The primary responsibility of the Research Associate will be to oversee the JEOL 2010F Field Emission TEM and auxiliary equipment, including an Oxford energy dispersive x-ray spectrometer (EDS), Gatan electron energy loss imaging filter and acquisition hardware and software. S/he will be responsible for coordinating and administering maintenance, access, training and use of the equipment by internal and external users. S/he will also be encouraged to develop sponsored research programs and collaborate with university faculty members. The qualified candidate will have a Ph.D. and practical experience in analytical electron microscopy and demonstrate good communication skills. The salary will be commensurate with qualifications and experience. The University of Kentucky offers comprehensive insurance and benefits packages and is an equal opportunity employer. Please send applications to:
We perform about 1000 exams/yr for viruses using EM. Below are some refs (check out especially 2, 5, 9, 11, 14). Also, at the bottom is a synopsis I give to our medical students; maybe it will be useful.
As for enteric viruses, EM is the best way to look for them. Many of them do not grow in tissue culture, and those that can be coaxed, do so under special conditions that are not employed in the routine culture lab. Biochemical identification (e.g., immunological kits, PCR, Western Blots) require a specific reagent to recognize the virus and if you don't use the right reagent, you will miss viruses (e.g., if you run a test for rotavirus, you will miss adenovirus, etc.) Furthermore, there aren't biochemical reagents for all viruses. With EM, you can see a wide variety of viruses by doing a negative stain of an aqueous extract of stool.
Good luck, Sara
REFERENCES ON VIRUS IDENTIFICATION BY ELECTRON MICROSCOPY
1. Cegielski JP, Msengi AE, Miller SE. 1994. Enteric viruses associated with HIV infection in Tanzanian children with chronic diarrhea. Ped AIDS HIV Inf 5:296-299.
2. Doane FW, Anderson N. 1986. Electron Microscopy in Diagnostic Virology: A Practical Guide and Atlas, Cambridge University Press, New York.
3. Lew JF, Glass, RI, Petric M, LeBaron CW, Hammond GW, Miller SE, Robinson C, Boutilier J, Riepenhoff-Talty M, Payne CM, Franklin R, Oshiro LS, and Jaqua M-J. 1990. Six year retrospective surveillance of gastroenteritis viruses identified at ten electron microscopy centers in the United States and Canada Pediatr Infect Dis J 9:709-714.
4. Mclean DM, Wong KK. 1984. Same Day Diagnosis of Human Virus Infections, CRC Press, Boca Raton, FL.
5. Miller SE. 1986. Detection and identification of viruses by electron microscopy. J Electron Microsc Tech 4:265-301.
5. Miller SE, Howell DN. 1988. Viral infections in the acquired immunodeficiency syndrome. J Electron Microsc Tech 8:41-78.
7. Miller SE. 1988. Diagnostic virology by electron microscopy. ASM News 54:475-481.
8. Miller SE. 1989. Electron microscopy in rapid viral diagnosis. EMSA Bull 19:53-59.
9. Hayat MA, Miller SE. 1990. Negative Staining: Applications and Methods. McGraw-Hill, New York.
10. Miller SE. 1991. Evaluation of electron microscopic information available from clinical samples. In de la Maza LM, Peterson EM (eds.), Medical Virology, Vol 10. Irvine, CA. pp 21-54.
11. Miller SE. 1995. Diagnosis of viral infection by electron microscopy. In Lennette EH, Lennette DA, Lennette ET (eds), Diagnostic Procedures for Viral, Rickettsial and Chlamydial Infections, American Public Health Assoc, Washington, DC. pp 35-76.
12. Oshiro LS, Miller SE. 1992. Application of electron microscopy to the diagnosis of viral infections. In Lennette EH (ed), Laboratory Diagnosis of Viral Infections. Marcel Dekker, New York. pp 45-68.
13. Palmer E, Martin ML. 1982. An Atlas of Mammalian Viruses, CRC Press, Boca Raton, FL.
14. Palmer E, Martin ML. 1988. Electron Microscopy in Viral Diagnosis, CRC Press, Boca Raton, FL.
15. Murphy FA, Fauquet CM, Bishop DHL, Ghabrial SA, Jarvis AW, GP Martelli, Mayo MA, Summers MD. 1995. Virus Taxonomy. Sixth Report of the International Committee on Taxonomy of Viruses. Springer-Verlag, New York.
2, 5, 9, 11-14 = Methods and micrographs 2, 14 = Excellent atlases 7, 8 = Brief synopses of technique and basis of virus ID 15 = Good general reference, but don't memorize!! Lists all viruses, not just human.
TWO WAYS TO LOOK AT VIRUSES BY EM: Negative staining Used with liquid samples Shows whole virus (size, surface structure important) Thin sectioning Used with cells and tissues Shows a slice of an infected cell Shows relationship of viruses to cell organelles (internal structure and cell organelle association important) Location within the cell is a clue to nucleic acid type DNA viruses are usually seen in the nucleus (some exceptions). RNA viruses are usually seen in the cytoplasm (some exceptions). Exceptions: Naked viruses get out of the cell by lysis; if the cell is very sick, there may be virions everywhere, e.g., adenovirus (DNA). Poxviruses are DNA viruses, but are constructed in the cytoplasm. Hepatitis B core particles (DNA) may be in the nucleus and cytoplasm. Dane particles (complete viruses) are only cytoplasmic. Some paramyxovirus (RNA) nucleocapsids, not the complete virion, can be seen in the nucleus (e.g., measlesvirus). Enveloped viruses are associated with cell membranes; the type of cell membrane (plasma membrane, nuclear membrane, endoplasmic reticulum, vacuoles) can be a clue to virus identification.
SUMMARY OF VIRUS TYPES (based on morphology): Naked viruses All human naked viruses are icosahedral (spherical). There are 3 size ranges from 20-35 nm, 45-55 nm, 70-90 nm. Examples: Small: parvovirus, enterovirus, rhinovirus Medium: polyomavirus, papillomavirus Large: rotavirus/reovirus, adenovirus Enveloped viruses All have a lipoprotein membrane on the outside of the nucleocapsid. Most derive this layer by budding through cell membranes; this pliable layer makes the virion pleomorphic. Poxviruses synthesize their shell de novo; sometimes they pick up an extra covering by budding. They are not pleomorphic, but different varieties may be more ovoid, others, brick-shaped. Most range in size from 40-400 nm. (Filoviruses are 80 nm by up to 1400 nm.) The envelope has proteins on the outside that may look like spikes or fuzz (e.g., influenza virus, coronavirus). Sometimes the spikes are too short to be distinguished by EM, making the virion appear smooth (e.g., herpesviruses, rubella virus). The nucleocapsid shape can be a. icosahedral, like the naked viruses; however, some are morphologically nondescript as seen by EM; b. filamentous, like a "slinky." c. complex (poxvirus cores look like dumbbells with lateral bodies) d. morphologically nondescript (just dense without any particular shape). Enveloped DNA viruses (except pox) construct their nucleocapsids in the nucleus. Enveloped RNA viruses construct their nucleocapsids in the cytoplasm; (measles nucleocapsids can sometimes be seen in the nucleus).
On Sat, 26 Feb 2000, Ronald R. Matias wrote:
} Date: Sat, 26 Feb 2000 08:19:49 +0800 } From: Ronald R. Matias {rrm-at-skyinet.net} } To: Microscopy-at-sparc5.microscopy.com } Subject: Need help on EM negative staining } } ------------------------------------------------------------------------ } 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 and Help! } } We are a hospital based research lab. starting to use the electron } microscope to identify or confirm pathogens that cause gastrointestinal } disturbances. We would like to know if someone out there could recommend } appropriate specific EM protocols (negative staining, processing of the } gastrointestinal contents, etc....) to visualize these pathogens (ranging } from viruses to bacteria and protozoans). We have a Jeol 1010 EM and perform } the routine EM preparations designed to examine tissues. } } In the interest of bring quality healthcare to our people, we will be very } much obliged, } } ronnie matias } }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
We perform about 1000 exams/yr for viruses using EM. Below are some refs (check out especially 2, 5, 9, 11, 14). Also, at the bottom is a synopsis I give to our medical students; maybe it will be useful.
As for enteric viruses, EM is the best way to look for them. Many of them do not grow in tissue culture, and those that can be coaxed, do so under special conditions that are not employed in the routine culture lab. Biochemical identification (e.g., immunological kits, PCR, Western Blots) require a specific reagent to recognize the virus and if you don't use the right reagent, you will miss viruses (e.g., if you run a test for rotavirus, you will miss adenovirus, etc.) Furthermore, there aren't biochemical reagents for all viruses. With EM, you can see a wide variety of viruses by doing a negative stain of an aqueous extract of stool.
Good luck, Sara
REFERENCES ON VIRUS IDENTIFICATION BY ELECTRON MICROSCOPY
1. Cegielski JP, Msengi AE, Miller SE. 1994. Enteric viruses associated with HIV infection in Tanzanian children with chronic diarrhea. Ped AIDS HIV Inf 5:296-299.
2. Doane FW, Anderson N. 1986. Electron Microscopy in Diagnostic Virology: A Practical Guide and Atlas, Cambridge University Press, New York.
3. Lew JF, Glass, RI, Petric M, LeBaron CW, Hammond GW, Miller SE, Robinson C, Boutilier J, Riepenhoff-Talty M, Payne CM, Franklin R, Oshiro LS, and Jaqua M-J. 1990. Six year retrospective surveillance of gastroenteritis viruses identified at ten electron microscopy centers in the United States and Canada Pediatr Infect Dis J 9:709-714.
4. Mclean DM, Wong KK. 1984. Same Day Diagnosis of Human Virus Infections, CRC Press, Boca Raton, FL.
5. Miller SE. 1986. Detection and identification of viruses by electron microscopy. J Electron Microsc Tech 4:265-301.
5. Miller SE, Howell DN. 1988. Viral infections in the acquired immunodeficiency syndrome. J Electron Microsc Tech 8:41-78.
7. Miller SE. 1988. Diagnostic virology by electron microscopy. ASM News 54:475-481.
8. Miller SE. 1989. Electron microscopy in rapid viral diagnosis. EMSA Bull 19:53-59.
9. Hayat MA, Miller SE. 1990. Negative Staining: Applications and Methods. McGraw-Hill, New York.
10. Miller SE. 1991. Evaluation of electron microscopic information available from clinical samples. In de la Maza LM, Peterson EM (eds.), Medical Virology, Vol 10. Irvine, CA. pp 21-54.
11. Miller SE. 1995. Diagnosis of viral infection by electron microscopy. In Lennette EH, Lennette DA, Lennette ET (eds), Diagnostic Procedures for Viral, Rickettsial and Chlamydial Infections, American Public Health Assoc, Washington, DC. pp 35-76.
12. Oshiro LS, Miller SE. 1992. Application of electron microscopy to the diagnosis of viral infections. In Lennette EH (ed), Laboratory Diagnosis of Viral Infections. Marcel Dekker, New York. pp 45-68.
13. Palmer E, Martin ML. 1982. An Atlas of Mammalian Viruses, CRC Press, Boca Raton, FL.
14. Palmer E, Martin ML. 1988. Electron Microscopy in Viral Diagnosis, CRC Press, Boca Raton, FL.
15. Murphy FA, Fauquet CM, Bishop DHL, Ghabrial SA, Jarvis AW, GP Martelli, Mayo MA, Summers MD. 1995. Virus Taxonomy. Sixth Report of the International Committee on Taxonomy of Viruses. Springer-Verlag, New York.
2, 5, 9, 11-14 = Methods and micrographs 2, 14 = Excellent atlases 7, 8 = Brief synopses of technique and basis of virus ID 15 = Good general reference, but don't memorize!! Lists all viruses, not just human.
TWO WAYS TO LOOK AT VIRUSES BY EM: Negative staining Used with liquid samples Shows whole virus (size, surface structure important) Thin sectioning Used with cells and tissues Shows a slice of an infected cell Shows relationship of viruses to cell organelles (internal structure and cell organelle association important) Location within the cell is a clue to nucleic acid type DNA viruses are usually seen in the nucleus (some exceptions). RNA viruses are usually seen in the cytoplasm (some exceptions). Exceptions: Naked viruses get out of the cell by lysis; if the cell is very sick, there may be virions everywhere, e.g., adenovirus (DNA). Poxviruses are DNA viruses, but are constructed in the cytoplasm. Hepatitis B core particles (DNA) may be in the nucleus and cytoplasm. Dane particles (complete viruses) are only cytoplasmic. Some paramyxovirus (RNA) nucleocapsids, not the complete virion, can be seen in the nucleus (e.g., measlesvirus). Enveloped viruses are associated with cell membranes; the type of cell membrane (plasma membrane, nuclear membrane, endoplasmic reticulum, vacuoles) can be a clue to virus identification.
SUMMARY OF VIRUS TYPES (based on morphology): Naked viruses All human naked viruses are icosahedral (spherical). There are 3 size ranges from 20-35 nm, 45-55 nm, 70-90 nm. Examples: Small: parvovirus, enterovirus, rhinovirus Medium: polyomavirus, papillomavirus Large: rotavirus/reovirus, adenovirus Enveloped viruses All have a lipoprotein membrane on the outside of the nucleocapsid. Most derive this layer by budding through cell membranes; this pliable layer makes the virion pleomorphic. Poxviruses synthesize their shell de novo; sometimes they pick up an extra covering by budding. They are not pleomorphic, but different varieties may be more ovoid, others, brick-shaped. Most range in size from 40-400 nm. (Filoviruses are 80 nm by up to 1400 nm.) The envelope has proteins on the outside that may look like spikes or fuzz (e.g., influenza virus, coronavirus). Sometimes the spikes are too short to be distinguished by EM, making the virion appear smooth (e.g., herpesviruses, rubella virus). The nucleocapsid shape can be a. icosahedral, like the naked viruses; however, some are morphologically nondescript as seen by EM; b. filamentous, like a "slinky." c. complex (poxvirus cores look like dumbbells with lateral bodies) d. morphologically nondescript (just dense without any particular shape). Enveloped DNA viruses (except pox) construct their nucleocapsids in the nucleus. Enveloped RNA viruses construct their nucleocapsids in the cytoplasm; (measles nucleocapsids can sometimes be seen in the nucleus).
On Sat, 26 Feb 2000, Ronald R. Matias wrote:
} Date: Sat, 26 Feb 2000 08:19:49 +0800 } From: Ronald R. Matias {rrm-at-skyinet.net} } To: Microscopy-at-sparc5.microscopy.com } Subject: Need help on EM negative staining } } ------------------------------------------------------------------------ } 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 and Help! } } We are a hospital based research lab. starting to use the electron } microscope to identify or confirm pathogens that cause gastrointestinal } disturbances. We would like to know if someone out there could recommend } appropriate specific EM protocols (negative staining, processing of the } gastrointestinal contents, etc....) to visualize these pathogens (ranging } from viruses to bacteria and protozoans). We have a Jeol 1010 EM and perform } the routine EM preparations designed to examine tissues. } } In the interest of bring quality healthcare to our people, we will be very } much obliged, } } ronnie matias } }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
We perform about 1000 exams/yr for viruses using EM. Below are some refs (check out especially 2, 5, 9, 11, 14). Also, at the bottom is a synopsis I give to our medical students; maybe it will be useful.
As for enteric viruses, EM is the best way to look for them. Many of them do not grow in tissue culture, and those that can be coaxed, do so under special conditions that are not employed in the routine culture lab. Biochemical identification (e.g., immunological kits, PCR, Western Blots) require a specific reagent to recognize the virus and if you don't use the right reagent, you will miss viruses (e.g., if you run a test for rotavirus, you will miss adenovirus, etc.) Furthermore, there aren't biochemical reagents for all viruses. With EM, you can see a wide variety of viruses by doing a negative stain of an aqueous extract of stool.
Good luck, Sara
REFERENCES ON VIRUS IDENTIFICATION BY ELECTRON MICROSCOPY
1. Cegielski JP, Msengi AE, Miller SE. 1994. Enteric viruses associated with HIV infection in Tanzanian children with chronic diarrhea. Ped AIDS HIV Inf 5:296-299.
2. Doane FW, Anderson N. 1986. Electron Microscopy in Diagnostic Virology: A Practical Guide and Atlas, Cambridge University Press, New York.
3. Lew JF, Glass, RI, Petric M, LeBaron CW, Hammond GW, Miller SE, Robinson C, Boutilier J, Riepenhoff-Talty M, Payne CM, Franklin R, Oshiro LS, and Jaqua M-J. 1990. Six year retrospective surveillance of gastroenteritis viruses identified at ten electron microscopy centers in the United States and Canada Pediatr Infect Dis J 9:709-714.
4. Mclean DM, Wong KK. 1984. Same Day Diagnosis of Human Virus Infections, CRC Press, Boca Raton, FL.
5. Miller SE. 1986. Detection and identification of viruses by electron microscopy. J Electron Microsc Tech 4:265-301.
5. Miller SE, Howell DN. 1988. Viral infections in the acquired immunodeficiency syndrome. J Electron Microsc Tech 8:41-78.
7. Miller SE. 1988. Diagnostic virology by electron microscopy. ASM News 54:475-481.
8. Miller SE. 1989. Electron microscopy in rapid viral diagnosis. EMSA Bull 19:53-59.
9. Hayat MA, Miller SE. 1990. Negative Staining: Applications and Methods. McGraw-Hill, New York.
10. Miller SE. 1991. Evaluation of electron microscopic information available from clinical samples. In de la Maza LM, Peterson EM (eds.), Medical Virology, Vol 10. Irvine, CA. pp 21-54.
11. Miller SE. 1995. Diagnosis of viral infection by electron microscopy. In Lennette EH, Lennette DA, Lennette ET (eds), Diagnostic Procedures for Viral, Rickettsial and Chlamydial Infections, American Public Health Assoc, Washington, DC. pp 35-76.
12. Oshiro LS, Miller SE. 1992. Application of electron microscopy to the diagnosis of viral infections. In Lennette EH (ed), Laboratory Diagnosis of Viral Infections. Marcel Dekker, New York. pp 45-68.
13. Palmer E, Martin ML. 1982. An Atlas of Mammalian Viruses, CRC Press, Boca Raton, FL.
14. Palmer E, Martin ML. 1988. Electron Microscopy in Viral Diagnosis, CRC Press, Boca Raton, FL.
15. Murphy FA, Fauquet CM, Bishop DHL, Ghabrial SA, Jarvis AW, GP Martelli, Mayo MA, Summers MD. 1995. Virus Taxonomy. Sixth Report of the International Committee on Taxonomy of Viruses. Springer-Verlag, New York.
2, 5, 9, 11-14 = Methods and micrographs 2, 14 = Excellent atlases 7, 8 = Brief synopses of technique and basis of virus ID 15 = Good general reference, but don't memorize!! Lists all viruses, not just human.
TWO WAYS TO LOOK AT VIRUSES BY EM: Negative staining Used with liquid samples Shows whole virus (size, surface structure important) Thin sectioning Used with cells and tissues Shows a slice of an infected cell Shows relationship of viruses to cell organelles (internal structure and cell organelle association important) Location within the cell is a clue to nucleic acid type DNA viruses are usually seen in the nucleus (some exceptions). RNA viruses are usually seen in the cytoplasm (some exceptions). Exceptions: Naked viruses get out of the cell by lysis; if the cell is very sick, there may be virions everywhere, e.g., adenovirus (DNA). Poxviruses are DNA viruses, but are constructed in the cytoplasm. Hepatitis B core particles (DNA) may be in the nucleus and cytoplasm. Dane particles (complete viruses) are only cytoplasmic. Some paramyxovirus (RNA) nucleocapsids, not the complete virion, can be seen in the nucleus (e.g., measlesvirus). Enveloped viruses are associated with cell membranes; the type of cell membrane (plasma membrane, nuclear membrane, endoplasmic reticulum, vacuoles) can be a clue to virus identification.
SUMMARY OF VIRUS TYPES (based on morphology): Naked viruses All human naked viruses are icosahedral (spherical). There are 3 size ranges from 20-35 nm, 45-55 nm, 70-90 nm. Examples: Small: parvovirus, enterovirus, rhinovirus Medium: polyomavirus, papillomavirus Large: rotavirus/reovirus, adenovirus Enveloped viruses All have a lipoprotein membrane on the outside of the nucleocapsid. Most derive this layer by budding through cell membranes; this pliable layer makes the virion pleomorphic. Poxviruses synthesize their shell de novo; sometimes they pick up an extra covering by budding. They are not pleomorphic, but different varieties may be more ovoid, others, brick-shaped. Most range in size from 40-400 nm. (Filoviruses are 80 nm by up to 1400 nm.) The envelope has proteins on the outside that may look like spikes or fuzz (e.g., influenza virus, coronavirus). Sometimes the spikes are too short to be distinguished by EM, making the virion appear smooth (e.g., herpesviruses, rubella virus). The nucleocapsid shape can be a. icosahedral, like the naked viruses; however, some are morphologically nondescript as seen by EM; b. filamentous, like a "slinky." c. complex (poxvirus cores look like dumbbells with lateral bodies) d. morphologically nondescript (just dense without any particular shape). Enveloped DNA viruses (except pox) construct their nucleocapsids in the nucleus. Enveloped RNA viruses construct their nucleocapsids in the cytoplasm; (measles nucleocapsids can sometimes be seen in the nucleus).
On Sat, 26 Feb 2000, Ronald R. Matias wrote:
} Date: Sat, 26 Feb 2000 08:19:49 +0800 } From: Ronald R. Matias {rrm-at-skyinet.net} } To: Microscopy-at-sparc5.microscopy.com } Subject: Need help on EM negative staining } } ------------------------------------------------------------------------ } 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 and Help! } } We are a hospital based research lab. starting to use the electron } microscope to identify or confirm pathogens that cause gastrointestinal } disturbances. We would like to know if someone out there could recommend } appropriate specific EM protocols (negative staining, processing of the } gastrointestinal contents, etc....) to visualize these pathogens (ranging } from viruses to bacteria and protozoans). We have a Jeol 1010 EM and perform } the routine EM preparations designed to examine tissues. } } In the interest of bring quality healthcare to our people, we will be very } much obliged, } } ronnie matias } }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
The University of Kentucky invites applications for a Research Associate in the area of electron microscopy. The Electron Microscopy Facility houses two SEMs, two TEMs one AFM and extensive sample preparation equipment that are used by undergraduate and graduate students in support of research and education. The primary responsibility of the Research Associate will be to oversee the JEOL 2010F Field Emission TEM and auxiliary equipment, including an Oxford energy dispersive x-ray spectrometer (EDS), Gatan electron energy loss imaging filter and acquisition hardware and software. S/he will be responsible for coordinating and administering maintenance, access, training and use of the equipment by internal and external users. S/he will also be encouraged to develop sponsored research programs and collaborate with university faculty members. The qualified candidate will have a Ph.D. and practical experience in analytical electron microscopy and demonstrate good communication skills. The salary will be commensurate with qualifications and experience. The University of Kentucky offers comprehensive insurance and benefits packages and is an equal opportunity employer. Please send applications to:
The University of Kentucky invites applications for a Research Associate in the area of electron microscopy. The Electron Microscopy Facility houses two SEMs, two TEMs one AFM and extensive sample preparation equipment that are used by undergraduate and graduate students in support of research and education. The primary responsibility of the Research Associate will be to oversee the JEOL 2010F Field Emission TEM and auxiliary equipment, including an Oxford energy dispersive x-ray spectrometer (EDS), Gatan electron energy loss imaging filter and acquisition hardware and software. S/he will be responsible for coordinating and administering maintenance, access, training and use of the equipment by internal and external users. S/he will also be encouraged to develop sponsored research programs and collaborate with university faculty members. The qualified candidate will have a Ph.D. and practical experience in analytical electron microscopy and demonstrate good communication skills. The salary will be commensurate with qualifications and experience. The University of Kentucky offers comprehensive insurance and benefits packages and is an equal opportunity employer. Please send applications to:
I don't know if you have received an answer since you posted this message, but if you'd like, we could have a look at your images and try to help you. What we need is of course the image(s) and a description of what you actually want to measure. Please be as specific as you can. Please send the material to my email address below.
Thanks.
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== 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: Exchange Administrator Sent: Wednesday, February 23, 2000 8:35 PM To: Michael Bode
Dear all, I am doing the image analysis of SEM micrographs. The micrographs contain both dense (light) and air cell (dark) areas. But two areas are not discrete. I'm having a difficulty of quantifying those separated fractions. I wonder if anyone has any ideas of how to solve the problem. Looking forward for your suggestions. Regards, Supanee
Occassionally someone sends a question to the listserver and there is no response. It doesn't happen often so I thought I would send this question again.
Several weeks ago, I was given some inner membranes from E. coli to be run up to Epon and sectioned for TEM. I treated the sample like tissue culture and pelleted the membranes. Fixed in 3% glut in PBS and post-fixed in 1% OsO4. Routine dehydration in alcohols. Embedded in EMbed 812 with BDMA in Beem Capsules. Left in oven 48 hours. When I tried to section, the tissue was too soft and appeared to not be infiltrated well. Put back into the oven for 2-3 more days. No difference. Thinking I had a neural meltdown while processing the samples, I tried again on fresh samples, paying particular attention to details. I also used new bottles of embedding media. Same results. The part of the block without the tissue is its usual hardness. The part of the block with the tissue (the tip of the beem capsule) is soft and tacky.
Has anyone experienced anything like this? Any suggestions on how I can get the sample into epon?
Thanks in advance for any ideas and/or suggestions.
George Lawton Chief Electron Microscopist Molecular and Cellular Imaging Facility UT Southwestern Medical Center at Dallas 5323 Harry Hines Blvd. Dallas, Tx 75390-9039 Phone: 214-648-7291 eMail: George.Lawton-at-email.swmed.edu
If you were working with a pellet in the bottom of a tube or BEEm cap, it is possible that you were not getting good exchange of reagents at the tip of the tube during dehydration and embedding. Or is you were centifuging in the final resin mixture before putting into the oven it is possible that your sample was packed so tightly that there was not enough resin in it to polymerize properly.
At 12:48 PM 02/28/2000 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Gregory W. Erdos, Ph.D. Ph. 352-392-1295 Assistant Director, Biotechnology Program PO Box 110580 Fax: 352-846-0251 University of Florida Gainesville, FL 32611
We have an old Leitz Orthoplan microscope and are looking for the Ultrapak incident light illuminator (housing with fixed ring mirror and interchangeable UO objectives of 4X to 60X primary magnification). These components have not been produced in many years. If anyone has this equipment, we would be pleased to pay for it and for the shipping. Thank you very much!
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Dear Doug,
We have some experience in using micro-PIXE (proton induced X-ray emission) and the so-called nuclear microprobe in toxicological research. It is more sensitive than EDS and WDS, is very easy to quantify (standardless technique). In principle you can have detection limits of the order of 1 ppm in point analysis mode. Specimens can be of any thickness and their actual thickness is assessed by the simultaneous use of the BS technique (proton backscatering spectrometry). What in my opinion makes it really attractive for this purpose is its very good scanning capability. Scan sizes are from ca. 2.5 mm x 2.5 mm down to the beam resolution, which is of the order of few micrometer (can be down to 1 micrometer or even less, but in most of our uses few micrometers is good enough). The sample preparation is similar as for EDS, using cryotechniques to avoid elemental losses and redistribution. We have so far done studies on Ni, Mo, Cd. The detection limits for As are better than for Mo and Cd. Of course, this technique cannot indicate the state of oxidation, you can really treat it as a "more sensitive EDS".
If you want more information, I can refer you to our earlier work and perhaps you might be interested in contacting me.
Best regards
xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx Dr Wojciech J. Przybylowicz Materials Research Group National Accelerator Centre PO Box 72 Faure 7131 South Africa E-mail: przybylowicz-at-nac.ac.za Fax: +27-21-8433543 Phone: +27-21-8433820 http://www.nac.ac.za/IBA/ http://www.nac.ac.za/materials/ xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx
I seem to recall a discussion of software that was available to keep track of printing jobs submitted to a network printer for billing purposes? An argument here for not installing a networked dyesub printer for digital images is who is going to be responsible for watching who's using it and how many prints they're making. We have had some problems like this with the Epson 850 which is on the network and somebody has ran through two color cartridges in a short time. Thanks for any advice.
We are looking for a program (preferably Macintosh-based) that will automatically measure the diameter of spheres that have been imaged in a TEM.
Typically, the spheres measure about 3-5 mm on the TEM negative and often are touching or slightly overlapped. I know that NIH Image should do this, but I have not been able to get reproduceable data using this program.
Someone told me about a program called "Bolero", written by some Spanish university researchers but it is an $8-10K program. Too rich for our tastes.
Any suggestions?
We do appreciate your suggestions.
John
#################################################################### John J. Bozzola, Ph.D., Director Micro-Imaging and Analysis Center 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 ####################################################################
I agree with Greg: sounds like an infiltration problem. Your best bet is to encase the bugs in agar. One way is to fix briefly (2-5 min) in glut, then pellet and let sit for a while (1 hr). This glues them together.
Don't resuspend them in the glut or it will fix the walls of the bugs so that they won't stick together.
Cut off the bottom of the tube if plastic or scrape out the cells if tube is glass, onto Parafilm and drain with wedge of filter paper. Divide into 1 cubic mm (or smaller) portions and cover with molter agar. Cut away any excess. Wash. Osmicate and embed as you would a piece of tissue.
Other folks have suspended bugs in molter agar and pelleted them through the agar. You have to do this before the agar cools. Then cut off the end with the cells.
Don't fix the agar/cells in glut; it will cross-link the agar so that the subsequent solutions can't get in. The glut left in the bugs after they're fixed and before they're encased is OK and will wash out.
Hope this helps.
Sara
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
In a message dated 2/28/00 11:44:37 PM, bozzola-at-siu.edu writes:
} Typically, the spheres measure about 3-5 mm on the TEM negative and } often are touching or slightly overlapped. I know that NIH Image } should do this, but I have not been able to get reproduceable data } using this program.
If none of the spheres have been cut off in the section so that you have stereological corrections to make for the missing parts, and you aren't having problems with thresholding, etc., then I don't see what your problem is. Use a watershed to separate the touching features, and use the maximum feret's diameter to estimate the sphere diameter, not the area (which may be reduced due to overaps). Or you can use the maximum values of the euclidean distance map (the ultimate eroded point) as the sphere radii and not even bother with the watershed.
Hello, Is there a web site where I can find more information about quantitative techniques in microscopy on SEM's? In particular the XPP correction procedure?
Many thanks Rachel
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You may want to try Triad Scientific Inc. in Edison, NJ. , triadsci-at-webspan.net or (800) 867-6690. When I was there last month to purchase a vacuum oven, a whole truck lot of optical microscopes and parts (Leitz, etc.) was being unloaded. Most were still in the original boxes. J. Roy Nelson Material Testing Laboratory Pennington, NJ mtl-at-njcc.com
Margie Bryant 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 } -----------------------------------------------------------------------. } } Date: Feb.28, 2000 } } Hello, } } We have an old Leitz Orthoplan microscope and are } looking for the Ultrapak incident light illuminator (housing with fixed } ring mirror and interchangeable UO objectives of 4X to 60X primary } magnification). These components have not been produced in many years. } If anyone has this equipment, we would be pleased to pay for it and for } the shipping. Thank you very much! } } Sincerely, } } Margie Bryant } } From: Margie } Bryant {mbryant-at-com1.med.usf.edu
I've found the same problem sometimes with Epon/Araldite in Eppendorf tubes. I think the pellet is so tightly bound to the bottom of the tube by the time 100% resin infiltration is reached (especially if you centrifuge along the way) that the resin doesn't have good access to the cells or material at the 'bottom'. I now lift (ease gently) the pellet off the bottom of the tube with a toothpick to ensure that the media are totally surrounding the pellet. Another choice is to cut away all the gooey resin and hope that the material at the 'top' of the pellet is well infiltrated and polymerized.
Regards, Marilyn
George Lawton 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 } -----------------------------------------------------------------------. } } Occassionally someone sends a question to the listserver and there is no response. It doesn't happen often so I thought I would send this question again. } } Several weeks ago, I was given some inner membranes from E. coli to be run up to Epon and sectioned for TEM. I treated the sample like tissue culture and pelleted the membranes. Fixed in } 3% glut in PBS and post-fixed in 1% OsO4. Routine dehydration in alcohols. Embedded in } EMbed 812 with BDMA in Beem Capsules. Left in oven 48 hours. When I tried to section, the tissue was too soft and appeared to not be infiltrated well. Put back into the oven for 2-3 more days. No difference. Thinking I had a neural meltdown while processing the samples, I tried again on fresh samples, paying particular attention to details. I also used new bottles of embedding media. Same results. } The part of the block without the tissue is its usual hardness. The part of the block with the tissue } (the tip of the beem capsule) is soft and tacky. } } Has anyone experienced anything like this? Any suggestions on how I can get the sample into epon? } } Thanks in advance for any ideas and/or suggestions. } } George Lawton } Chief Electron Microscopist } Molecular and Cellular Imaging Facility } UT Southwestern Medical Center at Dallas } 5323 Harry Hines Blvd. } Dallas, Tx 75390-9039 } Phone: 214-648-7291 } eMail: George.Lawton-at-email.swmed.edu
Two things come to mind. First, if processed in a microfuge tube, loosen the pellet from the tube and make sure all fluids and epoxy mixtures come into contact with all surfaces (as already recommended). Second, I always make it a practice to preheat the BEEM capsules and labels in the embedding oven all day or overnight before use to drive off any moisture. This really seems to help in our humid environment!
Good luck and aloha, Tina
Sunny and 80F but with mosquitoes.
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Rick: What you want to do is fairly easy if you are using Windows NT. It will maintain a log of the user name, number of prints, date, name of print job, etc. I posted instructions to this group in 1997. These instructions were published in Microscopy and Microanalysis Vol.3, number 6, p. 569. If you don't have access to this, let me know and I will send the directions. Stanley L. Flegler, Acting Director Center for Advanced Microscopy Michigan State University
Question: Currently studying for a project on HREM of semiconductors, namely GaN; I would like to know where I can obtain some good information on this subject. My lecturer does not have much insight in this area, I need to study its structure, and I am currently using the Cerius(2) software package to simulate images of GaN.
The Centre for Microscopy & Microanalysis (CMM) at The University of Queensland will demonstrate its web-based interactive Virtual Electron Microscopy service this Thursday (March 2, 2000) between 11am and 1pm. (Queensland Time; GMT +10)
CyberSTEM (WWW accessible Scanning and Transmission Electron Microscopy) is a service that became officially permanent on the web after Centre Directors approved funding last week.
URL: http://www.uq.edu.au/nanoworld/online.html
The CMM's videoconference service - begun in 1995 - has evolved into a webcam-based service that allows unlimited live connections to microscopy sessions conducted at the Centre. Visitors can interact with staff during organised sessions via telephone, by chat programs or via email. (Please note: Live interaction is only available during specifically organised sessions - direct all other inquiries to the address/email noted in the signature below.)
Though primarily intended to service paying customers who cannot physically attend sessions, video streaming from two of their microscopes (more later they hope) will be regularly accessible by the general public.
Sincerely,
Duncan
-- ************************************************************ Duncan Waddell (BSc) Senior Scientific Officer Centre for Microscopy and Microanalysis The University of Queensland, St. Lucia, Qld, 4072 Australia Telephone: +61-7-3365-4216 Facsimile: +61-7-3365-4422 WWW: http://www.uq.edu.au/nanoworld/nanohome.html ************************************************************ Any opinion expressed is that of the writer, and not necessarily that of CMM or of the University. ************************************************************
Dear George, In all my experience working up bacteria and bacterial fragments, I have always, and by the way this is the simplest, filtered all suspended cells on Millipore filters and processed the samples on the filters. This yielded excellent results. The fragments, cells stayed on the filters and never detached. Filter your suspended fragment solution using a 0.2um filter unit system attached to a vacuum system. The aspirator of a faucet works great. Rinse in buffer while on the filter system. Then remove the filter and gently place in a plastic or glass petri dish. Again, gently drop fixative over the sample enough to cover your filter and carefully slice it into your appropriate sizes and process from there. You will have success. If you have further questions, just give me a call.
Blessings, Maria
Maria Fazio-Zanakis Bioimaging and Molecular Histology Aventis Pharmaceuticals 1-908-231-3357 Fax: 1-908-231-3962 Email: Maria.Fazio-Zanakis-at-Aventis.com
-----Original Message----- } From: George Lawton [mailto:George.Lawton-at-email.swmed.edu] Sent: Monday, February 28, 2000 1:49 PM To: microscopy-at-sparc5.microscopy.com
Occassionally someone sends a question to the listserver and there is no response. It doesn't happen often so I thought I would send this question again.
Several weeks ago, I was given some inner membranes from E. coli to be run up to Epon and sectioned for TEM. I treated the sample like tissue culture and pelleted the membranes. Fixed in 3% glut in PBS and post-fixed in 1% OsO4. Routine dehydration in alcohols. Embedded in EMbed 812 with BDMA in Beem Capsules. Left in oven 48 hours. When I tried to section, the tissue was too soft and appeared to not be infiltrated well. Put back into the oven for 2-3 more days. No difference. Thinking I had a neural meltdown while processing the samples, I tried again on fresh samples, paying particular attention to details. I also used new bottles of embedding media. Same results. The part of the block without the tissue is its usual hardness. The part of the block with the tissue (the tip of the beem capsule) is soft and tacky.
Has anyone experienced anything like this? Any suggestions on how I can get the sample into epon?
Thanks in advance for any ideas and/or suggestions.
George Lawton Chief Electron Microscopist Molecular and Cellular Imaging Facility UT Southwestern Medical Center at Dallas 5323 Harry Hines Blvd. Dallas, Tx 75390-9039 Phone: 214-648-7291 eMail: George.Lawton-at-email.swmed.edu
Can anyone help me? I am doing research on the plant Achillea millefolium (Yarrow)and I need to find out the ploidy level of 2 seed sources. I am looking for information on companies/institutions that will do this work for me and a rough estimate of the cost. Many thanks, Fiona Russell.
-- Sent through Global Message Exchange - http://www.gmx.net
I recently tried to help out Dr. M. Rohde who was embedding in Spurr's formulation and attempting to locate an antigen.
I said that Spurr's formulation has no place in immunocytochemistry for all the reasons I listed then. This is an oversimplification (I am always harassed for time when trying to answer questions completely.) Let me tell you that I, myself, worked with a graduate student who used Spurr's medium, etched for one hour in sodium metaperiodate, and then got wonderful Au label. The reason for this success was that she had an antigen that was so sturdy and such plentiful supply that it was a candidate for the prozone effect. Her same exact protocol (actually mine) when tried by us in our laboratory was a spectacular failure with our antigen which we soon discovered was scarce and delicate and needed amplification up to the point where I wondered whether we were actually increasing the thickness of the section by amplification methods! Sort of like painting a wall until the room gets smaller! At any rate - yes, Spurr's can be used for immunocytochemistry. However, if at first you don't succeed, don't try again. Go to another epoxy, and then to an acrylic. I should not make absolute statements as I did. I did not mean to discourage those who successfully use Spurr's to abandon it. Besides, compared to what there is to be known about immunocytochemistry, my amount of knowledge can be dropped into a thimble with room left to stuff an entire thumb in after it.
I always reserve the right to be wrong!
Hildy Crowley Sr. Electron Microscopy Specialist Dep. of Biol. Sciences University of Denver Denver, CO
I remember seeing posts over the years regarding web-based equipment sign-up systems, but didn't save any of them. Can anyone direct me to an archived discussion thread, or have current suggestions on a very simple and easy way to set this up? I want to put this under a dept. web page on our intranet only, so security is not a major issue. All I want is a simple calendar that users can view and sign up for time slots. No retribution if time not used, but must require user's name so that they can be tracked down for questioning if needed. I have no experience in HTML programming but am not afraid to learn.
Thanks as always, Karen
-- Karen S. Zaruba kszaruba-at-mmm.com 3M Company, St. Paul, MN
Any suggestions on a really good video printer to include with an SEM purchase? We've had trouble over the years with the standard roll printers getting paper jams, which required sending out for servicing (i.e. lots of down time). We're willing to sacrifice some speed for something reliable. Are there "fast" ( { 1 min. per print) video printers that use the cut sheet paper?
Thanks, Karen
P.S. vendors please respond by E-mail (not phone) or dare to face my wrath. -- Karen S. Zaruba kszaruba-at-mmm.com 3M Company, St. Paul, MN
We have 2 Tracor Northern/Noran TN 5500 EDX systems that need a new home. We are looking for a possible trade for comparable priced equipment/software. If anyone is interested, please contact me directly.
Harry Ekstrom Honeywell Materials Laboratory Phoenix, AZ 602-231-2744
Sorry for the inconvenience, but we mistakenly scheduled our Tripod Polisherš Workshop over Memorial Day Weekend. While I'm sure that you would all love to spend your holiday weekend here with us, we felt it necessary to move the workshop to the following week. The workshop is now scheduled for June 2- 3, 2000 in San Clemente, CA.
For those of you who have already registered, you should have already been contacted and given the option of re-booking for the new date or canceling your reservation. Again, I apologize for the error. Certainly, if anyone wishes to come and visit us over the Memorial Day Weekend, we will welcome you with open arms - but you'll have to wait until the following weekend for the workshop!
David Henriks Vice President 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
Evex Analytical ( www.evex.com ) has a Jeol 840 with an Evex =3D Microanalysis system ( http://www.evex.com/prod2.htm ) and Active Digital Imaging System that we would like to replace with a new microscope.
Claudio Tarquinio Evex Analytical Microanalysis and Digital Imaging 609-252-9192 T 609-252-9091 F www.evex.com info-at-evex.com
---------- } From: Karen Zaruba {kszaruba-at-MMM.COM} } To: MSA Listserve {microscopy-at-sparc5.microscopy.com} } Subject: General: Video Printers } Date: February 29, 2000 6:45 PM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Any suggestions on a really good video printer to include with an SEM } purchase? We've had trouble over the years with the standard roll } printers getting paper jams, which required sending out for servicing } (i.e. lots of down time). We're willing to sacrifice some speed for } something reliable. Are there "fast" ( { 1 min. per print) video } printers that use the cut sheet paper? } } Thanks, } Karen } } P.S. vendors please respond by E-mail (not phone) or dare to face my } wrath. } -- } Karen S. Zaruba kszaruba-at-mmm.com } 3M Company, St. Paul, MN }
Karen -
We've always been happy with our little Sony UP-811 video printer. It doesn't give a very big print (about 5 inches wide), rather like a Polaroid, and not particularly high resolution or anything, but useful as a "working copy", with the actual image saved to disk. It's dead reliable, however, having been in continuous service since about 1993, with no problems that I know of. No connection with Sony, etc. (wouldn't mind owning some stock, though;-)
F.C. Thomas MicroAnalysis Facility Geological Survey of Canada Dartmouth, Nova Scotia
I had a Mitsubishi that was reliable, but in general, my feeling is that for reproducing EM images there are NO good video printers. Between the (typical) paper cost and poor resolution, I prefer to print captured digital images. Though slow, you can get an inkjet printer these days for 1/10 the price. Faster ones for a bit more...
Colleagues.... Can anyone help this guy? It's not my area.
Nestor Your Friendly Neighborhood SysOp ------------------------------------------
Dear Dr Zaluzec
I'd be very grateful if you could help me with a problem I'm having.
I'm writing an essay in low temperature scanning electron microscopy in biology, but I'm finding it difficult to track down up-to-date source material.
If you could suggest any papers, books or book chapters related to the subject, it would be a great help. Thanks.
The following web site, http://www.emitech.demon.co.uk/, has a downloadable technical brief on low temperature EM. It talks about Emitech's products, but also has a discussion of low-temp EM in more general terms. A more dated, but very useful, reference is the book "Low Temperature Methods in Biological Electron Microscopy", which is part of the "Practical Methods in Electron Microscopy" series edited by Audrey Glauert. My copy is dated 1985. I don't know if later editions exist.
Hope this is useful.
Disclaimer: We have no financial interest in Emitech's products, other than being users of them.
Regards, 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: DARRYL GRAY [mailto:ddg99-at-aber.ac.uk] Sent: Wednesday, March 01, 2000 7:53 AM To: Microscopy-at-sparc5.microscopy.com
Colleagues.... Can anyone help this guy? It's not my area.
Nestor Your Friendly Neighborhood SysOp ------------------------------------------
Dear Dr Zaluzec
I'd be very grateful if you could help me with a problem I'm having.
I'm writing an essay in low temperature scanning electron microscopy in biology, but I'm finding it difficult to track down up-to-date source material.
If you could suggest any papers, books or book chapters related to the subject, it would be a great help. Thanks.
Sorry I deleted the original question so I don't remember who posted it. Then I came across a couple of papers by John E. Scott on staining for PG's using Cupromeronic Blue. Never tried this myself, so I don't know how well it works. Here's one reference:
J.E. Scott and M. Haigh, "Identification of specific binding sites for keratan sulphate proteoglycans and chondroitin-dermatan sulphate proteoglycans on collagen fibrils in cornea by the use of Cupromeronic Blue in 'critical-electrolyte-concentration' techniques." Biochem J. (1988) 253:607-610.
-- Karen S. Zaruba kszaruba-at-mmm.com 3M Company, St. Paul, MN
Does anybody know a recipe for staining the amorphous part of poly(vinylidene fluoride) and poly (e-caprolactone)? We need to view the crystalline lamellae of these polymers by TEM.
Thank you in advance for your help.
Antoine
====================== Dr Antoine GHANEM SOLVAY Research and Technology Rue de Ransbeek 310 1120 Brussels Belgium Phone 32-2-2643422 Fax 32-2-2642055 antoine.ghanem-at-solvay.com ======================
Someone in the lab needs help with hand sectioning Arabidopsis seedling stems. They are small, less than a mm in dia. and very delicate. She needs cross sections of the stem from a specific zone to do microscopy and pigment localization.
I tried it using traditional techniques, but without success. She must have sections of fresh material for her stains, she says frozen sections don't work. Any ideas?
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
Suspend them in some stiff agarose and try a vibratome. There is also the old elder pith trick, if you haven't already tried it. There are only a few of us ancient enough to have ever heard of that one.
At 10:57 AM 03/01/2000 -0800, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Gregory W. Erdos, Ph.D. Ph. 352-392-1295 Assistant Director, Biotechnology Program PO Box 110580 Fax: 352-846-0251 University of Florida Gainesville, FL 32611
} } Someone in the lab needs help with hand sectioning Arabidopsis seedling } stems. They are small, less than a mm in dia. and very delicate. She needs } cross sections of the stem from a specific zone to do microscopy and } pigment localization. } } I tried it using traditional techniques, but without success. She must have } sections of fresh material for her stains, she says frozen sections don't } work. Any ideas? } We have made some headway in a related problem by encasing the fresh tissue in low melting point agarose, around 3%. The idea here is to make a cube of agarose with the stem oriented in side. You can then cut the whole cube with a razor. The agarose doesn't impede or interfere with most stains. This didn't work perfectly, but it was better than nothing.
Hope this helps, Tobias Baskin _ ____ ^ __ ____ Tobias I. Baskin / \ / / \ / \ \ University of Missouri / | / / \ \ \ Biological Sciences /___/ /__ /___ \ \ \__ 109 Tucker Hall / / / \ \ \ Columbia, MO 65211-7400 USA / / / \ \ \ voice: 573-882-0173 / /____ / \ \__/ \____ fax: 573-882-0123
Here are the most recent posts. I plan to use the calendar from brownbear but have not actually implemented it yet.
http://www.brownbearsw.com
Date: Thu, 26 Aug 1999 14:45:39 -0400 Reply-To: Confocal Microscopy List {CONFOCAL-at-LISTSERV.ACSU.BUFFALO.EDU} Sender: Confocal Microscopy List {CONFOCAL-at-LISTSERV.ACSU.BUFFALO.EDU} } From: Kristi DeCourcy {decourcy-at-MAIL.VT.EDU}
At 12:04 PM -0600 3/1/0, Karen Zaruba wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
************************ You can also try any number of cationic stains (ruthenium red, alcian blue. safranin-O, etc)at concentration of about 0.1-0.5%in the primary fixation step. These dyes are electron dense and will bind to the negatively charged PG's. You can also track down references from the 1970's & early 1980's by Simeonescu & Simeonescu who did a lot of work with these types of dyes. I have used them (years ago) to look at the extracellular matrix in heart muscle.
Lee
Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Confocal Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
I have a set of samples giving me fits here. This person is trying to visualize the ruthenim doped into a polymer. I have not had much luck working with this stuff and was wondering if any of you had any suggestions to pass along. A description of the sample follows.Here is what I have tried so far.
She originally brought this in on a glass slide and it is about 100µm thick. I tried to use backscattering to visualize the Ru metal in the FESEM. She thought it might be in little islands. No luck. It was too thick to shoot a beam through with the TEM so I embedded some in cold LR-White resin and sectioned. It offered enough support to get some areas thin enough to view through, but I did not see any metal. I suggested she take the grids to the Materials EM unit here on campus as the have EDS and are better equipped for this, but they saw nothing either.
I then had her try to spin coat slids,grids,and mica to float off the film, but she cannot get it to work so now I am back to the slides, scraping material off, and cold embedding. The last set was too rubbery though and even in epoxy on a good diamond I was unable to obtain a section of the material. It just moved out of the way and popped back up with nice resin sections floating away. I don't get any polymers here and we are almost exclusively a biologic unit so do any of you have any suggestions??
} Date: Wed, 1 Mar 2000 11:28:21 -0500 (EST) } From: Joanne Bedlek {bedlek-at-chem.ufl.edu} } To: Scott Whittaker {sdw-at-biotech.ufl.edu} } Subject: Re: your mail } } Scott, } } The polymer is a polydimethylsiloxane. The metal (Ru) is dissolved in } methylene chloride and added to the silicone fluid. As the polymer cures } it "hardens" to a rubbery material. } } The TEM images you took give us an idea of the polymer distribution. With } the new films I have, we are interested in the polymer distribution again. } I } suspect we will see the same droplet pattern as before. This will be a } characterization technique for a paper submission. } } As for the dispersion technique, MAIC tried EDS. They were not able to } detect the Ru. They did tell me that there is a higher content of Si on } the outer sides of the droplets as opposed to the interior areas. This is } interesting. } } I will drop off the new samples either next week or the week after. It } depends when I get back next Friday. } } I redid the one sample that was giving you so much trouble in slicing. I } hope it has cured better this time. } } } } This may be more of a dispersion thing. Materials should be able to see it } } if it is in there. You might get with them and see if they have done } } anything similar as we had no luck with that last set you brought in. I } } could also post a discussion to the microscopy listserver and see what } } comes up. I just need to know more about the composition of the samples } and } } stuff like that } }
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall EM Technician Gainesville, FL 32610 University Of Florida ph 352-392-1184 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
I agree with using the agar. Try 3% and go up if you need to. If the stem is soft, you'll be able to cut it with a "Vibratome" or vibrating microtome. If it is hard, you won't.
Sara
On Wed, 1 Mar 2000, Jon Krupp wrote:
} Date: Wed, 1 Mar 2000 10:57:57 -0800 } From: Jon Krupp {jmkrupp-at-cats.ucsc.edu} } To: Microscopy-at-sparc5.microscopy.com } Subject: LM, help with hand sectioning } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Greetings: } } Someone in the lab needs help with hand sectioning Arabidopsis seedling } stems. They are small, less than a mm in dia. and very delicate. She needs } cross sections of the stem from a specific zone to do microscopy and } pigment localization. } } I tried it using traditional techniques, but without success. She must have } sections of fresh material for her stains, she says frozen sections don't } work. Any ideas? } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } } } }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
Question: I read a lot of information about Naturpaths using Dark Field Microscopy for live blood analysis and blood crystallization testing. They claim they can see nutritional deficiencies as well as disease process through this type of testing. Is this possible and how accurate are these tests? Thanks.
We have the opportunity to purchase a used Zeiss 960 SEM for use by undergraduate biology students in a number of courses and for research.
I would be interested in any comments, advice, etc. about the instrument and its appropriateness for this type of application, such as maintenance, resistance to abuse, and ease of use.
Elder pith being in short supply you might try supporting it in a fresh carrot. I have had some luck with that in a hand microtome.
I came across the method in a book from the 30's.
Gordon
Gordon Couger gcouger-at-couger.com
Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00 } From: "Greg Erdos" {gwe-at-biotech.ufl.edu} } } } Suspend them in some stiff agarose and try a vibratome. There is also the } old elder pith trick, if you haven't already tried it. There are only a } few of us ancient enough to have ever heard of that one. } } } } Someone in the lab needs help with hand sectioning Arabidopsis seedling } } stems. They are small, less than a mm in dia. and very delicate. She needs } } cross sections of the stem from a specific zone to do microscopy and } } pigment localization. } } } } I tried it using traditional techniques, but without success. She must have } } sections of fresh material for her stains, she says frozen sections don't } } work. Any ideas? } } } } Jonathan Krupp } } Microscopy & Imaging Lab } } University of California } } Santa Cruz, CA 95064 } } (831) 459-2477 } } jmkrupp-at-cats.ucsc.edu } } } } } } Gregory W. Erdos, Ph.D. Ph. 352-392-1295 } Assistant Director, Biotechnology Program } PO Box 110580 Fax: } 352-846-0251 } University of Florida } Gainesville, FL 32611 } } }
Jon, it seems that project is beyond hand sectioning. There is a whole series of instruments available that use a oscillating or vibrating knife and the best can section down to 10 micrometers. Buy, beg, borrow don't steal one of those instruments. The mid to lower price range should do your job. Disclaimer: ProSciTech sells these instruments Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Thursday, March 02, 2000 4:58 AM, Jon Krupp [SMTP:jmkrupp-at-cats.ucsc.edu] wrote: } } Greetings: } } Someone in the lab needs help with hand sectioning Arabidopsis seedling } stems. They are small, less than a mm in dia. and very delicate. She needs } cross sections of the stem from a specific zone to do microscopy and } pigment localization. } } I tried it using traditional techniques, but without success. She must have } sections of fresh material for her stains, she says frozen sections don't } work. Any ideas? } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } }
} Question: I read a lot of information about Naturpaths using Dark Field } Microscopy for live blood analysis and blood crystallization testing. They } claim they can see nutritional deficiencies as well as disease process } through this type of testing. Is this possible and how accurate are these } tests? Thanks. } } --------------------------------------------------------------------------- }
You CAN see red cells. But the Naturopaths are only using this as a hook to give (pseudo)scientific validity to their diagnosis. It all COMPLETE HOKUM. } } }
We are looking for a high voltage measurement cable (cable which links the high voltage transformer to the measurement resistances) in working order and dedicated to a Siemens Elmiskop 102 TEM (manufacturing model number 2810)
Here are the specifications of the cable we are looking for : 125 kV - 1.5 m length
Would it be possible to also give us a cost estimation ?
Thanks in advance
Best regards
\\ _// X-FERT ! -(-at- -at-)- -------------------oOO--(_)--Ooo------------ Philippe Drouillon Solvay Research and Technology Electron Microscopy and Image Analysis - IT Coordinator Rue de Ransbeek, 310 B-1120 Brussels (Belgium) phone : (00 32) 2 264 24 47 mailto:philippe.drouillon-at-solvay.com
A modern substitute for elder pith is expanded polystyrene foam. The dense EPS strips sold by Diatome for cleaning diamond knives are excellent for sectioning very small items like Arabidopsis stems. Chris Jeffree
} From: Gordon Couger {gcouger-at-rfdata.net} To: Jon Krupp {jmkrupp-at-cats.ucsc.edu} , Microscopy-at-sparc5.microscopy.com, Greg Erdos {gwe-at-biotech.ufl.edu}
Dear Jon, I have had great success using LR White Medium grade methacrylate for such specimens as iris ovules, mistletoe/host tissues, and a variety of young plant material. Although time consuming, these tissue were processed with osmium tetroxide, rinsed in buffer, and exposed to a 0.025% tannic acid buffer to retain lipids, starch granules, and other items frequently lost during dehyration in EtOH.
One other advantage is the variety of staining procedures one can use with material embedded in LR White. Furthermore, in addition to lipid stabilization, osmium and tannic acid provide additional contrast to the sections.
Hope this helps. Any questions? Email me at tiekotte-at-up.edu
Cheers! -Ken
---------- Ken Tiekotter, Adjunct Professor The University of Portland Department of Biology 5000 N. Willamette Blvd. Portland, OR 97303
On Wed, 1 Mar 2000, Jon Krupp 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 } -----------------------------------------------------------------------. } } } Greetings: } } Someone in the lab needs help with hand sectioning Arabidopsis seedling } stems. They are small, less than a mm in dia. and very delicate. She needs } cross sections of the stem from a specific zone to do microscopy and } pigment localization. } } I tried it using traditional techniques, but without success. She must have } sections of fresh material for her stains, she says frozen sections don't } work. Any ideas? } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } } } }
} trying to } visualize the ruthenim doped into a polymer. I have not had much luck... } Scott, Sounds like a job for cryo-ultramicrotome thin-sectioning at approximately the glass transition temperature of the polymer. This then to be followed by TEM imaging and EDS analysis of thin-sections, or FESEM-EDS examination of the "faced-off" polymer block cross-section.
Many rubbery polymers can be stained "enblock" before sectioning, for example by submersion in a dilute ruthenium tetroxide (RuO4) solution. This "staining" often aids the sample prep, by hardening the polymer (actually it initiates some cross-linking) and allowing better sectioning and sometimes actually enabling ambient microtomy. It sounds like you will not be able to make use of this technique since you are looking for the Ru distribution in the polymer.
A great polymer reference is the book, Polymer Microscopy, by Linda Sawyer and David Grubb (Chapman and Hall). Good Luck! Brad Huggins BP Amoco, Naperville, IL Microscopy and Microanalysis Lab 630 420-3668
} ---------- } } From: Scott Whittaker[SMTP:sdw-at-biotech.ufl.edu] } Sent: Wednesday, March 01, 2000 4:12 PM } To: microscopy-at-sparc5.microscopy.com } Subject: TEM polymer help } } ------------------------------------------------------------------------ } 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 set of samples giving me fits here. This person is trying to } visualize the ruthenim doped into a polymer. I have not had much luck } working with this stuff and was wondering if any of you had any } suggestions } to pass along. A description of the sample follows.Here is what I have } tried so far. } } She originally brought this in on a glass slide and it is about 100µm } thick. I tried to use backscattering to visualize the Ru metal in the } FESEM. She thought it might be in little islands. No luck. } It was too thick to shoot a beam through with the TEM so I embedded some } in } cold LR-White resin and sectioned. It offered enough support to get some } areas thin enough to view through, but I did not see any metal. I } suggested } she take the grids to the Materials EM unit here on campus as the have EDS } } and are better equipped for this, but they saw nothing either. } } I then had her try to spin coat slids,grids,and mica to float off the } film, } but she cannot get it to work so now I am back to the slides, scraping } material off, and cold embedding. The last set was too rubbery though and } even in epoxy on a good diamond I was unable to obtain a section of the } material. It just moved out of the way and popped back up with nice resin } sections floating away. I don't get any polymers here and we are almost } exclusively a biologic unit so do any of you have any suggestions?? } } } } Date: Wed, 1 Mar 2000 11:28:21 -0500 (EST) } } From: Joanne Bedlek {bedlek-at-chem.ufl.edu} } } To: Scott Whittaker {sdw-at-biotech.ufl.edu} } } Subject: Re: your mail } } } } Scott, } } } } The polymer is a polydimethylsiloxane. The metal (Ru) is dissolved in } } methylene chloride and added to the silicone fluid. As the polymer cures } } it "hardens" to a rubbery material. } } } } The TEM images you took give us an idea of the polymer distribution. } With } } the new films I have, we are interested in the polymer distribution } again. } } I } } suspect we will see the same droplet pattern as before. This will be a } } characterization technique for a paper submission. } } } } As for the dispersion technique, MAIC tried EDS. They were not able to } } detect the Ru. They did tell me that there is a higher content of Si on } } the outer sides of the droplets as opposed to the interior areas. This } is } } interesting. } } } } I will drop off the new samples either next week or the week after. It } } depends when I get back next Friday. } } } } I redid the one sample that was giving you so much trouble in slicing. I } } hope it has cured better this time. } } } } } } } This may be more of a dispersion thing. Materials should be able to } see it } } } if it is in there. You might get with them and see if they have done } } } anything similar as we had no luck with that last set you brought in. } I } } } could also post a discussion to the microscopy listserver and see what } } } comes up. I just need to know more about the composition of the } samples } } and } } } stuff like that } } } } } } } } } } {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { } GO GATORS } Scott D. Whittaker 218 Carr Hall } EM Technician Gainesville, FL 32610 } University Of Florida ph 352-392-1184 } ICBR EM Core Lab fax 352-846-0251 } sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ } The home of " Tips & Tricks " } } } } } }
We used High- and Low-iron diamine staining in our lab for looking at glycosaminoglycans several years ago. This is principally EM level. They can be enhanced with Thiocarbohydrazide-silver proteinate staining. I still have the protocols that we used. My reference database is at another location but the ones below should help you get an idea of what is involved and who did the work. For HID/LID check into the work by Sam Spicer, I believe he pioneered it. For TCH-SP, the work is by Thiery and speaking French would be a plus or Sannes.
Hope this helps
Chuck
REFERENCESREFERENCESREFERENCES FOR IRON DIAMINE AND TCH-SP STAINING:
IRON DIAMINE STAININGIRON DIAMINE STAININGIRON DIAMINE STAINING:
Gad, Adel, and B. Sylven. On the nature of the high iron diamine method for sulfomucins. J Histochem Cytochem. 17:156-60 (1968).
Lev and Spicer. Am. J. Pathol. 46:23 (1965).
Sorvari, Tapani E. Histochemical observations on the role of ferric chloride in the high-iron diamine technique for localizing sulphated mucosubstances. Histochem. J. 4:193-204 (1972a).
Sorvari, Tapani E. Binding of ferric ions to nuclei and other tissue sites in sections stained for sulfated mucosubstances by the high-iron diamine methods. Stain Technol. 47:245-48 (1972b).
Sorvari, T. E. and H. S. Arvilommi. Some chemical, physical, and histochemical properties of three diamine fractions obtained by gel chromatography from the high-iron diamine staining solution used for localizing sulphated mucosaccharides. Histochem. J. 5:119-30 (1973).
Spicer, S. S. The use of various cationic reagents in histochemical differentiation of mucopolysaccharides. Am. J. Clin. Pathol. 36:393-407 (1961).
Spicer, S. S. Diamine methods for differentiating mucosubstances histochemically. J Histochem Cytochem. 13:211-34 (1965).
Spicer, S. S., et al. Ultrastructural visualization of sulphated complex carbohydrates in blood and epithelial cells with the high-iron diamine procedure. Histochem. J. 1O:435-52 (1978).
Spicer, S. S. and M. H. Jarrel. Histochemical reaction of an aromatic diamine with acid groups and periodate engendered aldehydes in mucopolysaccharides. J Histochem Cytochem. 9:368-79 (1961).
Takagi, Minoru, et al. Ultrastructural localization of acidic glycoconjugates with the low iron diamine method. J Histochem Cytochem. 30:471-6 (1982).
Denys, Francis R., et al. An improved technique to enhance high-iron diamine reactive sites with thiocarhohydrazide-silver proteinate. J. Nihon Univ. Sch. Dent. 25(2):103-6 (1983).
Sannes, P.L., et al. Ultrastructural localization of sulfated complex carbohydrates with a modified iron diamine procedure. J. Histochem. Cytochem. 27:1108-11 (1979).
Takagi, M., et al. J. Histochem. Cytochem. 30:1179 (1982).
Thiery, J.P. Mise in evidence des polysaccharides sur coupes fines en microscopie electronique. J. Microscop. 6:987-1018 (1967).
------------------------------------- Name: Charles Gilbert VOC: (704) 355-5261 Carolinas Medical Center FAX: (704) 355-8424 Dept of Pediatric Research digPager: (704) 355-4088 : 2058 PO Box 32861 Charlotte, NC 28232-2861
Does anyone know of a way to label for peptidoglycan at the light or electron microscope level?
William McManus Supervisor Electron Microscopy Facility Department of Biology Utah State University Logan UT 84322-5305
I am new in this discussion groud, my background is in scanning probe microscopy and mass spectroscopy. At this time I am interested in analysing plastic materials such as those used in semiconductor packaging. I would like to use FT-IR. Is this an appropriate direction to take given that some of my samples are going to be small (tens micrometers). What are the material limitations of the technique. Thank you in advance Jocho
Jochonia N. Nxumalo Material Science Engineer
Chipworks Inc. 3685 Richmond Road, Suite 500, Ottawa, Ontario, K2H 5B7 Tel:(613) 829-0414 Fax:(613) 829-0515 mailto:jnxumalo-at-chipworks.com
Dear Professor Luchtel My apologies for the confusion - I was writing without reference to Diatome's literature. EPS is just short for Expanded PolyStyrene but their polystyrol rods would be the same thing. I have found these Diatome rods useful for a number of applications, and that their density, which is a little higher than usual for commercial polystyrene foam, provides good support to the specimen when hand sectioning. However, if you want to try it out on the cheap, why not cut strips off an insulated polystyrene box or insulation tile.
By the way, the best type of blade for hand sectioning used to be the Blue Gillette double-edged carbon steel razor blades, which many of you may be too young to know about. The new, doubtless improved, stainless steel type never seems to be so sharp, and their edges dull faster. Is there still a manufacturer of carbon steel razor blades left in the world, or am I doomed to mourn forever?
Good luck Chris Jeffree
Date sent: Thu, 2 Mar 2000 07:56:53 -0400 To: "Chris Jeffree" {cjeffree-at-srv0.bio.ed.ac.uk} } From: Dan Luchtel {dluchtel-at-u.washington.edu}
} } Someone in the lab needs help with hand sectioning Arabidopsis seedling } stems. They are small, less than a mm in dia. and very delicate. She needs } cross sections of the stem from a specific zone to do microscopy and } pigment localization. } } I tried it using traditional techniques, but without success. She must have } sections of fresh material for her stains, she says frozen sections don't } work. Any ideas? } } Jon -
After you get the stem supported (you've gotten lots of good suggestions), try taping 2 razor blades together, with a thin spacer (thick paper, filecard, etc.). Cheaper than a vibratome...
Caroline
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
Dear Ken, You've spurred a question that is related, yet not exactly on the same track. Does tannic acid affect the antigenicity of the tissue? I am just about to embed some tissue in LR White for immunocytological studies, and though it is established that osmium is a problem for immuno work, I have heard nothing about tannic acid. I would love to find a way to enhance the contrast of my tissue, as it often has a surreal, ghostly appearance. Thanks, Kristen
At 01:20 AM 3/2/2000 -0800, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I received a sample of DNA microarray on a glass slide and I was asked to do SEM because the investigator wants to see single and double stranded DNA. Does anyone know if this is possible. My previous exposure to EM of DNA samples was in TEM. I would appreciate any help, comments or suggestions.
Thank you,
Corazon D. Bucana ******************************************************* Corazon D. Bucana, Ph.D. Department of Cancer Biology U.T. M.D. Anderson Cancer Center 1515 Holcombe Blvd. Box 173 Houston, Texas 77030 Phone: (713) 792-8106 FAX: (713) 792-8747 Email:bucana-at-audumla.mdacc.tmc.edu FAX: (713) 792-8747
The following equipment is surplus to requirements and is available on the basis that you are responsible for all removal and transport costs. If there are competing bids they will be sold to the highest bidder. There is no guarantee that the equipment is in operational or serviceable condition although, to the best of our knowledge, it was functional when last operated.
----- Original Message ----- } From: "Chris Jeffree" {cjeffree-at-srv0.bio.ed.ac.uk} } By the way, the best type of blade for hand sectioning used to be } the Blue Gillette double-edged carbon steel razor blades, which } many of you may be too young to know about. The new, doubtless } improved, stainless steel type never seems to be so sharp, and } their edges dull faster. Is there still a manufacturer of carbon steel } razor blades left in the world, or am I doomed to mourn forever?
I miss the old blades as well. I cured the shaving problem I quit. For hand sections I have better luck with a more rigid blade. Razor blades and straight razors tend to dig in for me. I have a lot better luck with a 2.5 inch wood chisel sharpened on glass with silicone carbide grit. It is a lot of trouble to get sharp but it works better for me than razor blades.
Gordon
Gordon Couger gcouger-at-couger.com
Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
} ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } Yes, carbon blades are still made - one of the makers is a Japanese company - I have a box in my hand, but unfortunately I can't read Japanese - the only English words on it is "Feather". There may also be many other makers of these in Asia and Eastern Europe.
Grazyna Tokarczyk gmtokarc-at-is.dal.ca Dalhousie University Halifax, Canada } } ----- Original Message ----- } } From: "Chris Jeffree" {cjeffree-at-srv0.bio.ed.ac.uk} } } By the way, the best type of blade for hand sectioning used to be } } the Blue Gillette double-edged carbon steel razor blades, which } } many of you may be too young to know about. The new, doubtless } } improved, stainless steel type never seems to be so sharp, and } } their edges dull faster. Is there still a manufacturer of carbon steel } } razor blades left in the world, or am I doomed to mourn forever? } } I miss the old blades as well. I cured the shaving problem I quit. } For hand sections I have better luck with a more rigid blade. } Razor blades and straight razors tend to dig in for me. I have } a lot better luck with a 2.5 inch wood chisel sharpened on glass } with silicone carbide grit. It is a lot of trouble to get sharp but } it works better for me than razor blades. } } Gordon } } Gordon Couger gcouger-at-couger.com } } Stillwater, OK www.couger.com/gcouger } 405 624-2855 GMT -6:00 } } } }
Dear all, We have a complete Balzers 301 freeze fracture unit with thickness monitor, many ancillary accessories and spare parts that is in need of a new home. No guarantees that the equipment is in operational or serviceable condition but, to the best of our knowledge, it was functional when last used. Please contact me directly for additional information. Cheers, Mark **************************************************** Mark A. Sanders University of Minnesota Program Director Twin Cities Campus Imaging Center St. Paul, MN 55108 23-35 Snyder Hall ph: 612-624-3454 1475 Gortner Ave. fax: 612-624-1799 http://biosci.umn.edu/imagingcenter President: Minnesota Microscopy Society http://resolution.umn.edu/MMS/
Dear colleagues, We are in the process to get a confocal microscope for a multi-user facility. We are not decided yet between different apparatus such as Wallac ultraview, zeiss LSM 510 or Biorad. We would very much appreciate any advice helping us for this choice.
Many thanks.
Pierre-Yves Sizaret Electron Microscopy Facility University of Tours France
Twice now, I have had customers in Europe refer to a technique as REM. The sample prep sounds the same as SEM, and the first time I passed it off as a typo. Now, with this second reference, I'm not so certain. Is it possible that these customers are referring to SEM, possibly with their native language? Or is it a whole different technique/instrument? Could someone please help me clear up this confusion?
Thanks in advance,
David Bell Scientist Electron Microscopy Lab Millipore Corporation 80 Ashby Road Bedford, MA 01730 (781) 533-2108
Raster Elektronen Mikroskopie = Scanning Electron Microscopy
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Robert J. Palmer Jr., Ph.D. Oral Infection and Immunity Branch National Institute of Dental and Craniofacial Research Bldg. 30, Room 308 National Institutes of Health Bethesda MD 20892 Ph 301-594-0025 FAX 301-402-0396
Has anyone heard of a fluorescent agar or a way to make agar fluorescent? We want to make sure the agar can be fluorescently imaged, but the dye cannot diffuse out into the surrounding media. Thanks- Dave
Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 75 N. Eagleville Rd. U-125 Storrs, CT 06269 Knecht-at-uconnvm.uconn.edu 860-486-2200 860-486-4331 (fax) home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html ************************************************************
Dear Colleagues In a microstructurally uniform alloy, can one expect to get the chemical composition of the alloy through microprobe (assuming all the corrections are applied correctly)? Would the composition tally with the one determined by chemical analysis? TIA Anita
Confocal issues are discussed on another listserver--see below
The best solution is to get one of each manufacturer's confocal systems! Each one has problems and each one has solutions. In my experience the Wallac system is not sensitive enough for some of our samples but is great for live imaging. The BioRad 1024 is difficult because of their interface to the Nikon TE300 and their use of an old OS2 operating system (the migration to Windows NT is still in the future) but they have a good service organization in our area and there are many other users that can help with advice. I do not have hands on experience with the Leica, Zeiss, Noran and other systems. They each have attractive features. A recent discussion of Zeiss and Leica will be sent directly to you. Make a list of features/advantages/disadvantages and weight them--the results may surprise you and guide your decision.
to subscribe to the confocal newslist send an email to:
{LISTSERV-at-LISTSERV.ACSU.BUFFALO.EDU}
in the body of the message write:
SUBSCRIBE CONFOCAL your name
Larry D. Ackerman Lily & Yuh Nung Jan Laboratories Howard Hughes Medical Institute UCSF, Box 0725, Rm U226 533 Parnassus Ave. San Francisco, CA 94143
Scanning Electron Microscopy (English) RasterElektronenMikroskopie (German)
(It's 3 long words in English, one monumental word in German. I once gave a talk titled: "Rasterelektronenmikroskopische Untersuchungsmethoden fuer prozessinduzierte Halbleiterdefekte".... German is not for the timid ;-)
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
} ---------- } From: "David_Bell-at-Millipore.com"-at-sparc5.microscopy.com[SMTP:"DAVID_BELL-at-MILLIP ORE.COM"-at-SPARC5.MICROSCOPY.COM] } Sent: Friday, March 03, 2000 9:02:50 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Question Re: REM } Auto forwarded by a Rule } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello Colleagues,
Twice now, I have had customers in Europe refer to a technique as REM. The sample prep sounds the same as SEM, and the first time I passed it off as a typo. Now, with this second reference, I'm not so certain. Is it possible that these customers are referring to SEM, possibly with their native language? Or is it a whole different technique/instrument? Could someone please help me clear up this confusion?
Thanks in advance,
David Bell Scientist Electron Microscopy Lab Millipore Corporation 80 Ashby Road Bedford, MA 01730 (781) 533-2108
For those who are 'hand sectioning', try the teflon coated single edged blades. I believe Ted Pella and EMS carry them. They are alittle more money, but they slide through tissue with great ease.
Cheers, Ken
On Thu, 2 Mar 2000, Gordon Couger 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 } -----------------------------------------------------------------------. } } } } ----- Original Message ----- } } From: "Chris Jeffree" {cjeffree-at-srv0.bio.ed.ac.uk} } } By the way, the best type of blade for hand sectioning used to be } } the Blue Gillette double-edged carbon steel razor blades, which } } many of you may be too young to know about. The new, doubtless } } improved, stainless steel type never seems to be so sharp, and } } their edges dull faster. Is there still a manufacturer of carbon steel } } razor blades left in the world, or am I doomed to mourn forever? } } I miss the old blades as well. I cured the shaving problem I quit. } For hand sections I have better luck with a more rigid blade. } Razor blades and straight razors tend to dig in for me. I have } a lot better luck with a 2.5 inch wood chisel sharpened on glass } with silicone carbide grit. It is a lot of trouble to get sharp but } it works better for me than razor blades. } } Gordon } } Gordon Couger gcouger-at-couger.com } } Stillwater, OK www.couger.com/gcouger } 405 624-2855 GMT -6:00 } } } }
The issue might only be are all the elements "visible" to the x-ray analyzer. I don't trust the C or N numbers that my system spits out. Since I am using EDS, I will usually trust my careful analyses to a few tenths of a percent. I can see down to about a tenth of a percent, but the relative errors are large. But it is usually well sufficient for determining the alloy.
Now just because the alloy is microstructurally uniform, do not count on the composition to be uniform. I have seen zoning in grains and changes in composition with length within single crystal ingots. And if you have multiphase microstructure, it can be challenging to determine to overall composition.
Warren S.
At 01:55 PM 3/3/2000 -0500, you wrote:
} Dear Colleagues } In a microstructurally uniform alloy, can one expect to get the chemical } composition of the alloy through microprobe (assuming all the corrections } are applied correctly)? Would the composition tally with the one } determined by chemical analysis? } TIA } Anita
In reply to David Bell's inquiry, I know some Eujropeans refer to an SEM as a Raster Electron Microscope.
Sam Purdy National Steel Technical Center Trenton MI
} ---------- } From: "David_Bell-at-Millipore.com"-at-sparc5.microscopy.com } Sent: March 2000 11:02 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Question Re: REM } } ------------------------------------------------------------------------ } 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 Colleagues, } } Twice now, I have had customers in Europe refer to a technique as REM. } The } sample prep sounds the same as SEM, and the first time I passed it off as } a } typo. Now, with this second reference, I'm not so certain. Is it } possible } that these customers are referring to SEM, possibly with their native } language? Or is it a whole different technique/instrument? Could someone } please help me clear up this confusion? } } Thanks in advance, } } David Bell } Scientist } Electron Microscopy Lab } Millipore Corporation } 80 Ashby Road } Bedford, MA 01730 } (781) 533-2108 } }
I have in my hot little hand a box of carbon steel single edge razor blades from Ted Pella. For the few times that I needed to to make a transparent section (being a LM metallographer), I found these razor blades to be effective. Usual disclaimers
Sam Purdy } ---------- } From: Grazyna M Tokarczyk } Sent: March 2000 926 } To: Gordon Couger } Cc: c.jeffree-at-ed.ac.uk; Dan Luchtel; microscopy-at-sparc5.microscopy.com } Subject: Re: hand sectioning - carbon blades } } ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } } } On Thu, 2 Mar 2000, Gordon Couger 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 } } -----------------------------------------------------------------------. } } Yes, carbon blades are still made - one of the makers is a Japanese } company - I have a box in my hand, but unfortunately I can't read Japanese } - the only English words on it is "Feather". There may also be many other } makers of these in Asia and Eastern Europe. } } Grazyna Tokarczyk } gmtokarc-at-is.dal.ca } Dalhousie University } Halifax, Canada } } } } ----- Original Message ----- } } } From: "Chris Jeffree" {cjeffree-at-srv0.bio.ed.ac.uk} } } } By the way, the best type of blade for hand sectioning used to be } } } the Blue Gillette double-edged carbon steel razor blades, which } } } many of you may be too young to know about. The new, doubtless } } } improved, stainless steel type never seems to be so sharp, and } } } their edges dull faster. Is there still a manufacturer of carbon steel } } } razor blades left in the world, or am I doomed to mourn forever? } } } } I miss the old blades as well. I cured the shaving problem I quit. } } For hand sections I have better luck with a more rigid blade. } } Razor blades and straight razors tend to dig in for me. I have } } a lot better luck with a 2.5 inch wood chisel sharpened on glass } } with silicone carbide grit. It is a lot of trouble to get sharp but } } it works better for me than razor blades. } } } } Gordon } } } } Gordon Couger gcouger-at-couger.com } } } } Stillwater, OK www.couger.com/gcouger } } 405 624-2855 GMT -6:00 } } } } } } } } } }
I am looking for information on an instrument that I recently acquired. The item is a McArthur Blood Cell Counter, which is a McArthur portable microscope with a number of mechanical additions and a special stage, apparently for doing blood count. It is in a neatly bundled case, very portable, but lacking instructions. If anyone one is familiar with the specific application of the set I would appreciate comments, particularly if you have ever used such an instrument.
There's also the technique of Reflection Electron Microscopy, which is the imaging analogue of RHEED (reflection high energy electron diffraction). I am sure your reference is to the SEM though, as the other respondents indicate.
Just for interest, REM (like RHEED) is a surface sensitive technique and can give beautiful images of monolayer surface steps, eg on the sapphire surface, and this can all be done in the TEM. The sample is simply rotated 90 degrees from the usual TEM imaging position and so electrons are 'reflected' from the surface at glancing angle as they pass through the sample chamber.
My best wishes to all,
Mark
%%%%%%%%%%%%%%%%%%%%%%%%%%% Mark Yeadon Senior Research Fellow Institute of Materials Research and Engineering 3 Research Link Singapore 117602
Assistant Professor Department of Materials Science National University of Singapore Singapore 119602
The original question has been answered well. However, I would like to add the historical footnote that von Ardenne's seminal 1938 paper was entitled "Das Elektronen-Rastermicroskop". A raster is a pattern of horizontal lines. The German word is derived form the Latin "rake". I guess a rake's pattern is a raster, a bit coarse for an SEM though. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Saturday, March 04, 2000 2:03 AM, "David_Bell-at-Millipore.com"-at-sparc5.microscopy.com [SMTP:"David_Bell-at-Millipore.com"-at-sparc5.microscopy.com] wrote: } } } Hello Colleagues, } } Twice now, I have had customers in Europe refer to a technique as REM. The } sample prep sounds the same as SEM, and the first time I passed it off as a } typo. Now, with this second reference, I'm not so certain. Is it possible } that these customers are referring to SEM, possibly with their native } language? Or is it a whole different technique/instrument? Could someone } please help me clear up this confusion? } } Thanks in advance, } } David Bell } Scientist } Electron Microscopy Lab } Millipore Corporation } 80 Ashby Road } Bedford, MA 01730 } (781) 533-2108 }
I have recently encountered several mentions of this type of thing recently, in the oddest places. One was a presentation by the motivational speaker, Tony Robbins, who, in front of 13,000 people at the Hartford Civic Center, described how his recent feelings of fatigue and lack of energy had been diagnosed by darkfield microscopy as due to chlamydia in his blood. He cited work done by Dr. Robert Young at InnerLight.
As for crystal testing, I am still a strong supporter of Polarized light work for that. (Interestingly, it looks a lot like darkfield and, perhaps, is mis-cited by Naturpaths who might not know the difference).
Hope this is helpful.
Caveat: MME has no connection with either Mr. Robbins, Dr. Young, or InnerLight.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
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We did a bit of market and applications research into this area for the 3 years before it emerged (as well as launching a company). These microarrays are typically analyzed on larger scale using instruments which are derivatives of confocal. If you are looking for individual DNA molecules first make sure that they are really likely to be there (i. e, get a full history of how this microarray was prepared and later used). I would then suggest looking at it with AFM. ThermoMicroscopes, Digital, and Molecular Imaging have all had experience in this area. Let me know if you need specific contact info.
Best regards Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education}
At 01:19 PM 3/2/00 -0600, Corazon Bucana wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Not only have I had my hands on several McArthurs, I had the privilege of visiting the inventor at home. Your comment about the "neat bundle" is very accurate. Dr. McArthur showed me a number of accessories (including fluorescence!) for this system. Although I did not have a chance for an extended use, my recollection was that all of them worked passably well.
There were several generations of McArthur microscopes. The early ones were made of metal and were very durable and usable. A somewhat later generation was made of white plastic, I think for the BBC Open University. The general opinion of some of my colleagues who knew this system indicated that the earlier generation was a better bet.
I was not aware that anyone was commercially making this microscope today. Dr. McArthur was advanced in years when I visited him over a decade ago and, at that time, their construction was very much a cottage industry. Is this a new system or are you buying it second hand? (I suspect the latter, since you mentioned that the instructions are missing). It is hard to comment on the bits and pieces without further information. I am planning to attend a number of meetings (Experimental Bio, M&M, Cell Bio, Neuro as well as Semicon West, Materials Solutions, and ISTFA) over the next 6 months. If you are going to any of them or will be in the area, I would be delighted to take a look at what you have and see if I can dredge up any old memories of how it works.
Finally, if you can send pictures, I can forward them to colleagues in the UK who own McArthurs.
Hope this is helpful.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education}
At 07:39 PM 3/3/00 EST, MICROFAB-at-aol.com"-at-sparc5.microscopy.com wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have recently encountered several mentions of this type of thing recently, in the oddest places. One was a presentation by the motivational speaker, Tony Robbins, who, in front of 13,000 people at the Hartford Civic Center, described how his recent feelings of fatigue and lack of energy had been diagnosed by darkfield microscopy as due to chlamydia in his blood. He cited work done by Dr. Robert Young at InnerLight.
As for crystal testing, I am still a strong supporter of Polarized light work for that. (Interestingly, it looks a lot like darkfield and, perhaps, is mis-cited by Naturpaths who might not know the difference).
Hope this is helpful.
Caveat: MME has no connection with either Mr. Robbins, Dr. Young, or InnerLight.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ******************************************************
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Amazing. Definitely a major feat of dark field light microscopy. To be able to see and distinguish an organism that is between 0.1u and 0.5u is truly amazing.
Did he say what magnification was realized in this DF system? The only DF condensers I know of for compound LMs are not aplanatic. So is begs the question of how one could resolve, much less see, such a tiny bacteria using LM.
Perhaps the sample was specially treated?
gary g.
At 01:09 PM 3/5/00 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
REM = 'Raster Elektronen Mikroskopie' in German (SEM in english) but also Reflection Electron Microscopy, which also needs not a lot of specimen preparation. It looks very much like preparation for SEM in deed. Regards Gerhard Frank
"David_Bell-at-Millipore.com"-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 } -----------------------------------------------------------------------. } } Hello Colleagues, } } Twice now, I have had customers in Europe refer to a technique as REM. The } sample prep sounds the same as SEM, and the first time I passed it off as a } typo. Now, with this second reference, I'm not so certain. Is it possible } that these customers are referring to SEM, possibly with their native } language? Or is it a whole different technique/instrument? Could someone } please help me clear up this confusion? } } Thanks in advance, } } David Bell } Scientist } Electron Microscopy Lab } Millipore Corporation } 80 Ashby Road } Bedford, MA 01730 } (781) 533-2108
-- Gerhard Frank UNIVERSITAET ERLANGEN-NUERNBERG Phone: +49 9131 85 28606 Institut fuer Werkstoffwissenschaften VII Fax: +49 9131 85 28602 Cauerstr. 6, D-91058 Erlangen, Germany frank-at-ww.uni.erlangen.de
Just a reminder that darkfield is detection limited, not resolution limited. It only takes 2-3 photons, scattered from a feature, to indicate that it is there.
Chlamydia is must be much bigger than the range you specified. We are in the process of sending out a special mailer for a new spectral imaging system, with chlamydia as the "cover shot". I think that this image was acquired with a 40x objective, but will check. Data is intentionally not provided on the card because we are encouraging recipients to go to the web page.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education}
At 04:03 PM 3/5/00 -0600, Dr. Gary Gaugler wrote: } Amazing. Definitely a major feat of dark field light microscopy. } To be able to see and distinguish an organism that is between } 0.1u and 0.5u is truly amazing. } } Did he say what magnification was realized in this DF system? } The only DF condensers I know of for compound LMs are } not aplanatic. So is begs the question of how one could } resolve, much less see, such a tiny bacteria using LM. } } Perhaps the sample was specially treated? } } gary g. } } } } At 01:09 PM 3/5/00 , you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Greetings, To respond to a comment made by Gary Gaugler, and without in ANY WAY seeking to validate or support the claims of naturopaths, darkfield microscopy has imaged individual microtubules, diam 25 nm. Same with Nomarski optics, and others. How is that possible? It is important to keep in mind what the famous "resolution limit" means, it means telling the difference between two and one object. It has nothing to say about the smallest absolute size of an object that can be detected.
If you imagine a perfectly black slide with a pinhole in it made by a perfect iris that can shrink right down to infinitely small (I did say a perfect iris), there is no limit in theory to how small that pinhole can be and still be imaged. Once the diameter of the pinhole becomes less than a certain length, set by diffraction, then the size of the image of the pinhole will no longer get any smaller. Instead, the contrast in the image will go down. But if you can put enough light through and have a good enough detector, you can see that image.
Going back to our microtubules, take a photo of the darkfield image and measure the diameter of the microtuble in the image: it will be around 250 nm or thereabouts.
For the record, the person who I think has published the most dark field images of microtubules is called Hotani, he works in Japan, I am not sure where.
Of course, doing this in practice is not easy, but it is certainly possible.
As ever, Tobias
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An earlier thread on this listserver (May-June 1999, and also summarized in the January/February 2000 issue of Microscopy and Microanalysis) asked about the availability of software for the processing and analysis of 16 bit per channel images, which are obtained from flat bed scanners, cooled digital cameras, and some microscopes (especially AFMs). Following the encouragement of members of this list, we've written a set of image processing and measurement routines for these images. Those interested can get information at http://members.aol.com/FoveaPro/
Disclaimer: Please note that Fovea Pro is a commercial product sold by Reindeer Games. My connection with the company is through my son, Chris, who wrote the programs, and my own role in writing the accompanying tutorial.
We have a Tracor Northern 5500 EDX unit that we would like to give away. It comes with a manual, mainframe, and monitor. If anyone is interested please contact:
Dr Brian Andrews NIH, NINDS Bethesda MD 301-435-2796
sba-at-helix.nih.gov
Thanks Chris :):):):):):):):):):):):):):):):):):):):):):):):):):):):):):)
Christine A. Brantner, Ph.D.
Biologist/Lab Manager
National Institutes of Health National Institute of Neurological Diseases and Stroke Lab of Neurobiology Section of Analytical Cell Biology Bldg 36, room 2A21 phone 301-435-2803 36 Convent Dr fax 301-480-1485 Bethesda, MD 20892-4062
Hello: I will add just a few comments to those of Tobias Baskin. When we first developed Video Microscopy in the early eighties, we were visulizing microtubules and other structures smaller than the ca. 200 nm resolution limit using DIC and computer image enhancement. This is discussed in the early papers from 1981 and 1983 (R.D. Allen and N.S. Allen 1983, J. of Microscopy),and more easily available in the Inoue and Spring second edition of Video Microscopy, Plenum Press. The important thing to remember is that you can visualize items smaller than 100 nm, but you are not resolving them. With the new method Polarization Modulation DIC we get very good images of microtubules and other fine structures (Holzwarth, G., Webb, S.J., Kubinski, D.J., N.S. Allen. 1997. Improving DIC Microscopy with Polarization Modulation. Jour. of Microscopy 188:249-254). Contrast is doubled with this method and it is worth a try. It is commercially available from Hamamatsu or you can build your own ,Nina Allen } Greetings, } To respond to a comment made by Gary Gaugler, and without in } ANY WAY seeking to validate or support the claims of naturopaths, } darkfield microscopy has imaged individual microtubules, diam 25 nm. } Same with Nomarski optics, and others. How is that possible? It is } important to keep in mind what the famous "resolution limit" means, } it means telling the difference between two and one object. It has } nothing to say about the smallest absolute size of an object that can } be detected. } } If you imagine a perfectly black slide with a pinhole in it } made by a perfect iris that can shrink right down to infinitely small } (I did say a perfect iris), there is no limit in theory to how small } that pinhole can be and still be imaged. Once the diameter of the } pinhole becomes less than a certain length, set by diffraction, then } the size of the image of the pinhole will no longer get any smaller. } Instead, the contrast in the image will go down. But if you can put } enough light through and have a good enough detector, you can see } that image. } } Going back to our microtubules, take a photo of the darkfield } image and measure the diameter of the microtuble in the image: it } will be around 250 nm or thereabouts. } } For the record, the person who I think has published the most } dark field images of microtubules is called Hotani, he works in } Japan, I am not sure where. } } Of course, doing this in practice is not easy, but it is } certainly possible. } } As ever, } Tobias } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- Nina Stromgren Allen Professor, Department of Botany; Director, Cellular and Molecular Imaging Facility Box 7612, Department of Botany North Carolina State University Raleigh, NC 27695-7612 Phone: 919-515-8382 (Office), 515-3525 (Lab), 515-2727 (Department Secretary) Fax: 919-515-3436
Is anyone familiar with a printer(s) that can provide equivalent detail to a polaroid print or to the images that can be collected from the scanning electron microscopes? I have looked at QMS Color Magic II printers that have 2400 dpi x 600 dpi, 257 shades of gray, with a cell size of 150 cells per inch (lines per inch). This is comparable to a Codonics 300 line dye-sub printer that I have seen. However, the dye-sub printers tend to smear fine details to the point of not being visible. I saw an add for an Alps printer with a dot size of 10 microns at 2400 dpi. The add did not indicate the shades of gray so I have no way of knowing what the numer of cells per inch are. I suspect that it would have 257 shades of gray which would give a cell size of 16x16 and a cells or lines per inch of 150. Does anyone have user information about these or any other printers that might work? No one talks about the cells per inch (lines per inch) in any of the documentation. For me, cells per inch has been the best measure of resolution. For the printers that we have, the extra dots (300 dpi matrix 4x4, 600 dpi matrix 8x8, 1200 dpi matrix 12x12, 2400 dpi matrix 16x16) serve to provide more dynamic shades of gray. A 300 dpi with 17 shades of gray and a 600 dpi with 65 shades of gray both have 75 cells per inch. The only difference is more contrast in the 600 dpi with 65 shades of gray. A simple formula for this is dpi/cell matrix size= cells (lines) per inch, 2400 dpi/16=150 cells per inch and a maximum # of shades of gray = 1+(dpi/lpi)squared. This tid bit of information came from The Image Processing Handbook, 2nd ed. by John C. Russ. I would appreciated any help that I can get in finding a printer that will do the job. Thanks.
In a message dated 3/6/00 5:55:31 PM, Vicki.L.Baliga-at-pmusa.com writes:
.."A 300 dpi with 17 shades of gray and a 600 dpi with 65 shades of gray both have 75 cells per inch. The only difference is more contrast in the 600 dpi with 65 shades of gray. A simple formula for this is dpi/cell matrix size= cells (lines) per inch, 2400 dpi/16=150 cells per inch and a maximum # of shades of gray = 1+(dpi/lpi)squared. This tid bit of information came from The Image Processing Handbook, 2nd ed. by John C. Russ...."
I guess it is worth pointing out that the calculation Vicki has quoted here is a bit optimistic. Most printers have dots that overlap somewhat, and the dots may spread a bit on the paper. Plus the dots are large enough to be resolved by the human eye, which creates interesting effects when they start to touch or overlap. And the shades of grey that perfect dots might produce would be linear rather than log, as human vision perceives brightness. It would be conservative to estimate that a printer that seems technically capable of producing (e.g.) 65 shades of grey in a given printing cell size will actually produce images with about half that number of really useful shades of grey. But that is still OK since that is about all the distinct shades that a person can detect on a print (and more than most Polaroid prints have, although the negatives are much better).
Halftone printing is never going to be as good as something that really covers the resolution cell with a uniform shade of grey (or color). Dye sub and a few ink jet printers do a much better job than laser printers in that regard.
I sent an email to LISTSERV-at-LISTSERV.ACSU.BUFFALO.EDU as you suggested with SUBSCRIBE CONFOCAL in the body of the message.
I got this message back: No LISTSERV list by the name of "CONFOCAL," is known to exist. Note that lists can be marked "confidential" and that the existence of such lists is usually known only to the server that is actually hosting it.
I'm very interested in finding a venue that is more oriented toward confocal microscopy. Any further suggestions?
Catherine Ripley ***************************************** Albert Einstein College of Medicine 1300 Morris Park Avenue Bronx, NY 10461 ***************************************** *****************************************
If you take a drop of blood from a pinprick and put it on a slide, you will see (at rather lower magnification) what the "live cell" analysts purport to see. You will see red cells starting to agglutinate. Thats all. It is nearly impossible to find a white cell. If you find a bacterium you are probably near death. Using dark field adds mystery but no more useful information.
Check out these websites for three different perspectives on live cell microscopy
1. http://www.hcrc.org/faqs/livecell.html
Critical evaluation pointing out the supposed features naturopaths claim as showing pathological change are natural variation.
Gushing plug for the technique including claiming ability to detect poor nutrition, vitamin deficiency etc. See the extract below.
None of these claims that illness can be diagnosed by observing coverslipped blood have any scientific validity at all.
"Live cell analysis can determine cellular nutritional status. Nutritional status is essential to aid in curbing and reversing the free radical cascade of destructive, deteriorating cell structures.
This unique analysis of peripheral blood can reveal a good prospectus of the immune system. By observing the morphology of the white corpuscles and their activity in contrast to the extent of active foreign antigens and microbes within the serum and red blood cells, the strength of the immune system can be judged. Weakness and dysfunction in any one of these three major components and influence the strength and function of the other two. This can be indicative of a progressive disease process."
A photographic negative and/or print is continuous tone. Digital printers (ink jet, laser, etc.) attempt to reproduce continuous tone via modulation of the size of the dots or the number of dots in an area. Continuous tone digital printers typically use dye sublimation to produce photographic quality results. In this regard, there really isn't a true correspondence to dpi. Alternatively, some ink jets use CMYK or even 5 color jets rather than RGB or RGB+B. Some of the new generation do a rather good job of color printing.
But since you are talking about SEM images, these are gray scale images rather than color. So, what are the options for b/w?
600dpi and 1200dpi laser printers do a rather good job of printing SEM images. One must adjust the toner density to achieve maximum quality results. But regardless, they are not continuous tone prints. The closest thing to continuous tone b/w is the Orion (I think that is the name) printer. It is an effective 300dpi and prints on expensive paper--which is par for the course, considering the cost of the printer (about $25K). And the printer is sloooooow.
I use a HP Laserjet 4M Plus and am very pleased with the results. But the printed images are not archival and are not continuous tone. But they are very inexpensive.
If you can describe what your actual end purpose is, perhaps others could offer some good options for you to consider. Are you talking about proofing, archiving, presentations, etc?
gary g.
At 11:20 AM 3/6/00 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Sir Could you please supply me details on "transmission electron microsocopy" for my 16yr old daughter at high school, or alternatively where to look on the Internet. Thanking you Peter Jupe
Splitting hairs on detecting microtubules vs. detecting pathogenic organisms...
I guess I will have a go at this and muddy the waters...
As a background...The inability of Chlamydiae to produce metabolic energy restricts these obligate 'parasites' to an intracellular existence.
The infectious component (elementary body) is about 3um in diameter, well within the resolvable range of the OLM. Once acquired in the host cell through phagocytosis, a vaculated 'initial body' (0.5 -1.0um) is formed: huge by OLM standards!
'Elementary bodies' and 'initial bodies' possess distinctive staining properties using Giemsa staining and Macchiavello's staining techniques, not be mention detection by immunocytochemistry.
One can perhaps differentiate Chlamydia induced inclusion bodies juxtaposed to the nucleus of the host cell using Darkfield illumination. However, differential diagnosis of Chylamydia by Darkfield alone is questionable.
If one considers Darkfield and the ability of a 'substance' of a given refractive index to scatter light with respect to the R.I. of the adjacent material, we must accept the notion of "detection vs. resolution' (B. Foster ' Optimizing Light Microscopy for Biological and Clinical Laboratories', pg. 57): therefore, we may detect an inclusion body, but can we identify the pathogen of the inclusion body? Perhaps Phase Contrast would be more sensitive to refractive differences?
As the devil's advocate, I see the issue of using Darkfield as a definite diagnostic tool being dilute to 'mine is smaller than yours'. Not being up on the naturopathic literature, I question how only using Darkfield Illumination can definitely dx Chlamydia?
However, comparing various organisms and the use of Darkfield Illumination, 'Treponema pallium' can easily be detected by Darkfield; an organism of 0.2 um wide, but 5-15 um in length! Furthermore, the inclusion body of Rabies can easily be detected by Darkfield and Brightfield; however, we i.d. the total sum of virons: an example of 'detection vs. resolution'.
Yes, we as microscopists scoff at unconventional claims. Perhaps, Mr. Tim Robbins should present a blood sample for us to examine under the light of mutual cooperation and enlightenment'!!
I present my premise for your delicate evisceration...
I also tried to subscribe to the confocal list when Larry suggested it and it worked like a charm. I cut and pasted the email address right from his posting.
LISTSERV-at-LISTSERV.ACSU.BUFFALO.EDU
Try again. Cheers, John
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-- C. John Runions, Ph.D. Department of Plant Sciences University of Cambridge Cambridge CB4 1YE UK
email at work cjr41-at-cam.ac.uk email at work runions-at-ntlworld.com Phone (01223) 766 545
In the earlier days {25 years ago} of medical diagnosis, before the advent of the rapid, highly specific methods of identifying pathogenic organisms, darkfield microscopy was routinely applied to rule out specific organisms. In particular, the diagnosis of syphilis was aided with this mode of imaging. When fluids from a suspect lesion were applied to a slide and imaged by darkfield, the characteristic shape of the spirochaete organisms was clearly seen. More recently, darkfield examination of synovial fluids from arthritic joints was used to rule out or diagnose Lyme disease, before specific testing became available.
Is there a short, general reference, like a review, that explains immuno EM, it's uses, major techniques, and shortfalls? I would like to use it to send to investigators that are thinking about using the technique but don't know much about labeling at the EM level. I have a lot of specific references and it would take a lot of time to reference or copy them. If the investigator wants to know more I can direct them to those, otherwise a general one would get them and me on the same page when we start talking specifics on their project. Thanks
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Rick Vaughn writes: Is there a short, general reference, like a review, that explains immuno EM, it's uses, major techniques, and shortfalls? I would like to use it to send to investigators that are thinking about using the technique but don't know much about labeling at the EM level. I have a lot of specific references and it would take a lot of time to reference or copy them. If the investigator wants to know more I can direct them to those, otherwise a general one would get them and me on the same page when we start talking specifics on their project. Thanks Rick Vaughn rlvaughn-at-unmc.edu
The best reference you can ask for is the 1993 book by Gareth Griffith: "Fine Structure Immunocytochemistry." Springer Verlag. In it you will find all the major techniques, pitfalls, antibody use, fixatives and even quantitative methods for estimating antigen number. My copy has been replaced many times, I think because someone else needed it more than I did!
by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id SAA01398 for dist-Microscopy; Wed, 8 Mar 2000 18:16:54 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id SAA01395 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Wed, 8 Mar 2000 18:16:24 -0600 (CST) Received: from [206.69.208.21] (mac21.zaluzec.com [206.69.208.21]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id SAA01388 for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 8 Mar 2000 18:16:13 -0600 (CST) X-Sender: zaluzec-at-ultra5.microscopy.com Message-Id: {v03007801b4ec9c2e23bd-at-[206.69.208.21]} Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
I know this has been covered periodically, but as I've not needed the technique, I've not paid attention. I need some methods of embedding cells cultured on glass coverslips. Obviously, the most important issue is removing the coverslip from the polymerized block.I've looked through my own reference materials and the only method I found involved using Thermanox coverslips (upon which these particular cells will not grow, I'm informed).Please reply dircectly to my email address as well as to the Listserver (I haven't received postings for several days). Thank you,Jaclynn M. Lett, Research Assistant jmlett-at-cid.wustl.eduFay and Carl Simons Center for Biology of Hearing and DeafnessCentral Institute for the Deaf4560 Clayton AvenueSt. Louis, MO 63110voice: 314-977-0257 fax: 314-977-0030
I need to fix cells for viewing with SEM. However, the fixation method needs to be consistent with my experimental methods:
1) I need to fix the cells in less than 1 minute and due to this time limitation, I cannot spin the cells into a pellet and must add the cell suspension to the fixative; and
2)I am looking for "effects" to the cell membrane caused by my experiment, so I need to keep any possible fixative damage/effects at a minimum.
I have fixed cells using plunge freezing and viewed them with TEM, this is time consuming and I ran into a lot of freeze damage. I would like to try chemical fixation and SEM. It has been suggested that I try glutaraldehyde + buffers, then postfix with osmium tetroxide and dehydrate in an ethanol series.
Is this the best method? I am open to suggestions. I am using suspensions of prostate cancer cells in RPMI (+serum) growth media; I can perform the experiment in buffered saline and use a high concentration of cells.
Thanks, Robyn
Robyn K. Schlicher, M.S. Laboratory for Drug Delivery Institute for Bioengineering and Biosciences Georgia Institute of Technology 315 Ferst Drive Atlanta, GA 30332 rkslick-at-yahoo.com
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hi } i am doing research at university of massachusetts lowell.i am working on } charaterization of Polytetrafluoroethylene(PTFE).While doing TEM i have } stained the PTFE with osmium tetroxide.Can any one enlighten me on the } mechanism oF OsO4 with PTFE? } thanks } ________________________________________________ } Amit A.Kaulgud } Research Assistant. } University of Massachusetts Lowell } Department of Chemical/Materials Engineering. } 170, Riverside Street,#3 } Lowell MA -01854. } Phone/Voice:978-459-3248. } email: amit_kaulgud-at-hotmail.com
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Below is a question from one of my former students who is now working in a hopsital histotech lab. It concern the selflife of glutaraldehyde fixative. I always mix my fix fresh, so some other perspectives might be useful!! Here's the Question:
} My boss wants to know how long a working solution of glutaraldehyde } remains stable (2.5% in cacodyalate buffer). How long can we keep it in } the refrigerator? } Bob Robert J. Schmitz Electron Microscope Lab Department of Biology, CNR Building University of Wisconsin, Stevens Point Stevens Point, WI 54481 phone (715) 346-2420 FAX (715)346-3624 email rschmitz-at-uwsp.edu http://biology.uwsp.edu/faculty/RSchmitz/home.html
In addition, I would point that Paul's web site at http://www.hei.org/htm/cryo.htm is not bad source, too.
} Date: Wed, 08 Mar 2000 00:40:11 -0800 } From: Paul Webster {pwebster-at-hei.org} } Subject: TEM - ImmunoEM reference } To: MSA listserver submission {Microscopy-at-sparc5.microscopy.com} } Reply-to: Paul Webster {pwebster-at-hei.org} } X-Mailer: QuickMail Pro 1.5.4 (Mac) } X-MIME-Autoconverted: from quoted-printable to 8bit by ultra5.microscopy.com id } CAA01345 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
At the Fall MRS this year there will be a symposium dedicated to environmental SEM and low vacuum SEM, invited speakers include Eric Doehne from Getty Conservation Institute, David Joy from the University of Tennessee and Matthew Phillips from The University of Technology in Sydney. Contributed papers from all environmental SEM and low vacuum SEM users are encouraged, the Preliminary Call for Papers follows:
Preliminary Call for Papers , Session for MRS 2000 (Boston) http://www.mrs.org/meetings/fall2000/cfp/symposia.html
Low Vacuum SEM/ESEM in Materials Science "Wet SEM: the Liquid Frontier of Microscopy"
Abstract Deadlines: June 5th for abstracts sent by mail and June 19th for abstracts submitted via the MRS Web Site (http://www.mrs.org).
Scanning electron microscopy and x-ray microanalysis can now be performed at pressures as high as 1 - 5000 Pa (~0.01 to 40 torr) in the class of instruments known variously as "low vacuum", "elevated pressure", and "environmental" SEMs. This represents an increase of 3 to 6 orders of magnitude in pressure compared to the operating conditions of ordinary high vacuum SEMs. At the upper end of this pressure range, water can be maintained in the liquid state at temperatures from 3 - 10 deg. C. This remarkable situation has opened up broad new areas of opportunity in many fields, especially materials science. Dynamic processes can be observed with many of the familiar strengths of conventional SEM: high resolution, large depth of focus, and a variety of imaging signals that reveal compositional, topographic, electrical and other specimen properties. X-ray microanalysis for elemental characterization is also possible, although somewhat compromised. Chemical, physical, and mechanical experiments can readily be performed in user-specified environments. The microscope can thus be thought of as an appendage to an experimental chamber. Papers are solicited in all aspects of materials science in which this new microscopy is used.
Co-organizers:
Dr. Brad Thiel Polymers & Colloids Group Cavendish Laboratory University of Cambridge Madingley Road Cambridge, CB3OHE, UK 44 1223 337 272 44 1223 337 000 (fax) bt202-at-cam.ac.uk
Dr. John Mansfield North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 (734) 936-3352 (734) 763-2282 FAX jfmjfm-at-engin.umich.edu
Dr. Dale Newbury Surface and Microanalysis Science Division National Institute of Standards and Technology Gaithersburg, MD 20899-8371 USA 301-975-3921 301-417-1321 (fax) dale.newbury-at-nist.gov
--
Dr. John Mansfield CPhys MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 763-2282 Cellular Phone: (734) 358-7555 (Leaving a phone message at 936-3352 is preferable to 358-7555) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42#161# 16' 48" Long. 83#161# 43' 48"
I have used fluoronanogold for a couple of times and it did not work on the EM level. Then I realized that this batch has been already expired. Does anyone know if upon aging fluoronanogold could lose gold leaving only FITC molecule attached to Ig? I did see a fluorescence signal before running silver enhancement though. Currently I will try regular ultrasmall gold probe to clear up this problem but I wonder what was the trick.
thanks for your help
Kyrill
********************** Dr. Kyrill Ukhanov Institute for Zoophysiology, University of Potsdam, Lennestrasse 7a, D-14471 Potsdam, Germany phone +49-0331-9774859 fax +49-0331-9774861
Dear Jaci, The problem of cell embedding after their cutivation on plastic or galss is rather simple. You do not need to use Thermanox. You can use glass. The trick is that immediately after polymerization you have place glasses at -20 degree C and your samples will be detached. If thsi scheme does not work you can use liquid nitrogen. However, do not insert smplaes into it (only glass). If this scheme does not work you can use commercial solution of HF (acid). It dissolve glass efficiently. For instance, coverslips are dissolved in 30 min. Then you have to wash samples in buffer (pH7.4) for a long time and water (several hours) otherwise you will have problems with contrasters. We usually glue samples after detachment from glass and work with this sandwich. For this you have to use incomplete polymerization of Epon on chamber slides (12 hours). Then samples can be detached at -20 and then glued with the fresh resin for 24 hours. If you use full polymerization time on glass or any polymerization on plastic you will not be able to glue samples.
Sincerely yours, Alexander Mironov Italy
On Wed, 8 Mar 2000, Jaci Lett 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 know this has been covered periodically, but as I've not needed the } technique, I've not paid attention. I need some methods of embedding } cells cultured on glass coverslips. Obviously, the most important } issue is removing the coverslip from the polymerized block.I've looked } through my own reference materials and the only method I found involved } using Thermanox coverslips (upon which these particular cells will not } grow, I'm informed).Please reply dircectly to my email address as well as } to the Listserver (I haven't received postings for several days). Thank } you,Jaclynn M. Lett, Research Assistant } jmlett-at-cid.wustl.eduFay and Carl Simons Center for Biology of Hearing and } DeafnessCentral Institute for the Deaf4560 Clayton AvenueSt. Louis, } MO 63110voice: 314-977-0257 fax: } 314-977-0030 } } } }
A few moments in liquid nitrogen and the coverslip will come off. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Thursday, March 09, 2000 10:10 AM, Jaci Lett [SMTP:jmlett-at-cid.wustl.edu] wrote: } } } I know this has been covered periodically, but as I've not needed the } technique, I've not paid attention. I need some methods of embedding } cells cultured on glass coverslips. Obviously, the most important } issue is removing the coverslip from the polymerized block.I've looked } through my own reference materials and the only method I found involved } using Thermanox coverslips (upon which these particular cells will not } grow, I'm informed).Please reply dircectly to my email address as well as } to the Listserver (I haven't received postings for several days). Thank } you,Jaclynn M. Lett, Research Assistant } jmlett-at-cid.wustl.eduFay and Carl Simons Center for Biology of Hearing and } DeafnessCentral Institute for the Deaf4560 Clayton AvenueSt. Louis, } MO 63110voice: 314-977-0257 fax: } 314-977-0030 } }
I presume all microscopists use somewhat arbitrary shelf-life assumptions, because we know from experience that within those constraints there normally is no trouble. UA in water } 1 month, made up Spurr's in freezer } 3 months, Lead Citrate } 1 year. For working strength GA I used quite conservatively 1 week.
GA forms an undesirable polymer with time and temperature. So freezing the bulk stock (ampoules can stand a fridge-freezer) results in a very long life, certainly a couple of years and just refrigerated its several months. This is arbitrary too since there is no definite cut-off when the stuff suddenly becomes useless.
I saw a publication many years ago that claimed the "undesirable polymer" protected cells against osmotic shock and in structural studies some polymer may be desirable (the amount can be determined simply in a spectrometer, please don't ask for the peak locations. I don't carry that info!) Apparently its different for immunological studies, for that the freshest GA should be used.
Soon after Sabatini advocated GA for TEM use, I guess now over 30 years ago, as noted, some studies related polymer to storage time/temperature. I don't think that these looked at working strength solutions, but I expect that there would be no difference in relative polymer concentrations.
The reason for shorter shelf-life for working solutions was assumed and probably originated with the common use of additives like sucrose or phosphate buffers. Sucrose in an Os fixatives leads to obvious oxidation within hours. I doubt that cacodylate in GA would be a problem and expect that it would remain useable for several months when refrigerated. I have never bothered to test this, however, and stuck with an assumed short shelf-life for working strengths GA. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Thursday, March 09, 2000 10:09 AM, Schmitz, Robert [SMTP:rschmitz-at-uwsp.edu] wrote: } } } Below is a question from one of my former students who is now working in a } hopsital histotech lab. It concern the selflife of glutaraldehyde fixative. } I always mix my fix fresh, so some other perspectives might be useful!! } Here's the Question: } } } My boss wants to know how long a working solution of glutaraldehyde } } remains stable (2.5% in cacodyalate buffer). How long can we keep it in } } the refrigerator? } } } Bob } Robert J. Schmitz } Electron Microscope Lab } Department of Biology, CNR Building } University of Wisconsin, Stevens Point } Stevens Point, WI 54481 } phone (715) 346-2420 } FAX (715)346-3624 } email rschmitz-at-uwsp.edu } http://biology.uwsp.edu/faculty/RSchmitz/home.html } }
Email: puusimak-at-paju.oulu.fi Name: Päivi Maria Susanna Uusimäki
School: the university of Oulu
Question: What is a suitable fixation-method for my specimens? I am trying to find antigen-antibody interactions on the bacterial membrane. My bacteria are different strains of LABs, my special interests are lipoteichoic acids and I am using confocal and fluorescence microscopes. The big problem has been : specimens tend to "float away" under was- hings !
I have recently been asked to participate in an effort at Los Alamos concerned with Electron Tomography of polymers. I haven't done the required image reconstruction before. Does anyone know of any commercially available software that will reconstruct several 2D images taken at various tilt angles into a single 3D reconstruction of an object? I suppose it would be analogous to the software used for CAT scans? Also, if you've done this before, what kind of hardware requirements are there? I've got a JEOL 2010 configured at the moment to allow only ±30 degrees of tilt. Is that enough? I assume I also need some way of automating the specimen tilting process and will need beam blanking to avoid specimen damage. Can anyone confirm or deny all this?
Thanks all. It was just some anomaly. It went through the second time.
Catherine Ripley ***************************************** Albert Einstein College of Medicine 1300 Morris Park Avenue Bronx, NY 10461 ***************************************** *****************************************
EuroFE 2000 is a forum to bring together interested parties in the field of Field Emission Technologies. This includes microscopists working with materials used for field emission sources (DLC, Nanotubes etc.) and those developing field emission based sources for SEM & TEM.
There is also a strong emphasis on links with other display technologies such as LCD's, Light Emitting Polymers and Phosphors.
The conference and workshop sessions will build on the success of EuroFE '99 & will be held in the UNESCO World Heritage city of Segovia.
The aim of this conference will be to focus on the European dimension of Field Emission, on Industrial Problems and to bring together in a Forum the various groups (from Industry and Public Institutions) working in this area. In addition to the normal program of keynote speakers, oral presentations and poster sessions, it is also planned to hold workshops on topics such as "overcoming obstacles to industrial production".
In common with EUROFE'99 conference, there will be an emphasis on the applications of field emission technologies, as well as basic scientific research. A crucial issue during a conference is time allowed to discussions between participants in order to exchange ideas and therefore possibly define strategies in order to solve real problems. During EUROFE'2000, following each session, large breaks will be set-up in order to allow these discussions.
One of the major sessions will be related to Field Emission applications such as future big FE flat panel displays (1m2). Companies such as Motorola, Samsung, PixTech, Candescent and Saint-Gobain will participate actively in this session. The "BigFED" project coordinated by EUROFE will be presented at the conference in order to define strategies to be able to present projects to the EU within this objective.
The key topics of interest will be:
Industrial applications (Flat panel displays, etc.) and related problems (reliability, lifetime, etc.) Field Emission from diamond, DLC and nanotubes FE simulation and modelling Understanding Field Emission Space applications Device characterisation (surface analysis, etc.) Novel cathodes - technology and fundamentals Other related areas: (Phosphors, LCDs, OLEDs, CRTs, New materials, Novel devices, etc.).
An important issue during conferences is to make possible student participation in order to form them and allow initiation of contacts with either industry or public institutions. For this reason, this year, the EUROFE2000 organisation set-up a special reduced registration fee for students including also the accommodation.
Regards
Tim
****************************************************************** Tim E. Harper EuroFE Network Co-Chairman EuroFE is a network of the European Science Foundation Phone +34 91 640 71 85 Fax +34 91 640 71 86
Responding to the message of {v03007805b4ec9d6d6eae-at-[206.69.208.21]} from R schlicher {rkslick-at-yahoo.com} :
} I need to fix cells for viewing with SEM. However, } the fixation method needs to be consistent with my } experimental methods: } } 1) I need to fix the cells in less than 1 minute and } due to this time limitation, I cannot spin the cells } into a pellet and must add the cell suspension to the } fixative; and
snip!
I would like to try chemical } fixation and SEM. It has been suggested that I try } glutaraldehyde + buffers, then postfix with osmium } tetroxide and dehydrate in an ethanol series.
Robyn,
You could try simultaneous glutaraldehyde + osmium fixation, which should work well for cells in suspension in particular. Mix up seperate solutions of each, such that when mixed in equal amounts, you get the concentrations of fixes and buffers you want. Its usually recommended that you do this at lower temperatures, like about 4C, so pre-cool the solutions and vials first, to prevent the glut and osmium from fighting with each other, but even for "less than a minute", as you put it, maybe you could still work at room temperature. Try mixing the two fixatives together without cells first, to determine the maximum time until discoloration occurs, which means the osmium is precipitating out or whatever goes on there.
Your first rinse could just be a large dilution, to seperate the warring fixatives, then spin down and rinse a few more times before going into the dehydration series.
Give me a few hours to dig out references for this technique, will send them out later.
Gib
Gib Ahlstrand Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN. USA. 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.cdl.umn.edu http://biosci.umn.edu/MIC/consortium.html
Here are two references for the simultaneous glut/osmium technique I mentioned earlier.
1. Franke, W.W., Krien, S. & Brown, J.R.M. (1969) Simultaneous glutaraldehyde osmium tetroxide fixation with post osmication. Histochemie, 19, 162-164.
2. Hirsch, J.G. & Fedorko, M.E. (1968) Ultrastructure of human leukocytes after simultaneous fixation with glutaraldehyde and osmium tetroxide and post fixation in uranyl acetate. J. Cell Biol. 38, 615-627.
Good luck!
Gib Ahlstrand
Responding to the message of {v03007805b4ec9d6d6eae-at-[206.69.208.21]} from R schlicher {rkslick-at-yahoo.com} :
} Hello, } } I need to fix cells for viewing with SEM. However, } the fixation method needs to be consistent with my } experimental methods: } } 1) I need to fix the cells in less than 1 minute and } due to this time limitation, I cannot spin the cells } into a pellet and must add the cell suspension to the } fixative; and } } 2)I am looking for "effects" to the cell membrane } caused by my experiment, so I need to keep any } possible fixative damage/effects at a minimum. } } I have fixed cells using plunge freezing and viewed } them with TEM, this is time consuming and I ran into } a } lot of freeze damage. I would like to try chemical } fixation and SEM. It has been suggested that I try } glutaraldehyde + buffers, then postfix with osmium } tetroxide and dehydrate in an ethanol series. } } Is this the best method? I am open to suggestions. } I am using suspensions of prostate cancer cells in } RPMI (+serum) growth media; I can perform the } experiment in buffered saline and use a high } concentration of cells. } } Thanks, } Robyn } } } Robyn K. Schlicher, M.S. } Laboratory for Drug Delivery } Institute for Bioengineering and Biosciences } Georgia Institute of Technology } 315 Ferst Drive } Atlanta, GA 30332 } rkslick-at-yahoo.com
Gib Ahlstrand Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN. USA. 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.cdl.umn.edu http://biosci.umn.edu/MIC/consortium.html
At 6:09 PM -0600 3/8/0, Jaci Lett wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Your are left with a mirror-finish block face that is perfectly intact. Even with all the safety precautions, I find this method preferable and more reliable in my hands than the "heat and snap-off" method that many people use.
Just don't let your students do it!
Good luck, Lee
Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Confocal Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
Dear microscopists, Can anyone offer advice on doing immunofluorescence at the light level? I have cryopreserved plant tissue that I am contemplating embedding and using for immunolocalization. I have a protocol that uses butyl, methyl methacrylate. I am wondering several things. Why butyl, methyl methacrylate? Are there other resins/media that are sufficient? I'd appreciate any advice anyone has. Kristen Lennon Kristen A. Lennon Cell, Molecular & Developmental Biology Group Department of Botany & Plant Sciences University of California Riverside, CA 92521 kalen-at-citrus.ucr.edu
I have a question that is not a typical microscopy question but I thought I would call upon the extensive knowledge of the readers of this list server.
Does anyone know who manufacturers elemental boron?
We are using boron as a substrate for particle microanalysis (especially carbonaceous particles). Elemental boron can be purchased in the form of lumps or nuggets several centimeters in dimension from chemical supply houses such as Alfa Aesar and Aldrich. These companies however will not reveal their suppliers and so any questions that I want to ask the manufacturer have to go through the third party supply house. In addition to being silly, this procedure is extremely slow and inefficient. I would prefer to bypass the third party altogether.
So if anyone has any idea who manufacturers elemental boron I would greatly appreciate hearing from you.
Disclaimer: Any opinions expressed are my own and not those of my employer, The Federal Government.
Eric Windsor
Eric S. Windsor Physical Scientist NIST 100 Bureau Dr. Stop 8371 Gaithersburg, MD 20899-8371 (301) 975-3930 Fax: (301) 417-1321 Eric.Windsor-at-NIST.Gov
One of the researchers here is trying to do silver and/or gold enhancement of an immunolabel in brain slices, but is getting amazing background. We switched to the gold enhancement because everyone was saying how much cleaner it is....didn't help. I've seen one report in which the researchers pre-washed their samples in EDTA, because they felt that they were getting background from Mg in the tissue that was nucleating silver deposition - anyone else have any thoughts/experience on this? Anyone else doing enhancement in tissue slices?
He is using kits to do the enhancement - I'm not sure which companies are the suppliers, but they aren't supposed to be light-sensitive (my first culprit). The samples are washed extensively in water before enhancement, and even the no-antibody controls are "lighting up".
I do not have the information you need but I am old enough to remember seeing one of the first papers on tomography. It chose a cute title, something like: "Algorithm for Reconstructive Tomography or ART" Which was fine until the next issue of the journal had a paper (giving an improvement on the method) called: "Fast Algorithm for Reconstructive Tomography."
a transmission electron microscope is in principle the same as a transmission light microscope, except that it uses high energy electrons instead of light. This allows a much higher resolution (it is possible to see atoms or cyrstalline structures), but necessitates much bigger instruments, as there is vacuum technology, high voltages and X-ray generation involved. Many different techniques are available on these machines.
Rather than explaining all to you, you could do a search on the web looking for "transmission electron microscopy" or "TEM". Another starting point would be to go to
http://ncmi.bcm.tmc.edu/
and select one of the sites under "Links". Or go to the MSA site
http://www.msa.microscopy.com/
and find your way from there. I am sure people will help you here if you have specific questions.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
} ---------- } From: JUPE Peter[SMTP:JUPE.PETER-at-HDH.COM.AU] } Sent: Monday, March 06, 2000 8:35:46 PM } To: 'Microscopy-at-MSA.Microscopy.Com' } Subject: Transmission electron microsocopy } Auto forwarded by a Rule } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Sir Could you please supply me details on "transmission electron microsocopy" for my 16yr old daughter at high school, or alternatively where to look on the Internet. Thanking you Peter Jupe
Dear Jaci. If you wish to have access to all the cells on the coverslip, the best way is to flat embed the whole coverslip in a Chang monolayer mold (20mm or 10mm) available from EMS. Be sure to drain the coverslips well and wipe excess resin from the bottom of the coverslip. Invert over filled mold and polymerize overnight. Remove from oven and use metal file to file down edges of embedded coverslip, then immerse in concentrated hydrofluoric acid (use plastic forceps, work in fume hood, wear nitrile glove sand rinse everything well with water). It will dissolve the glass in 15-30 mins. It works every time and you have a complete embedded coverslip. You can then stain it with toludine blue, find the identified cell, mark it with a Ladd marking objective. I then punch out the cell(s) of interest using a hole punch and attach the 5mm circles to blank stubs using quick setting epoxi-patch bond (hardens in 5-10 minutes). This is available from Dexter Corporation 1-603-474-5545 and works extremely well. I sometimes have low contrast so Alexander Mironov's washing method may solve this problem. Otherwise, stain for 30 mins in 5% aqueous UA and 3 mins in Lead citrate for good results. Good luck, JoAnn Buchanan
When I stepped outside of the science building tonight, I ran into, literally, our university president who said he was just walking over to try to find me. He cautioned me about overreacting and then proceeded to tell me to go ahead and get my best quote on a new SEM and service contract!!!!!!
Now that I've got both feet back on the ground, I need any help that any of you can provide, based on your experience and knowledge. We're looking for a new SEM that will support undergraduate courses, be a good training machine for students, support student and faculty research and have a good track record for reliability and being reasonably user friendly. It should be state of the art, but not be so expensive to operate and keep up that it would limit how much students and faculty could use it.
I was trained on an Amray in the 80's and ran our TEM until it became cost ineffective a few years ago. But I know things have changed considerably since my pre-computer controlled everything SEM's and would appreciated the benefit of any experience any of you might have had with new SEMS.
Anything to recommend? Models? Features to insist on or avoid? Reliability, service costs and frequency? Companies to go with or stay away from? What new models do you have and would you recommend them to others? Why or why not?
I'd really appreciate any and all insight you can provide. When your president walks to YOUR office and says the equipment grant proposal HE made to a major foundation was just approved and that he needs me to get the best quote on an SEM to him ASAP, I want to have something to go on, and reasons to go along with it based on user input, without delay.
Thanks in advance for any assistance you can provide.
Lee Hadden Professor and Chair, Department of Biology Wingate University Wingate, NC 28174
I am only using distilled Glutaraldehyde - and only up to 7 days after making up the fixative (Karnovsky's) and storing it in the fridge.
UV Spectra of pure Glutaraldehyde show a single absorbance peak at 280 nm (monomeric ) - whereas the commercial material shows an additional stronger peak at 235nm. This is assumed to be due to the formation of oligomer and polymer. The ratio of absorbance at 235 and 280 nm can be used as a measure of the quality of the GA.
Monomeric-polymeric mixtures yield good ultrastructural preservation when the ratio of monomeric to polymeric forms is 1:1 or 1:2. Ratios outside this limits may prove unsatisfactory. (Weakley 1974) (unfortunately I do not have the exact source/ Paper handy where I got that from.)
Dr. C. Hayward-Costa School of Life Sciences Kingston University Penrhyn Road, Kingston upon Thames Surrey KT1 2EE, UK 44(0)181 547 2000 x 2240 Email: c.hayward-at-kingston.ac.uk
I have 95% success detaching glass coverslips embedded with epon inverted on beem capsules described by Leona by simply placing the polymerized coverslip on a hotplate ( } 85.C) with the beem capsule up and gently prying them apart with a straigtedge razor. Cells stay where you want them. I found by that instead of using the beem capsules I just use the lid of one or the lid of a microfuge tube ( makes a better platform). Once separated put the top on a slide and the cells are easily visualized with brightfield (osmicated) or phase (non osmicated). Hank Adams Integrated Microscopy Core Molecular and Cellular Biology Baylor College of Medicine Houston, TX
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Reply to: Re: methods of embedding cells cultured on glass coverslips Growing cells on coverslips, fixing them in situ and processing them into epoxy resin is fairly straight forward. Remember that the cells are a monolayer and are only about 30 microns thick so fixation, rinsing, dehydration and infiltration times can be cut down considerably.
The final step of removing the coverslip from the polymerized block is not, however, as trivial as everyone makes it out to be. Yes, dipping the warm block in liquid nitrogen and then re-warming it, will eventually take the glass off the plastic (leaving the cells in the block), but only if the thin layer of plastic that always seems to appear on the back of the glass is removed. The block and glass are also more easily removed if the resin is polymerized with the glass resting cell-side down on resin. The alternative way, of immersing the coverslip cell-side up, and letting the resin cover the cells, is safer, but leads to problems in removing the glass.
If there is a problem with removing the glass, making scratches with a diamond scribe can help and sometimes bending the whole block and glass coverslip can be useful ( if this is possible).
Don't worry about immersing the block in the nitrogen either, there really doesn't seem to be just one way that this happens. If the process is able to remove the glass, the glass may shoot off so violently you may wish you had put on your safety glasses. The next block may not work.
One other method I heard of was to put the block, glass-side down, on a hot plate. Wait until the glass gets hot and I'm told it will easily slide off. I never tried it. Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm
Alexander Mironov wrote: } } Dear Jaci, } The problem of cell embedding after their cutivation on plastic or galss } is rather simple. You do not need to use Thermanox. You can use glass. The } trick is that immediately after polymerization you have place glasses at } -20 degree C and your samples will be detached. If thsi scheme does not } work you can use liquid nitrogen. However, do not insert smplaes into it } (only glass). If this scheme does not work you can use commercial solution } of HF (acid). It dissolve glass efficiently. For instance, coverslips are } dissolved in 30 min. Then you have to wash samples in buffer (pH7.4) for a } long time and water (several hours) otherwise you will have problems with } contrasters. } We usually glue samples after detachment from glass and work with } this sandwich. For this you have to use incomplete polymerization of Epon } on chamber slides (12 hours). Then samples can be detached at -20 and then } glued with the fresh resin for 24 hours. If you use full polymerization } time on glass or any polymerization on plastic you will not be able to } glue samples. } } Sincerely yours, Alexander Mironov } Italy } } } On Wed, 8 Mar 2000, Jaci Lett wrote: } } } I know this has been covered periodically, but as I've not needed the } } technique, I've not paid attention. I need some methods of embedding } } cells cultured on glass coverslips. Obviously, the most important } } issue is removing the coverslip from the polymerized block.I've looked } } through my own reference materials and the only method I found involved } } using Thermanox coverslips (upon which these particular cells will not } } grow, I'm informed).Please reply dircectly to my email address as well as } } to the Listserver (I haven't received postings for several days). Thank } } you,Jaclynn M. Lett, Research Assistant } } jmlett-at-cid.wustl.eduFay and Carl Simons Center for Biology of Hearing and } } DeafnessCentral Institute for the Deaf4560 Clayton AvenueSt. Louis, } } MO 63110voice: 314-977-0257 fax: } } 314-977-0030 } }
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm
by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id PAA04949 for dist-Microscopy; Thu, 9 Mar 2000 15:14:56 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id PAA04946 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Thu, 9 Mar 2000 15:14:26 -0600 (CST) Received: from newton.wadsworth.org (newton.wadsworth.org [199.184.18.6]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id PAA04939 for {microscopy-at-sparc5.microscopy.com} ; Thu, 9 Mar 2000 15:14:15 -0600 (CST) Received: from wadsworth.org (ithaca [172.16.1.89]) by newton.wadsworth.org (8.8.8/8.8.8) with ESMTP id QAA20303 for {microscopy-at-msa.microscopy.com} ; Thu, 9 Mar 2000 16:09:09 -0500 (EST) Sender: tivol-at-wadsworth.org Message-ID: {38C8115C.5905598E-at-wadsworth.org}
Chris Adams wrote:
Dear Chris,
} I have recently been asked to participate in an effort at Los Alamos } concerned with Electron Tomography of polymers. I haven't done the } required image reconstruction before. Does anyone know of any } commercially available software that will reconstruct several 2D } images taken at various tilt angles into a single 3D reconstruction } of an object?
Yes. SPIDER is commercially available, and will do 3D reconstructions by any of several methods. Check it out at www.wadsworth.org; click on Resources (pretty far down on the left-hand side); then click on SPIDER.
} I suppose it would be analogous to the software used } for CAT scans?
Some of the reconstruction methods, at least, are used in CAT scans, but others (e.g., projection onto convex sets) may not be--I don't know the latest software for CAT scans.
} Also, if you've done this before, what kind of } hardware requirements are there?
We run on an Onyx using unix; I don't know all the available versions of SPIDER.
} I've got a JEOL 2010 configured at } the moment to allow only ±30 degrees of tilt. Is that enough?
No. +/- 50 deg is adequate, +/- 60 deg is better.
} I assume I also need some way of automating the specimen tilting } process and will need beam blanking to avoid specimen damage. Can } anyone confirm or deny all this? }
Automation is optional, but beam blanking is mandatory-- especially for cryo-specimens. If your goniometer is sufficiently accurate, you can change the tilt setting with the beam blanked, then adjust the position and focus quickly after the beam is back on. If you don't have a sufficiently good goniometer, you need to have either a low-dose kit (assuming you can recognize both your area of interest and a nearby area for focusing) or a very sensitive video-rate camera, such as the intensified CCD we have on the HVEM, so you can find and focus the area of interest with minimal exposure of the specimen.
Hi Kristen, I have published several papers on using butylmethylmethacrylate for immuno at the light level, including a paper where cryofixation and freeze substituion were used, on plant material. Perhaps the protocol you have is by me?
To answer your questions, a mixture of butyl and methyl methacrylate gives nice blocks and preserves the tissue well. I have tried using pure butyl or pure methyl and the sectioning or preservation was worse. These resins, alone or in combination, have the crucial advtange of being mostly removable post embedment with a brief incubation in actetone. This improves the access of your antibody to your antigen greatly. It is important when doing immuno work at the light level on sections to remember that if an antibody can penetrate say 15 nm into the plastic (I am guessing at this number), this is most of the thickness of an ultra thin section but only a teeeny bit of a semithin section. So, for light work especially, being able to remove the embedment is crucial. The usual way to do this is to embed in paraffin and this is fine if you just want tissue level distinctions. But if you want subcellular localization or for other reasons you want your stuff to be better preserved, then plastic gives much nicer results. DOes this help? I will be quite happy to help you further with butylmethyl methacrylate, as I have a professional fondness for the stuff. Good luck, Tobias
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America _ ____ ^ __ ____ Tobias I. Baskin / \ / / \ / \ \ University of Missouri / | / / \ \ \ Biological Sciences /___/ /__ /___ \ \ \__ 109 Tucker Hall / / / \ \ \ Columbia, MO 65211-7400 USA / / / \ \ \ voice: 573-882-0173 / /____ / \ \__/ \____ fax: 573-882-0123
Dear Tamara Is he washing in de-ionized water? Brain is always absorbent, so timing and reagent volumes may need adjustment. Gold and silver enhancement are tricky so I would protect from heat and light as much as possible. Good Luck. Dana
We are looking for someone to provide analytical services on analyzing surface topography on biomedical implants specifically using a TOPSCAN confocal microscope. It needs to be performed on this instrument for comparison with data previously taken on such an instrument. Does anyone know of anyone who has this instrument and who might entertain performing such an analysis? Your help on this will be greatly appreciated.
Charles R. Anderson, Ph.D. President Anderson Materials Evaluation, Inc. 1450 South Rolling Road Halethorpe, MD 21227 (410) 455-5698 Fax (410) 455-5679
An independent materials analysis laboratory for failure analysis, quality control, product and process development. We offer small-spot XPS, Auger microprobe, AFM, SEM, optical metallographic microscopy, white light interference microscopy, mass spectroscopy, and thermal analysis (TGA, DSC, TMA, DMA) services.
To All: We have a Jeol 100CX II and experiencing a constant moving of the Objective Aperture moving when scanning grids, especially in Posistion # 2 Grid. Any solution or ideas? We changed apertures,cleaned holder etc. Thank you, Peter Stolzenberg, PESTO INC. pestoem-at-aol.com
We just bought a new top-end type microscope (Field emission semi-inlens SEM). I evaluated four companies. I would have been happy with any of the microscopes that I looked at. In my opinion, all of the microscopes available today are damn good throughout the product range. You will probably be happy with any microscope that you buy. If your budget is limited and defined as ours was, that is going to limit you as to what machine that you will buy. You can use that to see what the manufacturers will put into your package. There are other intangibles that go into the decision. The most important is service, in particular, the service in your area. Ask around! I did and got some interesting comments. Then there are some of the others that you have to judge for yourself. Things like the user interface -you will be training students to use it, you want a versatile machine, but one that is easy to learn to use; output capabilities; digital/film or both; networkability; etc.
If performance is the issue, then the best thing that you can do is to take a number of your samples and take them to the application labs and run them while you are there. You should take samples that are representative of the types of samples that you will be working with. Make sure that you compare the same conditions on all your samples and that the output that you get back can be compared at the same magnifications. Standardize the output that you want the images to be in, for example 8 bit grayscale TIFF images or whatever. You should also do the samples in the same order from lab to lab. Print the results from digital images on the same printer. Then get a group of experienced users to help you judge the results.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
--
} -----Original Message----- } From: hadden-at-wingate.edu [ mailto:hadden-at-wingate.edu {mailto:hadden-at-wingate.edu} ] } Sent: Thursday, March 09, 2000 2:51 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: suggestions for new SEM?? } } } -------------------------------------------------------------- } ---------- } 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 {http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html} } -------------------------------------------------------------- } ---------. } } } When I stepped outside of the science building tonight, I ran into, } literally, our university president who said he was just } walking over to } try to find me. He cautioned me about overreacting and then } proceeded to } tell me to go ahead and get my best quote on a new SEM and service } contract!!!!!! } } Now that I've got both feet back on the ground, I need any } help that any } of you can provide, based on your experience and knowledge. We're } looking for a new SEM that will support undergraduate } courses, be a good } training machine for students, support student and faculty } research and } have a good track record for reliability and being reasonably user } friendly. It should be state of the art, but not be so expensive to } operate and keep up that it would limit how much students and faculty } could use it. } } I was trained on an Amray in the 80's and ran our TEM until it became } cost ineffective a few years ago. But I know things have changed } considerably since my pre-computer controlled everything } SEM's and would } appreciated the benefit of any experience any of you might } have had with } new SEMS. } } Anything to recommend? Models? Features to insist on or avoid? } Reliability, service costs and frequency? Companies to go } with or stay } away from? What new models do you have and would you } recommend them to } others? Why or why not? } } I'd really appreciate any and all insight you can provide. When your } president walks to YOUR office and says the equipment grant } proposal HE } made to a major foundation was just approved and that he } needs me to get } the best quote on an SEM to him ASAP, I want to have } something to go on, } and reasons to go along with it based on user input, without delay. } } Thanks in advance for any assistance you can provide. } } Lee Hadden } Professor and Chair, } Department of Biology } Wingate University } Wingate, NC 28174 } } hadden-at-wingate.edu } http://www.wingate.edu {http://www.wingate.edu} } 704-233-8238 } } }
Dear Tamara: To figure out what went wrong with your investigator's experiment, we need more detail about how his experiment was conducted. Would you ask him to contact me and email his protocol to me? I believe that I can help him.
Hong ============== Hong Yi Emory Neurology hyi-at-emory.edu
On Thu, 9 Mar 2000, Tamara Howard 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 } -----------------------------------------------------------------------. } } } Greetings! } } One of the researchers here is trying to do silver and/or gold enhancement } of an immunolabel in brain slices, but is getting amazing background. We } switched to the gold enhancement because everyone was saying how much } cleaner it is....didn't help. I've seen one report in which the } researchers pre-washed their samples in EDTA, because they felt that they } were getting background from Mg in the tissue that was nucleating silver } deposition - anyone else have any thoughts/experience on this? Anyone else } doing enhancement in tissue slices? } } He is using kits to do the enhancement - I'm not sure which companies are } the suppliers, but they aren't supposed to be light-sensitive (my first } culprit). The samples are washed extensively in water before enhancement, } and even the no-antibody controls are "lighting up". } } Any suggestions will be greatly appreciated! } } Tamara Howard } CSHL } }
The topic of reducing background has been raised a number of times by our customers, and we have collected together several approaches and their references, and I hope some of the following are helpful:
One possible mechanism for "background" signal is hydrophobic interactions between gold particles and tissue components, and if your samples can tolerate them, additives which break these interactions down and solubilize hydrophobic species might help in reducing background. Washes which effectively solubilize gold compounds and other hydrophobic species include:
* 0.6 M triethylammonium bicarbonate buffer (prepared by bubbling CO2 into an aqueous suspension of triethylamine with stirring; for a reference, see Safer, D.; Bolinger, L., and Leigh, J. S.; J. Inorg. Biochem., 26, 77 (1986)).
* A small amount of detergent, such as Tween-20 or Triton X-100, or an amphiphile such as benzamidine or 1,2,3-trihydroxyheptane.
* If you are using the gold enhancer, raising the ionic strength of the solution is sometimes helpful - we have found that raising the sodium chloride concentration in the actual gold enhancement mixture to 0.5 M can actually reduce the background. Alternatively, lowering the pH of the gold enhancement mixture by about 0.5 pH units may help.
Acetonitrile coordinates quite well to silver ions, so if your background problem is due to the interaction of silver from the enhancement reagents with a component of your tissues, treatment with acetonitrile or a wash solution containing a proportion of acetonitrile may help (provided the sample can withstand it).
To reduce the background for silver enhancement, one procedure which has been found to be effective is to wash with 0.02 M sodium citrate buffer, pH 7.0, several times immediately before silver enhancement - this has been used to reduce background in experiments with the combined fluorescein and gold probe FluoroNanogold when used with HQ Silver (Nanoprobes). When the Danscher formulation of silver enhancer was used instead, 0.02 M sodium citrate buffer at pH 3.5 was found to be most effective. In immunoblots, we have also observed that washing with 0.05 M disodium EDTA, pH 4.6, before silver enhancement also results in low background. (Reference: Powell, R. D.; Halsey, Carol M. R.; Spector, D. L.; Kaurin, S. L.; McCann, J.;, and Hainfeld, J. F. A covalent fluorescent-gold immunoprobe: "simultaneous" detection of a pre-mRNA splicing factor by light and electron microscopy. J. Histochem. Cytochem., 45, 947-956 (1997)).
A number of methods have been described for stopping the silver enhancement reaction or for "back-developing" to remove extraneous deposited silver. These prevent the continuation of the reaction in the specimens after development is complete (for example, if the silver is only slowly removed from the tissue), and may help reduce your background signal.
Sodium thiosulfate (1 % aqueous solution, freshly made) is a good "stop" reagent for both silver and gold deposition (Van Driel, D. 1997. Gold toning for silver enhanced immunogold reacted tissue. Micros. Today, 97-7, 28), so it is reasonably safe to assume that there will be no further deposition after it is applied. However, one caveat: you should use caution with gold enhancement because there have been reports that sodium thiosulfate can remove the enlarged gold particles. This might be a good choice to begin with (1-2 minute incubation after enhancement and washing with water; after the sodium thiosulfate, rinse thoroughly again with deionized water).
Other reaction stop and back development methods include:
(i) 1% acetic acid (Scopsi, L. 1989. Silver-enhanced colloidal gold method., p. 260. In M. A. Hayat (ed.), In Colloidal Gold: Principles, Methods, and Applications, vol. 1. Academic Press, San Diego, CA).
(ii) 1% acetic acid followed by photographic fixer (Agefix, Agfa-Gevaert, or Ilfospeed 200, Ilford) (Scopsi, L. 1989., same reference as (i)).
(iii) direct photo fix, using those just mentioned (Burry, R.W. 1995. Pre-embedding immunocytochemistry with silver-enhanced small gold particles, p. 217-230. In MA Hayat (Ed.). Immunogold silver staining: Principles, methods and applications. CRC Press, Boca Raton).
(iv) brief rinse in 2.5% sodium chloride (Scopsi, L. 1989., same reference as (i)).
(v) 15-25% aqueous sodium thiosulfate plus 15% sodium sulfite (Danscher, G. 1981. Histochemical demonstration of heavy metals. A revised version of the silver sulphide method suitable for both light and electron microscopy. Histochemistry 71, 1-16).
(vi) 1% acetic acid, washes in acetate buffer, toning in 0.05% HAuCl4 3-10 min, with excess silver removed with 3% sodium thiosulfate. We found that Nanogold-labeled proteins run on a polyacrylamide gel kept low backgrounds when stopped with 10% acetic acid with 10% glucose in water, as opposed to just a water stop (Takizawa, T. and J.M. Robinson. 1994. Use of 1.4-nm immunogold particles for immunocytochemistry on ultra-thin cryosections. J. Histochem. Cytochem. 42, 1615-1623).
(vii) A modified Farmer's solution was used for the reversal (0.3 ml 7.5% potassium ferricyanide, 1.2 ml of 20% sodium thiosulfate, 60 ml water) (Danscher, G.: Histochemistry, 71, 1-16 (1981)). Application of this solution briefly to your sample before gold toning may help to remove background silver deposition.
Hope this helps,
Rick Powell
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Please help this gentleman. He needs inexpensive EDS system suitable for Philips TEM (horizontal entrance detector) complete, and old EDAX-9800 or similar, computer unit only. Reply to Bill Lawry at eht-at-stlnet.com or (314)531-9868. Thank you.
Vitaly Feingold Scientific Instruments and Applications (770)232-7785 ph. (770)232-1791 fax.
Dear Peter, I have a 7 holed, self-cleaning, gold foil aperture in my TEM. The hole diameters are 30um and 60um. At 60kV, the aperture does tend to wander abit until it gets warmed up. After an hour or so of operation, I generally have to tweak the xy alignment and then I am ready to continue.
I am always amazed when the instrument is used by other investigators. Frequently the aperture is halfway into the the beam, i.e., the crescent of the aperture is covering 1/3 of the field of view. As I stated, I am aware the gold foil aperture tends to drift slightly over time exposed to the beam, and consequently adjust accordingly.
I too would be keen to hear any additional comments on this 'trait' of gold foil apertures.
-Ken
On Thu, 9 Mar 2000 PESTOEM-at-aol.com-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 } -----------------------------------------------------------------------. } } } To All: } We have a Jeol 100CX II and experiencing a constant moving of the Objective } Aperture moving when scanning grids, especially in Posistion # 2 Grid. } Any solution or ideas? We changed apertures,cleaned holder etc. } Thank you, } Peter Stolzenberg, PESTO INC. } pestoem-at-aol.com } }
I have just started on a TEM-project investigating the microstructures of different tool steels. The steels contain various types of carbides with different compositions, shapes and sizes. I have started out by making thin foils using electrolytical jet polishing. I get nice thin foils where I can study and analyse the martensitic matrix, but the carbides are off course too thick to be analysed. To overcome this problem I have considered a combination of electropolishing and ion milling, but I have also heard about an extraction replication technique. However, I haven't been able to find any literature on this technique. Have any of you tried this technique and/or could you point out some literature describing the technique? Also, if you have other suggestions to deal with the described problem I would be pleased to hear about them.
Regards, Jesper
---------------------------------------------------- Jesper Vejloe Carstensen Research Scientist, M.Sc., Ph.D. Materials Research Department Risoe National Laboratory P.O. Box 49 DK-4000 Roskilde, Denmark Phone: +45 4677 5776 Fax: +45 4677 5758 E-mail: jesper.v.carstensen-at-risoe.dk Web: http://www.risoe.dk/AFM/Personal/Jeca/jeca.htm ----------------------------------------------------
The url of the conference site seems to have been stripped out by some mail servers. Just in case it's www.cmp-cientifica.com/eurofe
Regards
Tim
****************************************************************** Tim E. Harper EuroFE Network Co-Chairman EuroFE is a network of the European Science Foundation Phone +34 91 640 71 85 Fax +34 91 640 71 86
One accessory worth considering, especially for a SEM used by students would be a ChamberScope. This is a small IR video camera mounted to the chamber which will allow students to see the position of samples in the chamber in real time. Could avoid damaging crashes. Take a look at GW Electronics for one model....
We have Carl Zeiss LSM 510 2-photon microscope armed with a T:sapphire laser. We are successful with one color imaging, but we wpould like to try two-color two-photon imaging, and of course we have problems with finding a proper set of fluorophoes. My understanding is that they should have similar excitation and different emissions. Has anyone tried using two or more fluorophores ? Which combinations are the best? What wavelengths are recommended?
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Is there a short, general reference, like a review, that explains immuno EM, it's uses, major techniques, and shortfalls? I would like to use it to send to investigators that are thinking about using the technique but don't know much about labeling at the EM level. I have a lot of specific references and it would take a lot of time to reference or copy them. If the investigator wants to know more I can direct them to those, otherwise a general one would get them and me on the same page when we start talking specifics on their project. Thanks
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In the earlier days {25 years ago} of medical diagnosis, before the advent of the rapid, highly specific methods of identifying pathogenic organisms, darkfield microscopy was routinely applied to rule out specific organisms. In particular, the diagnosis of syphilis was aided with this mode of imaging. When fluids from a suspect lesion were applied to a slide and imaged by darkfield, the characteristic shape of the spirochaete organisms was clearly seen. More recently, darkfield examination of synovial fluids from arthritic joints was used to rule out or diagnose Lyme disease, before specific testing became available.
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Catherine,
I also tried to subscribe to the confocal list when Larry suggested it and it worked like a charm. I cut and pasted the email address right from his posting.
LISTSERV-at-LISTSERV.ACSU.BUFFALO.EDU
Try again. Cheers, John
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-- C. John Runions, Ph.D. Department of Plant Sciences University of Cambridge Cambridge CB4 1YE UK
email at work cjr41-at-cam.ac.uk email at work runions-at-ntlworld.com Phone (01223) 766 545
We are in the market for two relatively inexpensive flatbed scanners. One needs to have a USB attachment and drivers for a Mac. This one is pretty straightforward. Up to about $200 (US) should suit our needs.
For the second one I am considering one with an adapter to allow scanning of negatives and 35 mm slides. I have heard a bad report of a scanner not focussing correctly on the negative or transparency. Does anyone have any comments to support or refute the focusing problem? We are able to pay up to $400 if I can find one that performs well. Obviously, we are not considering high end scanners. I would prefer USB, but could cope with another type of connection. Thank you for any help you can provide.
Sincerely, Maureen Petersen Dept Plant Pathology University of FL Gainesville, FL 32611
It has been a number of years since I've been directly involved in operating and maintaining a JEOL 100CX, but as I recall both the specimen holder and the aperture are inserted into the rather limited space between the objective pole pieces, with the specimen rod fitting in above the aperture. If the aperture consistently moves when you move the specimen holder, then I'd strongly suspect that either the end of your specimen rod has been bent downward just a bit, or the end of the aperture holder has been bent upward slightly so that the specimen rod rubs the aperture holder when it is moved thus causing the aperture to move. The clearance between these two devices is only a fraction of a millimeter, and so it wouldn't take much bending to cause this problem.
Good luck,
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
} } Anita, } } I have been using the MultiPrep for TEM of semiconductors since December. It } } has made my life much much easier. I am responsible for the TEM } } preparation/imaging/data at ON Semi. Since I have started using the MultiPrep I } } have been able to consistently turn a job around in 1 day and less. I have only } } failed on 1 sample since December and that was due to my bad judgement. Once you } } get a routine for making the samples I have found the results to be very } } consistent. Do you have an Ion Mill? I use the PIPS and it contributes to } } the success of the samples I make. Although if you did not have a PIPS I think } } that you would still get very good results using the MultiPrep. } } For me the major differences between using the MultiPrep and the Tripod Polisher } } are: } } 1. Consistent amount of force placed on the samples using the MultiPrep. } } 2. Having the digital micrometer on the MultiPrep takes some of the guess work } } out of the sample prep. I am able to tell how much material I am removing } } 3. Piece of mind. Since I have developed a set routine I am able to make 2 } } samples a day and feel confident that they will not fail. Hand polishing has } } always been a crap shoot for me. } } From your signature line I am assuming you work on various materials. The work } } I have done is primarily Silicon, but I think that the results would be good for } } other materials also. I have submitted an abstract for the MSA conference this } } year. I would be happy to speak with you about the system if you will be } } attending. } } } } Leah L Dobbs } } ON Semiconductor } } CSAL TEM } } 602-244-4820 } } 1-888-637-9415 (6379415-at-skytel.com) } } s20260-at-onsemi.com } } } } My disclaimer " I do not work for Allied High Tech, and do not have vested } } interest in the companies financial success. I am merely a satisfied customer". } } } } } } ----- Original Message ----- } } From: "Anita Garg" {Anita.Garg-at-lerc.nasa.gov} } } To: {Microscopy-at-sparc5.microscopy.com} } } Sent: Thursday, February 24, 2000 5:55 AM } } Subject: Re: Allied Multiprep Polisher for TEM samples } } } } } } } ------------------------------------------------------------------------ } } } 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 } } } Does anybody have experience on the Allied MultiPrep Polishing } } } System, especially in terms of getting a good TEM sample? How well do } } } the Allied's Precision Cross-Section Tool and TEM Wedge Polisher work } } } on this semi automatic multiprep polisher? } } } Also, how does their TEM Wedge Polisher compare with the South Bay } } } Tripod Polisher? } } } TIA } } } Anita } } } ******************************************* } } } Dr. Anita Garg } } } NASA Glenn Research Center at Lewis Field } } } Advanced Metallics Branch } } } Mail Stop 49-3 } } } 21000 Brookpark Road } } } Cleveland, OH 44135 } } } Phone : (216) 433-8908 } } } Fax : (216) 977-7132 } } } E-mail : Anita.Garg-at-grc.nasa.gov } } } ******************************************* } } } } } } } }
The extraction replica technique was developed by Bob Fisher at the U. S. Steel Laboratory back in the early 1950's when we were all trying to identify the precipitates in various alloy systems for the first time. I don't seem to be able to lay my hands on the original reference; however, the technique is described in reasonable detail on pages 115-117 of the book 'Techniques for Electron Microscopy', Desmond Kay, Ed., Blackwell Scientific, 1961.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
Hi, we have some software freely downloadable for processing ED of protein crystals. It's at http://ncmi.bcm.tmc.edu/software.html. Look for packages auto and edp. The former is a front-end to a bunch of fortran programs for automated processing, whereas the latter is a GUI-based program.
Jaap
-- Jaap Brink, Ph.D., Biochemistry, One Baylor Plaza, Baylor College of Medicine, Rm. N420 Alkek Building, Houston, TX 77030 Phone: (713)798-6989 -- Fax: (713)796-9438 -- Email: jbrink-at-bcm.tmc.edu URL : http://ncmi.bioch.bcm.tmc.edu/~brink
On Fri, 10 Mar 2000, Chengge Jiao 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, } } Does anybody know where I can get free software for electron doffraction and } more... } } Can we download them free ? } } Chengge JIAO } } H.H.Wills Physics Lab } University of Bristol } c.g.jiao-at-bristol.ac.uk } }
Firstly, thanks to everyone who helped out with the earlier question on image analysis. We are using NIH Image 1.62 (which has a watershed filter to separate touching particles) to measure their diameters. (Worked like a charm -- thanks John Russ).
We need a NIST particle, sized 5 µm or so, to use as a standard. Unfortunately, the standard we recently purchased was sized using light scattering (which apparently gives a higher reading than electron microscopy) and our sizing does not agree with that figure.
Does anyone know were we can find 5.0 µm polystyrene particles that have been sized by TEM and are NIST certified or traceable?
Thanks,
#################################################################### John J. Bozzola, Ph.D., Director Micro-Imaging and Analysis Center 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 ####################################################################
Every year when I teach the liver lectures for my histology course, I explain to the students that up to 25% of the hepatocytes are binucleate and another 50% are polyploid. These textbook facts match up with what I see in the scope. But I don't understand why hepatocytes (or transitional epithelial cells) are frequently binucleate (or polyploid). Any one able to answer this structure - function question?
My second histology question concerns the gallbladder epithelium. I have a great prepared H&E 1.5 um paraffin slide from Carolina Biological. Most of the epithelial cells have a few fine red-stained granules in them. I assume these are glycoproteins and/or mucin-type glycoproteins. A few scattered cells in the epithelium have larger, intensely eosinophilic granules in them. I can't figure out what these cells are. None of the standard textbooks or atlases discuss them. Before anyone suggests they are mucin-filled goblet cells, I should point out that intestinal goblet cells are my main research focus and these are unlike any mucin secreting cell I have seen in the intestine, respiratory tract, stomach, conjunctiva, etc. They look, in fact, more like Paneth cells in the small intestine. Any ideas?
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)
It is my understanding that many instances of so-called aperture thermal drift are actually the result of hysteresis in either the condenser or objective lenses. This seems to be more of a problem in JEOL instruments than in others that I'm aware of. It is particularly pronounced when you spend time scanning large areas with little or no changes in magnification or brightness. When the current is changed to either lens, the periphery of an aperture may enter the field of view. By realigning the aperture, you actually misalign it along the optical axis. This is why a TEM is often in such abysmal alignment after use by students and less experienced users. Our JEM 1210 has 9 lenses, with each one potentially affected by hysteresis. Most of our users know when it is time to switch off the lens power temporarily and reload the alignment values. In the case of a less automated instrument such as the 100-C series, it will often suffice to run C2, Obj, and Int lenses through their entire ranges before aligning apertures.
The problem with extraction replicas for your carbides is that they will still be too thick to analyse! Extraction replicas work better the smaller the carbides (in my humble opinion!). I would recommend mechanical thinning followed by ion-milling (we typically don't bother with the intermediate jet-polishing step). Of course, if you also have fine carbides, you will need the extraction replicas in order to be able to examine them without interference from the matrix!
Good luck,
Tony Garratt-Reed.
At 10:31 AM 03/10/2000 +0100, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
** Anthony J. Garratt-Reed ** MIT Room # 13-1027 ** 77 Massachusetts Avenue ** Cambridge, MA 02139-4307 ** USA ** ** Phone: (+) 1-617-253-4622 ** Fax: (+) 1-617-258-6479 **
First of all you never indicated from which species this gall bladder epithelium came from. Goblet cells are characteristic in the bovine species. The cat epithelium may contain globule leukocytes. Also, endocrine cells have been reported in bovine.
Nancy A. Monteiro-Riviere, Ph.D., DABFE, DABFM Professor of Investigative Dermatology and Toxicology North Carolina State University College of Veterinary Medicine Department of Clinical Sciences Center for Cutaneous Toxicology and Residue Pharmacology 4700 Hillsborough St. Raleigh, NC 27606 Tel: 919-513-6426 FAX: 919-513-6358 email: Nancy_Monteiro-at-ncsu.edu CCTRP Homepage: http://cctrp.ncsu.edu
} ---------- } From: Tom Phillips[SMTP:PhillipsT-at-missouri.edu] } Sent: Friday, March 10, 2000 1:47 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: binucleate hepatocytes & granules in gallbladder: basic } histoquestions } } ---------------------------------------------------------------------- } -- } 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 } ---------------------------------------------------------------------- } -. } } } Fellow microscopists: } } Every year when I teach the liver lectures for my histology course, I } explain to the students that up to 25% of the hepatocytes are } binucleate and another 50% are polyploid. These textbook facts match } up with what I see in the scope. But I don't understand why } hepatocytes (or transitional epithelial cells) are frequently } binucleate (or polyploid). Any one able to answer this structure - } function question? } } My second histology question concerns the gallbladder epithelium. I } have a great prepared H&E 1.5 um paraffin slide from Carolina } Biological. Most of the epithelial cells have a few fine red-stained } granules in them. I assume these are glycoproteins and/or mucin-type } glycoproteins. A few scattered cells in the epithelium have larger, } intensely eosinophilic granules in them. I can't figure out what } these cells are. None of the standard textbooks or atlases discuss } them. Before anyone suggests they are mucin-filled goblet cells, I } should point out that intestinal goblet cells are my main research } focus and these are unlike any mucin secreting cell I have seen in } the intestine, respiratory tract, stomach, conjunctiva, etc. They } look, in fact, more like Paneth cells in the small intestine. Any } ideas? } } 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) }
I am searching for a Leitz Periplan photomicro eyepiece, 10X/18, 23mm diameter. The old Leitz part number was 519749. The current Leica part number is 11519749, although it is a discontinued item. This particular eyepiece has a threaded mount on the viewing end. The thread was used to mount eyecups on the eyepiece, but it was also used as a photoeyepiece.
If anyone has one of these photo eyepieces and would be willing to part with it, please contact me off-list.
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Your best bet is to embed in paraffin. However, if you still want to use resin for immunolocalization, you may try Immuno-Bed Embedding resin, available from Electron Microscopy Sciences. It is a fairly low viscosity media.
Soumitra
} Dear microscopists, } Can anyone offer advice on doing immunofluorescence at the light } level? I } have cryopreserved plant tissue that I am contemplating embedding and using } for immunolocalization. I have a protocol that uses butyl, methyl } methacrylate. I am wondering several things. Why butyl, methyl } methacrylate? Are there other resins/media that are sufficient? I'd } appreciate any advice anyone has. } Kristen Lennon } Kristen A. Lennon } Cell, Molecular & Developmental Biology Group } Department of Botany & Plant Sciences } University of California } Riverside, CA 92521 } kalen-at-citrus.ucr.edu
***************************************************************** Soumitra Ghoshroy Ph.D. Electron Microscopy Lab Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3600 Fax: 505-646-5665 e-mail: ghoshroy-at-nmsu.edu http://confocal.nmsu.edu/eml
Here are two references more recent than the two I sent earlier regarding simultaneous glut/osmium fixation methods for single cells in suspension:
Dentler, WL (1999) Fixation of Tetrahymena cells for electron microscopy. Methods in Cell Biology 62:323-331
but is modified from
Omoto, C.K, and Kung, C. (1980). Rotation and twist of the central pair microtubules in the cilia of Paramecium . J. Cell Biol 87:33-46
Responding to the message of {v03007805b4ec9d6d6eae-at-[206.69.208.21]} from R schlicher {rkslick-at-yahoo.com} :
} Hello, } } I need to fix cells for viewing with SEM. However, } the fixation method needs to be consistent with my } experimental methods: } } 1) I need to fix the cells in less than 1 minute and } due to this time limitation, I cannot spin the cells } into a pellet and must add the cell suspension to the } fixative; and } } 2)I am looking for "effects" to the cell membrane } caused by my experiment, so I need to keep any } possible fixative damage/effects at a minimum. } } I have fixed cells using plunge freezing and viewed } them with TEM, this is time consuming and I ran into } a } lot of freeze damage. I would like to try chemical } fixation and SEM. It has been suggested that I try } glutaraldehyde + buffers, then postfix with osmium } tetroxide and dehydrate in an ethanol series. } } Is this the best method? I am open to suggestions. } I am using suspensions of prostate cancer cells in } RPMI (+serum) growth media; I can perform the } experiment in buffered saline and use a high } concentration of cells. } } Thanks, } Robyn } } } Robyn K. Schlicher, M.S. } Laboratory for Drug Delivery } Institute for Bioengineering and Biosciences } Georgia Institute of Technology } 315 Ferst Drive } Atlanta, GA 30332 } rkslick-at-yahoo.com
Gib Ahlstrand Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN. USA. 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.cdl.umn.edu http://biosci.umn.edu/MIC/consortium.html
I am going to give a course on electron microscopy including TEM, SEM, STM, AFM, etc. to graduate students. Could you give me information on textbooks about these microscopy.
Thanks
**************************************************** Prof. Jianguo Wen
Department of Physics Boston College 140 Commonwealth Avenue Chestnut Hill, MA 02467 Tel: (617) 552-3586 Fax: (617) 552-8478 Email: wenji-at-bc.edu ****************************************************
Are there any Amray-trained service people out there that are independently servicing the 1910FE system on the West Coast? I'm in Sacramento CA. I am not looking for someone who knows how to spell Amray. I am looking for someone who knows what the 1910FE is all about. Big difference. Introductory questions:
1. Can you condition the emitter/gun? How and with what? 2. How do you handle the differential vacuum aperture? 3. what is your experience in working with the Windows control s/w? 4. What do you know about the IP8? 5. Do you have any knowledge about moving from 486 ISA to P-II or P-III systems for NibbleNet and frame capture?
Since the gobbling of Amray by KLA-Tencor, Amray SEM service is dismal--more so day by day. The service people are excellent--but they are being pulled in all directions, and mostly towards the KLA big semi systems.
The Amray SEM systems were/are really good--and that is why KLA bought Amray. But KLA is not a research SEM outfit like Amray was.
If I had to get something other than this 1910FE, I don't know what it would be. The Hitachi and Philips SEMs are great/nice but ridiculously expensive to buy and maintain. And I do not care for Jeol.
Me thinks that the research SEM options are shrinking daily.
Duke Scientific should produce something like this. I know that they used a TEM to perform their sizing at one time. I'm not sure if this is the still the case. If not, maybe they can point you in the right direction. Here's Duke's website as well as a few other particulate manufacturers:
================================================ Marc W. Helvey Accounts Manager VLSI Standards, Inc. 3087 North First Street San Jose, CA 95134-2006 phone:(408) 428-1800, ext. 108 FAX: (408) 428-9555 Mobile: (408) 307-3833 e-mail : marc.helvey-at-vlsistd.com {mailto:marc.helvey-at-vlsistd.com} internet: http://www.vlsistd.com {http://www.vlsistd.com/} ================================================
On Fri, 10 Mar 2000 19:11:40 -0500, Jianguo Wen wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I am going to give a course on electron microscopy including TEM, SEM, STM, } AFM, etc. to graduate students. Could you give me information on textbooks } about these microscopy. } } Thanks } } **************************************************** } Prof. Jianguo Wen } } Department of Physics } Boston College } 140 Commonwealth Avenue } Chestnut Hill, MA 02467 } Tel: (617) 552-3586 Fax: (617) 552-8478 } Email: wenji-at-bc.edu } **************************************************** } } Prof. Wen - In 1994, I took a course on electron microscopy (for biologists) at San Francisco State University given by Gregory Antipa (my former M.A. advisor and principal investigator). The textbook he used was titled "Electron Microscopy (:) Principles and Techniques for Biologists" by John J. Bozzola and Lonnie D. Russell (eds. Jones and Bartlett Publishers, Boston .. London). Although this textbook is intended for biology students, it was used in a class that any graduate student could sign up for given his/her interests. I found the textbook to be illuminating and interesting to read. It has a historical chapter on electron microscopy, and then discusses, among many topics, fixation uses / types of / .. embedding procedures and types - TEM; similar topics - SEM; microtomy; staining procedures, types, etc. ; TEM itself; SEM itself; immunocytochemistry and EM ; autoradiography; freeze-fracture ; analytical electron microscopy (e..g., x-ray microanalysis subchapter; EELS subchapter; SAD diffraction mode sub-subchapter), and many other topics relevant to electron microscopy. Unfortunately .. it does not cover STM (Scanning-Tunneling Microscopy) nor AFM (Atomic Force Microscopy). I personally do not know of any textbooks that cover those types of electron microscopy, but I would guess that someone in this list would have suggestions. For that matter, I am not even aware if this textbook is still being published, but perhaps Gregory Antipa (e-mail: antipa-at-sfsu.edu ) can tell you, or someone else. I hope that this information may serve as a useful starting point in your search for a textbook on electron microscopy for graduate students. Nelson Conti (former graduate student) San Francisco State University 1600 Holloway Avenue San Francisco, California (CA) 94132 URL: www.sfsu.edu
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Feather makes the blades we LM histotechs use most often in our everyday sectioning. They are distributed by Allegiance (used to Baxter, used to be SP....). Just FYI. Wanda Shotsberger (HT ASCP) -----Original Message----- } From: Grazyna M Tokarczyk [mailto:gmtokarc-at-is.dal.ca] Sent: Friday, March 03, 2000 8:26 AM To: Gordon Couger Cc: c.jeffree-at-ed.ac.uk; Dan Luchtel; microscopy-at-sparc5.microscopy.com
Dear John, I worked with characterisation of latex sphere size for many years and traced them to NIST standard. You may wish to contact Interfacial Dynamics Corp. as they manufacture spheres and no doubt have the capacity to trace to NIST. I do not have their address information at hand, but they are located in Portland, OR.
If you have other questions, please contact me at me University of Portland email address.
Dear Tamara, Without more information about the procedure that was followed it is hard to suggest a solution. But since the issue of background with immunogold and enhancement is becoming a regular topic in this List, it may be worthwhile to address it more in-depth. In troubleshooting I always find it a good idea to trace down the origin of the problem logically, rather than shooting in the dark. In all the years we have been involved in immunogold detection and silver enhancement, we have always benefited from simple approaches. These are documented in troubleshooting brochures we use during our workshops and which describe step-by-step how to proceed to have the best chance to pinpoint the problem. Anyone who is interested, please send me an e-mail.
Now just some comments on some of the things you mentioned:
- working on brain slices, you are probably doing pre-embedding. One of the issues in pre-embedding is penetration. Most of the time gold conjugates need a longer time to get inside thicker specimens, but under the proper conditions they do get there. But what often is easily forgotten is that when it takes a long time for reagents to penetrate into the slice, it also takes a long time for unreacted reagents to get washed out again. So washing should be appropriate.
- you mentioned that there is no difference between silver enhancement and gold enhancement. Have these specimens been incubated with gold conjugate? What if there is background even when the gold conjugate incubation step was left out? With any enhancement system there is a risk of getting background through (i) autonucleation or (ii) by the formation of metal particles in the specimen, formed as a result of the interaction of components in the specimen (e.g.aldehydes, borohydrid) with the noble metal salts in the enhancement mix. This may also be related to the type of metal salt in the enhancement solution: noble metal salts need only a little push to become reduced to the metal again and the more noble the metal the smaller the push it needs. As for judging if autonucleation has occurred, as long as the enhancement solution is as clear as it was when the enhancement started, this should not be a real problem.
- background from magnesium. Gallyas and Danscher (and many others) have looked into many aspects of (auto)metallography, I can give references if you are interested.
- light sensitivity of enhancement reagents: light sensitivity can never be reduced to zero. There are tricks that reduce light sensitivity to a degree where it no longer seriously interferes with enhancement on particles, but again, these are noble metal salts, and they need just so much.... In any case, if light would be causing a problem, which I don't think is very likely whatever brand you used, you should have the same phenomena as with autonucleation: the mixture should become turbid or colored after a while. If this is not the case, there is no reason to be worried about a possible light sensitivity.
Jan =========================== Jan Leunissen AURION http://www.aurion.nl Costerweg 5 6702 AA Wageningen phone: (31)-317-497676 fax: (31)-317-415955 You will find more tech info on our website.
Need help on PEELS } } To all, } } I am working on grain boundary chemistry measurement } with Digipeels. Try to find out the segregation } behavior of boron, which is about 0.5at% averagely. } However, I cannot find any edge for boron in the } spectrum. As I don't know what's the optimum setting } for PEELS to check the grain boundary segregation, if } you have any idea on this, please let me know. } } Thanks, } Lung __________________________________________________ Do You Yahoo!? Talk to your friends online with Yahoo! Messenger. http://im.yahoo.com
I'd like to point out that by extraction replica you will NOT overcome the problem of the carbides' thickness: you will get only a complete separation of them from matrix. So, according to my experience, the ion-milling would be a solution in that it could lead to a THINNING of your carbides.
In case there is a possibility to partially dissolve the carbides, by chemical means, after their extraction, then the extraction replica technique will work; but I don't know about such a chemical technique for dissolvind (partially!) the carbides.
Corneliu Sarbu, Ph.D. Dept.of Metallurgy and Materials Science Catholic University of Leuven Belgium
- - - - - - - - - - - - - - Original Message - - - - - - - - - - - - - - ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear all,
I have just started on a TEM-project investigating the microstructures of different tool steels. The steels contain various types of carbides with different compositions, shapes and sizes. I have started out by making thin foils using electrolytical jet polishing. I get nice thin foils where I can study and analyse the martensitic matrix, but the carbides are off course too thick to be analysed. To overcome this problem I have considered a combination of electropolishing and ion milling, but I have also heard about an extraction replication technique. However, I haven't been able to find any literature on this technique. Have any of you tried this technique and/or could you point out some literature describing the technique? Also, if you have other suggestions to deal with the described problem I would be pleased to hear about them.
Regards, Jesper
---------------------------------------------------- Jesper Vejloe Carstensen Research Scientist, M.Sc., Ph.D. Materials Research Department Risoe National Laboratory P.O. Box 49 DK-4000 Roskilde, Denmark Phone: +45 4677 5776 Fax: +45 4677 5758 E-mail: jesper.v.carstensen-at-risoe.dk Web: http://www.risoe.dk/AFM/Personal/Jeca/jeca.htm ----------------------------------------------------
- - - - - - - - - - - - End of Original Message - - - - - - - - - - - -
Dear Jesper: Extraction replicas are an old technique. I used it 30 years ago.
1. Prepare a lightly etched metallographic specimen. 2. Evaporate with carbon to form a thin film. 3. If you want increased contrast, shadow with Cr at an angle. 4. Score the surface with a razor to form 1 mm rectangles. 5. Etch again until the carbon squares float off. 6. Scoop up the rectangles and immerse in water.The square may flatten out. If so, it can be mounted on TEM grid and dried by touching the side of the grid to a piece of filter paper. 7. If the released square curls up when immersed in the rinse water. scoop it up with a grid and gently lower it into a bath of acetone. It should snap out into a flat sheet floating on top of the acetone. Again mount on a grid and dry. If this doesn't work, remove the replica from the acetone bath and lower it into the water bath so that surface tension flattens out the replica.
I'm surprised thnat you could not find a description of thsi technique in the literature. Try some of the older texts on TEM techniques.
Good luck, Sam Purdy National Steel Corp. Technical Center Trenton, MI, USA } ---------- } From: "Carstensen, Jesper Vejl¿" } Sent: March 2000 4:31 AM } To: 'Microscopy-at-MSA.Microscopy.Com' } Subject: TEM: Carbides in tool steels } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear all, } } I have just started on a TEM-project investigating the microstructures of } different tool steels. The steels contain various types of carbides with } different compositions, shapes and sizes. I have started out by making } thin } foils using electrolytical jet polishing. I get nice thin foils where I } can } study and analyse the martensitic matrix, but the carbides are off course } too thick to be analysed. To overcome this problem I have considered a } combination of electropolishing and ion milling, but I have also heard } about } an extraction replication technique. However, I haven't been able to find } any literature on this technique. Have any of you tried this technique } and/or could you point out some literature describing the technique? Also, } if you have other suggestions to deal with the described problem I would } be } pleased to hear about them. } } Regards, Jesper } } ---------------------------------------------------- } Jesper Vejloe Carstensen } Research Scientist, M.Sc., Ph.D. } Materials Research Department } Risoe National Laboratory } P.O. Box 49 } DK-4000 Roskilde, Denmark } Phone: +45 4677 5776 } Fax: +45 4677 5758 } E-mail: jesper.v.carstensen-at-risoe.dk } Web: http://www.risoe.dk/AFM/Personal/Jeca/jeca.htm } ---------------------------------------------------- } } } }
At 06:05 PM 3/10/00 , I wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
[snip]
Ability to spell "Amray" is/was not meant to be negative. I have had bad experience with service people who say they can work on systems but in reality cannot. I have replaced my Amray 1830 LaB6 system with the Amray 1910FE. The field emission system is much more complex and horribly more expensive to replace if not properly handled/serviced. There are special procedures especially for the FEI 305FE Zr/W gun assembly that must be followed or the emitter is zapped.
Bad experience was unfortunate with the LaB6 system. I fear it would be fatal on a field emission system. That is what I was clumsy in saying.
First you are going to have to do a bit of reading. I know of one person who studied the detection limits both experimentally and theoretically in PEELS recently ( {2 years) and that is Michael Natusch (formerly Cambridge). He found current detection limits of C in steel was only about two or three atomic percent (four to six times higher than your boron average). Check out Micron volume 30 (1999) pp173-183 (Natusch, Humphreys, Menon & Krivanek) as a starting point and for references.
Even if you do see a boron peak in the spectrum you will have to work out the signal to noise ratio (SNR) of the edge. The real problem is that you may see something that isn't there i.e. noise, or conversely you don't see something that you should see i.e. poor experimental execution.
To optimise the SNR you are going to need a lot of counts from the boundary itself. One way to see if your probe is on the boundary is to look for a possible chemical shift in the core loss edges of the matrix material since the bonding character of the material is different at the grain boundary.
Good luck. ******************************************************** Dr Jonathan Barnard
Analytical Materials Physics The Angstrom Laboratory, Uppsala University P O Box 534, SE-751 21 Uppsala, Sweden Phone: +46-(0)18-4716838 Fax: +46-(0)18-500131 http://www.angstrom.uu.se/analytical/home.html
Dear all on the Microscopy List, I am looking for good advice concerning surface analysis of metals using a microscope. The objects are prehistoric tools, jewellery, etc. made out of bronze, silver or gold (or combinations). I am interested in traces of the tools/techniques that was used on the object. The research technique should be non-destructive. The results will be used in archaeological research. I am looking for suggestions about what materials to use, what equipment is suitable/normally available and not too expensive. Is there anybody out there who deals with these subjects in a modern, industrial context - and who could tell me some more about, how they do it ? References for litterature, companies, etc.
Off hand, I would think that BSE imagine with a SEM might be useful. The difference between SE and BSE could be revealing. Without imaging the actual specimens, I can only speculate. But based on what you are analyzing, SE, BSE and perhaps EDX analysis can be insightful.
gary g.
At 12:29 PM 12/12/99 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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Please notice that extracting carbides from the matrix doesn't make them thinner. So, it seems that your idea about ion milling is right.
Best regards and good luck,
Witold
%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%% Witold Zielinski, Ph.D. Warsaw University of Technology Department of Materials Science and Engineering 02-507 Warszawa, Woloska 141 POLAND
We offer training in EM maintanance either through one of our "Monitoring and Maintaining the EM" short courses or on site as required by the client.
Steve Chapman Senior Consultant Protrain - for EM Consultancy and Training World Wide Tel 44+ 1280 814774 Fax 44+ 1280 814007 www.emcourses.com
----- Original Message ----- } From: electron microscope laboratory {emlab-at-udsm.ac.tz} To: {Microscopy-at-sparc5.microscopy.com} Sent: Friday, March 10, 2000 5:24 AM
a 45 degree laser line across the sample will tell a lot about the surface. Getting a fine enough beam would be fun.
If some one wants to try it I have a start on the code in C.
Gordon W5RED
G. C. Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger "You miss 100 percent of the shots you never take." - Wayne Gretzky
----- Original Message ----- } From: "Dr. Gary Gaugler" {gary-at-gaugler.com} To: "Sren Albek" {albek-at-dorit.ihi.ku.dk} Cc: "MSA listserver" {Microscopy-at-sparc5.microscopy.com} Sent: Sunday, March 12, 2000 7:53 PM
Dear Dr. Gauler,
The E.FJELD Co. has factory trained service engineers to handle conventional AMRAY SEM's (LaB6, tungsten). We offer routine service, spare parts and service agreements on AMRAY SEM's throughout the country.
Your requested is tough... It is extremely difficult, regardless of manufacturer (Hitachi, Philips, JEOL or KLA), to find experienced service people for FE technology. We do not supply such service.
If I find a source, I will keep you posted.
Regards,
Mark Reynolds
E.FJELD Co., Inc. 3 Executive Park Drive No. Billerica, MA 01862
TEL: (978)667-1416 x10 FAX: (978)667-9059
On Friday, March 10, 2000 7:06 PM, Dr. Gary Gaugler [SMTP:gary-at-gaugler.com] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Are there any Amray-trained service people out there that are } independently servicing the 1910FE system on the West Coast? } I'm in Sacramento CA. I am not looking for someone who knows } how to spell Amray. I am looking for someone who knows what } the 1910FE is all about. Big difference. Introductory questions: } } 1. Can you condition the emitter/gun? How and with what? } 2. How do you handle the differential vacuum aperture? } 3. what is your experience in working with the Windows control s/w? } 4. What do you know about the IP8? } 5. Do you have any knowledge about moving from 486 ISA to P-II } or P-III systems for NibbleNet and frame capture? } } Since the gobbling of Amray by KLA-Tencor, Amray SEM service } is dismal--more so day by day. The service people are excellent--but they are being } pulled in all directions, and mostly towards the KLA big semi systems. } } The Amray SEM systems were/are really good--and that is why KLA bought } Amray. But KLA is not a research SEM outfit like Amray was. } } If I had to get something other than this 1910FE, I don't know what it } would be. The Hitachi and Philips SEMs are great/nice but ridiculously } expensive to buy and maintain. And I do not care for Jeol. } } Me thinks that the research SEM options are shrinking daily. } } gary g. } }
I know little about liver, but if these are fairly large cells under high demands to produce [mRNAs], the polyploidy may be a response to a biological need. In a far simpler system, the nematode, there is good evidence that ploidy gets regulated in direct correspondance to the absolute size of the cell, across many different tissues, such that the ploidy perhaps matches the needs of the cell.
see Hedgecock and White (1985) Dev Biol 107: 128-133.
Though vertebrate cells may or may not be so closely regulated, the tendency might lie along a similar continuum?
Just a thought. Best regards, David H. Hall Center for C. elegans Anatomy Department of Neuroscience 1410 Pelham Parkway Albert Einstein College of Medicine Bronx, NY 10461
Check out my web page for examples of the different modes of scanning electron microscopy. It is a simple web page and has a section showing examples of the different modes of SEM. I recently quit my job and built a SEM lab in my basement and also have x-ray microanalysis for generating elemental spectra on micro-volumes. I am an expert in the different SEM contrast mechanisms, but not a metallurgist w/ any experience in the field that you mention. SEM is non-destructive provided you can get the material into the chamber. Some SEMs have large chambers, mine is on the smaller side (perhaps samples {4" and somewhat flat sample would be best). I would be glad to run 2-3 samples for free to show you what SEM can do as a feasibility study for SEM analysis of your samples. All my output can be in the form of jpg images so that you can send the data to experts in archaeology and they can comment on the significance.
Good Luck
Ric Felten www.semguy.com
-----Original Message----- } From: S¿ren Albek [mailto:albek-at-dorit.ihi.ku.dk] Sent: Sunday, December 12, 1999 1:29 PM To: Microscopy-at-sparc5.microscopy.com
Dear all on the Microscopy List, I am looking for good advice concerning surface analysis of metals using a microscope. The objects are prehistoric tools, jewellery, etc. made out of bronze, silver or gold (or combinations). I am interested in traces of the tools/techniques that was used on the object. The research technique should be non-destructive. The results will be used in archaeological research. I am looking for suggestions about what materials to use, what equipment is suitable/normally available and not too expensive. Is there anybody out there who deals with these subjects in a modern, industrial context - and who could tell me some more about, how they do it ? References for litterature, companies, etc.
In addition to the advice you have already received you might wish to consider contacting your local Forensic Science Laboratory and talk to the Firearm and Toolmark Section. What you wish to do is to determine the Class characteristics of the tools used, as you do not have the actual tool for comparison. There is also some Toolmark analysis done at certain types of machine shops but a really experienced Toolmark examiner at your local Forensic Science Lab would be much easier to locate and probably more interested in your project (I'm currently doing some work for a project on bullets from a sight, in my free evenings and I know others have in the past). Low power stereo microscopy can tell you a lot. The examiners can point you in the right direction and provide some literature sights etc., or may be interested in doing the work for you.
Jim
James L. Roberts Firearm and Toolmark Examiner Ventura Co. Sheriff's Lab (805) 654-2308 James.Roberts-at-mail.co.ventura.ca.us
} } } "S¿ren Albek" {albek-at-dorit.ihi.ku.dk} 12/12/99 10:29AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear all on the Microscopy List, I am looking for good advice concerning surface analysis of metals using a microscope. The objects are prehistoric tools, jewellery, etc. made out of bronze, silver or gold (or combinations). I am interested in traces of the tools/techniques that was used on the object. The research technique should be non-destructive. The results will be used in archaeological research. I am looking for suggestions about what materials to use, what equipment is suitable/normally available and not too expensive. Is there anybody out there who deals with these subjects in a modern, industrial context - and who could tell me some more about, how they do it ? References for litterature, companies, etc.
Hi, I need advice on double labeling (on-grid). I've had success doing each of the labeling experiments independently but now the researcher wants to see double labeling. I'm doing indirect labeling with a monoclonal antibody to fungal cell wall antigens followed by a gold labeled secondary. In addition, I want to label the sections with gold conjugated wheat germ agglutinin. Should I do the antibody labeling first and then the lectin? I've found one reference that I haven't had a chance to follow up on (Cailliez et al, 1988) in Immunogold Silver Staining Principles, Methods, and Applications by Hayat. Any advice would be greatly appreciated. Beth
we routinely use Lowicryl K4M. We normally buy it as a kit from EM science. They also offer a it as a MonoStep Single Component. We would like to try it. Has anybody done the comparison between the kit and the monostep stuff (quality, shelflive, handling, polymerisation)? Thanks for your time,
Dear all, thank you very much for all the kind answers, good ideas, and good suggestions. It has provided me with a good range of possibilities (I am a little overwhelmed by now :-). I must now evaluate the answers and I will most likely return with some more questions - some of these off-list, when appropriate.
Once again, thank you for the interest, and with many regards,
Dear Listers, I have a question about the cleaved glass blades used to cut cross sections of copolymers. Is there any way to quantify the sharpness of the blades? Do they have a radius of curvature, or anything like that? A client of ours has asked if the blade would be sharp enough to slice through the hard domain (polystyrene hardness) at -20 C or -80 C, or if it would just push it out of the way. This also applies to the disposable blades on a cryostat used to section the same sample at -20C prior to the cryo-ultramicrotoming. The sample is very brittle at -120C. Rosemary
Rosemary Walsh, Manager The Electron Microscope Facility for the Life Sciences, A Shared Technology Facility, The Life Sciences Consortium 1 South FrearLab Penn State University University Park, PA 16802 (814) 865-0212 rw9-at-psu.edu http://www.lsc.psu.edu/stf/em/home.html
Hello, We have an old semicaps 4000 for pc windows 3.1 system. I've been trying to use it to get scale bars burned into images, but I must say that it is very buggy. The images never burn scale bars correctly, calibration procedures are never fully written in the manual, and often minimizing and maximizing windows causes images to have very 'weird' color schemes.
After contacting semicaps I was told that there were no bug fixes to the software as it was bug free. Anyone out there have some experience with this system? 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
My post has generated several responses. All of which are very helpful. Thanks to all who responded.
From following up on what I have discovered from the response from Alex Greene, I'm rather stuck. It turns out that Amray has a lock on the FE gun/emitter that they developed and which incorporate the FEI W/Zr emitter (305FE). As such, no other party will support the Amray FE systems.
There are several folks who will and do support the Amray LaB6 systems. But none can or will support the FE systems. So for the time being, I am stuck with Amray. Be that as it may. As I write this, Amray (KLA-Tencor) is preparing to work on my FESEM under my existing maintenance contract. This is good. I guess that so long as this keeps up, I am OK. But I do expect that KLA will drop all Amray field support in the near future. Consequently, perhaps that will be a better time to seek alternative maintenance sources. I do expect at this future juncture, KLA/Amray will supply parts but will not supply warm bodies. If this is an opportunity for independents, who knows? If you are experiencing similar troubles, I would for one appreciate hearing about your situation. Otherwise...
Unless somebody has done that very same thing, one cannot know. However, I am optimistic that you could section that material with a glass knife. A new knife should run virtually to a molecular edge, although that finest edge would not survive the first section. The glass knife is sharp, the question concerns more its hardness and toughness. An LKB trainer years ago told me that a glass knife at liquid nitrogen temperature is almost as hard as is diamond at room temperature. So there is a good chance that glass could do. Try it. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Tuesday, March 14, 2000 9:05 AM, Rosemary Walsh [SMTP:rw9-at-psu.edu] wrote: } } } Dear Listers, } I have a question about the cleaved glass blades used to cut cross } sections of copolymers. Is there any way to quantify the sharpness of } the blades? Do they have a radius of curvature, or anything like that? } A client of ours has asked if the blade would be sharp enough to slice } through the hard domain (polystyrene hardness) at -20 C or -80 C, } or if it would just push it out of the way. This also applies to the } disposable blades on a cryostat used to section the same sample at } -20C prior to the cryo-ultramicrotoming. The sample is very brittle } at -120C. } Rosemary } } Rosemary Walsh, Manager } The Electron Microscope Facility for the Life Sciences, } A Shared Technology Facility, The Life Sciences Consortium } 1 South FrearLab } Penn State University } University Park, PA 16802 } (814) 865-0212 } rw9-at-psu.edu } http://www.lsc.psu.edu/stf/em/home.html } }
We would like to do immunohistochemistry on rat lung tissue and we are especially interrested in CD4, CD8 and some macrophage-markers such as ED1 and ED2. But we would like to have good morphology as well. I have found some articles about IHC (rat and mice) on tissue fixed in paraformaldehyde-lysine-peroidate and emdedded in low temp paraffin. But most of these articles are rather old (1985-1990). Is anyone familliar with these techniques? Is it difficult to repeat and to get similar results?
Joost Bruijntjes TNO Nutrition and Food Research Institute Zeist Holland E-mail: J.Bruyntjes-at-voeding.tno.nl
Dear Andrew In principle, sections can be orientated using any two features in the block that run parallel to each other and normal to the knife edge. It would be nice to have reference rods included in the block, and there are various possible approaches to this, but I cannot see a) a way of meeting both your criterion of providing a dimensional reference for the assessment of shrinkage, and of orientating the section b) a method which is generally applicable to all microtomy techniques.
To measure shrinkage, I think I would be looking at making the fresh specimen a standard size, following its dimensions through processing. Two ways of making specimens a standard size occur to me: 1) Cut slices of the tissue with razor blades mounted side- by side separated by a spacer. Re-cut the tissue slice to make strips with square cross section, or dice it into cubes 2) take a tissue core with a carefully sharpened stainless steel tube (cannula, large hypodermic needle) and follow the core diameter through the process.
For section orientation, reference cores or rods can be melted with a needle (wax, gelatine) or drilled (plastic) normal to the block face. The holes may be left as voids or filled with a contrasting embedding material (e.g. coloured wax, resin, gelatine). However this is all a bit of a fiddle, and becomes either very difficult to achieve or impossible in ultramicrotomy of small block faces, ultracryotomy or cryostat sectioning with Tissue-tek or other liquid embedment.
A general principle for section orientation and alignment that is applicable to almost all microtomy techniques is to cut a mesa on the block face of rhomboidal or trapezoidal shape. The mesa is cut by removing waste with the corner of the knife (which must be square) to a little more than the thickness of the serial sequence you wish to cut. Since the edge-facets of the mesa will be perfectly aligned normal to the block face the sections can be aligned using any two corners, or if they should be lost/obscured, by means of the surviving edges.
This may fail with whole-mounts. In this situation, usually there are features on the periphery of the specimen that can act as reference marks, and you still have the option to cut the block face into a trapezoid or make a mesa, or melt holes in the embedding material with a hot needle, etc.
Finally, a number of digital image analysis packages now offer fourier-based image correlation for registering images when making stereo-pairs, for example, and montages. AnalySIS is one of these:
www.soft-imaging.de
Chris
Date sent: Fri, 3 Mar 2000 09:01:49 +1300 To: c.jeffree-at-ed.ac.uk } From: Andrew McNaughton {andrew.mcnaughton-at-stonebow.otago.ac.nz}
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Please notice that extracting carbides from the matrix doesn't make them thinner. So, it seems that your idea about ion milling is right.
Best regards and good luck,
Witold
%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%% Witold Zielinski, Ph.D. Warsaw University of Technology Department of Materials Science and Engineering 02-507 Warszawa, Woloska 141 POLAND
To all, thank you for the many suggestions regarding the moving aperture, It is definitly not the distance between aperture & specimen holder. It was checked several times and the clearance is there. Also, the aperture was stable before and is now moving the most in the # 2 position. I would appreciate any further comments. Thank you . Peter A. Stolzenberg, Pesto Inc. " pestoem-at-aol.com"
hello, I have to prepare cross sectionning samples, and I do not know where to buy the small vise that I need to clamp the specimen when I have glued them (I have already the Mbond 610 adhesive). I would greetly appreciate any information about this thing (I am not even sure of the name of it) thank you in advance christine
} } Dear all } } We would like to do immunohistochemistry on rat lung tissue and we are } especially interrested in CD4, CD8 and some macrophage-markers such as ED1 } and ED2. But we would like to have good morphology as well. I have found } some articles about IHC (rat and mice) on tissue fixed in } paraformaldehyde-lysine-peroidate and emdedded in low temp paraffin. } But most of these articles are rather old (1985-1990). Is anyone familliar } with these techniques? Is it difficult to repeat and to get similar results? } } } Joost Bruijntjes } TNO Nutrition and Food Research Institute } Zeist } Holland } E-mail: J.Bruyntjes-at-voeding.tno.nl
********** I have had some luck with the PLP fixation you mention (McLean & Nakane, J Histochem Cytocem 22, 1974). I gives good structural preservation, and like most immuno techniques, you won't know if it works with your antigen until you try it. Are you sure you mean low temp paraffin and not resin? Lee
Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Confocal Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
I'm looking for a source for vibration isolation tables for ultramicrotomy. The marble tables that we are using now are not adequate (except during Spring Break when no one is in the building...) I'm thinking of a system that uses compressed gas (N2?) to float the tables.
Please respond offline (vendors welcome).
thanks, all
Ann Hein Lehman EM Facility Mgr Trinity College Hartford CT 860-297-4289 860-297-2538 ann.lehman-at-trincoll.edu
I have recently performed an immunoEM labeling procedure with myelinated nerve embedded in Lowicryl, K4M resin. Unfortunately the myelin is severely altered (it appears as large clealr blebs) because I am not able to osmicate this tissue due to the UV polymerization of this resin. Does anyone have any suggestions as to preservation of lipid laden myelin without the use of osmium? I used methanol in the dehydration procedure which has probably caused this artefact....
Thanks! Karen Jensen, M.S. Associate Scientist Electron Microscope Research Imaging Core University of Rochester Medical Center Rochester, NY
I lost contact w/ a certain person. All I can remember is that they were experienced in confocal microscopy and lived in North West Ct, I think Cornwall.
Dr. Gary Gaugler 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 } -----------------------------------------------------------------------. } } My post has generated several responses. All of which are very } helpful. Thanks to all who responded. } } From following up on what I have discovered from the response from } Alex Greene, I'm rather stuck. It turns out that Amray has a lock on } the FE gun/emitter that they developed and which incorporate the } FEI W/Zr emitter (305FE). As such, no other party will support the } Amray FE systems. } } There are several folks who will and do support the Amray LaB6 systems. } But none can or will support the FE systems. So for the time being, } I am stuck with Amray. Be that as it may. As I write this, Amray } (KLA-Tencor) is preparing to work on my FESEM under my existing } maintenance contract. This is good. I guess that so long as this } keeps up, I am OK. But I do expect that KLA will drop all Amray } field support in the near future. Consequently, perhaps that will } be a better time to seek alternative maintenance sources. I do } expect at this future juncture, KLA/Amray will supply parts } but will not supply warm bodies. If this is an opportunity for } independents, who knows? If you are experiencing similar } troubles, I would for one appreciate hearing about your situation. } Otherwise... } } Stay tuned. } } gary g. Gary, Your earlier comments on how good AMRAY is really puzzle me, but be that as it may, if any of their Federal Government customers are on the ball, Amray will be told, as was Perkin Elmer ETEC, that they must support the equipment for about 7 years or face lawsuits. There is an implied responsibility when one sells expensive equipment.
Perhaps supplying parts and detailed instructions will cover them legally, in which case we third party folks may be able to help out. If anyone has any more detailed info on Amray service, I would love to know because I have serviced them off and on over the years and would be willing to help out here in the Northeast/Mid-Atlantic area.
Ken Converse owner Quality Images third party SEM service 16 Creek Rd. Delta, PA 17314
Have a look at: Toda, Y et al: Application of tyramide signal amplification system to immunohistochemistry: A potent method to localize antigens that are not detectable by ordinary method. Pathology International 1999; 49: 479-483. They mention CD4 specifically in addition to other antigens and outline several antigen retrieval methods.
South Bay Technology, Inc. manufactures both a cross sectioning vise (formerly produced by VCR Group) and also an XTEM clamp. Both tools are used for the same purpose you describe although they do it a little bit differently. I am sending you a data sheet covering each one as an attachment to a separate e-mail. We also offer a complete cross section sample preparation kit which provides all of the bits and pieces that you would need to make the cross section.
David Henriks Vice President 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
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by Leroux christine } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
hello, I have to prepare cross sectionning samples, and I do not know where to buy the small vise that I need to clamp the specimen when I have glued them (I have already the Mbond 610 adhesive). I would greetly appreciate any information about this thing (I am not even sure of the name of it) thank you in advance christine
I am looking for a backscattered electron detector (for elemental contrast) to attach to a JEOL JSM-5800 SEM. This is for a university, so low-cost or donation would be most desirable. Thanks for your help!
Hi, Leroux: I think a negative tweezle will do, provided that your sample is small. Just make sure that you do not heat your sample too fast. Actually I designed a spring vice especially for cross section samples, it is very good for both big and small samples, the glue line can go thinner than 100nm easily, but normally I do not use it since almost all of my samples are small---those film growers are really stinge:) Anyway, if you need, I can send a schematic to you.
Good Luck.
Hao Li
On Tue, 14 Mar 2000, Leroux christine 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, } I have to prepare cross sectionning samples, and I do not know where to } buy the small } vise that I need to clamp the specimen when I have glued them (I have } already the } Mbond 610 adhesive). } I would greetly appreciate any information about this thing (I am not even } sure of the } name of it) } thank you in advance } christine } } ************************************** } Christine leroux } Lab. M.M.I } Universite de Toulon-Var, Bat.R } B.P.132 } F-83957 La Garde Cedex } tel: 00 33 (0) 4 94 14 25 07 } fax: 00 33 (0) 4 94 14 21 68 } } ************************************** } }
Does anyone have a protocol for making Toluidine blue stain to be used for staining thick sections of resin embedded biological samples ?
Thanks a lot,
Soumitra
***************************************************************** Soumitra Ghoshroy Ph.D. Electron Microscopy Lab Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3600 Fax: 505-646-5665 e-mail: ghoshroy-at-nmsu.edu http://confocal.nmsu.edu/eml
Can anyone suggest name of an individual or independent company in New Mexico, West Texas or Arizona who can fix/service research level compound microscopes ?
Thanks in advance,
Soumitra
***************************************************************** Soumitra Ghoshroy Ph.D. Electron Microscopy Lab Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3600 Fax: 505-646-5665 e-mail: ghoshroy-at-nmsu.edu http://confocal.nmsu.edu/eml
We have an old PGT System 4plus (no detector) sitting here that won't boot. Apart from that it was working fine. If anyone would be interested in it for parts (or whatever else you want), they are welcome to it. Not interested in cash, just pay for your shipping and its yours. We are located near Ottawa, Canada.
Please contact Mark Chambers at 599-6500 ext. 4269 for more info, or if you are interested.
You can buy a nice little vise from South Bay Technology. This vise was previously made by VCR. It can be put on a hot plate and has Teflon jaws and a Teflon slide base so that the epoxy doesn't stick. I have two of these. They work great. I put a small dab of epoxy on t he Teflon jaw to determine when the epoxy is cured and scraped it off with a glass microscope slide. SBT also has two spring loaded clamping devices that can be used, but I recommend the small vise. You can also buy a spring loaded clamping device from Fischione and from Gatan. -Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies." --
} -----Original Message----- } From: Leroux christine [mailto:leroux-at-univ-tln.fr] } Sent: Tuesday, March 14, 2000 9:45 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: TEM cross section preparation } } } -------------------------------------------------------------- } ---------- } 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 to prepare cross sectionning samples, and I do not } know where to } buy the small } vise that I need to clamp the specimen when I have glued } them (I have } already the } Mbond 610 adhesive). } I would greetly appreciate any information about this thing } (I am not even } sure of the } name of it) } thank you in advance } christine } } ************************************** } Christine leroux } Lab. M.M.I } Universite de Toulon-Var, Bat.R } B.P.132 } F-83957 La Garde Cedex } tel: 00 33 (0) 4 94 14 25 07 } fax: 00 33 (0) 4 94 14 21 68 } } ************************************** } }
Here is the web site for TMC: http://www.techmfg.com/vibrasolutions.html and http://www.techmfg.com/65floorplatfrm.html -Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
} -----Original Message----- } From: Lehman, Ann [mailto:Ann.Lehman-at-trincoll.edu] } Sent: Tuesday, March 14, 2000 9:37 AM } To: 'MSA Listserver' } Subject: Vibration isolation tables } } } -------------------------------------------------------------- } ---------- } 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, } } I'm looking for a source for vibration isolation tables for } ultramicrotomy. } The marble tables that we are using now are not adequate } (except during } Spring Break when no one is in the building...) I'm thinking } of a system } that uses compressed gas (N2?) to float the tables. } } Please respond offline (vendors welcome). } } thanks, all } } Ann Hein Lehman } EM Facility Mgr } Trinity College } Hartford CT } 860-297-4289 } 860-297-2538 } ann.lehman-at-trincoll.edu }
} Dear Microscopists, } } Does anyone have a protocol for making Toluidine blue stain to be used for } staining thick sections of resin embedded biological samples ? } } Thanks a lot, } } Soumitra } }
Toludine Blue Stain
100g Toludine Blue 100g Borax (sodium borate) 1000 mls ddw.
Older stain works better than newer. Filter before use.
Dr. Elaine Humphrey Biosciences Electron Microscopy Facility 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
Hello all, We are trying to get a project started that involves imaging the microstructure of mineral oils and polyalphaolefins as they solidify. The solid temperature range for materials like these is approximately -20 to -70 C. Does anyone have experience with these or similar materials. We are hoping to image the particle structure (crystals) by SEM. Any feedback from those with similar projects/experiences would be very welcome (on or off the list). Even better would be references to any published reports of this kind of material exam.
Thanks, Brad Huggins BPAmoco, Electron Microscopy Lab hugginbj-at-bp.com
At 01:13 PM 3/14/00 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
[snip]
} Gary, } Your earlier comments on how good AMRAY is really puzzle me, but be that } as it may, if any of their Federal Government customers are on the ball, } Amray will be told, as was Perkin Elmer ETEC, that they must support the } equipment for about 7 years or face lawsuits. There is an implied } responsibility when one sells expensive equipment. } } Perhaps supplying parts and detailed instructions will cover them } legally, in which case we third party folks may be able to help out. If } anyone has any more detailed info on Amray service, I would love to know } because I have serviced them off and on over the years and would be } willing to help out here in the Northeast/Mid-Atlantic area. } } Ken Converse } owner } Quality Images } third party SEM service } 16 Creek Rd. } Delta, PA 17314 } } 717-456-5491
I take it that you don't think that Amrays are very good. What aspect of them do you find lacking? I would tend to agree that the early models are no comparison to their generations of the last 6-10 years. The FE systems are particularly good. My 1910FE does a great job being an FE and having the flat lens. The 1800 series with conical lenses are not as good, but are required when doing IC wafers.
You have a great point about US Gov users. I know several of them. When does the 7 years start? From what event or timepoint? Amray has not stopped providing service. But the word is that they most likely will in about 2 years....based on their takeover by KLA-Tencor.
One fellow pointed out the proprietary nature of the FE gun. Without access to this, third party providers cannot work on the systems....at least not on the gun assembly.
My second topic for the day is FESEM or TEM imaging of the crystalline/amorphous microstructure of poly(ethylene terephthalate), PET. I have on many occasions had success achieving minimal contrast of such domains in certain unstained PET polymers in the SEM and FESEM. Recently we have "almost" been able to observe the crystalline domains with enough clarity to size them, this by direct examination of polished (microtomed) surfaces. I would like to pursue the microstructural characterization a step further by using staining or etching techniques (or some other trick that some one may have up their sleeve!). In Polymer Microscopy (L Sawyer, D Grubb) aminolysis appears to be (cautiously) recommended. Staining with PTA is also mentioned. I am looking to image crystalline domains in the range of approximately 50 to 150 nm.
Does anybody on the List have first hand experience with these or other techniques to enhance the direct (or indirect) observation of these domains in PET. I'm not looking to re-invent the wheel here, so any help will be greatly appreciated. Get back to me direct or on the List.
Again, Thanks in advance, Brad Huggins BPAmoco, Electron Microscopy Lab hugginbj-at-bp.com
} hello, } I have to prepare cross sectionning samples, and I do not know where to } buy the small } vise that I need to clamp the specimen when I have glued them (I have } already the } Mbond 610 adhesive). } christine
You haven't told us what equipment you will use to prepare your "cross sectionning samples", so it's hard to provide advice!
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/PCI/pci.html
I would suspect Elaine is giving a 10X stock solution?
We use 1% Toluidine Blue dissolved in 1% sodium borate. Filter before use. Diluting 10-fold with 1% sodium borate, to 0.1% toluidine blue, allows lightly counterstaining sections developed for autoradiography.
Refs. Mercer, E.H., J Royal Microscopy Society, 81:179 (1963) Burns, W.A., "Thick Sections: Technique and Applications", Diagnostic Electron Microscopy, Chapter 4, B.F. Trump and R.J. Jones, eds., John Wiley and Sons, New Yourk , 1978.
Regards, Glen
Glen MacDonald Research Scientist Hearing Research Laboratories of the Virginia Merrill Bloedel Hearing Research Center Box 35-7923 University of Washington Seattle, WA 98195-7923 (206) 616-4156 glenmac-at-u.washington.edu
} } Dear Microscopists, } } } } Does anyone have a protocol for making Toluidine blue stain to be used for } } staining thick sections of resin embedded biological samples ? } } } } Thanks a lot, } } } } Soumitra } } } } } } } Toludine Blue Stain } } 100g Toludine Blue } 100g Borax (sodium borate) } 1000 mls ddw. } } Older stain works better than newer. Filter before use. } } } Dr. Elaine Humphrey } Biosciences Electron Microscopy Facility } 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 } } } }
} We have an old semicaps 4000 for pc windows 3.1 system. } I've been trying to use it to get scale bars burned } into images, but I must say that it is very buggy. ...
First, I need to admit no experience with this system ... but even with my software which does produce accurate scale bars, I still prefer to annotate the images with an image editor like Photoshop.
If you have a magnification standard, then you should be able to determine an instrumental constant which can be assumed doesn't change with magnification, but may change with the SEM's working distance. You should be able to end up with this equation:
microns per pixel = constant/magnification
This may need to be determined for each fixed working distance, or you may be able to determine how to incorporate WD into the equation (my own software compensates with the objective lens setting so the mag is relatively accurate). If your image acquisition options include pixel dimentions (e.g., 256x256, 512x512, 1024x1024), then this needs to be a factor as well (in the denominator).
I then created a spreadsheet for the pixel dimentions of common scale bars for different magnifications, printed it, and then made it available at the Photoshop workstation.
I can provide an example of the spreadsheet to anyone interested.
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
Thanks Glen, you are quite right. It is easier to make up 10g Toluidine Blue, 10g sodium borate and 1000mls water. Elaine
} I would suspect Elaine is giving a 10X stock solution? } } We use 1% Toluidine Blue dissolved in 1% sodium borate. Filter before use. } Diluting 10-fold } with 1% sodium borate, to 0.1% toluidine blue, allows lightly } counterstaining sections developed for autoradiography. } } Refs. } Mercer, E.H., J Royal Microscopy Society, 81:179 (1963) } Burns, W.A., "Thick Sections: Technique and Applications", Diagnostic } Electron Microscopy, Chapter 4, B.F. Trump and R.J. Jones, eds., John } Wiley and Sons, New Yourk , 1978. } } Regards, } Glen } } } Glen MacDonald } Research Scientist } Hearing Research Laboratories of the } Virginia Merrill Bloedel Hearing Research Center } Box 35-7923 } University of Washington } Seattle, WA 98195-7923 } (206) 616-4156 } glenmac-at-u.washington.edu } } } } Dear Microscopists, } } } } } } Does anyone have a protocol for making Toluidine blue stain to be used for } } } staining thick sections of resin embedded biological samples ? } } } } } } Thanks a lot, } } } } } } Soumitra } } } } } } } } } } } } Toludine Blue Stain } } } } 100g Toludine Blue } } 100g Borax (sodium borate) } } 1000 mls ddw. } } } } Older stain works better than newer. Filter before use. } }
Dr. Elaine Humphrey Biosciences Electron Microscopy Facility 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
We have a FEI CM20 (TEM) and XL30 (FEGSEM) and are in the process of reviewing service contracts vs insurance. The service contract runs a min of 40K for both instruments. We currently have a tech that can do minor service, yearly maintenance, and alignments.
I'd appreciate any input as to how other organizations have dealt with this.
} } Dear Collegues } } I'm having a problem with LR-White polimerization. } } I use to polimerise the LR-white resin at 60ŒC without any aditives and it } usually works without problems. A new batch of the resin failed to } polimerise, and the manufacturer claims that it is necessary to use the } catalyst. } } I understand that the "accelerator" should not be used except for cold cure, } and even then may be harmfull since the specimen may be subject to too much } heat even under those conditions. } } I would like your opinion on: } 1. is it necessary to use the catalyst for heat cure? } } 2. If it is, is there any difference among the resins that may justify the } apparent success of the old protocol (good cure without catalyst?) } } Thanks in advance } } Dr. A.P. Alves de Matos } } }
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Weight 0.5 g Tolyidine Blue in some bottle (I am using 50 ml polypropylene tissue culture tubes), add 50 ml 1x PBS and heat it in boiled water for 20-30 min. When hot, filter it using Whatman filter paper (Cat No 1001 090).
Staining: Plastic sections up to 0.5 microns thick on the microscope slide cover with several drops of the stain and heat on heat-plate, 60oC for 5-10 min. Wash away stain with tap water, wash in distilled water, dry and enjoy the view.
Godd luck, Sergey.
} Date: Tue, 14 Mar 2000 13:18:28 -0700 } From: Soumitra Ghoshroy {ghoshroy-at-nmsu.edu} } Subject: Toluidine blue stain } X-Sender: ghoshroy-at-cnmailsvr.nmsu.edu } To: Microscopy-at-sparc5.microscopy.com } } ------------------------------------------------------------------------ } 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
We have a FEI CM20 (TEM) and XL30 (FEGSEM) and are in the process of reviewing service contracts vs insurance. The service contract runs a min of 40K for both instruments. We currently have a tech that can do minor service, yearly maintenance, and alignments. We have not had a service contract for the last 2 years and have had no problem getting FEI to come out ASAP when needed.
I'd appreciate any input as to how other organizations have dealt with this.
I have a (possibly) similar problem with JEOL 100SX microscope. Very often the beam keeps tilting in a motion that displaces it relative to the objective aperture. The effect is the same as moving the aperture. I suspect of circuit instability but the assistance was unable to find any trouble in that area. Another possibility is charge accumulation inducing electric fields and displacing the beam. Since you are lucki to have the problem mostly in one position, I would suspect of some kind of electric isolation of the aperture holder either in that hole or produced when the entire piece goes into that position. The charge effects could be also related to the specimen itself. Jeol seems to be particularly sensitive to this. If you are looking at epon sections try to coat them with a thin layer of carbon (after staining, evaporate directly into the sections) - it gives much improvement for me.
Dr. A.P. Alves de Matos
To all, thank you for the many suggestions regarding the moving aperture, It is definitly not the distance between aperture & specimen holder. It was checked several times and the clearance is there. Also, the aperture was stable before and is now moving the most in the # 2 position. I would appreciate any further comments. Thank you . Peter A. Stolzenberg, Pesto Inc. " pestoem-at-aol.com"
Most LR White is sold with catalyst already added. This will cure at 60 degrees and must be stored refrigerated. If you purchased uncatalysed (the only way we supply LR White because we are a long way from the UK) then prior to first use the catalyst must be added to the whole bottle and well mixed to dissolve. Thereafter store the catalysed LR White refrigerated.
If you want to cold cure you need to additionally add accelerator. Accelerator is not supplied with a bottle of LR White, but when you purchase a "kit". Instruction notes are linked online from the LR White. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Wednesday, March 15, 2000 11:27 AM, A.P. Alves de Matos [SMTP:apmatos-at-ip.pt] 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 Collegues } } } } I'm having a problem with LR-White polimerization. } } } } I use to polimerise the LR-white resin at 60oC without any aditives and it } } usually works without problems. A new batch of the resin failed to } } polimerise, and the manufacturer claims that it is necessary to use the } } catalyst. } } } } I understand that the "accelerator" should not be used except for cold } cure, } } and even then may be harmfull since the specimen may be subject to too } much } } heat even under those conditions. } } } } I would like your opinion on: } } 1. is it necessary to use the catalyst for heat cure? } } } } 2. If it is, is there any difference among the resins that may justify the } } apparent success of the old protocol (good cure without catalyst?) } } } } Thanks in advance } } } } Dr. A.P. Alves de Matos } } } } } } } }
I do not know what the objective aperture strip looks like on a 100 CX so I am sort of guessing.
I have seen some aperture rods where the end consists of say three or four holes of identical diameter and small Pt/Mo discs with different sized holes etched in. If the No2 aperture is moving the most then this could possibly explain it. This disc maybe more loosely held or has a worse connection to ground than the others. Can you check these with a multimeter or something?
What does the movement look like:- continuous slow/ slow with fast jumps/ rapid shaking/random direction/ one direction: along the rod or side to side? And does the total beam current affect the motion greatly?
******************************************************** Dr Jonathan Barnard
Analytical Materials Physics The Angstrom Laboratory, Uppsala University P O Box 534, SE-751 21 Uppsala, Sweden Phone: +46-(0)18-4716838 Fax: +46-(0)18-500131 http://www.angstrom.uu.se/analytical/home.html
I need to embed some complex glass mouldings which also contain components made of plastics (HDPE, PP, PC) in (for preference) clear resin for sectioning with a diamond saw. This has turned out to be much more problematical than I had expected, for two reasons:
1) Very considerable shrinkage of the resins occurs during curing (applies to acrylic, epoxy or polyester types) . This causes separation of the resin from the glass and plastics components inside partially-enclosed voids, so that the original spatial relationships are lost.
2) bonding of the resins to the glass is quite variable, but typically poor. Bonding to plastics is poor. When the bond-strength to glass is high, particularly with epoxies, the stresses induced by the shrinking resin during curing are large enough to fracture the glass.
Is there such a thing as a non-shrinking resin? How can I improve bond-strength between glass and resin? How can I improve bond-strength between resin and HDPE or PP?
Any tips gratefully received Yours Chris ===================================================================== DR CHRIS JEFFREE BIOSEM - BIOLOGICAL SCIENCES EM FACILITY UNIVERSITY OF EDINBURGH Daniel Rutherford Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JH, Scotland, UK Tel. #44 131 650 5345 FAX. #44 131 650 6563 Mobile 0410 585 401 email c.jeffree-at-ed.ac.uk SEM / TEM bookings sem-at-ed.ac.uk =====================================================================
To stain resin, e.g. Spurr's, toluidine blue must be prepared at high pH.
0.5% toluidine blue in 0.1%sodium carbonate (Na2CO3) (pH 11.1) works well. Stain sections on slides on a hot plate at 60C for 30sec. to 2 min. and rinse with water. Some resins and plastics are much easier to stain than Spurr's, e.g. JB-4, GMA etc. so I also use 0.1% tol blue in 0.01% sodium carbonate (pH 6.6). The basic protocol stays the same, you vary concentrations to adjust pH and therefore staining intensity. Hope this helps, John.
Soumitra Ghoshroy wrote:
} Dear Microscopists, } } Does anyone have a protocol for making Toluidine blue stain to be used for } staining thick sections of resin embedded biological samples ? } } Thanks a lot, } } Soumitra } } ***************************************************************** } Soumitra Ghoshroy Ph.D. } Electron Microscopy Lab } Box 3EML } New Mexico State University } Las Cruces, NM 88003 } Tel: 505-646-3600 } Fax: 505-646-5665 } e-mail: ghoshroy-at-nmsu.edu } http://confocal.nmsu.edu/eml
-- C. John Runions, Ph.D. Department of Plant Sciences University of Cambridge Downing St. Cambridge CB2 3EA UK
From your comments I would suspect that you have a charging problem. As it seems to be dependent on the aperture chosen it may be on the aperture mechanism. Try cleaning it, check that there is electrical continuity along the rod and that the mechanism is properly grounded. Do you have a touch switch? If so then ensure that it is grounded through the resistor properly.
Good luck, Ron
On Tue, 14 Mar 2000 PESTOEM-at-aol.com-at-sparc5.microscopy.com wrote:}
} thank you for the many suggestions regarding the moving aperture, It is } definitly not the distance between aperture & specimen holder. It was checked } several times and the clearance is there. Also, the aperture was stable } before and is now moving the most in the # 2 position. } I would appreciate any further comments. } Thank you . Peter A. Stolzenberg, Pesto Inc. " pestoem-at-aol.com" } }
=========================================================================== Mr. Ron Doole e-mail ron.doole-at-materials.ox.ac.uk Department of Materials, phone +44 (0) 1865 273701 University of Oxford, fax +44 (0) 1865 283333 Parks Road. Oxford. OX1 3PH. UK. ============================================================================
I know oil based fuels have been imaged at low temperature. Suggest you read (better still, Buy) my book "Low Temperature Microscopy and Analysis" Plenum Press New York 1992. I would modestly suggest it has a lot of information about how to prepare, observe and analyse specimens at low temperature. My own current interests revolve around low volyage, low temperature x-ray microabalysis of bio-organic materials. Let me know if I can be of any further help
Patrick Echlin University of Cambridge UK
pe13-at-cus.cam.ac.uk
On Tue, 14 Mar 2000, 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 } -----------------------------------------------------------------------. } } } Hello all, } We are trying to get a project started that involves imaging the } microstructure of mineral oils and polyalphaolefins as they solidify. The } solid temperature range for materials like these is approximately -20 to -70 } C. Does anyone have experience with these or similar materials. We are } hoping to image the particle structure (crystals) by SEM. Any feedback from } those with similar projects/experiences would be very welcome (on or off the } list). Even better would be references to any published reports of this } kind of material exam. } } Thanks, } Brad Huggins } BPAmoco, } Electron Microscopy Lab } hugginbj-at-bp.com } } }
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You have fallen into the same trap I did some time back. Some suppliers ship the LR White resin with the catalyst already included. Others do not, sending the catalyst along with instructions to add it prior to using the resin. The reason to not mix the ingredients prior to shipping is that if the resin is subject to hot temperatures or prolonged shipping time, the resin mix could be compromised. This is a good point which is certanly valid when shipping resin in the summer or to remote locations.
However, most of us do not run into these problems and are used to receiving the resin in the complete mixed form. In my case,I ordered from a different supplier than normal. The labeling of the bottles was not clear and I did not realize that the enclosed "extra" ingredient was the catalyst, not the accelerator used for cold curing polymerization, until I also ran into the problem of the resin not polymerizing as expected.
Moral of the story....always read the fine print...both in the catalog and after receiving the product, especially when ordering from a different supplier than in the past.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
On Tuesday, March 14, 2000, A.P. Alves de Matos {apmatos-at-ip.pt} wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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I have recently performed an immunoEM labeling procedure with } myelinated nerve embedded in Lowicryl, K4M resin. Unfortunately the } myelin is severely altered (it appears as large clealr blebs) because I } am not able to osmicate this tissue due to the UV polymerization of } this resin. Does anyone have any suggestions as to preservation of } lipid laden myelin without the use of osmium? I used methanol in the } dehydration procedure which has probably caused this artefact....
Karen,
It sounds like the myelin is being partially extracted during dehydration because of lack of OsO4 stabilization. You might try acrolein...it is the only fixative other than OsO4 that will preserve and retain lipids. Make sure that you read all the precautions about it...its pretty nasty stuff.
-=W.L. Steffens=- Department of Veterinary Pathology College of Veterinary Medicine University of Georgia Athens, GA 30602 USA http://www.vet.uga.edu/vpp/wls/steffens.html
At 1:18 PM -0700 3/14/0, Soumitra Ghoshroy wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
After your sections have been dried onto the slide, cover them with a drop of stain for 30 sec - 1 min. and wash off with water. Dry.
If its too light, repeat.
Lee
Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Confocal Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
} We are trying to get a project started that involves imaging the } microstructure of mineral oils and polyalphaolefins as they solidify. The } solid temperature range for materials like these is approximately -20 to -70 } C. Does anyone have experience with these or similar materials. We are } hoping to image the particle structure (crystals) by SEM. Any feedback from } those with similar projects/experiences would be very welcome (on or off the } list). Even better would be references to any published reports of this } kind of material exam. } }
Dear Bradley, Doug Dorset has done extensive work with electron crystallography of waxes and lipids. He does electron diffraction with a TEM, but perhaps his results will be useful to you. Yours, Bill Tivol
for methacrylate sections we use 0.5 % Toluidinblue in 0.1 mol phosphate buffer (it should also work just with water) and for epoxide sections 0.1% Toluidinblue in 2% Na-bicarbonate. staining time depends on temperature, thickness, and material. After staining wash with water and dry the sections.
regards, Anne
Soumitra Ghoshroy schrieb:
} ------------------------------------------------------------------------ } 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 Microscopists, } } Does anyone have a protocol for making Toluidine blue stain to be used for } staining thick sections of resin embedded biological samples ? } } Thanks a lot, } } Soumitra } } ***************************************************************** } Soumitra Ghoshroy Ph.D. } Electron Microscopy Lab } Box 3EML } New Mexico State University } Las Cruces, NM 88003 } Tel: 505-646-3600 } Fax: 505-646-5665 } e-mail: ghoshroy-at-nmsu.edu } http://confocal.nmsu.edu/eml
-- Dr. Anne Heller, AG Elektronenmikroskopie, Institut fr Botanik (210), Universitt Hohenheim, Garbenstra§e 30 D-70593 Stuttgart
I forgot another cheaper alternative that I have used in the past. You can use a small, medium, or large binder clip to clamp samples together depending on their size. You can use Teflon tape on the surface to prevent sticking to the clamp. If you make a little Teflon jig to hold the sample it works quite well. -Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies." --
} -----Original Message----- } From: Leroux christine [mailto:leroux-at-univ-tln.fr] } Sent: Tuesday, March 14, 2000 9:45 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: TEM cross section preparation } } } -------------------------------------------------------------- } ---------- } 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 to prepare cross sectionning samples, and I do not } know where to } buy the small } vise that I need to clamp the specimen when I have glued } them (I have } already the } Mbond 610 adhesive). } I would greetly appreciate any information about this thing } (I am not even } sure of the } name of it) } thank you in advance } christine } } ************************************** } Christine leroux } Lab. M.M.I } Universite de Toulon-Var, Bat.R } B.P.132 } F-83957 La Garde Cedex } tel: 00 33 (0) 4 94 14 25 07 } fax: 00 33 (0) 4 94 14 21 68 } } ************************************** } }
Ann, I'm also looking for vibration isolation tables for ultramicrotomy. I've been looking at Vibraplane (http://www.kineticsystems.com) for isolation tables and bench-top isolation platforms, which are also sold through stereo equipment companies (i.e. http://www.soundsofsilence.com http://www.audioodyssey.com). I need a height-adjustable isolation table - other vibration sensitive equipment in our lab is successfully being used on the isolation platforms on top of separate height-adjustable tables. I would appreciate hearing of any information you receive offline about the isolation tables.
Teresa Boes Analytical Services Lab Hewlett-Packard Corvallis, OR 541-715-7055 teresa_boes-at-hp.com
-----Original Message----- } From: Lehman, Ann [mailto:Ann.Lehman-at-trincoll.edu] Sent: Tuesday, March 14, 2000 6:37 AM To: 'MSA Listserver'
Dear Listers,
I'm looking for a source for vibration isolation tables for ultramicrotomy. The marble tables that we are using now are not adequate (except during Spring Break when no one is in the building...) I'm thinking of a system that uses compressed gas (N2?) to float the tables.
Please respond offline (vendors welcome).
thanks, all
Ann Hein Lehman EM Facility Mgr Trinity College Hartford CT 860-297-4289 860-297-2538 ann.lehman-at-trincoll.edu
} Date: Wed, 15 Mar 2000 11:15:59 -0500 } To:Soumitra Ghoshroy {ghoshroy-at-nmsu.edu} } From:sherwood-at-HELIX.MGH.HARVARD.EDU (Peggy Sherwood) } Subject:Re: Toluidine blue stain } } } Soumitra, } } We use a 0.5% Toluidine Blue in Borate Buffer (1 gram Toluidine Blue, 1 } gram sodium borate and 200ml water). Tousimis (and maybe other suppliers) } sells a Methylene Blue- Toludine Blue stain for epoxy stain (Cat. # 4166B) } which we have used: (consists of 0.2% methylene blue & 1.0% toluidine } blue certified stains in Na-Borate buffer). Either way--we always filter } our stains (using a syringe with filter unit attached) right onto } sections. Then we put slide ( super frost plus, so sections will adhere) } on hot plate (temp. -at- 60 decrees C). We rinse sections with water and } then check stain--time is arbitrary: depending upon tissue type & section } thickness as to how dark or light your stain result. } } When I worked in Ophthalmology research, I used a wonderful dichromatic } stain: methylene blue-azure II-basic fuchsin stain. It was similar to H } & E but for epoxy sections. (made great 2X2 slides). The reference is: } Humphrey, C.D. and Pittman, F.E. (1974) Stain Technology 42:9-14. I } actually have a copy that came as an LKB application note 303 which I } could fax to you. Feel free to contact me off line. Good luck! } } Peggy Sherwood } Wellman Labs of Photomedicine } Lab Associate-Photopathology Lab } Massachusetts General Hospital } 50 Blossom Street } Boston, MA 02114 } 617-726-6983 } 617-724-4839 (voice mail) } 617-726-3192 (fax) } sherwood-at-helix.mgh.harvard.edu } } } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thanks to all of you who responded to my question about the stain.
This is a great microscopy resource.
Best wishes.
Soumitra
***************************************************************** Soumitra Ghoshroy Ph.D. Electron Microscopy Lab Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3600 Fax: 505-646-5665 e-mail: ghoshroy-at-nmsu.edu http://confocal.nmsu.edu/eml
Soumitra, We use the following recipe for toluidine blue:
These stock solutions can be made in large volumes and stored. .5 % toluidine blue in distilled water .25 % sodium borate in distilled water
Mix equal parts of the above. Stain dried resin sections on hot plate (low setting) until edge of stain begins to evaporate (5-10 secs) Rinse, blot excess water from bottom of slide and dry on hot plate.
Mary Gail Engle Electron Microscopy & Imaging Facility University of Kentucky
Patrick, Are you telling me that in your book, you have a reference to some work where some oil based fuels were cooled to a solid, and them imaged in that frozen state? If so, this is precisely the kind of information that I would like to see. If not, do you have a reference? I do not have a copy of your book, but I have seen it, some years ago. In any case, thanks for the reminder on that resource.
And even more specifically, if anyone has knowledge of any similar work done on mineral oil and like materials... it sure would save me some work on the development side...
Thanks for the feedback, Brad
} ---------- } From: Dr P. Echlin[SMTP:pe13-at-cus.cam.ac.uk] } Sent: Wednesday, March 15, 2000 6:43 AM } To: Huggins, Bradley J } Cc: Microscopy listserver } Subject: Re: Cryo-microscopy of solid mineral oils } } Dear Bradley: } } I know oil based fuels have been imaged at low temperature. Suggest you } read (better still, Buy) my book "Low Temperature Microscopy and } Analysis" Plenum Press New York 1992. I would modestly suggest it has a } lot of information about how to prepare, observe and analyse specimens } at low temperature. My own current interests revolve around low volyage, } low temperature x-ray microabalysis of bio-organic materials. Let me } know if I can be of any further help } } Patrick Echlin } University of Cambridge UK } } pe13-at-cus.cam.ac.uk } } } On Tue, 14 Mar 2000, 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 } } -----------------------------------------------------------------------. } } } } } } Hello all, } } We are trying to get a project started that involves imaging the } } microstructure of mineral oils and polyalphaolefins as they solidify. } The } } solid temperature range for materials like these is approximately -20 to } -70 } } C. Does anyone have experience with these or similar materials. We } are } } hoping to image the particle structure (crystals) by SEM. Any feedback } from } } those with similar projects/experiences would be very welcome (on or off } the } } list). Even better would be references to any published reports of this } } kind of material exam. } } } } Thanks, } } Brad Huggins } } BPAmoco, } } Electron Microscopy Lab } } hugginbj-at-bp.com } } } } } } }
I want to observe pure iron by means of TEM (Jeol 1200 EX II) in order to study the dislocations substructure. But I have some problems, for example, I can not focus the image due to the magnetism of the ferrite.
I am looking for suggestions to avoid this problem. What could I do?
Thank you in advance.
Dr. J. M. Manero Universidad Politcnica de Catalunya. E.T.S.I. Industriales de Barcelona. Spain. e-mail: manero-at-cmem.upc.es
Ben, If you have other microscopes on campus you might consider forming a consortium of all the users and hiring your own on-site service engineer to take care of all the instruments. We've been doing this since 1981 and it's worked very well. One man's salary is considerably less than service contracts on many instruments. You also have the luxury of instant service. Mary Gail Engle Electron Microscopy & Imaging Facility University of Kentucky
1) Start (before final thinning) with the thinnest sample you can support ( {50 microns). Pure iron will be quite soft so you'll have to be very careful not to bend it - especially since you're looking at dislocation structure. You could support it on a copper key-grid to minimize damage. If you have to electropolish while it is on the copper key-grid, you'll need to lacquer the copper prior to electropolishing and then clean it off before putting it into the microscope. 2) If you still can't focus, try taking it out of the microscope and rotating it in the holder. Sometimes a different orientation really helps. 3 Plan on doing alignment every time you move or tilt. It is a given with magnetic samples. You'll need to adjust the beam tilt and the astigmatism continually. I always used the "dark-field" beam tilts as they are stronger than the bright field set. You'll be amazed at how good you get at aligning after a few months of magnetic work. Later work with non-magnetic samples will be a breeze. 4) One final warning. The objective lens can actually suck your sample out of the holder during insertion of the sample. If you have a really thin sample, it can break off chunks. These chunks/sample will ALWAYS stick to the lens and make every other user miserable. Your service guy will be forced at some point to clean off the lens of your sample (or sample bits) and you will be reviled. A simple solution is to always turn off the objective lens when you are inserting and removing your sample.
Good luck - magnetic work is possible and fun once you get the hang of it.
Robin Griffin
-----Original Message----- } From: Jose Maria Manero [mailto:manero-at-cmem.upc.es] Sent: Wednesday, March 15, 2000 11:59 AM To: Microscopy-at-sparc5.microscopy.com
Dear all,
I want to observe pure iron by means of TEM (Jeol 1200 EX II) in order to study the dislocations substructure. But I have some problems, for example, I can not focus the image due to the magnetism of the ferrite.
I am looking for suggestions to avoid this problem. What could I do?
Thank you in advance.
Dr. J. M. Manero Universidad Politcnica de Catalunya. E.T.S.I. Industriales de Barcelona. Spain. e-mail: manero-at-cmem.upc.es
We've been having stability problems with our mercury vapor lamps on our Reichert Polyvar microscope. We have been told by one source that the power supply is likely the problem and another source tells us that the bulbs are the problem. The source illumination, when imaged, can be seen to flicker at random intervals and causes the illumination to vary. Does anyone have experience with this microscope, and if so which brand of lamps do you use? This microscope uses a 220W/4 L1 lamp. Thanks.
David O'Neil National Research Council of Canada Institute for Marine Biosciences 1411 Oxford Street Halifax, NS B3H 3Z1 ph: (902)426-8258 fax: (902)426-9413 e-mail: david.o'neil-at-nrc.ca
I certainly agree that EM of magnetic samples is quite doable, but I'm not sure I'd agree that it is "fun".
Perhaps the best tip I can give is to reduce the amount of magnetic material within the polepiece. I have cut my magnetic samples down to less than the standard 3mm size and glued them securely onto a Cu slot grid. The magnetic effects are much less of a problem.
Cheers, Henk
At 12:13 PM 3/15/00 -0600, rgriffin-at-eng.uab.edu"-at-sparc5.microscopy.com wrote: } {snip}
} Good luck - magnetic work is possible and fun once you get the hang of it. } } Robin Griffin
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://web.ceof.ohio-state.edu Murphy's Law: "If anything can go wrong, it will." Commentary: "Murphy was an optimist."
Soumitra, We use a Toluidine Blue and Methylene blue staining solution:
Dissolve 0.5gm Borax in 95 ml DH2O, add 0.5gm of methylene blue, mix then add 5 ml of a 2% Toluidine blue solution in H2O.
Filter the stain just before staining. Stain for 1 minute on a hot plate set slightly lower than where the stain starts to bubble. Rinse with DH2O and dry.
We like this stain because it is fast, very clean, and very pretty. We prefer this stain to other Toluidine Blue solutions.
George
George Lawton Chief Electron Microscopist Molecular and Cellular Imaging Facility UT Southwestern Medical Center at Dallas 5323 Harry Hines Blvd. Dallas, Tx 75390-9039 Phone: 214-648-7291 eMail: George.Lawton-at-email.swmed.edu
A lab on our floor is having trouble with their B & L spectrophotometer. does anyone know who, hopefully in New England, might be able to service the unit?
1. Reduce the sample size as much as possible, glue a tiny Fe piece on a Cu or other nonmagnetic grids, for example. 2. Turn off the objective lens as you load your sample. 3. Do beam alignment as many times as necessary. -cy Rodel Inc. 451 Bellevue Road Newark, Delaware 19713 USA
-----Original Message----- } From: Jose Maria Manero [mailto:manero-at-cmem.upc.es] Sent: Wednesday, March 15, 2000 12:59 PM To: Microscopy-at-sparc5.microscopy.com
Dear all,
I want to observe pure iron by means of TEM (Jeol 1200 EX II) in order to study the dislocations substructure. But I have some problems, for example, I can not focus the image due to the magnetism of the ferrite.
I am looking for suggestions to avoid this problem. What could I do?
Thank you in advance.
Dr. J. M. Manero Universidad Politcnica de Catalunya. E.T.S.I. Industriales de Barcelona. Spain. e-mail: manero-at-cmem.upc.es
A simple construction we used for years with a heavy microscope was a wooden frame standing on a tennis balls. The microscope was on the 2nd floor of a building by a busy road. Never a shake was recorded on film!
Diana
Diana van Driel Department of Clinical Ophthalmology Sydney University GPO Box 4337 Sydney NSW AUSTRALIA 2001
As an independent service provider of several SEM manufacturers for over 20 years, I think I can also comment & agree with Ken on several fronts concerning Amray. However, I don't want to get into a "bashing" of any given manufacturer, at least online. I will say that every Company (SEM or otherwise) has their own way of doing things. Each Company is different, depending upon the people running the operation.
Many manufacturer's think they have a "lock" on the customer and on the service because of their FE guns or perhaps on another part. This reminds me of ETEC just before they got out of the SEM service business. ETEC had an epoxy encapsulated high voltage power supply that only ETEC manufactured. I inquired with ETEC about the cost of one of these units. ETEC responded with a price tag of $50,000.00. Yes $50,000.00 in 1982! ETEC service also let it be known that they were the only source for this part. Furthermore, customers considering a third party for service were informed (and threatened) by ETEC about this situation. Customers were stuck paying ETEC's exorbitant maintenance costs.
The ETEC customers were hesitant to go to a third party service until the high voltage issue had been resolved. I spent many days (and nights) re-engineering another high voltage power supply until I had it perfectly duplicated. When customers would ask about the high voltage issue, I would show them my version & invite them to use it in their system. Within two years I had over 50% of ETEC's service business at 60% of ETEC contract costs.
Today several manufacturers still think they have a "lock" on service. I have been informed by one manufacturer that this is their way of "putting you (third party maintenance) out of business". If I had the time and the inclination, I would spend it duplicating the FE gun some of these manufacturer's think they have a "lock" on. I have considered such a move because no one has a "lock" on technology.
I have had several requests but have not responded because the time & effort necessary is not viable for only three customer requests.. Fortunately for the manufacturer, I am busy and, quite frankly, not as "hungry" as I used to be. Maybe it comes with age. If I ever get "unhungry" or not as busy, I would take these manufacturer's up on their smugness and beat them at their own game by duplicating their FE guns. The ultimate winner is the customer.
The only manufacturer that has a great gun that can be serviced by the customer is Hitachi. Hitachi has the patent on an internal heater for their gun which makes exchanging FE guns obsolete. Hitachi FE guns can be re-conditioned by the customer in-house at a cost of $750.00 instead of $7,000.00 for Amray or $17,000.00 for JEOL. Hitachi is doing well and doesn't need to worry about "locking" third party maintenance or anyone else out. They have too many sales to worry about & don't have the time to be petty.
Great thing about the free enterprise system is that real talent always has customers "beating a path" to their door. Marginal talent needs to cajole customers to their way of thinking. Great talent has customers calling on them.
Regards,
Earl Weltmer Scanservice Corporation Third Party Maintenance
} Gary, } Your earlier comments on how good AMRAY is really puzzle me, but be that } as it may, if any of their Federal Government customers are on the ball, } Amray will be told, as was Perkin Elmer ETEC, that they must support the } equipment for about 7 years or face lawsuits. There is an implied } responsibility when one sells expensive equipment. } } Perhaps supplying parts and detailed instructions will cover them } legally, in which case we third party folks may be able to help out. If } anyone has any more detailed info on Amray service, I would love to know } because I have serviced them off and on over the years and would be } willing to help out here in the Northeast/Mid-Atlantic area. } } Ken Converse } owner } Quality Images } third party SEM service } 16 Creek Rd. } Delta, PA 17314 } } 717-456-5491
I take it that you don't think that Amrays are very good. What aspect
of them do you find lacking? I would tend to agree that the early models are no comparison to their generations of the last 6-10 years. The FE
systems are particularly good. My 1910FE does a great job being an FE and having the flat lens. The 1800 series with conical lenses are not as good, but are required when doing IC wafers.
You have a great point about US Gov users. I know several of them. When does the 7 years start? From what event or timepoint? Amray has not stopped providing service. But the word is that they most likely will in about 2 years....based on their takeover by KLA-Tencor.
One fellow pointed out the proprietary nature of the FE gun. Without access to this, third party providers cannot work on the systems....at
We are trying to examine oil inclusions trapped in quartz under UV light. The oil fluoresces and the quartz does not. Sounds easy. Unfortunately the epoxy that the sample is embedded in also fluoresces quite strongly making identification difficult. So I was wondering if there are any non-fluorescent epoxies available on the market, or alternatively if there are any pigments/dyes that can be added to the epoxy to reduce its fluorescence. If not then perhaps an alternative to embedding in epoxy might be the best option....
Any suggestions warmly welcomed. :-)
Michael Joss Quantitative Microscopist CSIRO Division of Petroleum Resources Fluid History Analysis Group Phone: 9490 8148 Fax: 9490 8467
} We are trying to examine oil inclusions trapped in quartz under UV light. } The oil fluoresces and the quartz does not. Sounds easy. Unfortunately the } epoxy that the sample is embedded in also fluoresces quite strongly making } identification difficult. So I was wondering if there are any } non-fluorescent epoxies available on the market, or alternatively if there } are any pigments/dyes that can be added to the epoxy to reduce its } fluorescence. If not then perhaps an alternative to embedding in epoxy } might be the best option.... } } Any suggestions warmly welcomed. } :-) } } Michael Joss } Quantitative Microscopist } CSIRO Division of Petroleum Resources } Fluid History Analysis Group } Phone: 9490 8148 } Fax: 9490 8467 }
} We are trying to examine oil inclusions trapped in quartz under UV light. } The oil fluoresces and the quartz does not. Sounds easy. Unfortunately the } epoxy that the sample is embedded in also fluoresces quite strongly making } identification difficult. So I was wondering if there are any } non-fluorescent epoxies available on the market, or alternatively if there } are any pigments/dyes that can be added to the epoxy to reduce its } fluorescence. If not then perhaps an alternative to embedding in epoxy } might be the best option.... } } Any suggestions warmly welcomed. } :-) } } Michael Joss } Quantitative Microscopist } CSIRO Division of Petroleum Resources } Fluid History Analysis Group } Phone: 9490 8148 } Fax: 9490 8467 }
The tennis balls will isolate vibration nearly perfect. We used this approach a lot in Russia simply because we had no other choice. But tennis balls have tendency to deflate under pressure of the weight of the machine. The deflation will become noticeable in a few moths. Then the entire frame has to be lifted in order to replace tennis balls. Much better approach is to use pneumatic antivibration mounts by Barry Controls. They come in different sizes for loads from 100 lb. to several tons, and available from a number of suppliers including McMaster-Carr. Unless, of course, Australian tennis balls are superior to Russian ones... Cheers.
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax -----Original Message----- } From: Diana van Driel {dianavd-at-eye.usyd.edu.au} To: MicroscopyList {microscopy-at-sparc5.microscopy.com}
I have read the previous suggestions and would agree with them - the message is get as much of that magnet out of your microscope as possible. However other tips that you may find useful:
If you have a side entry stage you will find it very difficult to set the eucentric height with a magnetic specimen. Either note the objective lens current at eucentric focus and set that or focus a non magnetic specimen at eucentric and then insert your secimen. Having done that focus the specimen using the eucentric height control.
Try to have as small a hole as possible in your specimen. This will reduce the effect of having a magnet one side of the beam and not the other. Of course the best specimen would not have a hole but just uniform thin area.
You will have to realign the beam every time you move the specimen.
When out of focus you may see the Fresnel image of Domain walls, when they change contrast from black to white (or vice versa) then you are in focus.
Good luck, Ron
On Wed, 15 Mar 2000 18:59:22 +0100 Jose Maria Manero {manero-at-cmem.upc.es} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear all, } } I want to observe pure iron by means of TEM (Jeol 1200 EX II) in order } to study the dislocations substructure. But I have some problems, for } example, I can not focus the image due to the magnetism of the ferrite. } } I am looking for suggestions to avoid this problem. What could I do? } } Thank you in advance. } } Dr. J. M. Manero } Universidad Politécnica de Catalunya. } E.T.S.I. Industriales de Barcelona. Spain. } e-mail: manero-at-cmem.upc.es } } }
---------------------- 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've read most of the replies but found no mention of differentiation. This is one of the most attractive features of the toluidine stain as it can give a two colour, pink and blue rendition. This shows cell features similar to a double staining with Haematoxylin & Eosin.
After staining on a hotplate add a drop of acid alcohol and then rinse the slide with distilled water. The acid alcohol can destain partially or even completely, but a brief application brings forth the two colours.
Make acid alcohol by adding to 25ml of 50% Ethanol a couple of drops of 1N HCl. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Wednesday, March 15, 2000 6:18 AM, Soumitra Ghoshroy [SMTP:ghoshroy-at-nmsu.edu] wrote: } } Dear Microscopists, } } Does anyone have a protocol for making Toluidine blue stain to be used for } staining thick sections of resin embedded biological samples ? } } Thanks a lot, } } Soumitra } } ***************************************************************** } Soumitra Ghoshroy Ph.D. } Electron Microscopy Lab } Box 3EML } New Mexico State University } Las Cruces, NM 88003 } Tel: 505-646-3600 } Fax: 505-646-5665 } e-mail: ghoshroy-at-nmsu.edu } http://confocal.nmsu.edu/eml
We are examining Staphylococcus epidermidis on the surface of acrylic bone cement. We have problems with the marking of bacteria. Artefacts of particles from the Polymethylmethacrylate (bone cement) look nearly the same than the bacteria. We fix with glut, dehydrate, do critical point drying and coat with gold afterwards. Does anyone have experience with marking bacteria in SEM to surely identify them? Thank you in advance, Christian Heisel, M.D. University of Heidelberg Dep.of Orthopedics Schlierbacher Landstra§e 200 A 69118 Heidelberg Germany Phone:0049-6221-965- Fax:0049-6221-966121 e-mail:christian.heisel-at-ok.uni-heidelberg.de
} I have recently performed an immunoEM labeling procedure with } } myelinated nerve embedded in Lowicryl, K4M resin. Unfortunately the } } myelin is severely altered (it appears as large clealr blebs) because I } } am not able to osmicate this tissue due to the UV polymerization of } } this resin. Does anyone have any suggestions as to preservation of } } lipid laden myelin without the use of osmium? I used methanol in the } } dehydration procedure which has probably caused this artefact....
Karen,
Try to switch to cryosections. This reference could help in solving your problem. Liou W, Geuze HJ, Slot JW Histochem Cell Biol 1996 Jul;106(1):41-58 -- Alexander Mironov Jr. Division of Tumor Biology, H4 The Netherlands Cancer Institute Plesmanlaan 121 1066 CX Amsterdam
One concern raised by a granting agency about our proposal for new equipment for a multi-user SEM lab was our lack of a management plan (primarily for time allocation among users and for conflict resolution). We have been operating under a first come-first served policy, and conflicts have not been a problem. I would greatly appreciate insights from managers of multi-user facilities of any sort as to your approaches to allocation of access to instrumentation, and methods of resolving conflicts among researchers.
fernando =================================================== Fernando D. Balducci Laboratorio de Microscopia Electrnica Facultad de Ingeniera - Bioingenieria Universidad Nacional de Entre Rios Argentina e-mail: microsc-at-fi.uner.edu.ar tel: 54 43 975100 fax: 54 43 975077
The technique we now use on Fe and ferritic steels allows getting an excellent behaviour in the TEM:
-A 1 mm hole is punched in the centre of a 3 mm TEM disk made out of stainless steel (e.g. SS 316) about 300 micron thick, -A 1 mm disk is punched from our ferritic specimen (about 300 micron thick), -This 1mm disk is inserted - slightly forced - in the hole of the 3 mm disk, -A drop of glue (epoxy) is applied (to make sure there is no leak during the electropolish that follows). The 3 mm disk holding the 1 mm disk can then be used as a normal TEM disk: -Mechanical polish down to about 50-80 microns, -Electropolishing with a 10 % vol. perchloric/methanol solution, 18V, 0C.
The final specimen is strong and the remaining magnetism is minimal in the TEM: we can now move and tilt around without too much realignment. Note that a puncher of high quality is essential. The one from 'Eckert' doesn't introduce visible deformation in the centre of the specimen; we tested that in annealed pure Fe.
Dear David: We have 15 year old Reichert Polyvar which, so far, has given us little trouble. We use Osram HLX Xenophot bulbs, 12V, 100 W. Oddly enough, we also have a Reichert MeF3 metallograph, about 10 years old.The light on this instrument flickers erraticly. We replaced the lamp control board with no success, switched lamps, etc. To no avail.Our service man thinks the trouble lies in the transformer, which is no longer available. We are buying a new separate light source but have not received it yet. We are not happy with Reichert/Leica concerning assistancein this problem. They have been less than helpful. On the other hand the service man(Dave Olney) from W. Nuhsbaum, McHenry IL, has been very helpful. I would be interesrted in whatever solution to this problem that you may come up with. How old is your instument?
I have financial interest in either Reichert/Leica or W. Nuhsbaum. } ---------- } From: O'Neil, David } Sent: March 2000 1:56 PM } To: Microscopy Listserver } Subject: LM: Fluorescence lamp instability } } ------------------------------------------------------------------------ } 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've been having stability problems with our mercury vapor lamps on } our Reichert Polyvar microscope. We have been told by one source that the } power supply is likely the problem and another source tells us that the } bulbs are the problem. The source illumination, when imaged, can be seen } to } flicker at random intervals and causes the illumination to vary. Does } anyone have experience with this microscope, and if so which brand of } lamps } do you use? This microscope uses a 220W/4 L1 lamp. Thanks. } } } David O'Neil } National Research Council of Canada } Institute for Marine Biosciences } 1411 Oxford Street } Halifax, NS B3H 3Z1 } ph: (902)426-8258 } fax: (902)426-9413 } e-mail: david.o'neil-at-nrc.ca }
} } Hi, folks; } } One concern raised by a granting agency about our proposal } for new equipment for a multi-user SEM lab was our lack of a } management plan (primarily for time allocation among users and for } conflict resolution). We have been operating under a first } come-first served policy, and conflicts have not been a problem. } I would greatly appreciate insights from managers of } multi-user facilities of any sort as to your approaches to allocation } of access to instrumentation, and methods of resolving conflicts } among researchers. } } Thanks for any information! } } Leslie Eibest } Zoology Dept. } Duke University } Leslie -
My method, FWIW, is simplicity itself. I have a scheduling calendar posted on the outer door of the lab, with a pen provided. Clients simply sign themselves up in whatever day/time slot they wish, provided it's still open. They can even sign up for days/times in future months, if they so desire. Haven't caught anyone erasing a previous booking yet....but then that's why I use a pen..... This method certainly favours the folks who have their act together far enough in advance so that they can schedule the instruments at their most convenient times. Clients who show up at the last minute looking for openings at their prefered time may well be out of luck. That's what we call a learning experience. I'm sure there is software available that can be mounted on your local network which would do exactly the same thing, but, you know, I think it makes more of a commitment for the client to actually make a trip down to the lab to physically sign up, and then they're less likely to forget about the scheduled session later. It's worked pretty well for me for about 5 years now, and haven't seen a good screaming match yet.
Frank Thomas MicroAnalysis Facility Geological Survey of Canada (Atlantic) Bedford Institute of Oceanography Dartmouth, Nova Scotia
} Hi, folks; } } One concern raised by a granting agency about our proposal } for new equipment for a multi-user SEM lab was our lack of a } management plan (primarily for time allocation among users and for } conflict resolution). We have been operating under a first } come-first served policy, and conflicts have not been a problem. } I would greatly appreciate insights from managers of } multi-user facilities of any sort as to your approaches to allocation } of access to instrumentation, and methods of resolving conflicts } among researchers. } } Thanks for any information! } } Leslie Eibest } Zoology Dept. } Duke University
***************************** Dear Leslie, I have run a multi-user EM facility for 12 years, a now also manage an optical microscopy facility. Both are core facilities for the medical school. For both facilities, users must be trained in the operation of the instruments before they are given free access (we have coded locks). During training, people set up appointments with me. Once "cleared fro soloing" they are given an idivualized code and are then free to reserve the instruments by means of a weekly sign-up sheet. For now, the sheet for the following week is posted on Thursday or Friday. Our in-house computing people keep promising us an on-line sign up. I can't wait since the 2 facil/lab group may reserve on a given day. This has worked well for us so far. It also seems to satisfy the granting instituions. For more details, you can check out the school's web site (http://www.med.cornell.edu/research/cores/index.html)select either electron microscopy or optical microscopy to get the operating procedures.
Lee
Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Confocal Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
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Hello everyone, We are having to justify using cervical dislocation without anesthesia on a mouse prior to removing the liver for fixation for our EM class. Does anyone have a reference on hand that we can cite to show that direct cervical dislocation is better (or not) for best preservation of tissue under the circumstances? We won't be doing perfusion, which would obviously provide better fix. Please respond directly to me at: jpshield-at-arches.uga.edu
Background: The class is held once a year, and one mouse is used (usually an extra, "too old" mouse from the monoclonal facility). The animal-use officer wants us to hike down to another building with the class and use CO2 anesthesia, prior to euthanasia, so the mouse suffers less (?). Then hike back up the hill with the tissue in fixative. We've never had a problem with our animal-use request until this year and the guy is busting our chops.
Thanks in advance, john ******************************************** John P. Shields Center for Ultrastructural Research 151 Barrow Hall University of Georgia Athens, GA 30602-2403 (706)542-4080 jpshield-at-arches.uga.edu
Hi Leslie We are a multi-user facility, used by the Faculty of Medicine and the Faculty of Science. Last fiscal year we had 653 individual users, the majority frequent users. We have TEM, SEM, confocal, fluorescent/brightfield, darkrooms and digital imaging equipment.
We use the first-come, first-served approach for many years and so far there hasn't been a problem with sign-up conflicts with the instruments. Most people around here are reasonable, thinking people.
We do not give open access after hours (9-5). But if a user is experienced with the instrument, and I know he/she is not going to mess it up for the next user, and he/she will take a responsibility lecture from me, there is a floating key. I used to be more lax with letting users in after hours, but after a week of just trouble shooting, I got tough. Since then, it has worked well. I remember when I did my PhD, how much more work I got done when there were no interuptions or distractions. So I have a sympathy for allowing responsible researchers in when I am not there, especially as this is a busy facility during the week day.
If it ever came to a conflict of access to the instruments, I am in charge. I would think poorly of researchers who didn't play by the rules. The only time, any problem has occured has been when someone has a crutial deadline. Then when the situation has been put to the person who has it booked, they can usually see themselves in that situation and have always been accommodating. Maybe it is the Canadian way, but I should think it is far more universal!
Good luck with your grant. Let me know if you need more information. Elaine
} } Hi, folks; } } One concern raised by a granting agency about our proposal } for new equipment for a multi-user SEM lab was our lack of a } management plan (primarily for time allocation among users and for } conflict resolution). We have been operating under a first } come-first served policy, and conflicts have not been a problem. } I would greatly appreciate insights from managers of } multi-user facilities of any sort as to your approaches to allocation } of access to instrumentation, and methods of resolving conflicts } among researchers. } } Thanks for any information! } } Leslie Eibest } Zoology Dept. } Duke University
Dr. Elaine Humphrey Biosciences Electron Microscopy Facility 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
At 08:22 AM 3/16/00 -0500, Leslie Eibest wrote: } Hi, folks; } } One concern raised by a granting agency about our proposal } for new equipment for a multi-user SEM lab was our lack of a } management plan (primarily for time allocation among users and for } conflict resolution). We have been operating under a first } come-first served policy, and conflicts have not been a problem. } I would greatly appreciate insights from managers of } multi-user facilities of any sort as to your approaches to allocation } of access to instrumentation, and methods of resolving conflicts } among researchers. }
Leslie,
I have been managing a multi-user facility since 1979. First it was sign up sheets. Now it is a web-based calendar. Have a look at out web site. This works fine for us. We use the web calendar because it is easier for people across campus to sign up this way than to trek over to the facility. Also, I don't have to answer the phone to make their appointments.
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 530 754 7536 fax http://katie.ucdavis.edu raharris-at-ucdavis.edu
Dear Jose, One other method that might work for you is the old "window-polishing" technique. This was the preferred method before the jet-polishing machines were in wide use. A sheet of the metal about one inch by two inches and as thin as possible is masked by stopping lacquer on the edges and electropolished in a bath of electrolyte. When it has polished down to the "lacey" point, a small protrusion of the metal is sliced off with a razor blade and sandwiched in a folding grid or between two grids. The edges, except for the sliced part, are thin enough to examine. It is a bit of an art and takes some practice to get right, but the resulting piece is small and securely held. -----Original Message----- } From: Jose Maria Manero [mailto:manero-at-cmem.upc.es] } To: Microscopy-at-sparc5.microscopy.com } Subject: [TEM] Problem with magnetic samples
} Dear all,
} I want to observe pure iron by means of TEM (Jeol 1200 EX II) in order } to study the dislocations substructure. But I have some problems, for } example, I can not focus the image due to the magnetism of the ferrite.
} I am looking for suggestions to avoid this problem. What could I do?
} Thank you in advance.
} Dr. J. M. Manero } Universidad Politcnica de Catalunya. } E.T.S.I. Industriales de Barcelona. Spain. } e-mail: manero-at-cmem.upc.es
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
Leslie, You can post a calendar on your internal web page for users to log onto, or if you do not have a web page post a calendar at each microscope. Let your users sign up as far in advance as they can, this eliminates most of the conflicts and the habitual "last minute guy" learns to get him/herself more organized. It also gives you a record of use, I also log in all down time due to maintenance service etc. The best part is this system is really easy once everyone buys into it. Good luck! } ---------- } From: Leslie Eibest } Sent: Thursday, March 16, 2000 5:22 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Multi-user facility managers } } ------------------------------------------------------------------------ } 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, folks; } } One concern raised by a granting agency about our proposal } for new equipment for a multi-user SEM lab was our lack of a } management plan (primarily for time allocation among users and for } conflict resolution). We have been operating under a first } come-first served policy, and conflicts have not been a problem. } I would greatly appreciate insights from managers of } multi-user facilities of any sort as to your approaches to allocation } of access to instrumentation, and methods of resolving conflicts } among researchers. } } Thanks for any information! } } Leslie Eibest } Zoology Dept. } Duke University } }
We have modified the basic sign up sheet due to very heavy demand on our microscope. Except for special cases (eg., time lapse studies, emergencies) we allow users to sign up for no more than two hour blocks of time and they cannot sign up for additional time until that block is completed. In addition, if they are more than 15 minutes late for their time the entire block of time is forfeited. (This has rarely happened but emphasizes the need for responsible behaviour.) This system was adapted from the one that Mei Li Wong uses for electron microscopes. Larry D. Ackerman Lily & Yuh Nung Jan Laboratories Howard Hughes Medical Institute UCSF, Box 0725, Rm U226 533 Parnassus Ave. San Francisco, CA 94143
Hi Leslie, Yikes! this reeks of infectious administratium but since you need to deal with the request... I have quite a few systems that are considered multiple user facilities. Over the last 9 years I can say I have never had a conflict to resolve or even heard about one... meaning the users work well together. Users make reservations on a calendar. That is their time. If the machine is down, that time is lost. There is not a ripple effect or compression of the schedule to squeeze them in Frankly if any user asked another for time due to there state of desperation I have no doubt that they would work together. I have a rule for my EMs that only one 4 hour period may be reserved at a time. I think this keeps the level of respect for someone else's time high. My good fortune aside, you could probably get an outline for conflict resolution from your HR dept. & append it to your proposal.
Bruce Brinson, Rice U.
Leslie Eibest 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, folks; } } One concern raised by a granting agency about our proposal } for new equipment for a multi-user SEM lab was our lack of a } management plan (primarily for time allocation among users and for } conflict resolution). We have been operating under a first } come-first served policy, and conflicts have not been a problem. } I would greatly appreciate insights from managers of } multi-user facilities of any sort as to your approaches to allocation } of access to instrumentation, and methods of resolving conflicts } among researchers. } } Thanks for any information! } } Leslie Eibest } Zoology Dept. } Duke University
We are working on a research project and we need an electron gun with 5-10 kV (wehneldt assembly) and a lens system (for electrons, not photons) that is typically found in SEM systems. Does anyone have a decommissioned or un-needed SEM or TEM equipment from which we could get these components at low or no cost (we are on a very tight budget)? We will even move out a complete system if it will help someone who needs the space.
I recently responded to a thread about Amray FE maintenance contracts. My response contained two errors: one numerical, one in perception. I wish to correct them at this time.
First the numerical correction: I stated that the JEOL FE gun exchange cost was $17,000.00: it is actually $7,000.00. The $17,000.00 figure was obtained from two JEOL customers which I presumed to be true as I am not in the habit to ask other competitors their pricing schedules.
Second the perception error: I stated that " Hitachi is doing well and doesn't need to worry about "locking" third party maintenance or anyone else out." I did not mean to imply that JEOL or any other manufacturer's business was poor -- just that Hitachi was doing well. I am sure that JEOL's business is excellent as they have an excellent product. Moreover, their service organization is one of the best. As their competitor we have a difficult time competing with JEOL.
Keep in mind that I do not have a "cozy" relationship with Hitachi. I just respect the technology & the professional co-operation I get from them & other manufacturer's like them. I can buy schematics, parts lists, & even service manuals from Hitachi that other manufacturers don't offer because they consider it proprietary. It makes my job easier.
I was using Hitachi as one example because in a pure free enterprise system the customer is the ultimate winner.
I apologize for the errors. I wish to thank JEOL for pointing out the error and their gracious attitude in dealing with this error.
We have a calendar that people fill in. As a courtesy to others, we ask that they not block more than 4 hours at a time. So far this has worked well. On the few occasions when another user was in a bind, scheduled users have been accommodating because they know that they could be in the same position.
Ron
-----Original Message----- } From: Leslie Eibest [SMTP:leibest-at-duke.edu] [} ] [} ] I would greatly appreciate insights from managers of multi-user facilities of any sort as to your approaches to allocation of access to instrumentation, and methods of resolving conflicts among researchers.
Place: IBM Microelectronics, E. Fishkill, NY, West Complex, Bldg 600.
The IBM Corporation has graciously agreed to be our host for this meeting at their facility in East Fishkill, NY (West Complex, Bldg 600). Attendees will be able to purchase lunch in the adjacent IBM cafeteria. Due to size and security issues IT IS ESSENTIAL THAT MEMBERS PRE-REGISTER so that an attendee list can be delivered to the IBM security folks to prepare guest badges and escorts. DUE TO THE TIGHT SECURITY CLEARANCE REQUIREMENTS, WALK-INS CANNOT BE ACCOMMODATED.
THE REGISTRATION DEADLINE IS MARCH 24th AND CAN BE ACCOMPLISHED ELECTRONICALLY. Please respond via email (or fax) to Evan Slow directly. A simple email note or a completed fax of the registration form is all that s required to register. You can then bring the required fee with you to the meeting. The meeting fee, which does not include lunch, is $15.00. ON-SITE REGISTRANTS WILL BE CHARGED $25.00 BUT, AGAIN, UNLESS YOU HAVE BEEN PREVIOUSLY CLEARED THROUGH IBM SECURITY, WE CANNOT ACCOMMODATE YOU.
9:45 - 10:45 : CHARACTERIZATION OF MICROSTRUCTURES IN MICROELECTRONIC INTERCONNECTS, Lynne Gignac, IBM T.J. Watson Research Center.
10:45 - 11:45 : IMAGING, DIFFRACTION AND SPECTROSCOPY WITH FIELD EMISSION GUN SEM (FEG SEM) AT LOW VOLTAGE, Vinayak Dravid, Northwestern U.
11:45 - 12:45 : DEEP-UV CONFOCAL MICROSCOPY OF SUB-MICRON FEATURES, Mike Torres, Metron Technology.
12:45 - 1:30 : LUNCH -- available for purchase in the IBM cafeteria.
1:30 - 2:30 : THE USE OF MONTE CARLO CALCULATIONS FOR QUANTITATIVE X-RAY MICROANALYSIS, Eric Lifshin, GE Corporate Research & Development.
2:30 3:30 : STUDIES OF SAMPLES HAVING SHALLOW SURFACE TOPOGRAPHY BY THE LOW-LOSS ELECTRON (LLE) METHOD IN THE SCANNING ELECTRON MICROSCOPE (SEM), Oliver Wells, IBM T. J. Watson Research Center.
by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id RAA11115 for dist-Microscopy; Thu, 16 Mar 2000 17:58:04 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id RAA11112 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Thu, 16 Mar 2000 17:57:33 -0600 (CST) Received: from sam.comms.unsw.EDU.AU (sam.comms.unsw.EDU.AU [149.171.96.20]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id RAA11099 for {microscopy-at-sparc5.microscopy.com} ; Thu, 16 Mar 2000 17:57:19 -0600 (CST) Received: from mel ([129.94.150.25]) by sam.comms.unsw.EDU.AU (8.8.8/8.8.8 Kenso-Central-NO-SPAM) with SMTP id KAA22727 for {microscopy-at-sparc5.microscopy.com} ; Fri, 17 Mar 2000 10:52:09 +1100 (EST) Message-Id: {3.0.1.32.20000317105504.00986dc0-at-pop3.unsw.edu.au} X-Sender: s7001031-at-pop3.unsw.edu.au X-Mailer: Windows Eudora Pro Version 3.0.1 (32)
At 08:22 16/03/00 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have run a first come-first served policy for 30 years. Its the way to go.
It now is self-administering through our equipment booking software.
VERY seldom we need to reason with excessive users. We can almost always sort problems out with time trading.
Use Internet explorer - Go to http://srv.emunit.unsw.edu.au/ - instrument booking and use login {guest} password {guest} to get its flavour.
} } Dr. Mel Dickson, Deputy Director, The Electron Microscope Unit, Adjunct Associate Professor, School of Microbiology & Immunology The University of New South Wales UNSW SYDNEY 2052 Australia. Phone +612 9385 6383 Fax +612 9385 6400
It helps to have a good management plan in place whether the facility is extremely busy or not. It avoids most conflicts and gives manager/coordinator a set of rules to use that can be applied to any situation that might pop up. You might not that busy currently on conflicts are occuring but later on the situation can quickly change. Granting agencies want to know if the equipment they are funding are accessible and not misused. We are a fairly busy facility and have been using calendars, lined up together, on a wall for each piece of equipment including work stations. We have one for the current week and below that one for the next week users can sign up. Depending on the instrument from two to three hour slots at a time during peak hours. After 6PM and before 8:30AM longer durations are possible. One can only cancel a day in advance on an instrument. No shows are charged. We cross out time slots for instrument maintenance as needed and as much inadvance as possible. Potential users first fill out an investigator profile form before they can cycle into the facility. This helps us target the instrument/technique they need. Based on several factors we prioritize users. All users must take a tutorial and demonstrate competency before they can solo. We do tutorials only one day a week unless there is an important reason that they cannot do it on that day. In a few weeks, I hope, we are going to our web based scheduler in which users have the a couple months to schedule in advance. It will automatically restrict a users' time usage to only peak hours or to peak and after hours depending on their competency, and other criteria. It will prevent users from making cancellations less then the day before. Down times, etc can be easily entered on the calender. The web based schedular will make it allot more convenient for users and core staff. And best of all, please god let it work smoothly, data from the scheduler will flow into our billing data base for semi automatic billing. So far, with the written calendar syst conflicts, however, usage is so high on some instruments not every user can get the slot they need and so are unhappy campers. In this case, we encourage users, if it is possible, to time their experiments based on when they can get on the instrument. I hope this helps.
Hank Adams Lab Manager Integrated Microscopy Core Molecular and Cellular Biology Baylor College of Medicine Houston, TX
At 08:22 16/03/00 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have run a first come-first served policy for 30 years. Its the way to go.
It now is self-administering through our equipment booking software.
VERY seldom we need to reason with excessive users. We can almost always sort problems out with time trading.
Use Internet explorer - Go to http://srv.emunit.unsw.edu.au/ - instrument booking and use login {guest} password {guest} to get its flavour.
} } Dr. Mel Dickson, Deputy Director, The Electron Microscope Unit, Adjunct Associate Professor, School of Microbiology & Immunology The University of New South Wales UNSW SYDNEY 2052 Australia. Phone +612 9385 6383 Fax +612 9385 6400
Christian, I have a few approaches you might consider: a. Where possible, I usually obtain a culture of the bacteria thought to adhere to a substrate and prepare it exactly the same way:fix, dehydrate, cpd and sputter-coat. b. Prepare two samples but after full prep, apply a cellulose acetate film (or duco cement) to one surface moistened with a drop of acetone and try to strip the bacteria from the bone cement -- I don't know about damage to the methacrylate. This technique produces a replica of the surface. I have used it to remove bacteria colonizing a planchet of hornblende. We were able to locate the pits made by the bacteria and image the bacteria removed, i.e., stripped from the mineral. c. If the above approach fails to produce the results you need, try making a replica with double sided C tabs, the type sold by EMS for X-ray microanalysis. d. If staph epidermidis has a glycocalyx you might try adding ruthenium red to the GA in the primary fix and prep as usual but instead of coating with Au or Au/Pd, evap C and use a backscatter detector to obtain a contrast BE image where the brighter areas should be the Ru stained bacteria and/or collect a spectrum to check for the presence of Ru. If there is a characteristic peak, you could map the Ru and localize the bacteria with an x-ray dot map. I would be interested in knowing how you resolved this problem. Rosemary Walsh The Electron Microscope Facility for the Life Sciences, A Shared Technology Facility, The Life Sciences Consortium 1 South FrearLab Penn State University University Park, PA 16802 (814) 865-0212 rw9-at-psu.edu {"http://www.lsc.psu.edu/stf/em/home.html" eudora="autourl"} http://www.lsc.psu.edu/stf/em/home.html
I sell the DruAedge blades. Same quality as the accu-edge, but at a better price. Please contact me for pricing and free samples. I carry both the High Profile and Low Profile Teflon coated blades.
Ford M. Royer Analytical Instruments, LLC 9921 13th Ave. N. Minneapolis, MN 55441 phone: (800) 565-1895, ext. 17 fax: (612) 929-1895 froyer-at-bitstream.net
Diana Papoulias 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 } -----------------------------------------------------------------------. } } Could someone please tell me where I can purchase accu-edge microtome } blades? } } Thank you. } } Diana Papoulias } USGS } 4200 New Haven Rd } Columbia, MO 65201 } } T:573 876 1902 } F:573 876 1876 } E:Diana_Papoulias-at-usgs.gov
I am trying to localise a beta-1,3-glucanase gene after Russian wheat aphid infestation. However I am getting quite heavy labeling in the chloroplasts and from the literature this haven't been previously documented. My control serum shows a small amount of labeling in cell walls but these seems to be random. The controls (before infestation) shows very little labeling. Work done with the same antibody on rust infested wheat plants have shown no, to very little labeling in the chloroplasts. My question is how can I make sure this is not artifacts and that the glucanase are for sure in the chloroplasts. Martin Wilding Department Botany & Genetics University of the Orange Free State P.O. Box 339 Bloemfontein 9300 South Africa
Tel +2751 4012818 Fax +2751 4488772 Email paam-at-rs.uovs.ac.za
E.A. Fischione Instruments, Inc. is seeking an individual for the position of a Sales Engineer. The successful candidate will be responsible for one or more states from both inside the office in Export, PA. and in the field. The candidate's responsibilities will include defining contacts, maintaining information in the sales database, qualifying leads and determining need, providing information and quotations for standard products, handling service/spare parts requests and coordinating the same with the Service Department. Must develop a thorough technical knowledge of standard products. Extensive travel required.
The candidate should have an Associates Degree in either Electron Microscopy technology, Material Science, Engineering, or Physics as a minimum; a B.S. or M.S. would be preferred.
Salary is commensurate with experience.
E.A. Fischione Instruments, Inc. is an equal opportunity employer. Please send your resume and salary requirements to:
Human Resources Director, MSA LS E.A. Fischione Instruments, Inc. 9003 Corporate Circle Export, PA 15632 Phone (724) 325-5444 FAX (724) 325-5443 E-mail: info-at-fischione.com www.fischione.com
This does not directly address the problem with your current lamp but you might want to consider looking at this new lamp assembly. We recently evaluated the system and there were no visible stability problems as well as several advantages over the standard mercury lamp housing.
The company is called EFOS. This unit replaces the standard mercury source for fluorescence and possibly transmitted light techniques. It uses an external 50W miniature arc lamp incorporated into an elliptical reflector which is connected to the microscope by a liquid light guide. Our unit was mounted on a Zeiss Axioscope.
You can visit the EFOS web site at http://www.efos.com/products/x-cite.htm
Thanks, Louie
At 2:56 PM -0400 3/15/00, O'Neil, David wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Louie Kerr Research and Education Support Coordinator Marine Biological Laboratory 7 MBL Street Woods Hole, MA 02543 508-289-7273 508-540-6902 (FAX) 508-292-0289 (Cell phone)
} I would greatly appreciate insights from managers of } multi-user facilities of any sort as to your approaches to allocation } of access to instrumentation, and methods of resolving conflicts } among researchers. }
Dear Leslie, Our facility is funded by NIH as a Biotechnological Resource, so we deal with both in-house and outside researchers. Like the other responders, we have not had any conflicts between users since we started. Because the outside users must spend con- siderable time arranging to travel here to use the instruments, we have had a policy which maximizes the utility of their stay here. We have a staff person whose function is outside user liason; she ascertains what the user wants, suggests ways to prepare the specimen to achieve that, examines trial grids, schedules the user, and stays with the user during the run. For the inexperienced user, she does almost everything except select the area of the grid to be photographed; for users who are familiar with our facility, she changes specimens, develops film, and is available for any other user needs. For the occasional user who can operate the HVEM on his own, she still changes the film and is available. The rest of the staff is responsible for starting the HVEM up in the mode the user wants, taking care of any problems which arise, and setting up for non-routine use--EDS, diffraction, etc. In-house users can be bumped to accomodate outside users and can only sign up a few weeks in advance. Outside users can sign up for as far in advance as they wish, and if any problems arise with the scope, our liason person calls to let them know so they can change plans if necessary. In periods of very high use, the staff arranges to keep the scope up for as long as 16 hours a day and will also arrange to be here after hours and/or on week- ends to accomodate users' schedules. Yours, Bill Tivol
How fast must the sequential image capture be to perform image ratios for dtermining something like calcium ion levels? If you have a fast interline or frame transfer camera, does a single filter wheel like a Ludl change filters fast enough?
SmithKline Beecham, a world class leader in Research and Development, continues to pioneer innovative pharmaceutical and healthcare products and services. We have the following opportunity available at our state-of-the-art suburban Philadelphia facility. Working in our Safety Assessment department you will provide technical support and scientific input into the design and execution of studies to elucidate the cellular mechanisms of drug-induced toxicities. You will prepare biological specimens for transmission and scanning electron microscopy, confocal microscopy, X-ray microanalysis and immunohistochemistry. You will also perform qualitative and quantitative data analysis using computer assisted image analysis systems. We require a BS/MS in a biological science, and 3-5 years of microscopy experience in cell biology, physiology, toxicology. SmithKline Beecham is dedicated to an innovative workplace and supports you with career long opportunities and learning. We offer a competitive benefits and compensation package. For confidential consideration, please forward your scannable resume to: SmithKline Beecham Attention: Human Resources AD CODE: 2K0325W c/o National Resume Processing P.O. Box 1070 Burlington, MA 01803 USA
Indicating ad code is essential. Principals only, no agencies, please. For a full listing of current opportunities, or to submit a resume online, visit our website at www.sb.com/careers.
Leslie, Just to add my two cents: I supervise a private Photopathology Lab for the Wellman Labs of Photomedicine at MGH. We have an Axiophot (Zeiss) photomicroscope that has an image analysis system attached. We have used a monthly sign-up calendar successfully for 10 years. The room is secured-- anyone wishing to use the system, must be checked out first by one of us. Once we are con- fident they know what they are doing, they then sign up ahead for a day and time. We allow after- hour use; many researchers work weekends and nights. I sign out a lab key to the individual and the room key is in a desk drawer. People are instructed to make sure the microscope is off, doors are locked, lights are off and room is secure. Other than the occasional light left on, the system has worked. The other advantage to having people sign-up is that if there is a problem with the micro- scope and/or room, we can go back to the last person who used it and advise them of the problem. We also use a sign-up system for a multi-headed teaching scope.
Whatever system you end up using--sign-up sheets or on-line, you have a record of use, which granting agencies are most interested in: making sure you get the most for your buck!
Good luck!
Peggy Sherwood Wellman Labs of Photomedicine-MGH 50 Blossom Street Boston, MA 02114 617-726-6983 (Photopathology Lab) 617-724-4839 (voice mail) 617-726-3192 (fax) e-mail: sherwood-at-helix.mgh.harvard.edu
Thanks so much to all of you who took the time to respond to my question. My management techniques are basically the same as most of yours, but my presentation was woefully inadequate. Thanks to you, I'm well-armed now!
We have had several instruments and many instrument users during each the last twenty five years. Originally, we used calendar sign-up and log book entries to compile data for accounting and annual reports. This became too time-consuming so we developed an in-house automated (computer-based) sign-up (registration) system in 1990. The system serves as an instrument reservation unit and an automated accounting system. We recently developed (continue to develop) a web-based system which allows researchers to reserve time from their offices or anywhere they can access the internet. You may wish to view it at the following URL: http://signu.la.asu.edu:8001/. (Login: guest Password: testit) You will not be able to view the accounting data. The sign-up program allows only qualified users to operate an instrument. Also, rules for successive time reservation are integrated into the program. Optional holders and spectrometers can be reserved at the time the instrument is signed for.
We presently have 90 research users. In addition, there are approximately sixty student users (grad and undergraduates) who are taking semester classes in microscopy or they have one or two sessions on individual instruments to better understand the role of electron microscopy for solving problems in Geology, solid state chemistry etc.
Each instrument has a keypad timer interfaced to either the filament or high voltage on/off switch, depending on ease of accessibility. The timer is interfaced to a 486 computer (obtained from surplus-there are lots of these available). The computer stores user time for each account as well as the number of films exposed for the instrument used. Because we have to pay for approximately 90% of our operations costs, we have to charge for use of the instruments. We also charge different hourly rates depending on user status (inside or outside user).
At the end of each month, the user time/film number data is dumped into another computer which compares the reserved time from the web-based reservation system to the actual use time and charges for the greater of the two. If the instrument develops problems during the use time and can't be used any more, the individual user is able to send a DOWN message to the reservation computer and the user is charged only for actual time used. The monthly use data is formatted into a spreadsheet which contains microscope use times and film numbers used. A principal investigator (PI) may have ten grad students who use five different instruments and three different grant account numbers during the month. He/she receives a statement at the end of the month which identifies the microscope user by name, the account number used, the hours used (rounded to the nearest minute)for each instrument and the total number of films exposed for each use time. The total costs for instrument use is given on the last line of the statement.
An advantage of this system is that within thirty minutes, one can sort data files and determine how many hours each microscope is used over any period of time; how many hours each user utilizes an instrument(s) over time; number of hours used for each account and total number of hours for all instruments over time. This info is useful for reports to administrators as well as for funding agency applications.
If you have questions about our system, please e-mail me directly or call at the number listed below.
John C. Wheatley Lab Manager Arizona State University Center for Solid State Science PSA-213 BOX 871704 Tempe, AZ 85287-1704
A short time ago there was a discussion about the different EMs RCA produced and when they were produced. The other day I was digging through my sfiles and came across a copy of the August 1967 issue of the RCA Scientific Instruments News which has pictures of all the models produced and the year of introduction:
EMA 1939 I don't think this model was ever sold commercially. I believe it was built in the RCA Labs for developmental uses
EMB 1940 This was the first commercial model
EMC 1944 This model had a horizontal column
EMT 1950 This was an inexpensive table model that was designed with use in high schools and small colleges in mind.. As I recall it had lenses made of permenant magnets to eliminate the need for lens power supplies and to hold the cost down.
EMU 1944 This was probably RCA's most popular model. It was on their leading EM for at least 10 years.
EMD 1948 This was an electron diffraction instrument, designed specifically for obtaining ED patterns by the reflection from solid surfaces at a grazing incidence.
EML 1954 This was the forerunner of the popular EM-U# and EM-U4 models, but provided an accelerating voltage of only 50kV. It was the model in which RCA pioneered the modern ergonomic layout of controls, with a desk-like design.
EMU-3 1955 Similar in appearance to the EML, but providing 100 kV.
EMU-4 1966 A refinement of the EMU-3, providing a double condenser lens, a specimen stage airlock system, etc.
RCA ceased marketing electron microscopes in 1969. I was President of the EMSA that year and discussed the matter with James Hillier, one of the pioneers in the field of electron microscopy, and a Vice President of RCA at that time. He said that it was determined that RCA could make more money selling records and related electronic devices than EMs, and so was phasing out of the EM business.
The issue of RCA Scientific Instruments News in question also has a picture of each model on the cover. I have had a copy run off in jpg format. I understand that it is not appropriate to transmit such info via this list server; however, if anyone would like a copy, let me know and I'll send it to you individually.
Happy St. Patrick's Day,
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
} Dear Listservers, } } I recently responded to a thread about Amray FE maintenance contracts. } My response contained two errors: one numerical, one in perception. } I wish to correct them at this time.
[snipped for brevity sake]
Thanks Earl for the update and correction. This is good info. And I see some room for additional discussion and inquiry to wrap up this topic.
The prospects for third party maintenance contracts are rather variable--depending mostly, I think, on what is being maintained under contract. This would be based on the availability of schematics, software, special parts, etc., etc. If some FESEM makers do not "guard" their FE guns relative to others, then of course, that will weigh heavily on third party options. For the record, here are some figures for Amray maintenance contracts. An 1830 LaB6 system runs $13,500 annually. This excludes "consumables" such as apertures, LaB6 cathode, pump oil, etc. But it does include on-site service, troubleshooting, travel, lodging, meals, rental car, things that break down, like Balzers 240 turbo pump or Edwards E2M8 mechanical pump, or the invariable broken wire. Actually, I have found the 1830 to be very reliable and a good performer.
The 1910FE runs $15,000 annually. Same terms as the 1830 but includes the computer control system. What I don't know is whether Amray will sell the FE gun assemblies to third party maintainers. I do know that for Amray users who are not under an Amray contract, the 305FE gun costs about $14,000 total to replace as a per-diem item. Again, I have found that this SEM also is very reliable. But what about performance?
Performance wise, I suppose we could have an "image shoot off" contest based on a standard specimen and different SEM instruments of different models and from different makers. But there are some basic differences among SEMs that would otherwise seem to be the same--but are not. Notable among these differences, for example, is the design and construction of the final lens pole piece. This is due, I think, to two or three driving forces. One is the need to image physically large specimens at various WDs. This means that one needs a large chamber with a stage that offers accommodation of large samples and wide WD range. The second force is the requirement to image whole semiconductor wafers ranging in diameters from 4" to 8" and to have a wide travel distance across the wafer and to operate well at low KV (photo resist examination, for example). The third force is the impact on the SEM when needing to do X-ray analysis.
The flat lens design is quite useable for large and small specimens when high tilt angles do not increase the WD to an unsatisfactory figure. In the case of IC wafers, the conical lens is required. These are available as 45 or 60 degree designs. Since most semiconductor qualification imaging is done at either zero or 45 degrees, the 60 degree lens is probably optimal. What Amray has done in regards to conical lenses is an evolutionary path. The 1830 has a conical lens to accommodate high tilt angles when working with wafers. It also can be configured with a large chamber. However, the design of the lens is such that high resolution is obtained at 10KV-15KV and above. It is not a high resolution SEM at low KV. Alternatively, with a 25KV or 30KV power supply, it has ample voltage to support the beam currents needed for x-ray work. I have no personal information on what the other makers have done and currently offer in regards to IC wafer inspection and analysis.
The Amray 3600LEAP was a major design departure. This system also has a 60 degree conical lens, a huge chamber but a specially designed lens and other features key to semiconductor work. The LEAP lens was designed for low KV imaging. At 2-3KV, the LEAP is virtually identical in resolution to the other lens types at 15KV and at the same WD. This is a key feature and requirement for imaging fine pitch and small feature size wafers and photoresist. The other important feature of the 3600LEAP SEM is the 5-place heated final aperture holder. Since the 3600 runs with 35u and 70u apertures (or less) as routine, these Pt apertures are continuously heated to keep the effects of contamination (photo resist again) at a minimum. Not only does this keep the apertures clean, but it also prevents accumulation of PR in the scan coil liner. This FESEM is a very capable instrument from my experience with it. It can easily image 0.15u wide gate poly at high resolution and 2.5KV at WD=6mm.
If one considers where I have wound up with this discussion, it is cause for pause to ask a fundamental question. That is, "What is a research SEM?" Different applications must surely lead one to select one type of instrument over another. Does it also direct a path to one manufacturer over another? Since I have not seen the myriad number of SEM models from all of the vendors, I cannot answer this question. But while there might be some disdain for one maker versus others, it seems tough for me to make any sort of quantitative decision when there are quite a few variables involved--solely based on application, much less vendors and maintenance options.
What do you think about this, based on your experience?
TOUR SPEAKER (Society for Applied Spectroscopy) - 4/12/2000 Meeting in Mountain View
"RAMAN SPECTROSCOPY AS A PROBE OF STRUCTURE/ORIENTATION IN POLYMERIC FIBERS"
by DR. BRUCE CHASE (DuPont Central Research, Wilmington DE)
at Michaels at Shoreline, Mountain View (Directions to follow)
Register 6:45 PM Dinner 7:30 PM Talk 8:30 PM
Cost: $30 for dinner ($10 students and teachers), free for talk.
Dinner Choices: (Indicate Choice when RSVP'ing)
Breast of Chicken, Piccata Chilean Sea Bass, Ginger Shallot Sauce Grilled Vegetable Brochette with Wild Rice
Reserve by April 10 to Steve Rabin, 650-564-5315 or steve.rabin-at-alza.com
(Please let us know if you're coming for dinner, or just the talk for headcount purposes)
Also, see our web site at www.sas-ncss.org
ABSTRACT Polymeric macroscopic properties such as tenacity, modulus, elasticity, etc. are determined by molecular level effects such as conformational state populations, orientation, and intermolecular interactions. Significant changes occur at the molecular level as a molten or solution phase polymer is spun into fiber form. The polymer goes from an unoriented, amorphous state to an oriented, semi-crystalline material via the deformations imposed by spinning and drawing. Raman spectroscopy can potentially provide information on both structure and orientation. Changes in band intensities can be related to conformational populations and to formation of crystalline regions. Changes in relative intensities as a function of incident and scattered polarization yields information on chain orientation. The use of polarized Raman scattering done in both 90 and 180 degree scattering geometries has been used to determine the second and fourth order orientation functions for both polyethylene and PET fibers. Raman measurements have been made in both the laboratory frame and on-line for 66 nylon and PET fibers. For both PET and 66 nylon, crystallinity levels and a qualitative measure of orientation can be obtained.
DIRECTIONS to Michaels (in Shoreline Park, Mountain View):
} From Anywhere in the Bay Area:
Get to Highway 101 toward Mountain View (from SF and the Peninsula, southbound; from San Jose and the East Bay via 237, northbound)
Go to the Shoreline Rd. exit and turn left at the end of the exit road. Follow Shoreline Rd. into (approx 1 mile) through Mountain View Park gate. Continue on the single lane road (golf course on your left) for approx 1 mile, turn left at Michaels restaurant sign into the parking lot.
Following up on the Hg burner problems, does anyone have a schematic of the "transformer" for these? There is assumed to be a true transformer plus a rectifier and filter. Without a circuit diagram, there is too much speculation for me about what is actually going on. It seems that there are some common problems out there. These ought to be readily solvable.
It really cannot be that difficult, can it? Maybe so.
Diana, These blades are available from Allegiance Scientific (used to be Baxter, used to be Scientific Products, used to be....). As far as I know, this is the only place that carries this brand. I have no financial interest in Allegiance, just passing along a fact. Wanda Shotsberger (HT ASCP)
-----Original Message----- } From: Diana Papoulias [mailto:Diana_Papoulias-at-usgs.gov] Sent: Thursday, March 16, 2000 2:53 PM To: microscopy-at-sparc5.microscopy.com
Could someone please tell me where I can purchase accu-edge microtome blades?
Thank you.
Diana Papoulias USGS 4200 New Haven Rd Columbia, MO 65201
I would like to get hold of an RCA model EMT for display for our museum collection. Ed Sharpe archivist for SMECC
{ { Subj: RCA EMs Date: 3/17/00 2:47:16 PM Pacific Standard Time From: bigelow-at-engin.umich.edu (Wil Bigelow) To: AAmy-at-dtsc.ca.gov, JRowe6427-at-aol.com, cgarber-at-2spi.com (Chuck Garber), john.mardinly-at-intel.com, microscopy-at-sparc5.microscopy.com (Microscopy Listserver), oshel-at-shout.net
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Hi all:
A short time ago there was a discussion about the different EMs RCA produced and when they were produced. The other day I was digging through my sfiles and came across a copy of the August 1967 issue of the RCA Scientific Instruments News which has pictures of all the models produced and the year of introduction:
EMA 1939 I don't think this model was ever sold commercially. I believe it was built in the RCA Labs for developmental uses
EMB 1940 This was the first commercial model
EMC 1944 This model had a horizontal column
EMT 1950 This was an inexpensive table model that was designed with use in high schools and small colleges in mind.. As I recall it had lenses made of permenant magnets to eliminate the need for lens power supplies and to hold the cost down.
EMU 1944 This was probably RCA's most popular model. It was on their leading EM for at least 10 years.
EMD 1948 This was an electron diffraction instrument, designed specifically for obtaining ED patterns by the reflection from solid surfaces at a grazing incidence.
EML 1954 This was the forerunner of the popular EM-U# and EM-U4 models, but provided an accelerating voltage of only 50kV. It was the model in which RCA pioneered the modern ergonomic layout of controls, with a desk-like design.
EMU-3 1955 Similar in appearance to the EML, but providing 100 kV.
EMU-4 1966 A refinement of the EMU-3, providing a double condenser lens, a specimen stage airlock system, etc.
RCA ceased marketing electron microscopes in 1969. I was President of the EMSA that year and discussed the matter with James Hillier, one of the pioneers in the field of electron microscopy, and a Vice President of RCA at that time. He said that it was determined that RCA could make more money selling records and related electronic devices than EMs, and so was phasing out of the EM business.
The issue of RCA Scientific Instruments News in question also has a picture of each model on the cover. I have had a copy run off in jpg format. I understand that it is not appropriate to transmit such info via this list server; however, if anyone would like a copy, let me know and I'll send it to you individually.
Happy St. Patrick's Day,
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
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Dear microscopist, Kindly give me information and websites available on adsorption, desorption and diffusion study of Chromium on Tungsten by field emission and field ion microscopy. Also nucleation and growth of Chromium on Tungsten. Thanking you,
Mangesh Bagade Govt College Of Engineering Pune Maharashtra, India.
____________________________________________________________________ Get free email and a permanent address at http://www.netaddress.com/?N=1
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Dear microscopist, Kindly give me information and websites available on adsorption, desorption and diffusion study of Chromium on Tungsten by field emission and field ion microscopy. Also nucleation and growth of Chromium on Tungsten. Thanking you,
Mangesh Bagade Govt College Of Engineering Pune Maharashtra, India.
____________________________________________________________________ Get free email and a permanent address at http://www.netaddress.com/?N=1
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H i Gary,
Your e-mail poses some interesting question & perspectives.
As far as the variation of Third Party maintenance contracts, Scanservice generally offers contracts that are exactly the same as the given manufacturer. The customer can then compare "apples against apples". Whenever there is a question regarding coverage we always look to see what the manufacturer's contract covers. The variation in contracts is only between the different manufacturer's. Cambridge (LEO) did not cover scan coils, ETEC did not cover rotary pumps or CRTs, etc. This eliminates the confusion for our customers. We can and do offer modified contracts where the customer is responsible for parts or a limited number of service calls but only at the customer's request.
All manufacturer's , SEM & otherwise, are required by Federal Law to sell parts to third party organizations. To withhold parts is considered a "restriction of trade" and has some hefty consequences. I have had accounts set-up for just about all the manufacturers. It is relatively easy process & most manufacturer's treat us like any other customer in need of parts.
The only exception is Amray that has a multi-step process for obtaining parts which is as follows:
1. Request part number. 2. Wait two to three days for part number. 3. Send credit references. 4. Wait two to three days for response. 5. Send banking references & information. 6.Wait two to three days for response.
The above process has occurred at least three times in the last three years. Finally, I requested that they send the parts COD or I could prepay. The response I got was that I "could have done that all along."
Bottom line: manufacturer's must sell to Third party organizations, most are very professional & co-operative.
As far as an "image shoot-off", I really don' think that would be very practical as there are significant variations even between the same model from a given manufacturer. I would expect the best performing SEM from each manufacturer involved, rather than an average of the industry.
When I worked at ETEC, we noticed differences between the different SEM columns. The demo unit always got the best column. Even today I recommend that customers purchase the "demo unit" as it probably has the best of everything & is tuned up. Moreover the "demo unit" can be sold at a lesser price as it is technically used. I also recommend that customers considering an SEM purchase use the model they are considering at another customer's site. In this way, the potential customer can evaluate the SEM model on their own terms using their own standards and without the applications engineer's influence. Most manucfacturer's want to use the standard "gold on carbon" to highlight their given SEM. I generally do not see many of my customers imaging "gold on carbon" on a daily basis. Customers has such a variety of samples: photo resist, uncoated teflon, etc.
This reminds me of a customer at Hughes who I recommended that she follow the above procedure of evaluating the SEM at a customer site & requesting that the demo unit be sold to her Company. She did neither. When she requested that the demo unit be sold to her Company, the manufacturer declined. No reason was given. She bought the FESEM anyway. After about three months I inquired about the status of her new FESEM. Instead of excitement she responded, "Oh it's OK. I guess I will just have to get used to it". She continued to use her standard tungsten SEM over the FESEM.
As far as your analysis of the SEM chambers & polepieces, historically the SEM industry has seen the large chamber sizes and conical polepieces you decribe for at least last twenty years. When I entered this industry in 1973, I remember the JEOL JSM-2 with a semi-conical lens. The lens was not a true conical lens as it did have a "flat tip" of about 4 cm.dia. but it did allow one to tilt samples at a higher angle. I was surprised to see JEOL had a JSM-1 (circa 1969) that had the same column as the JSM-2.
ISI had a model DS-130 that could be ordered with different final lenses: the standard semi-conical lens, wide bore (WB) for 0 working distances, or conical lens. The ISI conical lens had multiple angles on their design. It started with a 30 degree cone, the mid-section was 45 degree, and ended with a 70 degree cone. For it's day the DS-130 had pretty good optics. The electronics were junk in my opinion. The DS-130 had some extremly large chambers. One was known by the ISI engineers as a "turkey baster" chamber. It measured about 24 in. x 36 in. x 18 in high. Hitachi has had the S-806 and S-808 for semiconductor work. An full 8-inch wafer could be viewed as well as tilted 45 degrees. Hitachi also offerred an S-806C which is a conical lens. I believe the vintage for these machines was in the early eighties.
The JEOL JSM-U3 (circa 1970) had a heated final aperture strip that allowed the user to clean the aperture without removing it. This aperture design was replaced in the JEOL 35U and JEOL 35C (circa 1974?). The JEOL 35 series had a continuously heated final aperture. A smaller current (about 2 amps) was run through the aperture therby cleaning the aperture while using it.
I recently (last year) had the opportunity to attend a friend's/customer retirement party. Attending were many older & retired SEM engineers. They were reminiscing about the SEM industry and how the machine had evolved from their days at Westinghouse (yes Westinghouse) in the 1960's. The same questions and problems that we have today they had in the 1960's. Chamber sizes were initially large to accomodate their sample and stage sizes but vacuum was poor (10-4 torr). Chamber sizes were reduced to improve vacuum and lens designs were changed to accomodate tilt angles at the expense of resolution. And everyone could only dream of a stable FE gun. SEM nerds.
The upshot of all of this is that all of these issues has been historically addressed. Large & small chambers, different lenses, etc. will be with us as long as the application permits. When a manufacturer claims to have a "breakthough", it only remind me of what Steve Jobs (Apple Computer) said about Windows 95: " Windows 95: MacIntosh 1986." I think the next real breakthrough will be the lenses made from super-conductive wire or, better yet, the electrostatic columns being developed.
It is my understanding that the electrostatic columns are Shotky FE sources with backscatter dectectors fully integrated onto the "final lens". Future features include an integrated EDS detector. The columns are small & cheap enough to be "thrown away".
I really don't have a disdain for any given Company. I would only like to comment that every Company has a different "flavor" depending upon whom is running it. Most are a pleasure to work with, while others leave a "bad taste in your mouth".
Please don't take these comments personally as I respect your opinion & have throughly enjoyed the subjects you have brought up on this listserve & look forward to future your comments.
Regards,
Earl Weltmer
At 01:54 PM 3/16/00 , you wrote:
} Dear Listservers, } } I recently responded to a thread about Amray FE maintenance contracts. } My response contained two errors: one numerical, one in perception. } I wish to correct them at this time.
[snipped for brevity sake]
Thanks Earl for the update and correction. This is good info. And I see some room for additional discussion and inquiry to wrap up this topic.
The prospects for third party maintenance contracts are rather variable--depending mostly, I think, on what is being maintained under contract. This would be based on the availability of schematics, software, special parts, etc., etc. If some FESEM makers do not "guard" their FE guns relative to others, then of course, that will weigh heavily on third party options. For the record, here are some figures for Amray maintenance contracts. An 1830 LaB6 system runs $13,500 annually. This excludes "consumables" such as apertures, LaB6 cathode, pump oil, etc. But it does include on-site service, troubleshooting, travel, lodging, meals, rental car, things that break down, like Balzers 240 turbo pump or Edwards E2M8 mechanical pump, or the invariable broken wire. Actually, I have found the 1830 to be very reliable and a good performer.
The 1910FE runs $15,000 annually. Same terms as the 1830 but includes the computer control system. What I don't know is whether Amray will sell the FE gun assemblies to third party maintainers. I do know that for Amray users who are not under an Amray contract, the 305FE gun costs about $14,000 total to replace as a per-diem item. Again, I have found that this SEM also is very reliable. But what about performance?
Performance wise, I suppose we could have an "image shoot off" contest based on a standard specimen and different SEM instruments of different models and from different makers. But there are some basic differences among SEMs that would otherwise seem to be the same--but are not. Notable among these differences, for example, is the design and construction of the final lens pole piece. This is due, I think, to two or three driving forces. One is the need to image physically large specimens at various WDs. This means that one needs a large chamber with a stage that offers accommodation of large samples and wide WD range. The second force is the requirement to image whole semiconductor
wafers ranging in diameters from 4" to 8" and to have a wide travel distance across the wafer and to operate well at low KV (photo resist examination, for example). The third force is the impact on the SEM when needing to do X-ray analysis.
The flat lens design is quite useable for large and small specimens when
high tilt angles do not increase the WD to an unsatisfactory figure. In the case of IC wafers, the conical lens is required. These are available as 45 or 60 degree designs. Since most semiconductor qualification imaging is done at either zero or 45 degrees, the 60 degree lens is probably optimal. What Amray has done in regards to conical lenses is an evolutionary path. The 1830 has a conical lens to accommodate high tilt angles when working with wafers. It also can be configured with a large chamber. However, the design of the lens is such that high resolution is obtained at 10KV-15KV and above. It is not a high resolution SEM at low KV. Alternatively, with a 25KV or 30KV power supply, it has ample voltage to support the beam currents needed for x-ray work. I have no personal information on what the other makers have done and currently offer in regards to IC wafer inspection and analysis.
The Amray 3600LEAP was a major design departure. This system also has a 60 degree conical lens, a huge chamber but a specially designed lens and other features key to semiconductor work. The LEAP lens was designed for low KV imaging. At 2-3KV, the LEAP is virtually identical in resolution to the other lens types at 15KV and at the same WD. This is a key feature and requirement for imaging fine pitch and small feature size wafers and photoresist. The other important feature of the 3600LEAP SEM is the 5-place heated final aperture holder. Since the 3600 runs with 35u and 70u apertures (or less) as routine, these Pt apertures are continuously heated to keep the effects of contamination (photo resist again) at a minimum. Not only does this keep the apertures clean, but it also prevents accumulation of PR in the scan coil liner. This FESEM is a very capable instrument from my experience with it. It can easily image 0.15u wide gate poly at high resolution and 2.5KV at WD=6mm.
If one considers where I have wound up with this discussion, it is cause for pause to ask a fundamental question. That is, "What is a research SEM?" Different applications must surely lead one to select one type of instrument over another. Does it also direct a path to one manufacturer over another? Since I have not seen the myriad number of SEM models from all of the vendors, I cannot answer this question. But while there might be some disdain for one maker versus others, it seems tough for me to make any sort of quantitative decision when there are quite a few variables involved--solely based on application, much less vendors and maintenance options.
What do you think about this, based on your experience?
If you are looking for the traces of the tools which were used to make these surfaces, sounds like a job for an interferometer. There are several small ones which can be used for low to medium power (10x-50x objective mag) work. Hach used to carry a small Michelson; also, check with Nikon for both Michelson and Tolansky varieties. While the Tolansky is a contact method system, it is non-destructive and, since it is a multiple beam system, gives very high precision results. I would recommend photographing the interferograms then correlating with the "tooth marks" left by various tools. The old Polyvar also had an interferometer module.
For a more upscale, automated version, I'd suggest that you find a lab with either a Zygo New View or a Wyko RST. These devices are Scanning White Light Interferometers (SWLIs); both are automated, 3D imaging systems which might provide interesting insight but may also be overkill for your type of project. Zygo is just south of us, in central Connecticut, while Wyko is in Tucson. I believe that both have contract labs. (I have done some work in the Zygo facility)
Please contact me if you are interested in pursuing this approach.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education}
At 07:29 PM 12/12/99 +0100, S¿ren Albek wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
A client of ours makes a cut in a polymer block (I can check on the exact material, if you need) then looks at two parameters: 1) the angle made by the sides 2) the radius of curvature of the bottom
He uses a conventional image analysis system for both, using just the simple angle measurement function for the first and the diameter of a circle of best fit for the second. He had been using a stereo microscope but we took a look at the cut under a 10x objective/compound microscope on a recent visit and found that there was a lot to be learned about the quality of the cut from the shards and shredding left on the surface (I can't remember whether it was the upper or lower, so take a look at both).
Hope this is helpful.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education}
At 07:04 PM 3/13/00 -0400, Rosemary Walsh wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I think you must be referring to the famous Bill Miller, who was my "partner in crime" at Sarastro. You can reach him at Bill Miller {microbill-at-mohawk.net} .
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education}
At 11:12 AM 3/14/00 -0500, Ric Felten wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Raman is going to become as available as regular confocal. As a matter of fact, it is on the same price scale but can do both confocal and chemical imaging. It is an analytical technique which works well alongside fluorescence, but you need to collect the Raman signal separately. I just came back from PITTCON (THE major analytical chemical meeting) and some companies are collecting the signal in the near UV, others in the near IR.
A good starting point is Jack Koenig's book "Spectroscopy of Polymers" (Am. Chem. Soc.. Washington DC, 1992). I just saw him at PITTCON and he said that the new edition is either just coming out or about to come out.
The move to Raman is part of the merging of microscopy and spectroscopy. There are now several interesting true hybrid instruments which permit chemical as well as microscopy imaging. One is the Continuum from Spectra-Tech (Shelton, CT); the other is a series built on the DuraScope from SensIR (Danbury, CT). I had a chance to teach a microscopy class to the IR specialists from SpectraTech in January, so really had a chance to have my hands on the system. In addition to running full FT-IR chemical spectra, I was able to do low power darkfield. They also have regular phase and DIC objectives available (the Phase was a bit tricky to implement) and have just introduced a fluorescence module.
I will be doing a review for American Lab (Watch for Am Lab: "Focus on Microscopy") and will post pertinent excerpts for you within the next few weeks.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education}
At 03:00 AM 3/2/00 +0000, Jose Feijo wrote: } Since the subject was raised, could anyone point me out a good paper or internet source to understand Raman microscopy, and how to make it work? On the side, from the gurus, how much should we expect from it in the future? Is it like, it's going to substitute something already available, or otherwise it will complement specific aspects of visualisation of some special biological structure? What does it involve, special lasers, special optics, special computers? } } Thanks in advance } Jose } }
A propos of our earlier discussion, this just came across the wire from the Society for Appl. Spectroscopy:
TOUR SPEAKER (Society for Applied Spectroscopy) - 4/12/2000 Meeting in Mountain View
"RAMAN SPECTROSCOPY AS A PROBE OF STRUCTURE/ORIENTATION IN POLYMERIC FIBERS"
by DR. BRUCE CHASE (DuPont Central Research, Wilmington DE)
at Michaels at Shoreline, Mountain View (Directions to follow)
Register 6:45 PM Dinner 7:30 PM Talk 8:30 PM
Cost: $30 for dinner ($10 students and teachers), free for talk.
Dinner Choices: (Indicate Choice when RSVP'ing)
Breast of Chicken, Piccata Chilean Sea Bass, Ginger Shallot Sauce Grilled Vegetable Brochette with Wild Rice
Reserve by April 10 to Steve Rabin, 650-564-5315 or steve.rabin-at-alza.com
Also, see our web site at www.sas-ncss.org
ABSTRACT Polymeric macroscopic properties such as tenacity, modulus, elasticity, etc. are determined by molecular level effects such as conformational state populations, orientation, and intermolecular interactions. Significant changes occur at the molecular level as a molten or solution phase polymer is spun into fiber form. The polymer goes from an unoriented, amorphous state to an oriented, semi-crystalline material via the deformations imposed by spinning and drawing. Raman spectroscopy can potentially provide information on both structure and orientation. Changes in band intensities can be related to conformational populations and to formation of crystalline regions. Changes in relative intensities as a function of incident and scattered polarization yields information on chain orientation. The use of polarized Raman scattering done in both 90 and 180 degree scattering geometries has been used to determine the second and fourth order orientation functions for both polyethylene and PET fibers. Raman measurements have been made in both the laboratory frame and on-line for 66 nylon and PET fibers. For both PET and 66 nylon, crystallinity levels and a qualitative measure of orientation can be obtained.
I've been asked whether it is possible to look at a frozen powder using Cryo-SEM. The powder will be frozen and stored at -80 C and must be mounted whilst frozen (it may be possible to raise the temperature to -20 C) any ideas of an adhesive I could use at these low temperatures??
Thanks
Ian
Ian Hallett HortResearch Mt Albert Research Centre Private Bag 92 169 Auckland, New Zealand Fax 64-9-815 4201 Telephone 64-9-815 4200 EMail ihallett-at-hort.cri.nz
} As far as your analysis of the SEM chambers & polepieces, historically } the SEM industry has seen the large chamber sizes and conical polepieces } you decribe for at least last twenty years.
For those, like me, who were interested in your recollections about SEM chamber and lens design, and who may be considering attending ICEM-15 in South Africa in 2002, make a note that there will be an exhibition at the congress retracing the history of electron microscopy in South Africa. This will include a variety of old EM equipment, including some of the SEM's to which you referred.
Regards
Rob
Robin H Cross Chairman : 15th International Congress on Electron Microscopy (ICEM-15) tel: +27 46 603 8168/9 - fax: +27 46 622 4377 email: r.cross-at-ru.ac.za http://www.ru.ac.za/emu/icem-15.htm
** remember that ICEM-15 in 2002 is in Durban, South Africa **
Ian, I've used Duro-Takš 80-1061 to hold particles during analysis at liquid nitrogen temperatures . For more information see: http://www.fbi.gov/programs/lab/fsc/backissu/july1999/ward.htm
Dennis ________________________________________________________________________ Dennis C . Ward voice: 202-324-2982 FBI Laboratory fax: 202-324-4018 Microanalysis Laboratory email: DCWard-at-concentric.net
----- Original Message ----- } From: "IAN HALLETT" {ihallett-at-hort.cri.nz} To: {Microscopy-at-sparc5.microscopy.com} Sent: Sunday, March 19, 2000 7:45 PM
Hello Ian
} I've been asked whether it is possible to look at a frozen powder } using Cryo-SEM.
That shouldn't pose any problems. Choice of adhesive would depend on the characteristics of the powder.
Rob
Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa R.Cross-at-ru.ac.za tel: +27 46 603 8168 - fax: +27 46 622 4377 http://www.ru.ac.za/emu/index.htm
** remember that ICEM-15 in 2002 is in Durban, South Africa **
I would like to thank those who took the time to respond to my e-mail. I received a lot of valuable information and advice on different techniques to be used (and NOT to be used) when studying carbides in tool steels. I appreciate your help.
Kind regards, Jesper
---------------------------------------------------- Jesper Vejloe Carstensen Research Scientist, M.Sc., Ph.D. Materials Research Department Risoe National Laboratory P.O. Box 49 DK-4000 Roskilde, Denmark Phone: +45 4677 5776 Fax: +45 4677 5758 E-mail: jesper.v.carstensen-at-risoe.dk Web: http://www.risoe.dk/AFM/Personal/Jeca/jeca.htm ----------------------------------------------------
We are planning to buy a new diamond knive for sectioning biological samples. My question is if anybody has experience with using 35 degree angle diamond knives and if they have any disantvantages compared to 45 degree angle diamond knives. We are mainly sectioning unicellular algae embedded in LR-gold and hope to reduce compression problems by using a 35 degree angle knive. Our problem is that, especially algae with thick cell walls, tend to fall out of the sections. This problem seems to be caused by compressions, because using a brand-new 45 degree angle diamond knive I had much less algae falling out of the sections. Because we can effort only one new knive we have to use it as an all-round knive. Is a 35 degree knive suitable to replace a 45 degree knive or is it much more fragile and/or less durable?
Thanks in advance for your time and help.
Sincerely,
Stefan Geimer
*********************** Stefan Geimer University of Cologne Botanical Institute Gyrhofstr. 15 D-50931 Cologne Germany phone: +49-(0)221-470-3795 fax: +49-(0)221-470-5181 ************************
Apparently there is more equipment lying around that needs a good home. If anyone is interested, please respond directly to Mark Chambers via mark-at-semiconductor.com or at (613) 599-6500 ext 4269.
Marisa
} -----Original Message----- } From: Mark Chambers } Sent: Friday, March 17, 2000 4:18 PM } To: Marisa Ahmad } Subject: RE: Liquidation of assets } } Hi Marisa, } } Is the listserver you used for the PGT EDS announcement a suitable venue } for the plasma asher and the Mosaid tester? } } The barrel etcher is a Plasmaline model 411 and whoever wants it will need } to get a vacuum pump for it. The etcher was working but it was not an } efficient way to decap packages. } } The Mosaid tester is a model MS2300 and that's basically all I know about } it. } } Thanks } Mark
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
You can buy them from Electron Microscopy Sciences. 1-800-523-5874
Soumitra
***************************************************************** Soumitra Ghoshroy Ph.D. Electron Microscopy Lab Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3600 Fax: 505-646-5665 e-mail: ghoshroy-at-nmsu.edu http://confocal.nmsu.edu/eml
I have been asked to find a new home for a fully working JEOL 100CX tem which is currently in use in the south of England. It has the ASID stem attachment, and it could also be available with a Link AN10 analysis system.
Probably the only cost will be the dismantling & removal charges. If anyone is interested, please reply to me directly. My only commercial interest is that I might be involved with the dismantling etc.
Bob Phillips MicroServiS bob.phillips-at-microservis.co.uk
Please check out the web-site for ICHC 2000 (11th International Congress for Histochemistry and Cytochemistry) 3-8th Sept. York, UK.
We have put together a fabulous speaker list covering a range of the most important areas of development and application of cutting edge techniques in Cell Biology and Imaging today. Lead speakers include Roger Tsien, Stefan Hell, Alan Fine, Alan Bode, Hans Tanke, Jennifer Lippincott-Schwartz, Angus Lamond, Jim Smith, Richard Haugland, Paul Monaghan, Mike Ormerod, Simon Gilroy Nick White etc. etc. etc.
www.med.ic.ac.uk/external/ichc_2000
ABSTRACT DEADLINE 15TH MAY 2000.
Best wishes
Gary Coulton Dr. Gary Coulton Molecular Pathology Division of Biomedical Sciences Imperial College School of Medicine The Sir Alexander Fleming Building South Kensington London SW7 2AZ
tel 0044 (0)171 594 3190 fax 0044 (0)171 594 3022
PLEASE NOTE AS FROM APRIL 1ST 2000 THE FOLLOWING NUMBERS WILL REPLACE THOSE ABOVE AND SHOULD BE USED.
------------------------------------- Announcing the 11th International Congress of Histochemistry and Cytochemistry (ICHC 2000)
"Understanding Biocomplexity: The Post-Genome Challenge"
September 3-8, 2000, York, United Kingdom
ICHC 2000 will comprise 27 symposia addressing the latest developments and applications of histochemistry and cytochemistry in the life sciences including medicine.
Many leading experts to speak
On-line registration now open!!!!!!!!!!
For further details of the meeting and how to pre-register please visit our web-site at http://www.med.ic.ac.uk/external/ichc_2000
The Department of Materials Science and Engineering at the University of Pennsylvania is seeking an engineer or scientist for a technical staff position in its advanced characterization facility. This facility contains a number of state-of-the-art microscopes and scattering facilities as well as ancillary detectors, specimen preparation equipment, and data processing and computational hardware and software. The facility serves Penn research programs, as well as academic, industry and government users from the Delaware Valley region and beyond. The University of Pennsylvania is located in Philadelphia, one of the nation's most vibrant cities. The university, a member of the Ivy League, was founded by Ben Franklin and is the fourth oldest and first secular university in the US. The Laboratory for Research on the Structure of Matter was constructed to house one of the original three Materials Research Laboratories (MRLs) in the US. The university has a continuing tradition of leading materials research and has housed the MRL (now MRSEC) continuously for 40 years.
Under the direction of the facility manager, the successful candidate will be responsible for conducting experiments with users on facility equipment, training new users and maintenance of facility equipment not covered under service contract. The successful candidate must have the communication skills and self-confidence to interact with technical users with a wide range of expertise and background. Job performance will be assessed based on the success in experimental interactions with users, the operational state of equipment for which the staff member is responsible, progress in the acquisition of new skills, and facility appearance.
A Bachelors degree in physical science or engineering is required, although an advanced degree is preferred. The successful candidate must have experience with electronics, ultra-high vacuum technology and computer interfacing of equipment. Experience in the operation and use of electron microscopes and ion beam lines and associated end-stations is desirable. Experience in the use of Auger electron spectroscopy or scanned probe imaging is also desirable.
The text of the official job listing from the University of Pennsylvania web site follows. Please refer to the Penn Human Resources web site for the official hiring policy of the university at www.hr.upenn.edu. For information about the specific position, please contact the facility manager, Dr. Douglas Yates, at dmyates-at-lrsm.upenn.edu.
Text of web site listing:
Reference Number: 00034808DL Job Title: RESEARCH COORDINATOR SR School/Center: ENGINEERING & APPLIED SCIENCE Department: MATERIALS CHARACTERIZATION Date Posted: 3/9/00
Duties: Train or coordinate training of facility users; assist users performing experiments; compose monthly MCF users billing summary; organize purchasing of consumable laboratory materials; maintain or coordinate maintenance of equipment; maintain working environment & appearance of laboratory.
Qualifications: BA/BS in Physical Science or Engineering required, advanced degree preferred; 3 to 5 years experience with electronics, ultra-high vacuum technology & computer interfacing of equipment & in operation & use of electron microscopes, ion beam lines and/or scanned probe imaging.
******************************************************* Douglas M. Yates, Ph.D. Manager, Materials Characterization Facility
(215) 573-6123 dmyates-at-lrsm.upenn.edu
University of Pennsylvania Department of Materials Science & Engineering 3231 Walnut Street Philadelphia, PA 19104-6272 *******************************************************
One of your best sources of information is going to be Diatome USA. They have an actual lab set up to test cutting problems of a large variety. They also have access to the information of the company who makes these knives.
Please consider that your problem may not be with the knife or the angle at which the knife is sharpened. Most "breaking out" problems are related to the production of the block, especially 1) the infiltration 2) the match between the specimen and the formulation used. Contact Diatome, at EMS, and ask to speak with Stacy Kirsch, who is an expert with these problems. (I have no business interest in Diatome - I have just received the most excellent advice from them over the years)
Hildy Crowley Sr. Electron Microscopist University of Denver Denver, CO
P.S. Should processing be the basic problem, please contact me. I may be able to help you.
My specialists tell me, that Boron shows a plasma loss at 22.7 eV and a K-edge at 188 eV. They also tell me, that it is hard to see B in samples as it usually disappears rapidly. To see B, they suggest focusing on a different sample area and then only expose the area in question for the PEELS acquisition.
In case you are using our software on a LEO microscope for the acquisition, you should select the MDF option (Micro Dose Focusing).
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== 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: Exchange Administrator Sent: Saturday, March 11, 2000 12:35 PM To: Michael Bode
Need help on PEELS } } To all, } } I am working on grain boundary chemistry measurement } with Digipeels. Try to find out the segregation } behavior of boron, which is about 0.5at% averagely. } However, I cannot find any edge for boron in the } spectrum. As I don't know what's the optimum setting } for PEELS to check the grain boundary segregation, if } you have any idea on this, please let me know. } } Thanks, } Lung __________________________________________________ Do You Yahoo!? Talk to your friends online with Yahoo! Messenger. http://im.yahoo.com
We bought a JEOL 100SX for about $130,000 in 1987. The scope was for student use in an undergraduate course. It was used for 4 to 8 weeks per year for 7 years. It was used very little. About 1994, the water chiller failed and cooked the objective mini-lens and some other electronics. It was one year off the service contract. JEOL could not guarantee that they could fix it for less than $11,000 so it was moth balled. It was functional at 8000X and higher. We have to get rid of this in a real hurry. It someone wants some or all of this machine please contact me ASAP. Within about a week it will be in the dump. I recall that at the time it failed there was a user at Stanford (I think) who had the same model. If anyone knows who that was, please have him contact me.
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 530 754 7536 fax http://katie.ucdavis.edu raharris-at-ucdavis.edu
It seems that nowadays every SEM vendor is offering variable pressure models, which are conventional SEMs with a plumbing modification that allows the sample to be at pressures of up to about 4 torr (500 Pa) while the electron column operates at the conventional high vacuum. I am very interested in buying a variable pressure SEM with an EDS, but was recently warned that EDS has very poor spatial resolution in the variable pressure mode because of the large beam spread due to electrons scattering off the gas molecules in the sample chamber. Is this really a significant issue? Do any of you have experience using EDS with variable pressure SEMs? Thanks,
In short, it is an issue, but not intractable. For careful quantitative work we reduce the vacuum as much as the sample permits. We can usually get by with about 40 Pa of helium atmosphere on concrete samples. Even at 40 Pa we can pick up percent levels of elements away from the beam. For example, a Co chip in a standard mount can easily show 1% Fe from the stainless steel carrier. For qualitative work, that is not much of a problem, but if you are looking for trends in that element (Fe in Co), it would be impossible in low vacuum mode.
BTW, Helium greatly reduces the scattering compared to air at the same pressure.
I would encourage you to go ahead and get the VP SEM, but work through a few exercises with it to get a feel for its limits.
At 05:22 PM 3/20/2000 -0500, you wrote: } It seems that nowadays every SEM vendor is offering variable pressure } models, which are conventional SEMs with a plumbing modification that } allows the sample to be at pressures of up to about 4 torr (500 Pa) while } the electron column operates at the conventional high vacuum. I am very } interested in buying a variable pressure SEM with an EDS, but was recently } warned that EDS has very poor spatial resolution in the variable pressure } mode because of the large beam spread due to electrons scattering off the } gas molecules in the sample chamber. Is this really a significant issue? } Do any of you have experience using EDS with variable pressure SEMs? } Thanks,
---------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
It is a good point to contact DIATOME. It reminds to me, also, that they (DIATOME) may cut your samples on site and sent back the grids (if I understand them correctly) for free. So, you may try 45 and 35o knife and compare the results before you make final decision. As for 35o. 45o is more universal and durable. If you rich enough only for one diamond knife, it should be 45o, I believe. 35o supposed to be a "second knife" for "special occasions". I have no financial interest in DIATOME, but happy user of DIATOME knife
Sergey
} Date: Mon, 20 Mar 2000 12:00:52 -0700 (MST) } From: HILDEGARD CROWLEY {hcrowley-at-du.edu} } Subject: Re: 35 Degree Angle Diamond Knive } X-Sender: hcrowley-at-odin.cair.du.edu } To: "Stefan.Geimer" {stefan.geimer-at-uni-koeln.de} } Cc: Microscopy-at-sparc5.microscopy.com } } ------------------------------------------------------------------------ } 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
Diatome, for one, can supply you with clear ev idence for significantly reduced compression with a 35 degree knife over a 45 degree knife. No surprise there, since compression should be reduced with knife angle in materials which compress significantly. We are a 'materials science' lab, where we section all manner of metals and alloys, composites, powders, fibres, coatings, thin films, etc, etc. For us reduced compression is not a big deal, but durability obviously is. I can report that, for the last 5 years or so since their acquisition, our two Diatome 35 degree knives are used about 90% of the time, with no catastrophic failures or large increase in sharpening frequency.
I agree with Hildegard Crowley that breaking out is far more likely to be due to embedding/bonding problems. another possibility is simply a dull edge. Your increased success with a fresh (presumably demo) knife could indicate the latter. If the edge is quite rounded from use, the effective angle at the point of sectioning will shoot up incredibly, which will then increase the compressive forces that expose any embedding/bonding deficiencies.
That said, if you can only afford one knife, look in the mirror and ask yourself how careful are the people who might use the knife. A simple consideration of the forces on a knife edge during sectioning will tell you that a lesser angle should be slightly more susceptible to side forces. However, just the thought of a reduced angle might make an inexperienced operator sufficiently nervous to increase that risk greatly!
Tom Malis Group Leader - Characterization Materials Technology Laboratory Natural Resources Canada (Govt. of Canada) 568 Booth St., Ottawa, Canada ph. 613-992-2310 FAX 613-992-8735 email: malis-at-nrcan.gc.ca
} ---------- } From: Stefan.Geimer } Sent: Monday, March 20, 2000 9:59 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: 35 Degree Angle Diamond Knive } } ------------------------------------------------------------------------ } 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 planning to buy a new diamond knive for sectioning biological } samples. My question is if anybody has experience with using 35 degree } angle diamond knives and if they have any disantvantages compared to 45 } degree angle diamond knives. We are mainly sectioning unicellular algae } embedded in LR-gold and hope to reduce compression problems by using a } 35 degree angle knive. Our problem is that, especially algae with thick } cell walls, tend to fall out of the sections. This problem seems to be } caused by compressions, because using a brand-new 45 degree angle } diamond knive I had much less algae falling out of the sections. Because } we can effort only one new knive we have to use it as an all-round } knive. Is a 35 degree knive suitable to replace a 45 degree knive or is } it much more fragile and/or less durable? } } } Thanks in advance for your time and help. } } Sincerely, } } Stefan Geimer } } } } *********************** } Stefan Geimer } University of Cologne } Botanical Institute } Gyrhofstr. 15 } D-50931 Cologne } Germany } phone: +49-(0)221-470-3795 } fax: +49-(0)221-470-5181 } ************************ } } } }
Can anyone help Karen Maxwell?, reply to her as well as the list as she is not a subscriber.
Nestor Your Friendly Neighborhood SysOp
Below is the result of your feedback form. It was submitted by (maxwell-at-lec.med.utoronto.ca) on Wednesday, March 15, 2000 at 17:03:53 ---------------------------------------------------------------------------
Question: I have been looking at a paper from 1984 (Kochan et al) where they used EM to study the preconnector from bacteriophage lambda. They discovered that the axial hole in the ring shaped structure was 2.2-2.5 nm in diameter. They used negative staining with ammonium molybdate, uranyl acetate, and uranyl formate. In more recent studies (Valpuesta et al, 1999), the connector of phi29 was examined using a three-dimensional cryo-reconstruction, and the axial hole was found to be 3.3 nm. These are two different phage, but my question is, could the old techniques used in the case of the bacteriophage lambda preconnector (negative staining) have under-estimated the diameter of the axial hole? And if so, do you have an idea of the percent error that may be inherent in the technique? Is the cryo-reconstuction likely to give a value that is more accurate? Could you recommend any references that would address these questions?
At 04:22 PM 3/20/00 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have not had an opportunity to try these systems but from what I can glean, they are not totally similar to conventional SEMs. In particular, the VP SEMs do not use the E-T detector, but rather a gas discharge detector. There are some good examples of these images around but I do wonder what the actual difference is. Perhaps owners can tell us. Or warn us???
At 04:22 PM 3/20/00 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have not had an opportunity to try these systems but from what I can glean, they are not totally similar to conventional SEMs. In particular, the VP SEMs do not use the E-T detector, but rather a gas discharge detector. There are some good examples of these images around but I do wonder what the actual difference is. Perhaps owners can tell us. Or warn us???
Everett Ramer wrote: } It seems that nowadays every SEM vendor is offering variable pressure } models, which are conventional SEMs with a plumbing } modification that allows the sample to be at pressures of up to } about 4 torr (500 Pa) while the electron column operates at the } conventional high vacuum. I am very interested in buying a } variable pressure SEM with an EDS, but was recently warned that } EDS has very poor spatial resolution in the variable pressure } mode because of the large beam spread due to electrons } scattering off the gas molecules in the sample chamber. Is this } really a significant issue? Do any of you have experience using } EDS with variable pressure SEMs? } Thanks,
The fraction of primary electrons that are scattered by the gas in the specimen chamber is approximately given by:
1-I/Io = 1 - exp(-psL/kT)
where p is the pressure s the total scattering cross section for scattering of x keV electrons on the gas used L the distance between the last pressure limiting aperture and the sample k the Boltzmann constant and T the absolute temperature.
You can therefore reduce the contribution of X-rays excited by scattered primary electrons by reducing p (lowest possible pressure), s (use gas with low scattering cross section and the highest possible acceleration voltage balanced against overvoltage considerations) and L.
As Warren Straszheim wrote this will overcome the beam skirt problem to a large degree. In the situations where this is not sufficient, you can use an extrapolation method that has been developed here at Risoe National Laboratory. In short, you perform the analysis at various different pressures and use the above equation to extrapolate to the result you would get under zero pressure. A more detailed description can be found at the web-address
***************************** J. B. Bilde-Soerensen Senior Research Scientist, Ph. D. Materials Research Department Risoe National Laboratory DK-4000 Roskilde Denmark
I was wondering if anybody was using a GIF filter. We have one mounted on a 120 kV TEM and are trying to image small particles (5-10 nm) and doing some energy filtering to image different elements in the particles (for example gold, silicon, copper, cerium...) . This is not really what I would call trivial work and we are still working out the set up (window size and positionning for example) in a rather empiric way. The main problem is often to get a decent signal in the interesting energy region and still keep the windows fairly narrow. I would add that we often have to work at low dose because of the beam sensitivity of our samples, but I guess our GIF setups can be improved. Any suggestion ?
I am staining virus infected cell DNA (in cell culture) with Hoechst 33258 fluorescent stain. I need to make correlation of the fluorescent spots with cellular structure.
Do you know of any counter-stain that can be used without destroying the fluorescent Hoechst staining?
Thanks Dr. A.P. Alves de Matos biologist apmatos-at-ip.pt
Diana: TAAB Laboratories Equipment, Ltd, 3 Minerva Court House,Calleva Park Aldermaston, Berks,RG7 8NA, UK
They list single edge razor blades, with and without a bac k in stainless and carbon steels. They also list double edge razor blades, 0.004 in. thick, in stainless steel.
Best regards,
Sam Purdy } ---------- } From: Jeff & Wanda Gray } Sent: March 2000 10:12 PM } To: Diana Papoulias; microscopy-at-sparc5.microscopy.com } Subject: RE: accu-edge blades } } ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } Diana, } These blades are available from Allegiance Scientific (used to be Baxter, } used to be Scientific Products, used to be....). As far as I know, this is } the only place that carries this brand. } I have no financial interest in Allegiance, just passing along a fact. } Wanda Shotsberger } (HT ASCP) } } -----Original Message----- } } From: Diana Papoulias [mailto:Diana_Papoulias-at-usgs.gov] } Sent: Thursday, March 16, 2000 2:53 PM } To: microscopy-at-sparc5.microscopy.com } Subject: accu-edge blades } } } ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } Could someone please tell me where I can purchase accu-edge microtome } blades? } } Thank you. } } Diana Papoulias } USGS } 4200 New Haven Rd } Columbia, MO 65201 } } T:573 876 1902 } F:573 876 1876 } E:Diana_Papoulias-at-usgs.gov } } }
The extrapolation method (aka Variable Pressure Method) described by Dr. Bilde-Sorenson has been implemented by EDAX in a software feature called ViP-Quant. With this method the EDS analysis under Bad-Vacuum conditions can be done with nearly the same accuracy as under High-Vacuum conditions. Although this method was developed with the ESEM microscope in mind, it of course can be applied to all Bad-Vacuum scanning electron microscopes.
Please contact your local EDAX representative for more information and a copy of the new ViP-Quant brochure, or request one through www.edax.com.
} -----Original Message----- } From: Joergen Bilde-Soerensen 5802 [mailto:j.bilde-at-risoe.dk] } Sent: Tuesday, March 21, 2000 10:04 AM } To: Everett Ramer } Cc: Microscopy-at-sparc5.microscopy.com } Subject: Re: EDS in Variable Pressure SEM } } } ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } } Everett Ramer wrote: } } It seems that nowadays every SEM vendor is offering variable pressure } } models, which are conventional SEMs with a plumbing } } modification that allows the sample to be at pressures of up to } } about 4 torr (500 Pa) while the electron column operates at the } } conventional high vacuum. I am very interested in buying a } } variable pressure SEM with an EDS, but was recently warned that } } EDS has very poor spatial resolution in the variable pressure } } mode because of the large beam spread due to electrons } } scattering off the gas molecules in the sample chamber. Is this } } really a significant issue? Do any of you have experience using } } EDS with variable pressure SEMs? } } Thanks, } } The fraction of primary electrons that are scattered by the gas in } the specimen chamber is approximately given by: } } 1-I/Io = 1 - exp(-psL/kT) } } where p is the pressure } s the total scattering cross section for scattering of x keV } electrons on the gas used } L the distance between the last pressure limiting aperture and } the sample } k the Boltzmann constant } and T the absolute temperature. } } You can therefore reduce the contribution of X-rays excited by } scattered primary electrons by reducing p (lowest possible } pressure), s (use gas with low scattering cross section and the } highest possible acceleration voltage balanced against } overvoltage considerations) and L. } } As Warren Straszheim wrote this will overcome the beam skirt } problem to a large degree. In the situations where this is not } sufficient, you can use an extrapolation method that has been } developed here at Risoe National Laboratory. In short, you } perform the analysis at various different pressures and use the } above equation to extrapolate to the result you would get under } zero pressure. A more detailed description can be found at the } web-address } } { HYPERLINK http://www.risoe.dk/afm/news1new.htm }http://www.risoe.dk/afm/news1new.htm
Best regards, Joergen Bilde-Soerensen
***************************** J. B. Bilde-Soerensen Senior Research Scientist, Ph. D. Materials Research Department Risoe National Laboratory DK-4000 Roskilde Denmark
Dear Colleagues, I want to fix insects at particular moments of muscle activity. I plan to freeze the bugs in liquid nitrogen and section their heads, after including them in hard Durcupan. I have no experience in fixating and including frozen pieces. Note that hard resin should be used, provided the hardness of the insect cuticle. Does anybody know a simple method for the transference from liquid nitrogen to plastic? Thank your very much in advance, Claudio Dr. Claudio R.Lazzari lazzari-at-bg.fcen.uba.ar
------------------------------------------ Laboratorio de Fisiologa de Insectos Dpto. Cs. Biolgicas, Univ. Buenos Aires Ciudad Universitaria, 1428 Buenos Aires Argentina Tel.(54 11) 4576 3300, ext. 332, FAX (54 11) 4576 3384
Gabriel Adriano Rosa Centro de Imagenes y Microscopia Departamento de Ciencias Biologicas Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires Ciudad Universitaria, 4 piso, Pab. II, CP 1428, Buenos Aires, ARGENTINA TE -(54-11)-4576-3349 e-mail cimic-at-bg.fcen.uba.ar FAX (54-11)-4576-3384
I don't think 7-AAD will work - it is used as a live/dead stain, and I know that it works much better on detergent-permeabilized cells than on merely PFA-fixed ones.
If you are contemplating microinjection (ugh!), maybe try the fluorescent dextrans instead of UTPs...they won't mess up your nucleic acids. We use them as injection markers and they don't seem to harm the cells.
Tamara Howard CSHL
On Tue, 21 Mar 2000, Donald O'Malley wrote:
} Hi Folks, } } Thanks for the flurry of info about DNA/RNA staining. I've summarized some of this info below in case anyone else has needs related to ours. } What I neglected to mention is that we are trying to count nuclei inside of living mouse embryos. That's why we're searching for a non-invasive, membrane crossing, nuclear-selective dye. } } Summary of recent listserv messages: } } } Possible Dyes for live TISSUE, visible wavelength, } selective NUCLEAR staining: } } 7 aminoactinomycin. But does it cross membranes? } microinjected Alexa-UTPs
I just got back from PITTCON. When I was visiting the EDAX booth they showed me a new program through which they have resolved this problem. I would suggest at least inquiring.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
At 05:22 PM 3/20/00 -0500, Everett Ramer wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dr. Alves de Matos, Regarding your question about counter staining for cellular structure--have you considered scanning your Hoechst image and overlaying a Nomarski image? If this doesn't yield enough detail there are fluorescent dyes specifically for organelles such as mitochondria and golgi. Molecular Probes sells them.
Mary Gail Engle Electron Microscopy & Imaging Facility University of Kentucky
Karl E. Garsha, Doctoral Student AOP Fellow Chief Coordinator-Graduate Student Association Department of Biological Sciences University of Wisconsin-Milwaukee P.O. Box 413 Milwaukee, WI 53201 Office: 459 Lapham Hall Phone: (414) 229-4316 Mobile: (414) 617-4295 E-Mail: keg-at-csd.uwm.edu
---------- } From: "Karl E. Garsha" {keg-at-csd.uwm.edu} To: maxwell-at-lec.med.utoronto.ca
Firewire Camera Anyone? Does anyone know of, or have information on current or upcoming digital cameras for light microscopy that utilize the Firewire or USB interface for quick and easy importing of light microscope images through a color digital camera, for example to a Mac G4 (or even the iMac G3)? Does this technology exist in a microscope camera? (yet)
In a message dated 03/21/2000 5:15:50 PM US Mountain Standard Time, HUGGINBJ-at-BP.com writes:
{ { Firewire Camera Anyone? Does anyone know of, or have information on current or upcoming digital cameras for light microscopy that utilize the Firewire or USB interface for quick and easy importing of light microscope images through a color digital camera, for example to a Mac G4 (or even the iMac G3)? Does this technology exist in a microscope camera? (yet) } }
The only one I am aware of is the Optronics MagnaFire, which is a cooled camera with the IEEE-1394 FireWire interface.
Previous versions of the Optronics cameras used a parallel (printer) port interface so they had to be used on PC's only. It would be worth inquiring about Mac compatibility. Sorry I don't have the answer to this part...I only know the MagnaFire has the FireWire interface.
Visit {http://www.optronics.com} and click on "MagnaFire" for specs.
We are in the midst of writing a proposal for a new film scanner and I am hoping that someone out there can help with a recommendation for a top of the line film scanner. The major application would be TEM negatives. A model we are currently looking at is the Nikon LS-4500AF Multi Format Film Scanner.
If anyone has archived recommendations from previous discussions and could send them on to me, that would be wonderful.
Thank you, Valerie Leppert
Dept. of Chem. Eng. and Mat. Sci. U. of California, Davis
I do not work for Optronics, but know a Mac version of Drivers is due out very shortly for the Magnafire Camera. For more information, check out....
http://www.optronics.com
Also, Nikon has an SLR based 2.74 megapixel Digital Camera suitable for most Microscopy (including bright emission Fluorescence with higher ISO) called the D1, based on a N90s Body with a F-mount and Firewire connection. Approx. $5300
Nikon also has a much less expensive digital camera Coolpix 990 coming in the next few weeks. This is a 3.34 million pixel, true resolution camera with live NTSC out (for simultaneous real-time video) and USB connectivity with an Electronic Shutter Release built-in for around $1000. It too, can handle most Microscopy techniques, but is limited in fluorescence by its 100 ISO rating. It is a C-mount type camera.
**I have no corporate affiliation with/nor any financial agreement with Optronics, Inc. I do however have strong ties with Nikon (as you can see), but make very little money from either product ;)
} 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 } -----------------------------------------------------------------------.
} } In a message dated 03/21/2000 5:15:50 PM US Mountain Standard Time, } HUGGINBJ-at-BP.com writes: } } { { Firewire Camera Anyone? } Does anyone know of, or have information on current or upcoming digital } cameras for light microscopy that utilize the Firewire or USB interface for } quick and easy importing of light microscope images through a color digital } camera, for example to a Mac G4 (or even the iMac G3)? Does this } technology exist in a microscope camera? (yet) } } } } } The only one I am aware of is the Optronics MagnaFire, which is a cooled } camera with the IEEE-1394 FireWire interface. } } Previous versions of the Optronics cameras used a parallel (printer) port } interface so they had to be used on PC's only. It would be worth inquiring } about Mac compatibility. Sorry I don't have the answer to this part...I only } know the MagnaFire has the FireWire interface. } } Visit {http://www.optronics.com} and click on "MagnaFire" for specs. } } Cheers, }
Dear Karl I disagree with you, that "UA in particular, will positively stain proteins". Let start from definition: positively stain object is a dark object on the light background (therefore stain penetrate/interact with object making it "stained", dark in the terms of EM); "negative staining" (NS) is opposite. We have light (stain do not interfere with the sample) object on the dark background of stain (or dark ring of stain around the object, depends from stain and technique). The beauty of the NS is that stain (UA in particular) easily penetrates cavities in the object making visible fine details (for instance, I was able to recognize variable and constant domains of the Fab of the IgG molecules as well as domains of the Fc using UA staining). May be this effect you call "positively staining". But it is not.
Yes, UA may stain RNA and DNA positively. For this reason UA gives us "mixed stain" for ribosomes (negative for proteins and positive for RNA components). The disadvantage UA is its low pH.
As for subject of the current discussion. I think Cryo in general will give us more correct information than others EM techniques. But, we have to keep in mind, that in Cryo-technique, to make image relatively contrast, people should go to very high overfocusing (from half to couple microns, I believe). For this reason, the best Cryo-results I know, yielded only 2-3 nm resolution (and this is a huge problem in Cryo - to get better resolution): the same or even worse as for "negative staining" (I expect 1.5 nm resolution for UA). We have to count resolution when compare the data. I lost the original message from which this discussion was started, but it seems to me, that difference between NS and Cryo was not so dramatic and may be comparable if we will count the resolution. In general, I don't believe any EM data if resolution is not shown. Talking about resolution is complicated because of the difference between "resolution of the instrument" (0.14 nm for TEM currently), "physical resolution of the method" (resolution limit for overfocusing, for instance, radiation damage, drift, noise/signal ratio etc) and "resolution of the sample" (for biological samples they are not the same: preparation of the sample may affect sample's intactness/structure). Cryo is the best for sample preservation, but it is low in contrast. UA may affect sample's structure (may not) but the resolution limited only by granularity of stain (in theory it is a couple angstroms).
As for "averaging" of the images, I am a little bit skeptical about that. It is great deal if your sample is uniform in shape (symmetry is a great help too). If your biological sample is functionally flexible (IgG for instance), averaging will "hide" some interesting details even if you will distribute images in classes.
In my point of view, it is difficult to extract absolute numbers from EM. Inside one methodology you could easily compare the data, but it becomes difficult when you want to compare the data obtained by different techniques. This is reality of EM. I think Scanning Probe Microscopy is very promising to obtain the direct measurements on the samples under physiological conditions (EM is so far from "physiological conditions", unfortunately). I am so sorry, EM!
} Date: Mon, 20 Mar 2000 16:46:25 -0600 } From: "Karl E. Garsha" {keg-at-csd.uwm.edu} } Subject: FW: neg stain vs cryo EM plus image averaging } To: microscopy-at-sparc5.microscopy.com } X-Mailer: Microsoft Outlook Express for Macintosh - 4.01 (295) } } ------------------------------------------------------------------------ } 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 posted a previous message asking for a counterstain for Hoechst 33258, however I fail to mention that I want to use the counterstain in brignt field ilumination, switching to fluorescence as needed. Something that reveals celular structure like Giemsa would be OK. Giemsa however does not alow simultaneous staining with Hoechst as far as I can see.
Regarding EFTEM on small particles, this should actually work quite well. That is, elemental mapping works reasonably if you choose the right objective apperture. You should simulate the spatial resolution to see what can be attained under your conditions (HT, Cc, Cs etc). However it will be very difficult to get real concentration information out of these images. I'm trying EELS with nanoprobe but so far no succes (everything gets destroyed before I take a spectrum, allignment is very very difficult) Still, focussing remains a problem. Comon practice is to focus at 100eV and then suppose everything is OK for higher losses. I do not understand the theory of this technique (blurring by finite slitwidth & Cc?) nor thus it work well. Any comments on this are welcome.
Jo
************************************************************* * Jo Verbeeck * * University of Antwerp * * Dept. EMAT (Electron Microscopy for Materials Research) * * e-mail: joverbee-at-ruca.ua.ac.be * * tel: +32(0)3 218 02 49 * * fax: +32(0)3 218 02 57 * *************************************************************
On Tue, 21 Mar 2000, GuessWho wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear all, } } I was wondering if anybody was using a GIF filter. We have one mounted } on a 120 kV TEM and are trying to image small particles (5-10 nm) and } doing some energy filtering to image different elements in the particles } (for example gold, silicon, copper, cerium...) . This is not really what } I would call trivial work and we are still working out the set up } (window size and positionning for example) in a rather empiric way. The } main problem is often to get a decent signal in the interesting energy } region and still keep the windows fairly narrow. } I would add that we often have to work at low dose because of the beam } sensitivity of our samples, but I guess our GIF setups can be improved. } Any suggestion ? } } Any general comment on the GIF warmly welcome ! } } Thanks } } Olivier } } }
A few weeks ago, when our server was working correctly (*??!!*), there were e-mails talking about the purchase of a new SEM. Now we appear to be back on line I felt that our ideas may be of use to the mailers?
As part of the consultancy side of our business we purchase SEM for our clients. This means we are buying instruments regularly and are therefore probably one of the few professional SEM purchasing organisations?
In our team we have staff who have worked for most of the SEM manufacturers as demonstrators and application specialists, so we have seen the purchase game from both sides and of course try to use this to our clients advantage. Unlike the average microscopist, who probably only buys two instruments in their career, we have been able over the years to formulate a purchase plan, a purchasing criteria that we apply to each purchasing project. In our position we are unable to buy from the nicest salesman or the normal "gut" feeling, we must be absolutely certain that we are purchasing the best instrument for our clients applications.
Set out below are the headings from a lecture that I give on instrument purchase. Only the basic ideas are presented ( I guess if you need more I shall be attacked by Microscopy Today to give just that) but you will get the theme. It is important to realise that the specimen chamber of a SEM, the specimen-detector geometry, determines what we will see. For this reason you will find that one instrument will do a particular task better than any other in the price range, you need test specimens that will sort this out for you. If you have a multi instrument laboratory I always feel that it is a good idea to have instruments from different manufacturers optimising the laboratory for optimising a wider range of applications.
"Buying a New Microscope"
How Do You Start?
Collect ALL the brochures and prices Form a view of the new facilities being made available If you do not understand any features ASK the salesman Check through all the users desired facilities? Determine who will use the instrument now? Would there be others if you purchased particular accessories?
Formulate a Purchase Specification
Price range - we always look one level higher Set essential instrument specifications Set desired instrument specifications give them points and produce a "desirability assessment"
Why use a consultant?
Only he will without prejudice talk to all interested parties Interdepartmental feuds could spoil the case A different questioner provides different answers Their wide knowledge base may develop additional features They will have experience producing a "desirability assessment" which often brings surprises. They will be experienced with ways of dealing with the sales staff, they will keep them off your back
How to handle the demonstration? DO NOT LET THEM
Show you how wonderful the alignment procedures are - you will not spend all day aligning the instrument! Dominate YOUR demonstration Take you into totally uninteresting areas of the instrument - you should be buying because of the image quality Show their favourite gimmick Use their own favourite specimen Take you out of the room when about to load one of your specimens Take you to a two hour lunch with lots of drink
How to handle the demonstration before you set off
Select a maximum of three specimens that you know really well and that are important to your laboratory The specimens must be capable of meaningful low and high magnification imaging Develop a demonstration criteria for each specimen Have a tie break specimen available as suggested by the consultant Provide each company with an exact demonstration programme
What a consultant would do during a demo
Time the demonstrator to produce images at specific magnifications under very specific conditions Encourage the demonstrator (this is not a customer v demonstrator competition) allow them to try their own ideas with each specimen also, give them information that you have found to be of importance with your specimens Stop the demonstrator taking you away from your standard path - you do wish to compare each instrument under exactly the same conditions there is no time for other deviations
After the demo
Layout the results Compare performance instrument to instrument and time taken to obtain the results Produce a short list Compare the short list with the "desirability assessment" Research local and national service performance of the best two instrument manufacturers, and the manufacturers reputations
Finally Decide upon the instrument that you want, the specification and then haggle
Hope this helps?
Steve Chapman Senior Consultant Protrain for Training and Consultancy in EM World Wide Tel & Fax 44+ 1280 814774 e-mail protrain -at-emcourses.com web site www.emcourses.com
The ESEM with a cryostage has been used to observe such things as ice cream and asphalt/solvent mixtures. The ice cream work was published out of the Cavendish Labs [THiel, Donald]. The other was a lab experiment.
Hi! I have problem obtaining a good replica using a rotary shadow coating method. Samples are proteins suspended in a Tris, NaCl and CaCl2 buffer. They are absorbed onto a mica and dried in vacuum. I am using platinum wire (0.1 mm in diameter, 8 cm long) wrapped around tungsten filament followed by carbon coating. This replica is then transferred onto the grids. Coating is done in Hitachi HUS 5GB vacuum evaporator: vacuum 10-6 torr, 20 mA current through the electrode for 12 sec. The distance from the electrode to the plate with the samples is 10 cm. The problem is that I get very little coating (so little that there is no replica) and if I go higher then 20 mA tungsten brakes. I have no problem when I use palladium/gold wire with same parameters but that gives coarse granulation. This is the first time I used this method and I run out of ideas how to improve it. Thanks in advance for your replies. Dorota
I have just caught up on some of the questions on this server and I think I can contribute to the blade discussion. As far as I remember Accu-Edge Blades were re-branded Feather blades sold by Anglia Scientific, a UK microtome manufacturer in the 1970's and 80's. Anglia were taken over by Shandon Scientific and in turn they all disappeared into Life Sciences International. Phil Parker of Anglia I understand is still designing microtomes for LSI.
In short for Accu-Edge read Feather. We supply these Feather blades as do other microtome suppliers,
Best regards
Terry Cooper TAAB Laboratories Equipment Ltd 3 Minerva House, Calleva Park Aldermaston, Berks, RG7 8NA, England
Call for Papers Michigan Microscopy & Microanalysis Society Spring Meeting
The Michigan Microscopy & Microanalysis Society (Local Affiliate of MSA) will hold its Spring Meeting May 12th, 2000 at the Genoe Woods Conference Center in Brighton, Michigan. If you are interest in presenting a microscopy related paper or attending, please contact Deborah Rothe for more information, email: drrothe1-at-dow.com .
The society encourages student participation by providing free meeting registration for students whom present a paper. A cash award is sponsor by the society vendors for the best student paper.
Robert C. Cieslinski Michigan Microscopy & Microanalysis
Robert C. Cieslinski The Dow Chemical Company Microscopy & Microanalysis (517) 636-6875 email: rccieslinski-at-dow.com
} The other Firewire camera I know if currently is the MicroImager. } Check out http://www.qimaging.com/ Dave
} } The only one I am aware of is the Optronics MagnaFire, which is a cooled } camera with the IEEE-1394 FireWire interface. } } Previous versions of the Optronics cameras used a parallel (printer) port } interface so they had to be used on PC's only. It would be worth inquiring } about Mac compatibility. Sorry I don't have the answer to this part...I only } know the MagnaFire has the FireWire interface. } } Visit {http://www.optronics.com} and click on "MagnaFire" for specs. } } Cheers, } } Bob Chiovetti
Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 75 N. Eagleville Rd. U-125 Storrs, CT 06269 Knecht-at-uconnvm.uconn.edu 860-486-2200 860-486-4331 (fax) home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html ************************************************************
I would suggest that you look at UMAX Powerlook III. It is a flatbed scanner with built in transparency adapter, so you can scan reflective and transmissive media. Its resolution is 1200x2400 pixels. This should handle most anything you might have. I use this scanner all the time. They are about $1100 these days. For super high quality 35mm scanning, I use the Polaroid SprintScan 35+. It does 2700 dpi, D=3.4. It runs about $1500. For medium format and 4x5" transparent media, I use the Polaroid SprintScan 45. It runs about $4500 but will handle 35mm and 6x6cm media.
I would not recommend the Nikon.
gary g.
At 07:48 PM 3/21/00 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Valerie, I can't comment on the Nikon scanner but I have plenty to say about the Polaroid Sprintscan 45, most of which is probably best left unsaid! Let me summarize. We purchased the Polaroid Sprintscan 45 in late 1998 and planned to operate it on a PC with Windows 98 operating system. Our first unit was defective and was replaced, however, the trouble shooting process was very time consuming. Our second unit functioned intermittently. Again, after much trouble shooting we returned the unit to Polaroid Canada. After having it tested we were told that there were no problems with the Sprintscan 45. The unit was returned to us but continued to cause us grief with its erratic behavior. We spent more time trying to diagnose the problem and even called in a computer specialist who spoke directly with Polaroid's tech support personnel in an effort to resolve the problem. In the end we found that the Sprintscan appears not to like Windows 98. The scanner has been running sucessfully on an older PC with Windows 95. Why? Has there been an identified problem associated with Windows 98? If so why aren't customers alerted? Having this knowledge would have saved us an enormous amount of time and money. I have been waiting for a comment from Polaroid for well over a month.
I would be interested in hearing from other Sprintscan users who experienced similar problems.
Regards, Paul
Paul J. Gerroir Microscopy Materials Characterization Xerox Research Centre of Canada 2660 Speakman Drive Mississauga, Ontario L5K 2L1
} -----Original Message----- } From: Valerie Leppert [SMTP:vjleppert-at-ucdavis.edu] } Sent: Tuesday, March 21, 2000 8:48 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Film Scanner Recommendations? } } ------------------------------------------------------------------------ } 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 all- } } We are in the midst of writing a proposal for a new film scanner and I } am hoping that someone out there can help with a recommendation for a top } of the line film scanner. The major application would be TEM negatives. A } model we are currently looking at is the Nikon LS-4500AF Multi Format Film } Scanner. } } If anyone has archived recommendations from previous discussions and could } send them on to me, that would be wonderful. } } Thank you, } Valerie Leppert } } Dept. of Chem. Eng. and Mat. Sci. } U. of California, Davis } } vjleppert-at-ucdavis.edu }
Yes, there is a community of researchers involved in the use of EDS in ESEM (mostly with the Electroscan now Phillips microscope but the work is not exclusive of other low vacuum SEMs). See the Microscopy and Microanalysis Meeting Proceedings from the past several years (in 1999-see Wight pg 290, Carlton pg 292). EDS spatial resolution can be an issue depending on your conditions. Some general rules (to reduce the scattering and therefore improve the EDS spatial resolution) are to reduce the beam-gas-path length (shorten the distance the primary electrons need to travel thru the gas molecules), lower the chamber pressure (reduce the number of gas molecules in the path), use a less scattering gas (Helium has already been suggested), and use as high an accelerating voltage as reasonable. Several corrections have been suggested for removing the unwanted contributions to the EDS spectrum: the extrapolation method (already mentioned), beam stop method, spectral subtraction method, and continuum method. There are also simulation models that predict scattering by the gas, I am not aware of any that predict EDS spectra based on a multiphase or multicomponent system. I highly recommend that you take a typical sample and ask each of the microscope manufacturers demonstrate imaging and EDS in their scope.
I have no interest in any microscope or EDS system, {fontfamily} {param} Times {/param} Certain commercial software, equipment, instruments, or materials are identified in this report to specify adequately the experimental procedure. Such identification does not imply recommendation or endorsement by the National Institute of Standards and Technology, nor does it imply that the materials or equipment identified are necessarily the best available for the purpose. {/fontfamily}
} It seems that nowadays every SEM vendor is offering variable pressure models, which are conventional SEMs with a plumbing modification that allows the sample to be at pressures of up to about 4 torr (500 Pa) while the electron column operates at the conventional high vacuum. I am very interested in buying a variable pressure SEM with an EDS, but was recently warned that EDS has very poor spatial resolution in the variable pressure mode because of the large beam spread due to electrons scattering off the gas molecules in the sample chamber. Is this really a significant issue? Do any of you have experience using EDS with variable pressure SEMs?
} Thanks,
-------------------note: new mailing address----------------------
Scott Wight e-mail: scott.wight-at-nist.gov
NIST 222/A113 W voice: 301-975-3949
100 Bureau Dr STOP 8371 | fax: 301-417-1321
Gaithersburg, MD 20899-8371 \|/ disclaimer: Any opinion expressed is my own and does not represent those of my employer.
} We are in the midst of writing a proposal for a new film } scanner and I am hoping that someone out there can help } with a recommendation for a top of the line film scanner. } The major application would be TEM negatives. ...
I'll not knock the Nikon scanner, but I do believe your ultimate choice should be based on trial. Most film scanners anticipate normal films and normal exposures ... which addresses the film's optical density (... defined as Dmax minus Dmin ... approximately 3 f/stops for every OD unit ...). For normal negatives this generally doesn't exceed '3', and for normal positives, rarely exceeds '3.4'. However, the OD for x-ray films and electron induced exposures are reported to approach '4'. You should take one of your most problematic and representative films and test the scanners, paying particular attention to the detail/noise in the densest areas of the negative. I'll also mention ... you need not be limited to dedicated film scanners. I don't recall the PPI resolution of the Nikon scanner, but some of the newest flatbeds, together with their transparency options, I'm sure can approach the the resolution of the Nikon (e.g., 1600ppi for the new Epson) ... although I would tend to believe the film scanners are the most likely to deliver the best recognition for higher Dmax.
cheerios, shAf :o)
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
Hans, Could you please explain the main idea of the Vip-Quant algorithm. I cannot even imagine how quantitative analysis could have nearly the same accuracy as in high-vacuum mode without a priori knowledge of the composition of surrounding areas. Thank you,
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} -----Original Message----- } From: Hans Dijkstra [mailto:hans.dijkstra-at-edax.nl] } Sent: Tuesday, March 21, 2000 7:54 AM } To: Everett Ramer } Cc: Microscopy-at-sparc5.microscopy.com; Joergen Bilde-Soerensen 5802 } Subject: RE: EDS in Variable Pressure SEM } } } Dear Everett, } } The extrapolation method (aka Variable Pressure Method) } described by Dr. } Bilde-Sorenson has been implemented by EDAX in a software } feature called } ViP-Quant. With this method the EDS analysis under Bad-Vacuum } conditions can } be done with nearly the same accuracy as under High-Vacuum conditions. } Although this method was developed with the ESEM microscope } in mind, it of } course can be applied to all Bad-Vacuum scanning electron microscopes. } } Please contact your local EDAX representative for more } information and a } copy of the new ViP-Quant brochure, or request one through } www.edax.com. } } With best regards, } } Hans Dijkstra }
We are planning to upgrade our water supply from a reverse-osmosis, deionization system by adding a "polishing" system. Since our volume requirements are low, we're considering the Millipore Simplicity Ultrapure Water System. Specs for this countertop unit are a resistivity of 18.2 megaohms-cm, with total organic content of {15 ppb for the product water. The water would be used for routine EM use, i.e., making up reagants, staining, immunolabeling, etc.
My question is: does anyone have any experience with these systems and would you be willing to share it with us? Please feel free to respond off-line.
Thanks very much. 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/
We have a SprintScan 45, and while we don't work it to the bone, it has been relatively reliable(We've had it serviced once since it was purchased, and that was to have the internal motherboard replaced). I'm sorry to hear that you've had so many problems Paul. We've had ours since late 97, and it's been scanning consistently on a Win95 machine. It IS very fussy with SCSI--let this be a warning to everyone. It won't work with the new fast cards, but won't work with the really old cards either. We've currently got it on a Adaptec AHA 15XX series SCSI2 card with UW negotiation. We haven't been able to try it on a Win98 machine because all of our Win98 machines have SCSI cards which are too fast. I've heard good things about the Nikon(especially about Digital ICE). Minolta also has Digital ICE. The SprintScan isn't perfect, but it gets our job done. I would NOT recommend a flatbed with transparency adapter for dedicated film scanning tasks--in general graphic arts professionals agree that if you want to maximize scan quality from film, it's best to get the dedicated film scanner rather than the multi-option flatbed. Dedicated film scanners tend to have better bit depth too.
Of course, if money is no object, a drum scanner or an Imacon flextight scanner will bring you the best images.
Disclaimer--I have no financial stake in any of the aforementioned corporate entities blah blah blah
Sony makes 2 firewire cameras, the DFW-V300 and DFW-V500. Both are c-mount cameras and can be used on a light microscope. They are priced in a range of $2000 or so.
Seth Grotelueschen MIS, Inc. www.paxit.com sethg-at-paxit.com
Hans, Could you please explain the main idea of the Vip-Quant algorithm. I cannot even imagine how quantitative analysis could have nearly the same accuracy as in high-vacuum mode without a priori knowledge of the composition of surrounding areas. Thank you,
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} -----Original Message----- } From: Hans Dijkstra [mailto:hans.dijkstra-at-edax.nl] } Sent: Tuesday, March 21, 2000 7:54 AM } To: Everett Ramer } Cc: Microscopy-at-sparc5.microscopy.com; Joergen Bilde-Soerensen 5802 } Subject: RE: EDS in Variable Pressure SEM } } } Dear Everett, } } The extrapolation method (aka Variable Pressure Method) } described by Dr. } Bilde-Sorenson has been implemented by EDAX in a software } feature called } ViP-Quant. With this method the EDS analysis under Bad-Vacuum } conditions can } be done with nearly the same accuracy as under High-Vacuum conditions. } Although this method was developed with the ESEM microscope } in mind, it of } course can be applied to all Bad-Vacuum scanning electron microscopes. } } Please contact your local EDAX representative for more } information and a } copy of the new ViP-Quant brochure, or request one through } www.edax.com. } } With best regards, } } Hans Dijkstra }
Hi Ya'll: We are looking for a CCD camera for our Philips 430 TEM. We would like to get the best camera for the best price, e.g., either buying a used camera or a new non-Gatan camera (Gatan seems to be twice as expensive as the others). We would be using the camera for bright field and high resolution TEM of materials (semiconductors) rather than for biological specimens. Does anyone have a camera they would like to sell/donate or does anyone have recommendations as to a less-expensive camera that they know can be used for materials applications. Thanks, Mike Coviello Lab Manager University of Texas -at- Arlington
I probably have rotary pump oil in my oil vapor diffusion pump (long story, situation remedied, I hope). My question is, how well do I have to clean out the diffusion pump? Solvent clean and bake out, or only wipe down? What are the probable effects of contamination? Will it affect the vacuum quite a bit?
After overhauling the vacuum system on our Zeiss 10/A TEM because a faulty anti-suction device allowed (a LOT) of r.p. oil to migrate throughout the vacuum lines, I am only getting a vacuum of 6 X 10-4 when I should get 5 X 10-5. Either I've got a leak or a contaminated d.p. When I cleaned out the d.p. I only wiped it out well, but did not wash it with solvent and bake it out. I am trying to clean it in place rather than desolder the 13 wires (again) to get it all the way out of the scope, and then have to resolder (again) while lying on my stomach in the dark recesses of the instrument with glasses that are the wrong focal length... Call me lazy, if you wish, but I'm trying to find the easiest but effective way to do this!
Any tips and tricks appreciated!
Aloha, Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
The Sprintscan 45; some people swear by them, others swear at them.
I swear by mine. But only after much trouble, and what sounds identical to your experience. Here's what I found:
It really makes no difference as I see it whether one uses Win95 or Win98. The crux of the problem is the SCSI controller and how it talks to the scanner. The Sprintscan 35+ and the 45, the Agfa Arcus II, and several other scanners were totally flaky and crash prone on my system. I too sent back the Sprintscan 45 only to be told that it was fine. The SCSI adapter is a key item in this whole picture. Which adapter were/are you using?
At the time, I was running a FW SCSI system with an Adaptec AHA-2940UW. The rear plate connector on this host adapter is a 68-pin FW connector. The scanners are SCSI-I/II and are narrow, not wide....and definitely not fast. So, in order to run a scanner on a FW connector, one needs a FW--} SCSI-I cable and (this is the kicker) a high byte terminator on the host adapter external connector and a standard narrow SCSI terminator at the scanner. Good luck trying to get this high byte terminator. Adaptec has a part number for it but I gave up trying to get one from them. Maybe you will be luckier than I was.
The alternative is to connect to the narrow SCSI bus at the host adapter and snake that out the rear of the PC using a blank panel interface that accepts the 50-pin ribbon cable connector and presents a SCSI-II high density connector at the rear of the computer. This can work and it did with the Agfa scanner. But it did not work with the Sprintscans. Even when set for 10MB/s and no negotiation, it still would hang the system. The solution? I got a Mac G3/266. I still have it and I still use it for all scanning and photo input.
Very interesting, and totally successful. All scanners work flawlessly on the Mac's legacy external SCSI bus. I still am running FW SCSI inside for the disks, but using the external SCSI for scanners, CD-R, CD-RW and 8mm tape. The easiest way to run the scanners is via a common TWAIN interface plug-in in Photoshop. My current main flatbed is a UMAX Powerlook III. I have not tried it on my PC. My current PC is ATA-66 EIDE with an Adaptec AHA-2930 narrow SCSI adapter. It might run the UMAX and the Polaroid but everything is hooked up to the Mac. With DAVE on the Mac, all PCs, printers and the Mac talk to one another via TCP/IP on a 10BaseT LAN. Any node that needs external access gets it via an ISDN router. So there is no problem transporting files across platforms. Worst case is a Zip disk.
Any Mac system before the G4 should work fine. The pre-G3 systems are really cheap now. G3 systems can also be found rather inexpensively. A simple system is all that one needs. Heavy duty computing is done on the PC but photo input and scanning is done on the Mac. This solved all of my problems--and from your discussion, they were the same as you experienced.
Hope this helps.
gary g.
At 09:27 AM 3/22/00 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Sergey, I appreciate your feedback. It is obvious to me that you are pretty good at this. First, I was unclear about the message to which I was responding. I was responding to the subject "bacteriophage question" by way of Nestor. To my limited knowledge, UA will positively stain proteins-UA is used as a positive stain in thin section TEM. I understand the concept(s) of negative staining well, and I appreciate that UA is one of the best negative stains for high resolution work (small granularity+nice density). Totally asymmetrical molecules may be 3-D reconstructed. It just takes a brute force approach involving thousands of micrographs of the molecule of interest randomly orientated in vitreous ice. Although your point about symmetry makes good sense-a more symmetrical molecule should yield a higher resolution reconstruction (require fewer micrographs for the same level of confidence in a structure). Your point about intrument limitations is also well recieved. A quick concept: both TEM negative staining and Scanning Probe (AFM) studies of protein macromolecules usually involve the use of charged surfaces to facilitate adherance and/or orientation of the molecules (i.e. carbon+glow discharge). Some feel that this may distort the shape of the molecule (and indeed it will depending on the charge). In the case of Scanning Probe, there are those who would say that the "quasi-physical" interaction of the probe (presumably in tapping mode for molecules under physiological conditions) distorts the reconstructed image of the protein. I'ld like to say "sorry probe", but I don't have enough confidence in these statements to do so. Cheers, Karl G.
----- Original Message ----- } From: Sergey Ryazantsev {sryazant-at-ucla.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, March 22, 2000 1:09 AM
Dear Dorota
You have to use such amount of platinum that it will create a small drop of the melted metal at the end of the "V"-shape tungsten filament. If the drop will be large, it is very likely that filament will broke. It seems to me, 8 cm is a huge amount of platinum. Again, what is diameter of the filament? 20 mA is a tiny current, seems to me, for thermal evaporation. But, I never work on HUS 5GB, may be they have special setup. As for granulation, I don't think that platinum will dramatically improve the quality of your shadowing. Try platinum-palladium at least. May be, you have to try buffers, which are able to vaporize in vacuum: ammonium acetate or bicarbonate adjusted by CO2, for instance. For such applications I am using for many years 50-150 mM ammonium acetate buffer (with some additives if necessary, magnesium acetate, for instance) and tungsten shadowing by electron gun. I also use thickness monitor to control shadowing. I also use one or bi-directional shadowing: it resolves details better. I am using rotary shadowing only for DNA and not in all cases. If I have any problems with shadowing, I am using "test-object" - latex spheres to determine the problem. Sometimes the problem is just wrong angle of shadowing. If you have further questions, you may contact me off-listServer.
Sergey
} Date: Wed, 22 Mar 2000 10:13:39 -0400 } From: Dorota Wadowska {wadowska-at-upei.ca} } Subject: TEM-shadow coating } To: microscopy-at-sparc5.microscopy.com } Organization: University of P.E.I. } Priority: normal } } ------------------------------------------------------------------------ } 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
Our specimen was sprayed on to freshly cleaved mica, rotary shadowed with platinum, and carbon coated. We are having difficulty removing the carbon replica. Scoring around the edge did not help. Would anyone have a suggestion? Thankyou.
Donald Gantz Boston Univ School of Medicine gantz-at-med-biophd.bu.edu
Dear Listservers, Does anyone have a good method for skeletal muscle fixation for Transmission Electron Microscopy? We will not be able to use a perfusion method. A fixative method would also be greatly appreciated.
Many Thanks
Margery Stark Abbott Laboratories Department of Microscopy and Microanalysis D-45M/AP31 200 Abbott Park Rd. Abbott Park IL 60064-6202
Steve Chapman writes: } If you have a multi instrument laboratory I always feel } that it is a good idea to have instruments from different manufacturers } optimising the laboratory for optimising a wider range of applications. } Steve, You make some excellent points. However, I believe that, (just as you state) potential benefits can be had by choosing different manufacturers - at the same time you are missing some very significant opportunities if you create a facility with many different manufacturers. Two important, potential opportunities are: reduced training time and learning curves for multiple users, and, discounted service contracts from the manufacturer.
For example, we have in the recent past had the benefit of significantly reduced service contract costs by having multiple instruments from JEOL and Philips. At one time our research facility here, happened to have four JEOL EMs. These four SEMs covered three vastly different SEM instrumentation ranges (and eras) and serviced different applications. They were all very capable of handling the tasks for which they were purchased, and we were able to save big bucks each year over the cost of single instrument service contract prices. In addition we had multiple users who could go from one instrument to another without any significant additional training (or retraining) because of the relatively familiar user interface that accompanied these microscopes.
Just another perspective. Have Fun, Brad Huggins
} ---------- } From: Steve Chapman[SMTP:PROTRAIN-at-CompuServe.COM] } Sent: Wednesday, March 22, 2000 5:49 AM } To: American EM Soc } Subject: Buying a SEM } } ------------------------------------------------------------------------ } 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 } } A few weeks ago, when our server was working correctly (*??!!*), there } were } e-mails talking about the purchase of a new SEM. Now we appear to be back } on line I felt that our ideas may be of use to the mailers? } } As part of the consultancy side of our business we purchase SEM for our } clients. This means we are buying instruments regularly and are therefore } probably one of the few professional SEM purchasing organisations? } } In our team we have staff who have worked for most of the SEM } manufacturers } as demonstrators and application specialists, so we have seen the purchase } game from both sides and of course try to use this to our clients } advantage. Unlike the average microscopist, who probably only buys two } instruments in their career, we have been able over the years to formulate } a purchase plan, a purchasing criteria that we apply to each purchasing } project. In our position we are unable to buy from the nicest salesman or } the normal "gut" feeling, we must be absolutely certain that we are } purchasing the best instrument for our clients applications. } } Set out below are the headings from a lecture that I give on instrument } purchase. Only the basic ideas are presented ( I guess if you need more I } shall be attacked by Microscopy Today to give just that) but you will get } the theme. It is important to realise that the specimen chamber of a SEM, } the specimen-detector geometry, determines what we will see. For this } reason you will find that one instrument will do a particular task better } than any other in the price range, you need test specimens that will sort } this out for you. If you have a multi instrument laboratory I always feel } that it is a good idea to have instruments from different manufacturers } optimising the laboratory for optimising a wider range of applications. } } "Buying a New Microscope" } } How Do You Start? } } Collect ALL the brochures and prices } Form a view of the new facilities being made available } If you do not understand any features ASK the salesman } Check through all the users desired facilities? } Determine who will use the instrument now? } Would there be others if you purchased particular accessories? } } Formulate a Purchase Specification } } Price range - we always look one level higher } Set essential instrument specifications } Set desired instrument specifications give them points and produce a } "desirability assessment" } } Why use a consultant? } } Only he will without prejudice talk to all interested parties } Interdepartmental feuds could spoil the case } A different questioner provides different answers } Their wide knowledge base may develop additional features } They will have experience producing a "desirability assessment" which } often } brings surprises. } They will be experienced with ways of dealing with the sales staff, they } will keep them off your back } } How to handle the demonstration? DO NOT LET THEM } } Show you how wonderful the alignment procedures are - you will not spend } all day aligning the instrument! } Dominate YOUR demonstration } Take you into totally uninteresting areas of the instrument - you should } be } buying because of the image quality } Show their favourite gimmick } Use their own favourite specimen } Take you out of the room when about to load one of your specimens } Take you to a two hour lunch with lots of drink } } How to handle the demonstration before you set off } } Select a maximum of three specimens that you know really well and that are } important to your laboratory } The specimens must be capable of meaningful low and high magnification } imaging } Develop a demonstration criteria for each specimen } Have a tie break specimen available as suggested by the consultant } Provide each company with an exact demonstration programme } } What a consultant would do during a demo } } Time the demonstrator to produce images at specific magnifications under } very specific conditions } Encourage the demonstrator (this is not a customer v demonstrator } competition) allow them to try their own ideas with each specimen also, } give them information that you have found to be of importance with your } specimens } Stop the demonstrator taking you away from your standard path - you do } wish } to compare each instrument under exactly the same conditions there is no } time for other deviations } } After the demo } } Layout the results } Compare performance instrument to instrument and time taken to obtain the } results } Produce a short list } Compare the short list with the "desirability assessment" } Research local and national service performance of the best two instrument } manufacturers, and the manufacturers reputations } } Finally } Decide upon the instrument that you want, the specification and then } haggle } } Hope this helps? } } Steve Chapman } Senior Consultant } Protrain for Training and Consultancy in EM World Wide } Tel & Fax 44+ 1280 814774 } e-mail protrain -at-emcourses.com } web site www.emcourses.com }
Dear Karl, As my knowledge on thin sections, UA stains DNA and RNA, lead citrate - some proteins and OSO4 - some lipids. Sometimes UA (as well as any others stains) may just penetrate hydrophilic areas in the plastic sections (which correspondent, usually, with biological material areas). In this case, I agree, it may be named "positive staining". But in this case UA stain all areas, which accessible to water, therefore it is not specific "positive" stain for proteins.
As for 3D reconstruction. This is a subject for long and very interesting discussion. I know just a few examples of the complete 3D reconstruction of asymmetrical particles. Mostly, these works comes from Frank's (Spider program) and Marin van Heel (Imagic program) groups. The resolution on it is about 2 nm - not so impressive, if you count, that they did 3D on 26 nm ribosome. Frank's algorithm with random tilted particles (on the support film) has "dead angle" (between 60 and 90), therefore, the reconstruction generally speaking is not "complete". Van Hell's algorithm supposed to be free from such limitation, but it is not enough reconstructions performed to prove this technique. But, I am very optimistic about that in general. Both algorithms used "classification" of particles by type. Criteria for such classification determined by operator, therefore it is possible that in one class we will count slightly different particles (different shape or different orientation or both), averaging of these particles will enhance major features and lose fine details. Combination X-ray analysis and 3D reconstruction data (for phasing of the X-ray data set at the beginning) may help in this case. Marat Yusypov's grop did it for ribosome last year with great success. But, I have to point, using X-ray crystallography you studied some particular conformation under conditions of crystallization, which is not necessary correctly reflect the real conformation in solution.
As for Scanning Probe Microscopy, I agree that probe may distort the sample. To eliminate such problem, people scan the same area in different directions. If the images are the same, it means, that distortion from probe at least smaller than observed creatures. SPM may be very successful for membrane proteins (which are difficult for EM), pore complexes, etc. It is not necessary to use "modified" surfaces (glow-discharge, etc) for SPM or "negative staining" (I never use it for NS, the secret is a quality of the support carbon film). But, I agree with you, Karl that, as any "microscopy", SPM is limited in some way in their abilities. Sorry, STM.
Karl, thank you so much for such nice discussion.
Sergey.
} Date: Wed, 22 Mar 2000 15:39:32 -0600 } From: Karl Garsha {keg-at-csd.uwm.edu} } Subject: Re: neg stain vs cryo EM plus image averaging } To: Microscopy-at-sparc5.microscopy.com, Sergey Ryazantsev {sryazant-at-ucla.edu} } X-Mailer: Microsoft Outlook Express 5.00.2615.200 } X-MSMail-Priority: Normal } X-MimeOLE: Produced By Microsoft MimeOLE V5.00.2615.200 } } Dear Sergey, } I appreciate your feedback. It is obvious to me that you are pretty good at } this. First, I was unclear about the message to which I was responding. I } was responding to the subject "bacteriophage question" by way of Nestor. } To my limited knowledge, UA will positively stain proteins-UA is used as } a positive stain in thin section TEM. I understand the concept(s) of } negative staining well, and I appreciate that UA is one of the best negative } stains for high resolution work (small granularity+nice density). } Totally asymmetrical molecules may be 3-D reconstructed. It just takes } a brute force approach involving thousands of micrographs of the molecule of } interest randomly orientated in vitreous ice. Although your point about } symmetry makes good sense-a more symmetrical molecule should yield a higher } resolution reconstruction (require fewer micrographs for the same level of } confidence in a structure). } Your point about intrument limitations is also well recieved. } A quick concept: both TEM negative staining and Scanning Probe (AFM) } studies of protein macromolecules usually involve the use of charged } surfaces to facilitate adherance and/or orientation of the molecules (i.e. } carbon+glow discharge). Some feel that this may distort the shape of the } molecule (and indeed it will depending on the charge). } In the case of Scanning Probe, there are those who would say that the } "quasi-physical" interaction of the probe (presumably in tapping mode for } molecules under physiological conditions) distorts the reconstructed image } of the protein. I'ld like to say "sorry probe", but I don't have enough } confidence in these statements to do so. } Cheers, } Karl G. } } ----- Original Message ----- } From: Sergey Ryazantsev {sryazant-at-ucla.edu} } To: {Microscopy-at-sparc5.microscopy.com} } Sent: Wednesday, March 22, 2000 1:09 AM } Subject: FW: neg stain vs cryo EM plus image averaging } } } } ------------------------------------------------------------------------ } } 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 Karl } } I disagree with you, that "UA in particular, will positively stain } } proteins". Let start from definition: positively stain object is a dark } } object on the light background (therefore stain penetrate/interact with } } object making it "stained", dark in the terms of EM); "negative staining" } } (NS) is opposite. We have light (stain do not interfere with the sample) } } object on the dark background of stain (or dark ring of stain around the } } object, depends from stain and technique). The beauty of the NS is that } } stain (UA in particular) easily penetrates cavities in the object making } } visible fine details (for instance, I was able to recognize variable and } } constant domains of the Fab of the IgG molecules as well as domains of the } } Fc using UA staining). May be this effect you call "positively staining". } } But it is not. } } } } Yes, UA may stain RNA and DNA positively. For this reason UA gives us } } "mixed stain" for ribosomes (negative for proteins and positive for RNA } } components). The disadvantage UA is its low pH. } } } } As for subject of the current discussion. I think Cryo in general will } } give us more correct information than others EM techniques. But, we have } } to keep in mind, that in Cryo-technique, to make image relatively } contrast, } } people should go to very high overfocusing (from half to couple microns, I } } believe). For this reason, the best Cryo-results I know, yielded only 2-3 } } nm resolution (and this is a huge problem in Cryo - to get better } } resolution): the same or even worse as for "negative staining" (I expect } } 1.5 nm resolution for UA). We have to count resolution when compare the } } data. I lost the original message from which this discussion was started, } } but it seems to me, that difference between NS and Cryo was not so } dramatic } } and may be comparable if we will count the resolution. In general, I } don't } } believe any EM data if resolution is not shown. Talking about resolution } } is complicated because of the difference between "resolution of the } } instrument" (0.14 nm for TEM currently), "physical resolution of the } } method" (resolution limit for overfocusing, for instance, radiation } damage, } } drift, noise/signal ratio etc) and "resolution of the sample" (for } } biological samples they are not the same: preparation of the sample may } } affect sample's intactness/structure). Cryo is the best for sample } } preservation, but it is low in contrast. UA may affect sample's structure } } (may not) but the resolution limited only by granularity of stain (in } } theory it is a couple angstroms). } } } } As for "averaging" of the images, I am a little bit skeptical about that. } } It is great deal if your sample is uniform in shape (symmetry is a great } } help too). If your biological sample is functionally flexible (IgG for } } instance), averaging will "hide" some interesting details even if you will } } distribute images in classes. } } } } In my point of view, it is difficult to extract absolute numbers from EM. } } Inside one methodology you could easily compare the data, but it becomes } } difficult when you want to compare the data obtained by different } } techniques. This is reality of EM. I think Scanning Probe Microscopy is } } very promising to obtain the direct measurements on the samples under } } physiological conditions (EM is so far from "physiological conditions", } } unfortunately). I am so sorry, EM! } } } } } Date: Mon, 20 Mar 2000 16:46:25 -0600 } } } From: "Karl E. Garsha" {keg-at-csd.uwm.edu} } } } Subject: FW: neg stain vs cryo EM plus image averaging } } } To: microscopy-at-sparc5.microscopy.com } } } X-Mailer: Microsoft Outlook Express for Macintosh - 4.01 (295) } } } } } } ------------------------------------------------------------------------ } } } 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 } } } -----------------------------------------------------------------------. } } } } } } } } } } } } } } } _____________________________________________________ } } } } } } Karl E. Garsha, Doctoral Student } } } AOP Fellow } } } Chief Coordinator-Graduate Student Association } } } Department of Biological Sciences } } } University of Wisconsin-Milwaukee } } } P.O. Box 413 } } } Milwaukee, WI 53201 } } } Office: 459 Lapham Hall } } } Phone: (414) 229-4316 } } } Mobile: (414) 617-4295 } } } E-Mail: keg-at-csd.uwm.edu } } } } } } ---------- } } } } From: "Karl E. Garsha" {keg-at-csd.uwm.edu} } } } To: maxwell-at-lec.med.utoronto.ca } } } Subject: neg stain vs cryo EM plus image averaging } } } Date: Mon, Mar 20, 2000, 4:45 PM } } } } } } } } } Dear Karen, } } } My gut reaction: cryo-EM/3-D reconstruction will yield a much more } accurate } } } value with regard to measurement of macromolecular features. } } } Some "negative" stains, UA in particular, will positively stain } } } proteins, and so it would be expected that pore size might, in theory, be } } } over-estimated in neg. stain images. This is a shortcoming of neg. } staining } } } as opposed to cryo-EM (if the protein isn't glycosylated). I have no } doubt } } } that investigators more experienced than I would argue this point } however. } } } The real power of 3-D reconstruction is that it averages many images } and } } } determines a structure that is statistically probable...the derived } } } structure has a certain level of confidence. 3-D reconstruction, as well } as } } } 2-D averaging, may be performed on either neg. stained specimens } (Wilkens, } } } et. al., Journal of Biological Chemistry, 1999) or cryo-prepared } specimens } } } (many important citations...one of my favorites is Boisset, et. al., } Journal } } } of Structural Biology 109, 39-45 (1992). } } } Either way you need the computer software/hardware and theory } (Journal } } } of Structural Biology Vol. 116, Number 1, 1996). } } } My main point is that a (responsibly) digitally enhanced image should } } } provide more accurate measurement of macromolecular features than a raw } } } negative stain photo. } } } Best Regards, } } } Karl G. } } } } } } } } } _____________________________________________________ } } } } } } Karl E. Garsha, Doctoral Student } } } AOP Fellow } } } Chief Coordinator-Graduate Student Association } } } Department of Biological Sciences } } } University of Wisconsin-Milwaukee } } } P.O. Box 413 } } } Milwaukee, WI 53201 } } } Office: 459 Lapham Hall } } } Phone: (414) 229-4316 } } } Mobile: (414) 617-4295 } } } E-Mail: keg-at-csd.uwm.edu } } } } } } } } _____________________________________ } } } } Sergey Ryazantsev Ph. D. } } Electron Microscopy } } UCLA School of Medicine } } Department of Biological Chemistry } } Box 951737 } } Los Angeles, CA 90095-1737 } } } } Phone: (310) 825-1144 } } Pager: (310) 845-0248 } } FAX (departmental): (310) 206-5272 } } mailto:sryazant-at-ucla.edu } } http://www.bol.ucla.edu/~sryazant } } } } } } } } } } _____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
I thank you as well for your knowledge and insight, Dr. Ryazanstev. I've enjoyed this discussion. -Karl ----- Original Message ----- } From: Sergey Ryazantsev {sryazant-at-ucla.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, March 22, 2000 5:26 PM
At 05:59 PM 3/21/00 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
It exists. But...and this is a big but....what performance do you seek? Most of these, at least from what I have seen, are not industrial strength. They are more consumer- than research-grade units. But maybe this will satisfy your needs. The higher quality units tend to use SCSI from what I have seen and used.
Donald Gantz wrote: =============================================== Our specimen was sprayed on to freshly cleaved mica, rotary shadowed with platinum, and carbon coated. We are having difficulty removing the carbon replica. Scoring around the edge did not help. Would anyone have a suggestion? Thankyou. =============================================== There are probably as many different ways to approach this kind of problem as there are people doing platinum replicas! The most reliable method we have ever discovered is to use a 1% aqueous polyacrylic acid solution, deposited as one drop on the surface with the stubborn replica. Twenty four hours later, the drop has spread, the water has evaporated leaving a thin but very tough PAA film. Then with something sharp (e. g. scalpel blade, but wear eye protection for this!), slide the blade edge underneath the edge of the PAA film, and if you have the "art", it will literally just "pop off" . Now you have the tough PAA layer and the carbon coating, and where it once rested on the mica, should now be a corresponding area free of replica.
Next the PAA is floated on a surface of water, carbon side up, and again, do other things and come back 24 hours later. The PAA will have all dissolved into the water, leaving the carbon/Pt film floating on the surface of the water. The film is then picked up on grids as you would any other floating film.
Note: Patience of clearly a virtue, be sure to give it the full 24 hours or your grids will be PAA contaminated, with a major loss of contrast. Use a deep petri dish for this so that there is sufficient volume of water to efficiently dissolve the PAA.
If this sounds too complicated and you don't have patience, there is an alternative we also use, it involves the use of Victawet® for EM (see our URL http://www.2spi.com/catalog/spec_prep/victawet.html
The Victawet is evaporated from a tungsten basket (see website instructions) and a very thin layer of the release agent is deposited onto the mica (or glass slide). Then apply your samples, shadow with Pt/C and the replica now is almost guaranteed to float off on the first try.
One note: The "better" the grade of mica, I am told, the easier is the release of such films. Grade V1 mica supposedly releases easier and that might be because there are fewer cleavage steps. We have not tested that theory ourselves so on that there can be no guarantees.....but it does make sense. If you are not familiar with the different mica "grades", see URL http://www.2spi.com/catalog/submat/chart.html
Disclaimer: SPI Supplies is a supplier of Victawet for Electron Microscopy and mica substrate materials.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.spi.cc ######################## ============================================
I have one of these for sale. I may have two. These are specifically for microscopy. At highest resolution, they take a 28MB TIF file. This is as high of resolution I have ever seen. Interface is SCSI-II small sized narrow-SCSI connector.
For platinum: try 1-10% aqueous HF. Use it instead water to float replica. Insert mica slowly. Pickup replica on the grid and rinse with water. Use corrosion-resistant tweezers. Do not immerse grids into HF solution if it is copper. You have to do it fast to avoid corrosion of the grid. I was using copper grids with carbon perforated (holey) film. Perhaps, carbon protected grid form HF. This is easiest way for platinum shadowing.
You may use even 50% HF - it did not affect platinum shadowing as well as carbon. Be careful, HF is extremely corrosive, avoid direct contact and vapors.
If you have questions, you may contact to me off-line.
Good luck!
Sergey
} Date: Wed, 22 Mar 2000 17:31:48 -0400 (EDT) } From: "GANTZ-at-med-biophd.bu.edu"-at-sparc5.microscopy.com } Subject: Removal of Carbon Replica from Platinum-shadowed sample } To: MICROSCOPY-at-sparc5.microscopy.com } X-VMS-To: MICROSCOPY-at-MSA.MICROSCOPY.COM } } ------------------------------------------------------------------------ } 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 would like to thank those who have replied to my query about diff pump contamination. Most have asked for more details...
How long have I let the scope pump after cleaning it out and changing diff pump oil? Just shy of two weeks. The most improvement in vacuum came within 4 days, then leveled off.
Do I think I got oil into the viewing chamber or column? Yeah, lots. Lots. Most was on the anode plate, and quite a few droplets on the liquid nitrogen anticontaminator plate. And even a drop hanging off the filament! I can see all of you squirming... I cleaned the entire column, including a couple of parts I had never been into before, and all the vac lines. I was as thourough as possible, under the circumstances.
I presume I will have a fair amount of vapor around for years to come. However, the scope operation and resolution aren't bad, considering. The only thing that keeps me from slitting my wrist is that we have this great, new, fancy LEO 912 EFTEM that I have been trying to get all my users trained on, anyway. This means that the old Zeiss 10 can be my hobby scope. It reminds me of the Volkswagens I used to work on. Great German engineering!
So one friend thinks that small amounts of rotary pump oil in the diffusion pump will "burn off" and cease to be a problem, one thinks that it will only migrate up into the column, and a couple of third party service people don't have a clue what to expect. No "official" word from Zeiss/LEO yet. Hoping some of you clever microscopists will have either direct knowledge or a grand theory backed by thoughtful physics!
Aloha, Tina
Life's not all fun and Photoshop. **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
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As they say, "It's the simple ideas that make money." And this is SIMPLE & GREAT INCOME!
Our research has found that many people have tried one or more of the following...
Free Classifieds? They just don't work anymore Web Site? Takes thousands of surfers Banners? Expensive and losing their punch E-Zine? Hope they have a -huge- subscriber list Search Engines? Forget it, unless you're in the top 20
S O W H A T D O E S W O R K ?
Although often misunderstood, there is one method that has proven to succeed time-after-time.
E - M A I L M A R K E T I N G ! !
IT'S A FACT... It if you're not using your computer to generate GOOD INCOME, you're leaving money on the table.
Here's what the experts have to say about E-Mail Marketing:
"A gold mine for those who can take advantage of bulk e-mail programs" -The New York Times
"E-mail is an incredible lead generation tool" -Crains Magazine
"Blows away traditional Mailing" -Advertising Age
Here's a potential earnings example: Let's say you have a product or service that can bring a profit of around $30. Remember, on the Internet, you can make money 7 days a week, 24 hours a day... even while you sleep, orders come from all over the world!
The way to reach thousands of people, generate orders and build wealth is person-to-person direct.
1. How do you find the millions of people on the Internet?
2. What are you going to tell them when you do reach them?
HERE'S THE ANSWER TO QUESTION #1
M I L L I O N S V O L U M E 9
***New - 10 Million addresses - Just Released***
The cleanest, most comprehensive e-mail address list in the world! We're proud to offer it.
O N E O F A K I N D
This is a first. No one has gone to the work it takes to produce an e-mail address list of this quality.
Here's how we prepare our e-mail lists:
1. We clean and eliminate all duplicates.
2. Next, we use a filter list of 400+ words/phrases to clean even more. No address with inappropriate or profane wording survive!
3. Then we used our private database of thousands of known "extremists", opposed to commercial e-mail, and kicked off every one we could find.
4. And finally, we sorted the list into easy-to-manage packets of 20,000 addresses in a simple text (.txt) format.
5. All domains have been verified as valid.
WHAT DID WE END UP WITH?
Volume 9... 10 Million Addresses Strong!
An address list so clean you'll want to start mailing today!
N O B R A G - J U S T F A C T
With our super clean e-mail address lists you'll send less...and get better results...
* Y O U G E T W H A T Y O U P A Y F O R *
Our NEW 10 Million, Volume 9, address list CD will result in:
* Higher Response Rates * Higher Sales Conversion Ratios * More Receptive prospects; Less Flames & Non-Buyers. * Less Contact With Anti-Commerce Radicals & Extremists.
Remember that potential income chart at the beginning of this message? Can you imagine the kind of money you could make if you mailed one million pieces and sold only one tenth (.01%) of one percent? You do the math, you'll be amazed!
We've been in the list brokerage business for over 5 years and we've never compromised on quality. We won't release any address list until it passes our "high standards" test.
This is not a rental list that is restricted to a one-time mailing. You are purchasing an e-mail address list for your own personal mailings and may use it over-and-over.
DON'T HESITATE on this offer or you will miss out on the least expensive, legal and most effective way to market... PERIOD!
Order within 72 hours and we'll include the following FREE Bonuses... we call this our "BUSINESS ON A CD" bonus.
1. To help you get started we include basic proven Professional Mailing Software. This software has sold for as high as $499.00 in the past. No demo, but a full working version (SORRY, SINCE THE SOFTWARE IS FREE WE CANNOT OFFER ANY TECHNICAL SUPPORT, however set-up instructions are included).
2. Every survey has always indicated that the most profitable product to sell on the Internet is INFORMATION!
Our "BUSINESS ON A CD" gives you 650 reports/manuals/books that are yours to use and sell. With these "Special Reports" you may instantly start your "Information Product" business... plus a sample SALES LETTER is included to help you GET STARTED FAST!
3. "THE BULK E-MAIL SURVIVAL GUIDE" A manual/guide that addresses the Bulk E-Mail business. Especially useful for beginners. "THE BULK E-MAIL SURVIVAL GUIDE" will answer most of your questions and concerns about Bulk E-Mail. An exclusive for our customers... INCLUDED FREE.
4. "LISTMATE" - This is the software the Pro's use to manage their mailing lists. We've included two versions, both are fully functional demo's, the only limit is the file size.
5. "SCIENTIFIC ADVERTISING"! This book is responsible for untold millions of dollars in sales and profits. Many of today's internet "gurus" have used this powerful book as the foundation for marketing courses that they have written & sold for as much as $495. Marketeer's that have studied this book have been so deeply inspired, that it has changed their entire way of doing business, and they've gone on to make fortunes -- it's yours FREE with your order!
This "BUSINESS ON A CD" bonus is yours absolutely FREE if you order within the next 72 hours --- After that... . it's gone!
***SPECIAL BONUS*** Order within the next 72 hours and receive an additional 972,565 e-mail addresses as a prompt ordering bonus. Order Now!
D O N ' T H E S I T A T E E-Marketing is the most effective and fastest way to market anywhere... PERIOD!
O R D E R N O W . . . SAME DAY SERVICE (M-F) if your order is received before 2pm Pacific. 24hour fax service, just fax to: 1-435-806-3011
To order, via credit card simply cut/paste and print out the EZ ORDER FORM below and fax to our office today.
**** MILLIONS CD - Volume 9 ****
**** NOW ONLY $247! ****
This "Special Price" is in effect for the next 72 hours, after that we go back to our regular price of $299.00 ... Don't delay... you can be in business tomorrow!
We accept Visa, Mastercard, Amex and Checks by Fax. Fax your order to: 1-435-806-3011
----------------------Cut & Paste---------------------- ---------------------EZ Order Form---------------------
_____Yes! I want everything! I am ordering within 72 hours. Include my FREE "Business On A CD" bonus along with your 10 Million Vol. 9 E-Mail address CD (plus 972,565 bonus addresses) for the special price of only $247.00 + shipping as indicated below.
_____Oop's I missed the 72 hour "special". I am ordering Vol. 9 at the regular price of $299.00 + shipping.
***PLEASE SELECT YOUR SHIPPING OPTION***
____I would like to receive my package FedEx OVERNIGHT. I am including $15 for shipping. (Hawaii & Alaska $20 - Canada $25, all other International add an *additional* $25 [$40 total] for shipping)
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(Required) SIGNATURE:x_________________________________ I understand that I am purchasing the Millions Vol. 9 e-mail address CD, the addresses are not rented, but are mine to use for my own mailing, over-and-over. Free bonuses are included, but cannot be considered part of the financial transaction. I understand that it is my responsibility to comply with any laws applicable to my local area. As with all software, once opened the CD may not be returned, however, if found defective it will be replaced with like product at no charge.
You may fax your order to us at: 1-435-806-3011
CHECK BY FAX SERVICES!
Please Note: Sorry, we can only accept checks drawn on U.S. banks.
If you would like to fax a check, tape your check below and fax it to our office along with the EZ Order Form to: 1-435-806-3011
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This sounds very like the problem we had removing polymer single crystals deposited on mica. The heavy metal tends to "key" to the mica, as it also does if one is shadowing a bulk polymer specimen directly. What we do in both cases is to put on a tiny amount of carbon first, about one-tenth of the thickness of the main carbon film.
If the chemistry of your specimen allows, one might spray onto a polystyrene surface, and then dissolve the PS with butanone.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
On Wed, 22 Mar 2000 GANTZ-at-med-biophd.bu.edu-at-sparc5.microscopy.com wrote:
} Our specimen was sprayed on to freshly cleaved mica, rotary } shadowed with platinum, and carbon coated. We are having difficulty } removing the carbon replica. Scoring around the edge did not help. } Would anyone have a suggestion? Thankyou. } } Donald Gantz } Boston Univ School of Medicine } gantz-at-med-biophd.bu.edu }
More chemical can access the replica if the replica is scored into 3mm squares - which is required for TEM anyway. A backing of wax or plastic is useful if the replica tends to fall apart - I don't expect that this would happen on a mica surface. For separating and cleaning use a spotting plate or a small watchglass within a Petrie dish. I suggest to immerse the mica (or replicas) alternating in a solution of chromic acid (made up from Pot dichromate) and then concentrated household bleach - with a water rinse in between. Leave the mica and later the specimen in those solutions for several hours or over night. 2 periods in both solutions would be minimal for most tissues, but in your case, once the replica has separated, the actual cleaning should be minimal. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Thursday, March 23, 2000 7:32 AM, "GANTZ-at-med-biophd.bu.edu"-at-sparc5.microscopy.com [SMTP:"GANTZ-at-med-biophd.bu.edu"-at-sparc5.microscopy.com] wrote: } } Our specimen was sprayed on to freshly cleaved mica, rotary } shadowed with platinum, and carbon coated. We are having difficulty } removing the carbon replica. Scoring around the edge did not help. } Would anyone have a suggestion? Thankyou. } } Donald Gantz } Boston Univ School of Medicine } gantz-at-med-biophd.bu.edu
Doehne, E., A New Correction Method for High-Resolution Energy-Dispersive X-ray Analyses in the Environmental Scanniing Electron Microscope. Scanning 1997, Vol. 19, 75-78.
Carlton, R. A., Energy Dispersive X-ray Spectrometry in the Environmental Scanning Microscope. Microscope 1999, Vol. 47:1, 5-11.
Cheers,
Stephen M. Harmon Electron Microscopist United States Environmental Protection Agency M.S. 681 26 W. Martin Luther King Blvd. Cincinnati, OH 45268 513.569.7184 Harmon.Stephen-at-epa.gov
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It seems that nowadays every SEM vendor is offering variable pressure models, which are conventional SEMs with a plumbing modification that allows the sample to be at pressures of up to about 4 torr (500 Pa) while the electron column operates at the conventional high vacuum. I am very interested in buying a variable pressure SEM with an EDS, but was recently warned that EDS has very poor spatial resolution in the variable pressure mode because of the large beam spread due to electrons scattering off the gas molecules in the sample chamber. Is this really a significant issue? Do any of you have experience using EDS with variable pressure SEMs? Thanks,
Hi, I am interested in buying an ultramicrotome which is not necessary be a new one. Does anybody have idea where I should contact? Thank's in advances. Sincerely,
Young Ho Koh NSB program UMass Amherst, MA 01003 kohyh-at-bio.umass.edu
At 4:45 PM -0600 3/22/0, Stark,Margery wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
**************************
I've worked with muscle (skeletal and cardiac) for years. If you can't perfuse, you need to keep the muscles from contracting down on themselves. Depending on the size of the muscle to be fixed, it can be handled as a whole or blunt dissected into small (1-2mm diam) bundles and tied to applicator sticks before immersing it in fix. It helps if you've boiled the sticks in water for 10 minutes or so to extract any resins, etc. The smaller the bundle, the better your initial fixation. You may want to wash the bundles in bufer with "high" potassium (100mM KCl) to relax it. I fix in 2.5% glut, 4% paraform. + approx. 0.02% picric acid (2 ml of a sat'd sol'n in 40 ml of fix) in 0.1 M Na-cacodylate buffer(Ito & Karnovsky J Cell Bio 89(abstr. 418) 1968). You can keep the 100mM KCl in this too. I would suggest that you fix for 30 min or so at RT them overnight in the 'fridge. Wash well, then cut into smaller pieces before osmium. dehydrate, etc as usual. I like Spurr's resin, but feel free to use your favorite.
Good luck, Lee
Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Confocal Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
University of Central Florida Advanced Materials Processing and Analysis Center (AMPAC)
The Advanced Materials Processing and Analysis Center (AMPAC) is seeking candidates for a position as Assistant in Research to support the Materials Characterization Facility (MCF). The individual must have a Master degree or a Bachelor degree from an accredited institution in an appropriate area of study related to surface science and materials characterization, and have at least two years of experience in materials characterization or related areas. The person will be responsible for establishing and maintaining laboratory infrastructure, laboratory maintenance, equipment installation, operation and maintenance, and be an investigator in contracted research. The person must be able to conduct independent research and development in materials characterization with an emphasis in ion beam technology and have the ability to interact with students and provide instruction in the use and interpretation of data. The individual must have extensive knowledge and experience in the operation and maintenance of materials characterization equipment. AMPAC is particularly interested in individuals desiring an academic setting to further their professional goals. The applications will be reviewed beginning April 17, 2000 and will continue to be reviewed until the position is filled.
Applicants should send vitae and three references to Dr. Vimal Desai, Director, 12424 Research Parkway, Suite 408, Orlando, FL 32826. UCF is an equal opportunity/affirmative action employer. As an agency of the State of Florida, all application materials and selection procedures are available for public review.
******************************************************************* Lucille A. Giannuzzi, Ph.D.
Associate Professor, Mechanical, Materials, and Aerospace Engineering Director, UCF/Cirent Materials Characterization Facility University of Central Florida 12443 Research Parkway, Suite 305 Orlando, FL 32826 email lag-at-mail.ucf.edu phone (407) 275-4354,5,6 fax (407) 275-4321 -------------------------------------------------------------------- "Good judgement comes from experience.
Experience comes from making bad judgement."
Mark Twain ********************************************************************
Hi Ya''ll: In reference to my previous message: I'm sorry I might have given the impression that I feel the Gatan digital camera is overpriced...We have lots of Gatan equipment and are extremely happy with it. Maybe I should have said: we would like to know what are the lower-priced options for a CCD camera for a Philips 430 (without singling out Gatan). I am sorry for any offense this may have caused. Regards, Mike Coviello University of Texas -at- Arlington
The main idea of the ViP-Quant procedure is rather straight-forward and elegant. It is based on the phenomenon that as the pressure increases the size of the skirt effect remains fairly constant, although the number of skirt electrons increases, and thus the contribution of the skirt-effect to the EDS signal increases. This intensity increase is linear with the pressure.
This is valid for low- to medium pressures, or at a higher (ESEM) pressure with a very short free-path length of the skirt electrons, since in both cases you can assume the skirt electrons have scattered only once. At higher pressures with longer working distances these assumptions are no longer fully valid.
So to do an EDS analysis you take 2 measurements at different pressures, the low-pressure one at a pressure where you can just avaid charging, and a high-pressure one at at least twice that pressure. The measured intensities of all elements are then plotted as a function of pressure, and extrapolated to pressure zero, i.e. high vacuum. The extrapolated results will be very close to the results hat would have been obtained directly if the sample could have been analyzed at high vacuum conditions.
Of course anyone can do this plotting and extrapolation manually, that is: if your EDS software allows you to enter intensities manually! The only thing the EDAX ViP-Quant is offering as an extra is that the software does it automatically for you. There are no proprietary miracles involved, EDAX has just listened carefully to what science has to offer....
Best regards,
Hans Dijkstra
Disclaimer: The above is my personal opinion, and not necessarily EDAX's. ------------------------------------------------------------- EDAX Europe www.edax.com Ringbaan Noord 103 Tel.: +31-13-5364000 P.O. Box 4144 Fax.: +31-13-5356279 5004 JC Tilburg E-mail: hans.dijkstra-at-edax.nl the Netherlands -------------------------------------------------------------
} -----Original Message----- } } From: "Dusevich, Vladimir", INTERNET:DusevichV-at-umkc.edu } To: Everett Ramer, INTERNET:Everett.Ramer-at-netl.doe.gov } "'Hans Dijkstra'", INTERNET:hans.dijkstra-at-edax.nl } } CC: Joergen Bilde-Soerensen 5802, INTERNET:j.bilde-at-risoe.dk } [unknown], INTERNET:Microscopy-at-sparc5.microscopy.com } } Date: 3/22/00 4:35 PM } } RE: RE: EDS in Variable Pressure SEM } } } ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } Hans, } Could you please explain the main idea of the } Vip-Quant algorithm. I cannot even imagine } how quantitative analysis could have nearly the } same accuracy as in high-vacuum mode without } a priori knowledge of the composition of } surrounding areas. } Thank you, } } Vladimir } } } Vladimir M. Dusevich, Ph.D. } Electron Microscope Lab Manager } 3127 School of Dentistry } 650 E. 25th Street } Kansas City, MO 64108-2784 } } Phone: (816) 235-2072 } Fax: (816) 235-5524 } Web: http://www.umkc.edu/dentistry/microscopy } } } -----Original Message----- } } From: Hans Dijkstra [mailto:hans.dijkstra-at-edax.nl] } } Sent: Tuesday, March 21, 2000 7:54 AM } } To: Everett Ramer } } Cc: Microscopy-at-sparc5.microscopy.com; Joergen Bilde-Soerensen 5802 } } Subject: RE: EDS in Variable Pressure SEM } } } } } } Dear Everett, } } } } The extrapolation method (aka Variable Pressure Method) } } described by Dr. } } Bilde-Sorenson has been implemented by EDAX in a software } } feature called } } ViP-Quant. With this method the EDS analysis under Bad-Vacuum } } conditions can } } be done with nearly the same accuracy as under High-Vacuum conditions. } } Although this method was developed with the ESEM microscope } } in mind, it of } } course can be applied to all Bad-Vacuum scanning electron microscopes. } } } } Please contact your local EDAX representative for more } } information and a } } copy of the new ViP-Quant brochure, or request one through } } www.edax.com. } } } } With best regards, } } } } Hans Dijkstra } } } } } } } } } ----------------------- Internet Header -------------------------------- } Sender: Microscopy-request-at-sparc5.microscopy.com } Received: from ultra5.microscopy.com ([206.69.208.10]) } by spdmgaac.compuserve.com (8.9.3/8.9.3/SUN-1.9) with ESMTP } id LAA16645; } Wed, 22 Mar 2000 11:34:33 -0500 (EST) } Received: (from daemon-at-localhost) } by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id KAA01363 } for dist-Microscopy; Wed, 22 Mar 2000 10:27:23 -0600 (CST) } Received: from no_more_spam.com (sparc5 [206.69.208.10]) } by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id KAA01359 } for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Wed, 22 } Mar 2000 10:26:53 -0600 (CST) } Received: from UMKC-MAIL01.umkc.edu (email.exchange.umkc.edu } [134.193.71.1]) } by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id KAA01352 } for {Microscopy-at-sparc5.microscopy.com} ; Wed, 22 Mar 2000 } 10:26:41 -0600 (CST) } Received: by umkc-mail01 with Internet Mail Service (5.5.2650.21) } id {HJ6P423S} ; Wed, 22 Mar 2000 10:21:30 -0600 } Message-ID: {95A711A70065D111B58C00609451555C01CA7552-at-UMKC-MAIL02} } From: "Dusevich, Vladimir" {DusevichV-at-umkc.edu} } To: "'Hans Dijkstra'" {hans.dijkstra-at-edax.nl} , } Everett Ramer } {Everett.Ramer-at-netl.doe.gov} } Cc: Microscopy-at-sparc5.microscopy.com, } Joergen Bilde-Soerensen 5802 } {j.bilde-at-risoe.dk} } Subject: RE: EDS in Variable Pressure SEM } Date: Wed, 22 Mar 2000 10:21:28 -0600 } MIME-Version: 1.0 } X-Mailer: Internet Mail Service (5.5.2650.21) } Content-Type: text/plain; } charset="iso-8859-1" } Errors-to: Microscopy-request-at-sparc5.microscopy.com } }
I sometimes found this and never had a great explanation of why it happened. I would make sure that your bell jar is clean, i.e.free from oil. I did find that the thickness of the carbon sometimes mattered. If it was too thick it shattered. Too thin you cannot see it or it won't stay together. Sometimes in between it would not float off.
Good luck.
ML } } } Our specimen was sprayed on to freshly cleaved mica, rotary } shadowed with platinum, and carbon coated. We are having difficulty } removing the carbon replica. Scoring around the edge did not help. } Would anyone have a suggestion? Thankyou. } } Donald Gantz } Boston Univ School of Medicine } gantz-at-med-biophd.bu.edu
Mei Lie Wong Electron Microscope Laboratory Department of Biochemistry HHMI-UCSF Ph. 415-476-4441 Fax 415-476-1902 http://util.ucsf.edu/agard/wong/index.html email wong-at-msg.ucsf.edu
At 11:25 AM -0500 3/23/0, koh young ho wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
***************** for used instruments in the NY Metropoitan are you can try: M.O.C. at (914)268-6450 and Marcus Meyerhoff (I'lm not sure I've spelled that correctly) at (732) 747-6228
My name is George Laing from National Graphic Supply. NGS is a vendor of electronic imaging and traditional photographic products to scientific markets. Our customers have had excellent results scanning TEM negatives with the Agfa Duoscan T2500 scanner. The T2500 looks like a traditional "flatbed" scanner but utilizes a unique "Twin Plate" design similar to a negative carrier, that eliminates the use of glass in the film holder. This eliminates Newton rings,dust on the scanner glass, etc. Optical resolution is 2500x2500ppi(maximum resolution 5000x5000), Dmax is 3.5. The T2500 has Apochromatic optics and also includes glassless film holders for 35mm, 120/220 and 4x5 films. Also included is a glass drawer for transparent originals up to 8"x10". Agfa's Fotolook software is included for Mac and Windows, interface is SCSI. In addition, the T2500 will also scan reflective originals up to 8.5" x 14". There are three models in the Duoscan family ranging from $750 to $5175 list price. I can provide sample output and literature for anyone who wishes to contact me directly or visit http://www.agfahome.com/product/CatProd_DisplayPublic.html?id=3115
George Laing National Graphic Supply E-mail: scisales-at-ngscorp.com Phone: (800) 223-7130 X3109 USA
Valerie, I can't comment on the Nikon scanner but I have plenty to say about the Polaroid Sprintscan 45, most of which is probably best left unsaid! Let me summarize. We purchased the Polaroid Sprintscan 45 in late 1998 and planned to operate it on a PC with Windows 98 operating system. Our first unit was defective and was replaced, however, the trouble shooting process was very time consuming. Our second unit functioned intermittently. Again, after much trouble shooting we returned the unit to Polaroid Canada. After having it tested we were told that there were no problems with the Sprintscan 45. The unit was returned to us but continued to cause us grief with its erratic behavior. We spent more time trying to diagnose the problem and even called in a computer specialist who spoke directly with Polaroid's tech support personnel in an effort to resolve the problem. In the end we found that the Sprintscan appears not to like Windows 98. The scanner has been running sucessfully on an older PC with Windows 95. Why? Has there been an identified problem associated with Windows 98? If so why aren't customers alerted? Having this knowledge would have saved us an enormous amount of time and money. I have been waiting for a comment from Polaroid for well over a month.
I would be interested in hearing from other Sprintscan users who experienced similar problems.
Regards, Paul
Paul J. Gerroir Microscopy Materials Characterization Xerox Research Centre of Canada 2660 Speakman Drive Mississauga, Ontario L5K 2L1
} -----Original Message----- } From: Valerie Leppert [SMTP:vjleppert-at-ucdavis.edu] } Sent: Tuesday, March 21, 2000 8:48 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Film Scanner Recommendations? } } ------------------------------------------------------------------------ } 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 all- } } We are in the midst of writing a proposal for a new film scanner and I } am hoping that someone out there can help with a recommendation for a top } of the line film scanner. The major application would be TEM negatives. A } model we are currently looking at is the Nikon LS-4500AF Multi Format Film } Scanner. } } If anyone has archived recommendations from previous discussions and could } send them on to me, that would be wonderful. } } Thank you, } Valerie Leppert } } Dept. of Chem. Eng. and Mat. Sci. } U. of California, Davis } } vjleppert-at-ucdavis.edu }
hello, I was wondering if I could get a sampling of opinions on the different attachments to light microscopes, dissecting microscopes to display video to an external monitor. I've seen a few different products available, and would like to know any experiences people may have had with different systems. Any comments are greatly appreciated, and I will happily make a summary of the information.
Sincerely, Gordon Vrdoljak.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ 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
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The ongoing discussion about replica techniques continues to be very educational. I wish to suggest another approach to this analysis problem. Occasionally I jet polish foils of metals which contain precipitates. Certain acid mixtures seem more "agressive" and preferentially remove the precipitates. This seems prone to happen when using nitric or perchloric acid mixtures. However, a few years ago when I began to use "non-acid" electrolytes, precipitates were sometimes thinned beautifully and were still retained in the thinned foil. I once got a hole in the center of a large precipitate! For more information, see Ultramicroscopy, Vol.19,(1986). Perhaps more research should be funded along this "thread". A second approach has been to "design" less agressive electrolytes that rely on hydrochloric acid, for example, which sometimes works for some materials but is not in The general literature. Within safty considerations,worthwhile electrolyte improvements may be achieved with a bit of work! Good luck.
Bernie Kestel Materials Science Division Argonne National Laboratory 9700 South Cass Avenue Argonne, Il., 60439
by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id PAA05355 for dist-Microscopy; Thu, 23 Mar 2000 15:14:34 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id PAA05350 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Thu, 23 Mar 2000 15:14:04 -0600 (CST) Received: from UMKC-MAIL01.umkc.edu (email.exchange.umkc.edu [134.193.71.1]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id PAA05343 for {microscopy-at-sparc5.microscopy.com} ; Thu, 23 Mar 2000 15:13:52 -0600 (CST) Received: by umkc-mail01 with Internet Mail Service (5.5.2650.21) id {HJ6P40N0} ; Thu, 23 Mar 2000 15:08:43 -0600 Message-ID: {95A711A70065D111B58C00609451555C01CA7555-at-UMKC-MAIL02} "Dusevich, Vladimir" {DusevichV-at-umkc.edu} Cc: Microscopy Listserver {microscopy-at-sparc5.microscopy.com}
Hans, Thank you very much for explanations. But for me this method will not work - quite often I analyze wet specimens at pretty high pressure (5-6 Torr). Thank you again,
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} -----Original Message----- } From: Hans Dijkstra [mailto:hans.dijkstra-at-edax.nl] } Sent: Thursday, March 23, 2000 12:19 PM } To: DusevichV-at-umkc.edu } Cc: Microscopy Listserver } Subject: RE: EDS in Variable Pressure SEM } } } Dear Vladimir, } } The main idea of the ViP-Quant procedure is rather } straight-forward and } elegant. It is based on the phenomenon that as the pressure } increases the } size of the skirt effect remains fairly constant, although } the number of } skirt electrons increases, and thus the contribution of the } skirt-effect to } the EDS signal increases. This intensity increase is linear with the } pressure. } } This is valid for low- to medium pressures, or at a higher } (ESEM) pressure } with a very short free-path length of the skirt electrons, } since in both } cases you can assume the skirt electrons have scattered only } once. At higher } pressures with longer working distances these assumptions are } no longer } fully valid. } } So to do an EDS analysis you take 2 measurements at different } pressures, the } low-pressure one at a pressure where you can just avaid } charging, and a } high-pressure one at at least twice that pressure. The } measured intensities } of all elements are then plotted as a function of pressure, } and extrapolated } to pressure zero, i.e. high vacuum. The extrapolated results } will be very } close to the results hat would have been obtained directly if } the sample } could have been analyzed at high vacuum conditions. } } Of course anyone can do this plotting and extrapolation } manually, that is: } if your EDS software allows you to enter intensities } manually! The only } thing the EDAX ViP-Quant is offering as an extra is that the } software does } it automatically for you. There are no proprietary miracles } involved, EDAX } has just listened carefully to what science has to offer.... } } Best regards, } } Hans Dijkstra } } Disclaimer: The above is my personal opinion, and not } necessarily EDAX's. } ------------------------------------------------------------- } EDAX Europe www.edax.com } Ringbaan Noord 103 Tel.: +31-13-5364000 } P.O. Box 4144 Fax.: +31-13-5356279 } 5004 JC Tilburg E-mail: hans.dijkstra-at-edax.nl } the Netherlands } ------------------------------------------------------------- } } } -----Original Message----- } } } } From: "Dusevich, Vladimir", INTERNET:DusevichV-at-umkc.edu } } To: Everett Ramer, INTERNET:Everett.Ramer-at-netl.doe.gov } } "'Hans Dijkstra'", INTERNET:hans.dijkstra-at-edax.nl } } } } CC: Joergen Bilde-Soerensen 5802, INTERNET:j.bilde-at-risoe.dk } } [unknown], INTERNET:Microscopy-at-sparc5.microscopy.com } } } } Date: 3/22/00 4:35 PM } } } } RE: RE: EDS in Variable Pressure SEM } } } } } } } -------------------------------------------------------------- } ---------- } } 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 } } } -------------------------------------------------------------- } ---------. } } } } } } Hans, } } Could you please explain the main idea of the } } Vip-Quant algorithm. I cannot even imagine } } how quantitative analysis could have nearly the } } same accuracy as in high-vacuum mode without } } a priori knowledge of the composition of } } surrounding areas. } } Thank you, } } } } Vladimir } } } } } } Vladimir M. Dusevich, Ph.D. } } Electron Microscope Lab Manager } } 3127 School of Dentistry } } 650 E. 25th Street } } Kansas City, MO 64108-2784 } } } } Phone: (816) 235-2072 } } Fax: (816) 235-5524 } } Web: http://www.umkc.edu/dentistry/microscopy } } } } } -----Original Message----- } } } From: Hans Dijkstra [mailto:hans.dijkstra-at-edax.nl] } } } Sent: Tuesday, March 21, 2000 7:54 AM } } } To: Everett Ramer } } } Cc: Microscopy-at-sparc5.microscopy.com; Joergen Bilde-Soerensen 5802 } } } Subject: RE: EDS in Variable Pressure SEM } } } } } } } } } Dear Everett, } } } } } } The extrapolation method (aka Variable Pressure Method) } } } described by Dr. } } } Bilde-Sorenson has been implemented by EDAX in a software } } } feature called } } } ViP-Quant. With this method the EDS analysis under Bad-Vacuum } } } conditions can } } } be done with nearly the same accuracy as under } High-Vacuum conditions. } } } Although this method was developed with the ESEM microscope } } } in mind, it of } } } course can be applied to all Bad-Vacuum scanning electron } microscopes. } } } } } } Please contact your local EDAX representative for more } } } information and a } } } copy of the new ViP-Quant brochure, or request one through } } } www.edax.com. } } } } } } With best regards, } } } } } } Hans Dijkstra } } } } } } } } } } } } } } } } } ----------------------- Internet Header } -------------------------------- } } Sender: Microscopy-request-at-sparc5.microscopy.com } } Received: from ultra5.microscopy.com ([206.69.208.10]) } } by spdmgaac.compuserve.com (8.9.3/8.9.3/SUN-1.9) with ESMTP } } id LAA16645; } } Wed, 22 Mar 2000 11:34:33 -0500 (EST) } } Received: (from daemon-at-localhost) } } by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id KAA01363 } } for dist-Microscopy; Wed, 22 Mar 2000 10:27:23 -0600 (CST) } } Received: from no_more_spam.com (sparc5 [206.69.208.10]) } } by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP } id KAA01359 } } for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Wed, 22 } } Mar 2000 10:26:53 -0600 (CST) } } Received: from UMKC-MAIL01.umkc.edu (email.exchange.umkc.edu } } [134.193.71.1]) } } by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP } id KAA01352 } } for {Microscopy-at-sparc5.microscopy.com} ; Wed, 22 Mar 2000 } } 10:26:41 -0600 (CST) } } Received: by umkc-mail01 with Internet Mail Service (5.5.2650.21) } } id {HJ6P423S} ; Wed, 22 Mar 2000 10:21:30 -0600 } } Message-ID: {95A711A70065D111B58C00609451555C01CA7552-at-UMKC-MAIL02} } } From: "Dusevich, Vladimir" {DusevichV-at-umkc.edu} } } To: "'Hans Dijkstra'" {hans.dijkstra-at-edax.nl} , } } Everett Ramer } } {Everett.Ramer-at-netl.doe.gov} } } Cc: Microscopy-at-sparc5.microscopy.com, } } Joergen Bilde-Soerensen 5802 } } {j.bilde-at-risoe.dk} } } Subject: RE: EDS in Variable Pressure SEM } } Date: Wed, 22 Mar 2000 10:21:28 -0600 } } MIME-Version: 1.0 } } X-Mailer: Internet Mail Service (5.5.2650.21) } } Content-Type: text/plain; } } charset="iso-8859-1" } } Errors-to: Microscopy-request-at-sparc5.microscopy.com } } } } }
I've heard from several SEM/TEM independent maintenance companies that some of the manufacturer's have engaged in "price dumping" when negotiating a service contract.
In other words, if the manufacturer is aware that they have competition
for a service contract they will reduce the contract by as much as forty
percent.
I am all for the free enterprise system, but I thought that when a contract price is offered it is the same for all customers especially government contracts that must abide by the GSA rules. GSA rules states
that the price offered is the lowest price for this service. It would be illegal for the manufacturer (or anyone) to offer a contract at less than the cost offered to GSA customers.
I am surprised by this move as our contracts are the same as we abide by the GSA rules.
EDS in ESEM is a pretty similar thing to EDS in SEM if you do not need trace/minor (~1%) element analysis and use some standard precautions. Some vendors even sell as an option gaseous detectors which could be placed close (1mm) to a sample and greatly reduce a beam scattering. I routinely use EDS at water vapor pressure up to 6 Torr. I've put an X-ray map taken in environmental conditions at our web site http://www.umkc.edu/dentistry/microscopy If low concentrations are of no interest to you then you will not see real difference in performance of an EDS in VP and high vacuum modes, especially if you do not work with wet specimens which need really high pressure.
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} } } It seems that nowadays every SEM vendor is offering variable } pressure models, which are conventional SEMs with a plumbing } modification that allows the sample to be at pressures of up } to about 4 torr (500 Pa) while the electron column operates } at the conventional high vacuum. I am very interested in } buying a variable pressure SEM with an EDS, but was recently } warned that EDS has very poor spatial resolution in the } variable pressure mode because of the large beam spread due } to electrons scattering off the gas molecules in the sample } chamber. Is this really a significant issue? Do any of you } have experience using EDS with variable pressure SEMs? } Thanks, } }
Some of the K-12 classrooms we serve in our Bugscope (http://bugscope.beckman.uiuc.edu/) program have expressed interest in sending us aquatic or marine invertebrates to prepare for their sessions. What I'd like to know is how I should ask them to prepare the samples for me (e.g., should I ask them to overnight pondwater or seawater samples in plastic bottles?), and after that, what should I do with them? I can't really be sending glutaraldehyde and cacodylate to gradeschool kids, but I'm assuming I'll want to do some sort of standard fixation/dehydration/HMDS treatment, presumably on Millipore filters, once I get samples. Any tips?
} We are in the midst of writing a proposal for a new film scanner and I } am hoping that someone out there can help with a recommendation for a top } of the line film scanner. The major application would be TEM negatives. A } model we are currently looking at is the Nikon LS-4500AF Multi Format Film } Scanner. } } Valerie Leppert } Dept. of Chem. Eng. and Mat. Sci. } U. of California, Davis
We use a Polaroid SprintScan 45 for SEM, TEM and other large film but find that the negatives need to be somewhat tailored to the scanner's needs. We have great difficulty with very dense or very high contrast negatives, especially from the TEM. Staining and microscope settings need to be adjusted to produce negatives that are within the range of the scanner. It then produces very good results.
_____________________________________ Tom Gore, Advanced Imaging Laboratory Biology Department, University of Victoria Box 3020, Station CSC, Victoria, BC, V8W 3N5 Canada voice (250) 721-7134 fax (250) 721-7120 web: http://web.uvic.ca/ail/
We are looking for an independant service provider for our JEOL-880 SEM. They would need to be very familiar with JEOL equipment since the 880 is a rare bird (the only one in the States, I believe). I believe it's a 1200 TEM column married to 840 electronics. We are located in central Oklahoma. TIA.
Bill
-- ============================================================= Bill Chissoe III Electron Microscopist University of Oklahoma 770 Van Vleet Oval Norman, Ok. 73019 E-mail: wchiss-at-ou.edu Ph. (405)325-4391 =============================================================
I am looking for a vendor to purchase The Image Processing Tool Kit by John Russ. I placed an order with Amazon nearly a month ago but have yet to receive anything. Hmmmm.
Thank you,
John B.
#################################################################### John J. Bozzola, Ph.D., Director Micro-Imaging and Analysis Center 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 ####################################################################
-----Original Message----- From: Michael Coviello [SMTP:coviello-at-mae.uta.edu] Sent: Thursday, March 23, 2000 4:44 AM To: Microscopy-at-sparc5.microscopy.com Subject: TEM-Digital camera recommendations
------------------------------------------------------------------------ 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 Ya'll: We are looking for a CCD camera for our Philips 430 TEM. We would like to get the best camera for the best price, e.g., either buying a used camera or a new non-Gatan camera (Gatan seems to be twice as expensive as the others). We would be using the camera for bright field and high resolution TEM of materials (semiconductors) rather than for biological specimens. Does anyone have a camera they would like to sell/donate or does anyone have recommendations as to a less-expensive camera that they know can be used for materials applications. Thanks, Mike Coviello Lab Manager University of Texas -at- Arlington
MARCH 30TH MEETING AT GENENTECH TOPIC: Microscopy and Public Health.
Our two speakers come from the USDA, Agricultural Research Service in Albany, CA. Robert Mandrell's presentation is titled "Analysis of Human Pathogens on Food Surfaces by Stereo- and Confocal Micros-copy". Robert is the Research Leader of the Food Safety and Health Unit at the USDA facility. Also speaking is De Wood on "Immunolocalization using FESEM and a Backscatter Detector". De heads up the Microscopy & Imaging Lab at the USDA.
Please call Greg Lum at 415/338-1339 or Email: "glum-at-sfsu.edu" for dinner reservations and entree choice by Monday March 27th. The meeting starts at 5:00 p.m. with a beverage reception. The dinner will start at 6:00 pm.
DINNER RESERVATIONS Entree choice (select one) $20 nonmember, $15 regular members; $8 student members Menu choices for Genentech meeting on March 30th [ ] Chicken Parmesan [ ] Flank Steak with Mushroom Sauce [ ] Pasta Primavera
Please call Greg Lum at 415/338-1339 or Email: "glum-at-sfsu.edu" for dinner reservations and entree choice by Monday March 27th. The meeting starts at 5:00 p.m. with a beverage reception. The dinner will start at 6:00 pm.
There are two labs on campus with the Agfa Duoscan T2500 scanners. As stated in the message from National Graphics Supply, they have both a flat bed and a transparency drawer. They will scan at 16-bit gray scale as well as in 8-bit and RGB. Weve been very happy with ours, primarily using it for TEM and SEM negatives, and for 35 mm. One caveat, the 2500 dpi capability is limited to a 4 inch x 14 inch area (1/2 width of the scan area). Set-up was simple and no crashes (so far). The software is easy to learn with identical interfaces for both the standalone application and the Photoshop compatible plug-in.
We purchased ours locally from a professional photography retailer.
Glen
Glen MacDonald Research Scientist Hearing Research Laboratories of the Virginia Merrill Bloedel Hearing Research Center Box 35-7923 University of Washington Seattle, WA 98195-7923 (206) 616-4156 glenmac-at-u.washington.edu
Hum.... This is very interesting. Government customers are one thing, non-government customers are another. GSA rules and principles do not apply to non-government customers. If you would please cite the FAR that is the basis for your assertion, this could help a lot to clarify your statement.
I have never heard of "price dumping." But I have heard of competition. One must keep U.S. government business separate from others. Even then, how does one mingle them together? On what basis is this done?
How much of this "competition" is based on multiple unit discounts versus actual price reduction? And as a consumer, what real difference does it make? If the consumer can negotiate a good deal, all the better for the consumer, right?
Mixing GSA into free enterprise is like apples and oranges, so to speak.
What do you think?
gary g.
At 03:19 PM 3/23/00 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
You are right. The 880 is an "immersion lens" SEM, a very rare animal. There are only two that I know of: One at OU, the second was at IBM in France but is now in the back of my office sadly used for parts.
Earl Weltmer
Bill Chissoe 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 an independant service provider for our JEOL-880 SEM. } They would need to be very familiar with JEOL equipment since the 880 is } a rare bird (the only one in the States, I believe). I believe it's a } 1200 TEM column married to 840 electronics. We are located in central } Oklahoma. TIA. } } Bill } } -- } ============================================================= } Bill Chissoe III } Electron Microscopist } University of Oklahoma } 770 Van Vleet Oval } Norman, Ok. 73019 } E-mail: wchiss-at-ou.edu Ph. (405)325-4391 } =============================================================
The Company providing the service does certify to the GSA customer that the prices given are the same or lower than the prices given to any other entity: public, government or private.
Earl Weltmer
"Dr. Gary Gaugler" wrote:
} Hum.... This is very interesting. Government customers are one } thing, non-government customers are another. GSA rules and } principles do not apply to non-government customers. If you } would please cite the FAR that is the basis for your assertion, } this could help a lot to clarify your statement. } } I have never heard of "price dumping." But I have heard of } competition. One must keep U.S. government business separate } from others. Even then, how does one mingle them together? } On what basis is this done? } } How much of this "competition" is based on multiple unit discounts versus } actual price reduction? And as a consumer, what real difference } does it make? If the consumer can negotiate a good deal, all } the better for the consumer, right? } } Mixing GSA into free enterprise is like apples and oranges, so } to speak. } } What do you think? } } gary g. } } At 03:19 PM 3/23/00 , you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Hi All, } } } } I've heard from several SEM/TEM independent maintenance companies that } } some of the manufacturer's have engaged in "price dumping" when } } negotiating a service contract. } } } } In other words, if the manufacturer is aware that they have competition } } } } for a service contract they will reduce the contract by as much as forty } } } } percent. } } } } I am all for the free enterprise system, but I thought that when a } } contract price is offered it is the same for all customers especially } } government contracts that must abide by the GSA rules. GSA rules states } } } } that the price offered is the lowest price for this service. It would } } be } } illegal for the manufacturer (or anyone) to offer a contract at less } } than the cost offered to GSA customers. } } } } I am surprised by this move as our contracts are the same as we abide } } by } } the GSA rules. } } } } Regards, } } } } Earl Weltmer
OK....to a GSA customer. What about to other customers? And what precedence does one GSA contract or schedule have for other instantiations? They are, in my experience, single point events. They are not precedence events.
Where is the original thread that spawned this message?
Note, that in my experience, GSA contracts are distinct from other Federal government contracts. i.e., GSA is one thing, other gov contracts are another. GSA negotiates rates....they do not establish them. Therefore, for a particular GSA contract or rate structure, the rates are pre-negotiated for fed users to adopt as per their own contracting department. Or, they can use the GSA schedule and go with that vehicle and its added surcharge. (Yes, GSA contracts cost more than face value). GSA has two flavors: negotiated rates (the user does their own contracting) and negotiated contracts (the user buys into the GSA contract and pays a surcharge for doing so).
Which flavor are you talking about?
gg
At 09:19 PM 3/23/00 , you wrote: } The Company providing the service does certify to the GSA customer that the } prices given are the same or lower than the prices given to any other entity: } public, government or private. } } Earl Weltmer } } "Dr. Gary Gaugler" wrote: } } } Hum.... This is very interesting. Government customers are one } } thing, non-government customers are another. GSA rules and } } principles do not apply to non-government customers. If you } } would please cite the FAR that is the basis for your assertion, } } this could help a lot to clarify your statement. } } } } I have never heard of "price dumping." But I have heard of } } competition. One must keep U.S. government business separate } } from others. Even then, how does one mingle them together? } } On what basis is this done? } } } } How much of this "competition" is based on multiple unit discounts versus } } actual price reduction? And as a consumer, what real difference } } does it make? If the consumer can negotiate a good deal, all } } the better for the consumer, right? } } } } Mixing GSA into free enterprise is like apples and oranges, so } } to speak. } } } } What do you think? } } } } gary g. } } } } At 03:19 PM 3/23/00 , you wrote: } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } Hi All, } } } } } } I've heard from several SEM/TEM independent maintenance companies that } } } some of the manufacturer's have engaged in "price dumping" when } } } negotiating a service contract. } } } } } } In other words, if the manufacturer is aware that they have competition } } } } } } for a service contract they will reduce the contract by as much as forty } } } } } } percent. } } } } } } I am all for the free enterprise system, but I thought that when a } } } contract price is offered it is the same for all customers especially } } } government contracts that must abide by the GSA rules. GSA rules states } } } } } } that the price offered is the lowest price for this service. It would } } } be } } } illegal for the manufacturer (or anyone) to offer a contract at less } } } than the cost offered to GSA customers. } } } } } } I am surprised by this move as our contracts are the same as we abide } } } by } } } the GSA rules. } } } } } } Regards, } } } } } } Earl Weltmer
On the very low end A Snappy digitizer and a surveillance video camera produce what I would consider the minimal acceptable image. It is good enough for some work I would say it is comparable to a Polaroid image or slightly worse.
} From a quality per dollar point of view it can't be beat. I would say it is good enough for collaboration, web publishing and some publishing.
This set up and 35mm camera should meet all quality needs as long as you don't need real-time images. It does not compare to a top of the line dedicated system. But it does better than anything but a dedicated system.
My best effort to date is: http://www.couger.com/microscope/onion.jpg This is the raw image with no enhancement and the enhanced version is: http://www.couger.com/microscope/onionE.jpg This version was contrast enhanced and mapped to false color. The subject is a partially dehydrated onion cell. There are depth of focus problems and possibly some vibration problems. There is only 7.5 bits of information in the raw image. I attribute this to loss of dynamic range in the camera due to age.
This was taken with a Leitz darkfield condenser and a Leitz 63x 0.85 objective with a variable aperture for darkfield work. The camera was a monochrome RCA surveillance videocron camera and a Snappy digitizer.
I don't think the microscope end can be improved on much in the onion image. I still had dynamic range problems. The only solution I can think of is to take multiple pictures at different light levels and combine them. A high resolution CCD camera will give better resolution but I don't think the dynamic range problem will go away using video cameras. Maybe I will get around to exposing a 4X5 negative and see what is really there. But after using digital capture it is an awful lot of trouble.
I am strictly an amateur with a microscope but I do have professional experience in digital images.
Gordon
Gordon Couger gcouger-at-couger.com
Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
----- Original Message ----- } From: "Gordon Vrololjak" {gvrdolja-at-nature.Berkeley.EDU} To: {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, March 23, 2000 1:56 PM
Try going direct:
Reindeer Games, Inc. 235 S. Main St. #201 Gainesville, FL 32601 352.384.1850
http://members.aol.com/foveapro
jcr6-at-aol.com
gg
At 06:21 PM 3/23/00 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I am looking for a vendor to purchase The Image Processing Tool Kit by John Russ. I placed an order with Amazon nearly a month ago but have yet to receive anything. Hmmmm. } } Thank you, } } John B. } } #################################################################### } John J. Bozzola, Ph.D., Director } Micro-Imaging and Analysis Center } 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 } ####################################################################
} I am looking for a vendor to purchase The Image Processing Tool Kit by } John Russ.
I believe you can purchase it directly from John Russ.
Rob
Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa R.Cross-at-ru.ac.za tel: +27 46 603 8168 - fax: +27 46 622 4377 http://www.ru.ac.za/emu/index.htm
** remember that ICEM-15 in 2002 is in Durban, South Africa **
Scott Some aquatic invertebrates such as gastrotrichs are difficult to fix for SEM in a life-like relaxed state because aldehyde fixatives induce violent contraction. A paper by May+ advocates relaxing the organisms (rotifers in this instance) first by narcosis with procaine, or similar anaesthetics. However, in our experience gastrotrichs can sense these anaesthetics, and considerable distortion results. There is a considerable literature on this topic covering the properties of EDTA, menthol, osmium tetroxide, etc. etc. for this purpose, but mostly these solutions are ineffective at least on gastrotrichs.
We have found sodium azide at ~0.1% to be very effective at preventing distortion. Gastrotrichs seem not to be irritated by it, but simply go to sleep. We have usually mounted them for LM in glycerol, in which they can be preserved and collected for transit or for subsequent processing and examination. The fixation/ dehydration/ HMDS procedure you suggest would probably be very effective, but you might also try Ensikat & Barthlott's* approach of simply examining them in the SEM in the glycerol-wet state, or after drying them from glycerol under vacuum.
+May, L. (1985) The use of procaine hydrochloride in the preparation of rotifer samples for counting. Verh. Internat. Verein. Limnol. 22, 2897-2990 *Ensikat & Barthlott (1993) Liquid substitution - a versatile procedure for sem specimen preparation of biological-materials without drying or coating Journal of Microscopy 172, 195-203
Sodium azide is obviously a serious poison, and schoolkids cannot handle the powder, but would it be acceptable for them to use 0.1% solution under supervision by a teacher? I guess this depends on local legislation and attitudes, and also on the age and level of experience and responsibility of the students.
Hope this helps Chris
Date sent: Thu, 23 Mar 2000 16:16:36 -0600 To: Microscopy-at-sparc5.microscopy.com } From: Scott Robinson {sjrobin-at-itg.uiuc.edu}
Dear Vladimir,
Under the circumstances you describe you are probably on the edge of the condition where the virtual composition changes linearly with the pressure, but this depends on the working distance and the gas that you use. However, if you still have some pressure range to work with, then making several measurements at a range of pressures may still allow you to use a non-linear extrapolation.
The main problem might be that wet specimens often have a thin water film on the sample, which may adversely affect EDS analysis. Keeping exact control over sample temperature, pressure and humidity is required to get suitable EDS conditions.
} -----Original Message----- } From: Dusevich, Vladimir [mailto:DusevichV-at-umkc.edu] } Sent: Thursday, March 23, 2000 10:09 PM } To: 'Hans Dijkstra'; Dusevich, Vladimir } Cc: Microscopy Listserver } Subject: RE: EDS in Variable Pressure SEM } } } Hans, } Thank you very much for explanations. } But for me this method will not work - } quite often I analyze wet specimens at } pretty high pressure (5-6 Torr). } Thank you again, } } Vladimir } } Vladimir M. Dusevich, Ph.D. } Electron Microscope Lab Manager } 3127 School of Dentistry } 650 E. 25th Street } Kansas City, MO 64108-2784 } } Phone: (816) 235-2072 } Fax: (816) 235-5524 } Web: http://www.umkc.edu/dentistry/microscopy } } } -----Original Message----- } } From: Hans Dijkstra [mailto:hans.dijkstra-at-edax.nl] } } Sent: Thursday, March 23, 2000 12:19 PM } } To: DusevichV-at-umkc.edu } } Cc: Microscopy Listserver } } Subject: RE: EDS in Variable Pressure SEM } } } } } } Dear Vladimir, } } } } The main idea of the ViP-Quant procedure is rather straight-forward and } } elegant. It is based on the phenomenon that as the pressure increases the } } size of the skirt effect remains fairly constant, although the number of } } skirt electrons increases, and thus the contribution of the skirt-effect to } } the EDS signal increases. This intensity increase is linear with the pressure. } } } } This is valid for low- to medium pressures, or at a higher (ESEM) pressure } } with a very short free-path length of the skirt electrons, since in both } } cases you can assume the skirt electrons have scattered only once. At higher } } pressures with longer working distances these assumptions are no longer } } fully valid. } } } } So to do an EDS analysis you take 2 measurements at different pressures, the } } low-pressure one at a pressure where you can just avoid charging, and a } } high-pressure one at at least twice that pressure. The measured intensities } } of all elements are then plotted as a function of pressure, and extrapolated } } to pressure zero, i.e. high vacuum. The extrapolated results will be very } } close to the results hat would have been obtained directly if the sample } } could have been analyzed at high vacuum conditions. } } } } Of course anyone can do this plotting and extrapolation manually, that is: } } if your EDS software allows you to enter intensities manually! The only } } thing the EDAX ViP-Quant is offering as an extra is that the software does } } it automatically for you. There are no proprietary miracles involved, EDAX } } has just listened carefully to what science has to offer.... } } } } Best regards, } } } } Hans Dijkstra } } } } Disclaimer: The above is my personal opinion, and not necessarily EDAX's. } } ------------------------------------------------------------- } } EDAX Europe www.edax.com } } Ringbaan Noord 103 Tel.: +31-13-5364000 } } P.O. Box 4144 Fax.: +31-13-5356279 } } 5004 JC Tilburg E-mail: hans.dijkstra-at-edax.nl } } the Netherlands } } ------------------------------------------------------------- } } } } } -----Original Message----- } } } } } } From: "Dusevich, Vladimir", INTERNET:DusevichV-at-umkc.edu } } } To: Everett Ramer, INTERNET:Everett.Ramer-at-netl.doe.gov } } } "'Hans Dijkstra'", INTERNET:hans.dijkstra-at-edax.nl } } } } } } CC: Joergen Bilde-Soerensen 5802, INTERNET:j.bilde-at-risoe.dk } } } [unknown], INTERNET:Microscopy-at-sparc5.microscopy.com } } } } } } Date: 3/22/00 4:35 PM } } } } } } RE: RE: EDS in Variable Pressure SEM } } } } } } } } } } } -------------------------------------------------------------- } } ---------- } } } 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 } } } } } -------------------------------------------------------------- } } ---------. } } } } } } } } } Hans, } } } Could you please explain the main idea of the } } } Vip-Quant algorithm. I cannot even imagine } } } how quantitative analysis could have nearly the } } } same accuracy as in high-vacuum mode without } } } a priori knowledge of the composition of } } } surrounding areas. } } } Thank you, } } } } } } Vladimir } } } } } } } } } Vladimir M. Dusevich, Ph.D. } } } Electron Microscope Lab Manager } } } 3127 School of Dentistry } } } 650 E. 25th Street } } } Kansas City, MO 64108-2784 } } } } } } Phone: (816) 235-2072 } } } Fax: (816) 235-5524 } } } Web: http://www.umkc.edu/dentistry/microscopy } } } } } } } -----Original Message----- } } } } From: Hans Dijkstra [mailto:hans.dijkstra-at-edax.nl] } } } } Sent: Tuesday, March 21, 2000 7:54 AM } } } } To: Everett Ramer } } } } Cc: Microscopy-at-sparc5.microscopy.com; Joergen Bilde-Soerensen 5802 } } } } Subject: RE: EDS in Variable Pressure SEM } } } } } } } } } } } } Dear Everett, } } } } } } } } The extrapolation method (aka Variable Pressure Method)described by Dr. } } } } Bilde-Sorenson has been implemented by EDAX in a software feature called } } } } ViP-Quant. With this method the EDS analysis under Bad-Vacuum conditions can } } } } be done with nearly the same accuracy as under High-Vacuum conditions. } } } } Although this method was developed with the ESEM microscope in mind, it of } } } } course can be applied to all Bad-Vacuum scanning electron microscopes. } } } } } } } } Please contact your local EDAX representative for more information and a } } } } copy of the new ViP-Quant brochure, or request one through www.edax.com. } } } } } } } } With best regards, } } } } } } } } Hans Dijkstra
Thanks for the info. I really didn't want to go through & look up the FAR again.
The GSA wasn't my real inquiry anyway. I just wanted to find out how widespread this practice extends. The GSA issue seems to get in the way.
OK to rephase: How many have experienced an extreme reduction in service contract price (20% or more) when confronted with competition?
Thank You,
Earl Weltmer
"Allen R. Sampson" wrote:
} Actually, both are right here. Government stipulations have apparently } eased recently, but a part of the government contract procedure has in the } past been a certification by the service provider that the rates being } quoted are normal and appropriate. This has essentially required details } of a few other comparable contracts that were similar in price and } requirements. } } I have not had to go through that particular body cavity search for some } time. I assume the government learned that certifications of this sort are } basically worthless, as there are companies large and small who are willing } to do a little spin control in order to secure contracts. Government } agencies are probably unlikely to bother verifying compliance with their } regulations - not a problem in this field alone. } } My memory goes only so far, as does my desire to look up old records. } However, if you're really interested, you may want to start with the old } DAR (Defense Acquisition Regulation) 7-1903.41 - Service Contract Act of } 1965, also updated and known as FAR 52.222-40. In the late 80's, it seems } to have been a part of the small business set-aside program (FAR 52.219-04) } to require a price list or a verifiable listing of 3 similar items sold to } establish a market price. You might also look into the Walsh-Healey Public } Contract Act, FAR 52.222-20. } } DOD FAR Supplement (46 CFR Chapter 2) clause 11.111-10 states - "...The } Contractor agrees that the prices for the supplies or services furnished } under this contract are as low or lower than those charged the supplier's } most favored customer for comparable quantities under similar terms and } conditions, in addition to any discounts for prompt payment." } } Frankly each government contract I have generates about 1/2" of paper work } per year. I give little concern to pricing matters, either I win a } contract or not. The same goes for commercial customers. I try to price } my services to all customers based on the expenses I expect to incur and a } modest income for myself (perhaps way too modest, I reflect, as the time to } finish the tax year has come). There are many other regulations and laws } that I have to pay more attention to. } } Whether these, and other, regulations, laws and requirements have any } actual practical effect is open to debate. I can't say that I have noticed } any recent trend for manufacturers to discount their service contracts - } although I don't normally inquire about other bids submitted. I wouldn't } be surprised, though, if manufacturers are finding it advantageous to } discount service prices now to capture customers while the capital } expenditures are running high in the current economy. However, when those } capital budgets dry out in the next economic cycle, they will find that the } service end will have a larger effect on their profitability and they will } increase their profit margins as best they can. } } Consider this a normal result of the economic cycles that we are all slaves } to. As a third party service provider, you can generally think of me as an } independent car repair shop. In good times, people will go to } manufacturer's service, or buy a new car - in bad times they will make } every effort to stretch their capital investment. In good economic times, } I catch a good deal of lower end business from those who have instruments } orphaned by their manufacturer or are very price conscious. In bad } economic times, I'm the guy who can help you avoid laying off personnel by } reducing your costs. } } In either case, just be glad that there are independent service sources } available for these instruments and consider that they will only be there } as long as it is economically feasible for them to survive all economic } scenarios. That is even more true for manufacturers who would rather sell } you a new instrument than service a 3 year old instrument. That } short-sighted corporate position has proven to be a death knell in this } industry. In many cases it has only been the availability of third party } service that has allowed a company to survive in a given market for a } little longer. Public relations is a two-sided blade - it can cut real } deep when times are bad. } } Government laws, rules and regulations can have a significant effect on } manufacturers serving both government and commercial customers. As has } been stated before, manufacturers wishing to do business with the } government are often required to provide service for a stated period of } time and, as stated here, are required to provide service at a market rate. } A first year law student would recognize that, in this country, commercial } concerns benefit from the standards set by the government. It would } behoove any company to provide an even pricing structure across the board, } as any other position opens them up to legal action from either side. } } Since the government rules require comparison under similar "terms and } conditions", the question of multiple instrument discounts is appropriately } accounted for. We are not talking '"apples and oranges" here. Rather, we } are talking about a closed feedback loop where any change in either side } must affect the other. The only wiggle-room here is in the quality of } service provided. Yes, the consumers, both government and commercial, can } bid the service price down. But in the end, only the service quality will } be compromised. } } Allen R. Sampson, Owner } Advanced Research Systems, St. Charles, Illinois 60174 } voice 630.513.7093 fax 630.513.7092 } } -----Original Message----- } From: Dr. Gary Gaugler [SMTP:gary-at-gaugler.com] } Sent: Thursday, March 23, 2000 6:18 PM } To: Earl Weltmer } Cc: MSA listserver } Subject: Re: Dumping of Service Contracts costs } } ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } Hum.... This is very interesting. Government customers are one } thing, non-government customers are another. GSA rules and } principles do not apply to non-government customers. If you } would please cite the FAR that is the basis for your assertion, } this could help a lot to clarify your statement. } } I have never heard of "price dumping." But I have heard of } competition. One must keep U.S. government business separate } from others. Even then, how does one mingle them together? } On what basis is this done? } } How much of this "competition" is based on multiple unit discounts versus } actual price reduction? And as a consumer, what real difference } does it make? If the consumer can negotiate a good deal, all } the better for the consumer, right? } } Mixing GSA into free enterprise is like apples and oranges, so } to speak. } } What do you think? } } gary g. } } At 03:19 PM 3/23/00 , you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Hi All, } } } } I've heard from several SEM/TEM independent maintenance companies that } } some of the manufacturer's have engaged in "price dumping" when } } negotiating a service contract. } } } } In other words, if the manufacturer is aware that they have competition } } } } for a service contract they will reduce the contract by as much as forty } } } } percent. } } } } I am all for the free enterprise system, but I thought that when a } } contract price is offered it is the same for all customers especially } } government contracts that must abide by the GSA rules. GSA rules states } } } } that the price offered is the lowest price for this service. It would } } be } } illegal for the manufacturer (or anyone) to offer a contract at less } } than the cost offered to GSA customers. } } } } I am surprised by this move as our contracts are the same as we abide } } by } } the GSA rules. } } } } Regards, } } } } Earl Weltmer
We find that we get better silver staining of cross-sections of hair viewed bt TEM when embedded in LR White than Araldite resin.
We are looking for a way to allow us to orientate a bundle of hair fibres (about 5mm long) and keep the bundle intact whilst heat-curing the LR White anaerobically. We do not have the means to lower the temperature of the resin, nor use UV light for curing.
Can anyone suggest a means of keeping the bundle of hairs intact within the LR White whilst embedding and curing? Any support must be able to withstand the effects of acetone and LR White resin in liquid form.
Thanks for your attention
Jeremy Sanderson Please reply to jb_sanderson-at-yahoo.com
********************************************************************** Dr Kingsley Micklem * MEDICAL INFORMATICS UNIT Director, Medical Informatics Unit * CELLULAR SCIENCE University Department of Cellular Science * Room 5501, John Radcliffe Hospital, * OXFORD Headington, Oxford, OX3 9DU * Tel 44 (1865) 222039 * website: FAX 44 (1865) 221693 *http://phoenix.jr2.ox.ac.uk **********************************************************************
Here is a question for those of you who use sucrose to provide cryoprotection for tissues:
We are preparing perfusion fixed brain tissue by freeze substitution for post-embedding immuno gold. Following fixation we prepare 500 micron vibratome sections, which we then wish to infiltrate with 2 M sucrose in buffer prior to freezing. We have been transferring the brain sections to 1 M sucrose, then when they sink, transferring again to 2 M. The problem is that the sections never sink in the 2 M sucrose.
Do others have this problem? Do we have to cut the sections into small pieces first, or is there some other way to get them to go under? Or should we not worry about making them sink?
Thanks for your advice.
Marie
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology, U-2131 University of Connecticut Storrs, CT 06269-2131 Phone: 860-486-3588 Fax: 860-486-1936
I have done similar things in a manner such as this: Take the cap off a BEEM capsule and lay it down open side up. Lay your hair fibers flat in the cap and add LRW. Place a glass slide on top of the cap to seal it off during polymerization. Heat polymerize. Use a dremel moto-tool or jewelers hacksaw to trim out a rectangle of the LRW. Good luck, Tom
} } } We find that we get better silver staining of cross-sections of hair } viewed bt TEM when embedded in LR White than Araldite resin. } } We are looking for a way to allow us to orientate a bundle of hair fibres } (about 5mm long) and keep the bundle intact whilst heat-curing the LR } White anaerobically. We do not have the means to lower the temperature of } the resin, nor use UV light for curing. } } Can anyone suggest a means of keeping the bundle of hairs intact within } the LR White whilst embedding and curing? Any support must be able to } withstand the effects of acetone and LR White resin in liquid form. } } Thanks for your attention } } Jeremy Sanderson } Please reply to jb_sanderson-at-yahoo.com } } ********************************************************************** } Dr Kingsley Micklem * MEDICAL INFORMATICS UNIT } Director, Medical Informatics Unit * CELLULAR SCIENCE } University Department of Cellular Science * } Room 5501, John Radcliffe Hospital, * OXFORD } Headington, Oxford, OX3 9DU * } Tel 44 (1865) 222039 * website: } FAX 44 (1865) 221693 *http://phoenix.jr2.ox.ac.uk } **********************************************************************
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)
Jeremy, We have had good luck keeping roots straight by sanwiching them between films of formvar on wire loops. We make a loop with a 3mm diameter and a long stem out of very thin copper wire (36 gauge in usa). We then cast rectangles of formvar about 4 mm x 8 mm and line the loop up over the floating rectangle so that the plane of the loop is parallel to the short side of the rectangle and right in the middle. Then we quickly plunge the loop straight down onto the formvar and into the water and pull it back out again. This gives us a nice film on the loop. Give this at least a few hours to dry (but they will keep for ages--we plant the stem of the loop in some wax and cover with a beaker). Then when ready to go, place your sample, either before or after fixation, on the loop and repeat with a second formvar rectangle.Now you have the sample sandwiched. It is very nice for small samples because solution exchange becomes a snap. The formvar definitely stands up to actetone, butyl methyl methacrylate, Spurs, and I am 95% sure LR white. It is easy to dehydrate through and get other things through even antibodies go through (although they are slowed down a bit).
Hope this helps.
You got more questions, let me know.
Tobias Baskin
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We are facing a similar problem here and have just had some preliminary success with polymerizing LR White in a vacuum dessicator placed in an oven. We were using gelatin capsules full of LR White placed upside down over coverslips upon which cells have been grown. Standard polymerization did not adhere the gel caps to the coverslips, supposedly due the presence of oxygen (it was worth a try). However, the trial run in the dessicator seems to have worked well, and the cover slip detached cleanly when placed in liquid nitrogen.
You might try this with flat embedding molds to preserve the orientation of your bundles. If you do, please let us know the results.
Another approach might be to embed the hair bundles and later cut them out of the LR White in a piece of resin, orient them the way you want, and glue the resin piece onto the tip of a blank block for ultramicrotomy.
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/
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We find that we get better silver staining of cross-sections of hair viewed bt TEM when embedded in LR White than Araldite resin.
We are looking for a way to allow us to orientate a bundle of hair fibres (about 5mm long) and keep the bundle intact whilst heat-curing the LR White anaerobically. We do not have the means to lower the temperature of the resin, nor use UV light for curing.
Can anyone suggest a means of keeping the bundle of hairs intact within the LR White whilst embedding and curing? Any support must be able to withstand the effects of acetone and LR White resin in liquid form.
Thanks for your attention
Jeremy Sanderson Please reply to jb_sanderson-at-yahoo.com
********************************************************************** Dr Kingsley Micklem * MEDICAL INFORMATICS UNIT Director, Medical Informatics Unit * CELLULAR SCIENCE University Department of Cellular Science * Room 5501, John Radcliffe Hospital, * OXFORD Headington, Oxford, OX3 9DU * Tel 44 (1865) 222039 * website: FAX 44 (1865) 221693 *http://phoenix.jr2.ox.ac.uk **********************************************************************
off the top of my head: pack hairs into a small tube made out of thin paper - velin tissue or Rizla cigarette paper wrapped round a cocktail stick - the gum on the edge of a cigarette paper should survive the resin
Date sent: Fri, 24 Mar 2000 08:50:13 -0600 To: Microscopy-at-sparc5.microscopy.com } From: Kingsley Micklem {kingsley.micklem-at-cellular-science.oxford.ac.uk}
Hi, all,
Many suggestions have been given to me after my posting the Personal TEM Website http://syli.homepage.com in this list sever. Thanks a lot to these contributers!
Now the site has been updated according to these good suggestions. More journals are added into the TEM-related Journals. and an additional page for JOB LIST and RESUMES was included for head-huntings in TEM region. Now you may send me your head-hunting ads or your resume to me and I will place it in this part. Thanks again.
Shu-You Li ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Please visit my new homepage - a personal website on transmission electron microsocpy (TEM) - at http://syli.homepage.com at your convenience! It contains not only my current research work, but also LINKS to useful journal informations, instruction for authors, on-line EELS database, periodic table, physical constants, online TEM course and etc. It also contains JOB list and RESUMEs for head-hunting in TEM region. I am looking forward to your invaluable suggestions! ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ ************************************************** Shu-You Li, Dr. Institut fuer Physikalische Chemie Johannes Guttenberg Universitaet Jakob-Welder-Weg 11 D-55099 Mainz, Germany
OK - this is extremely low-tech, but it works (if none of the other suggestions you receive do the trick). Cotton thread! I've made bundles of fibres by tying them up with fine sewing thread. You can tie several places along the length of the bundle so it will be held more firmly if you need to. You can process, dehydrate, and embed. Just make certain that you have cotton thread - some of the polymer ones might melt in the solvents or resins.
I've also heard of using fishing monofilament for stuff that doesn't require nasty solvents - this is probably just the "guy version" of the thread technique :)
Tamara Howard CSHL
On Fri, 24 Mar 2000, Kingsley Micklem 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 find that we get better silver staining of cross-sections of hair } viewed bt TEM when embedded in LR White than Araldite resin. } } We are looking for a way to allow us to orientate a bundle of hair fibres } (about 5mm long) and keep the bundle intact whilst heat-curing the LR } White anaerobically. We do not have the means to lower the temperature of } the resin, nor use UV light for curing. } } Can anyone suggest a means of keeping the bundle of hairs intact within } the LR White whilst embedding and curing? Any support must be able to } withstand the effects of acetone and LR White resin in liquid form. } } Thanks for your attention } } Jeremy Sanderson } Please reply to jb_sanderson-at-yahoo.com } } ********************************************************************** } Dr Kingsley Micklem * MEDICAL INFORMATICS UNIT } Director, Medical Informatics Unit * CELLULAR SCIENCE } University Department of Cellular Science * } Room 5501, John Radcliffe Hospital, * OXFORD } Headington, Oxford, OX3 9DU * } Tel 44 (1865) 222039 * website: } FAX 44 (1865) 221693 *http://phoenix.jr2.ox.ac.uk } ********************************************************************** } } } }
Of course you should all consider that cutting or grinding ANY embedding resin should be undertaken with the knowledge that resin dust can cause serious health effects. I don't know if any of my allergies or other health quirks have been 'encouraged' from exposure to resin dust or solvents, but judging from the discussions on the listserver about some people's experiences with resins, I recommend that you treat that dust with the same respect as paraformaldehyde powder or asbestos!
Just thought I'd share that safety concern.
Gregg Sobocinski Parke Davis Pharmaceutical Research Ann Arbor, Michigan, USA Gregg.Sobocinski-at-wl.com
-----Original Message----- } From: Tom Phillips [mailto:PhillipsT-at-missouri.edu] Sent: Friday, March 24, 2000 10:28 AM To: Kingsley Micklem Cc: Microscopy-at-sparc5.microscopy.com
I have done similar things in a manner such as this: Take the cap off a BEEM capsule and lay it down open side up. Lay your hair fibers flat in the cap and add LRW. Place a glass slide on top of the cap to seal it off during polymerization. Heat polymerize. Use a dremel moto-tool or jewelers hacksaw to trim out a rectangle of the LRW. Good luck, Tom
} } } We find that we get better silver staining of cross-sections of hair } viewed bt TEM when embedded in LR White than Araldite resin. } } We are looking for a way to allow us to orientate a bundle of hair fibres } (about 5mm long) and keep the bundle intact whilst heat-curing the LR } White anaerobically. We do not have the means to lower the temperature of } the resin, nor use UV light for curing. } } Can anyone suggest a means of keeping the bundle of hairs intact within } the LR White whilst embedding and curing? Any support must be able to } withstand the effects of acetone and LR White resin in liquid form. } } Thanks for your attention } } Jeremy Sanderson } Please reply to jb_sanderson-at-yahoo.com } } ********************************************************************** } Dr Kingsley Micklem * MEDICAL INFORMATICS UNIT } Director, Medical Informatics Unit * CELLULAR SCIENCE } University Department of Cellular Science * } Room 5501, John Radcliffe Hospital, * OXFORD } Headington, Oxford, OX3 9DU * } Tel 44 (1865) 222039 * website: } FAX 44 (1865) 221693 *http://phoenix.jr2.ox.ac.uk } **********************************************************************
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)
hi earl- i've been considering the whole service contract routine lately since i have an older sem, a newer fesem, and a vanilla tem under contract. the sum of my contracts (from the vendors) is about $40K. i have a hard time justifying this level of expenditure, but have been burned in the past with replacement part costs under "limited" arrangements. in my role i have to recover all lab costs, and contracts are a substantial portion of them.
my main vendor (SEMs) has offered me a 5% "discount". this seems a bit light (to me) because i am limited in terms of my cost recovery mechanisms. a commercial or industrial lab would seem to have more flexibility than a shared facility at a small university. currently i'm negotiating with them to extend this discount. i'm not sure what will happen if i can't keep these costs under control...perhaps this is an opportunity for a crafty entrepreneur. at the very least it may be a signal to look for 3-rd party service, in-house methods, and/or go bare on older equipment.
brian ******************** } OK to rephase: How many have experienced an extreme reduction in service } contract price (20% or more) when confronted with competition? } } Thank You, } } Earl Weltmer
---------------------------------------------- Brian McIntyre Electron Microscopy Lab- River Campus Univ of Rochester Rochester, NY 14627 716-275-3058/4875
Does anyone have for sale, or know of a supplier of Au/Pd target for the older model Balzers-Union Sputtering Device Type 07120-A, serial number 235? It is an early version of that coater series and uses targets for an SCD010 type coater with serial numbers 101-317. The target is ~5 cm diameter with a threaded mounting hole. Thanks!
Hello Listers, We are trying to do some electron microprobe work on the Yucca Mountain Project for the DOE. However, we may need external verification of our standards by a laboratory that is on the Qualified Supplier List, specifically for the YMP. Therefore, unless a lab has done work directly for Yucca Mountain specifically, they don't count (even DOE run
national labs). This is all according to our QA people, and I am hoping
that someone out there may have direct knowledge of the YMP QA and/or know someone who does. Please respond offline to me. Many thanks in advance, Sarah
Sarah A.W. Lundberg Lab (EPMA) (702) 895-2660 or Electron Microanalysis and (SEM) (702) 895-2462 Imaging Laboratory Office (702) 895-1134 Department of Geoscience, UNLV Fax (702) 895-4064 4505 S. Maryland Parkway Box 454010 Las Vegas, NV 89154-4010 Dept Office (702) 895-3262
I am thinking of trying to put together a laser tweezer setup on my microscope. It doesn't seem too difficult for a simple system, but I am having trouble finding the appropriate laser so I don't have to build it myself from the module. Has anyone out there found a good source for a near IR laser appropriate for this application? Any other words of wisdom for a laser/laser tweezer novice appreciated. Dave Knecht --
Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 75 N. Eagleville Rd. U-125 Storrs, CT 06269 Knecht-at-uconnvm.uconn.edu 860-486-2200 860-486-4331 (fax) home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html ************************************************************
Micklem, I embed a variety of fibers for cross-sectioning using the following. Collect little round pieces of paper from your office paper hole punch. Using a sharp needle punch an appropriately sized hole in the center of several of them. Thread your fibers to be embedded through the center hole of the paper circles. The number required will depend on the length, diameter and mass of the fibers to be sectioned. I find that a fine pair of tweezers and a disectin microscope make this job easier. Gently place this assembly into your embedding capsule (I use BEEM 00). With a pipette, slowly add resin down the sides to fill the capsule (I use Spurrs with Z-6040 adhesion promoter). The fiber/paper assembly will self-center in the capsule and maintain this orientation through curing. Be careful with your choice of paper, it must be dry, and some porous paper will outgas bubbles. Test paper samples ahead of time to identify a suitable type. I'm sure you can adapt this to work with LR White.
Good luck,
Phil Swab Engineering Development Deposition Sciences Inc. Santa Rosa, CA 707-566-3718 phil.swab-at-depsci.com
-----Original Message----- } From: Kingsley Micklem [SMTP:kingsley.micklem-at-cellular-science.oxford.ac.uk] Sent: Friday, March 24, 2000 6:50 AM To: Microscopy-at-sparc5.microscopy.com
We find that we get better silver staining of cross-sections of hair viewed bt TEM when embedded in LR White than Araldite resin.
We are looking for a way to allow us to orientate a bundle of hair fibres (about 5mm long) and keep the bundle intact whilst heat-curing the LR White anaerobically. We do not have the means to lower the temperature of the resin, nor use UV light for curing.
Can anyone suggest a means of keeping the bundle of hairs intact within the LR White whilst embedding and curing? Any support must be able to withstand the effects of acetone and LR White resin in liquid form.
Thanks for your attention
Jeremy Sanderson Please reply to jb_sanderson-at-yahoo.com
********************************************************************** Dr Kingsley Micklem * MEDICAL INFORMATICS UNIT Director, Medical Informatics Unit * CELLULAR SCIENCE University Department of Cellular Science * Room 5501, John Radcliffe Hospital, * OXFORD Headington, Oxford, OX3 9DU * Tel 44 (1865) 222039 * website: FAX 44 (1865) 221693 *http://phoenix.jr2.ox.ac.uk **********************************************************************
A client asked me to photograph bacilli and endospores on a cotton thread (he supplied the sample already sputtered with au/pd). I took one really nice photo, then began experiencing movement of the thread fibers due to electrostatic charges. It seems the cells & endospores are concentrated at the unsupported ends of the fibers; when I look at thte interior, bundled fibers, which are better restrained, they are practically devoid of cells. I can only guess that the cells/spores were drawn to the outer edges of the thread as the buffer evaporated.
Any advice, insights or ideas will be rewarded with a big Thank You, Thank You, Thank You.
Cheers,
Paul Grover :o) Chief Microscopist and Bottle Washer Microvista Laboratory 1220 Cincinnati St. Lafayette, IN 47904
} From: "Sobocinski, Gregg" {Gregg.Sobocinski-at-WL.com} } } } Of course you should all consider that cutting or grinding ANY embedding } resin should be undertaken with the knowledge that resin dust can cause } serious health effects. I don't know if any of my allergies or other health } quirks have been 'encouraged' from exposure to resin dust or solvents, but } judging from the discussions on the listserver about some people's } experiences with resins, I recommend that you treat that dust with the same } respect as paraformaldehyde powder or asbestos! } } Just thought I'd share that safety concern.
The unrecalled products cause more reaction problems than the cured product.
But as one that has been too careless with air quality don't risk you health breathing dusts, chemicals or solvents any more than absolutely necessary.
I think the reaction products of an automobile my be worse than most things in the lab so you may need to wear you respirator on your drive to work. About the time the catalytic converter came along my asthma really flared. some how I don't thing H2SO3 is good for me.
Take good care of you health if the average age at death continues it present course you may have to live to 120 or more.
Take care Gordon
G. C,. Couger 624 Cheyenne Stillwater, OK 74075-1411 405 624 2855 between 3:00 pm to 1:00 am CST 405 742 2758 cell
We solved the resin dust/ Dremel tool problem by rigging a small vacuum cleaner with the intake very close to where we would be grinding, which was usually under a disecting scope. I got the idea while watching a cast being cut from a broken foot. The cast cutter had a vacuum attached. Our rig takes care of the fine particles, while larger ones generally fall to the earth and can be cleaned up later. This more of an issue with epoxies, I suspect. LR White is a dental resin, I believe, and the dentist will grind in your mouth or over you using your chest as a table. I don't know if that means it is safe.
Greg Erdos University of Florida Greg Erdos 5410 SE 185th AVe Micanopy, FL 32667 352-466-0843
You might be able to buy a metal foil of a suitable size and fix it to what you already have (or a custom made holder) using silver-loaded epoxy?
Goodfellow Metals (England) used to have such foils. I could check this further if you wish, plus send you their contact details when I am back in the lab.
Regards - Keith
_______________________________ Dr. Keith Ryan Marine Biological Association of the UK The Laboratory Citadel Hill Plymouth Devon PL1 2PB England
Tel. ++44 (0)1752 633249 Tel. ++44 (0)1752 633279 The 279 number has an answering machine
Hi, I had done an evapouration of cobalt on a quartz sample. Now I want to etch the quartz sample to remove the substrate so that I can do Transmission Microscopy for charactrization. My problem is that If I etch the substrate with HF will it be harmful to carbon? If it is so what are the other materials that can be used to etch quartz. I will be very pleased if you can answer me. Thanking you Ramu, Senior student in M.Sc, Dept of Physics, IIT Bombay.
meet me at ramu8sp-at-ccs.iitb.ernet.in suvee-at-wowmail.com, srisuvee-at-yahoomail.com *******************************************************************************
Does anyone know of some sort of vacuum vessel that would hold a good number of boxed specimens under vacuum? I have several mechanical pumps (dual stage) but have not seen any sizable, sturdy containers. I would like to store my specimens under a vacuum at all times unless loading into the SEM. The purpose is to keep any fungi, moisture and dust from contaminating the specimens.
The problem with large vacuum containers is strength and therefore cost. Its those 14.7 pounds for every sq inch the atmosphere exerts on vacuum vessels, a fact EMists know and understand. Some don't realize that another order of magnitude in high vacuum makes no real difference to that pressure and a very poor vacuum still exerts over 14 lbs. on every sq inch of surface area.
The practical and economic solution is the use airtight cabinets with drying agents or a trickle of dry nitrogen gas. Afterall its mostly moisture that impedes pumping in the SEM or causes specimens and coatings to fall apart with time. Disclaimer: Proscitech ships world-wide a large of desiccating cabinets and vacuum desiccators. See Online } Contents } Page E7 Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Sunday, March 26, 2000 11:05 AM, Dr. Gary Gaugler [SMTP:gary-at-gaugler.com] wrote:} } Does anyone know of some sort of vacuum vessel that } would hold a good number of boxed specimens under } vacuum? I have several mechanical pumps (dual stage) } but have not seen any sizable, sturdy containers. I would } like to store my specimens under a vacuum at all times } unless loading into the SEM. The purpose is to keep any } fungi, moisture and dust from contaminating the specimens. } } Vendor responses are welcome. } } gary g. }
I am trying to help a new SEM user w/ a tough resolution shot. For some reason he was not able to find the quoted resolution for the SEM (JEOL JSM 6400) he was using. If anybody out there happens to know its exact or approximate quoted resolution, that would be very helpful. His electron source is W.
thanks to the many responders about this post. Let me add some more info to help zero in on a solution--if there is one.
The reason I am seeking a vacuum container is two-fold. First, I have very limited N2 availability. I use industrial grade bottled N2 and dry it with a molecular sieve for venting my SEM chamber during specimen exchange. I vent at about 5psi. The bottles are at 1900 psi and hold 130cu ft of N2. One bottle lasts about two months. I have two bottles--on-line and standby. A "trickle" of N2 through a dessicator would work but for how long per bottle? I don't know.
The other factor for a vacuum container is that if a specimen is under a roughing pump vacuum, if it is at all defective, the body of the specimen will implode. Thus saving the time of fooling with it in the SEM. If it does not implode, then it will be dry and not require very much pump down time before opening the column isolation valve. Thus ensuring minimal effect on the column ion pump.
Steve D'Angelo showed a very nice metal dessicator unit, which would be great if I had a lot of N2....I presume a lot of N2. If I take one of my 130 cu ft cylinders and leak N2 into the dessicator, about how long will it last? And what is the minimum leak pressure needed to make the dessicator function as a drying unit? I could get another cylinder of N2 and another sieve for this unit if the gas volume was sufficient to run the box for two to three months.
University of Central Florida Advanced Materials Processing and Analysis Center (AMPAC)
The Advanced Materials Processing and Analysis Center (AMPAC) is seeking candidates for a position as Assistant in Research to support the Materials Characterization Facility (MCF). The individual must have a Master's degree or a Bachelor's degree from an accredited institution in an appropriate area of study related to surface science and materials characterization, and have at least three years of experience in materials characterization or related areas. The person will be responsible for establishing and maintaining laboratory infrastructure, laboratory maintenance, equipment installation, operation and maintenance, and be an investigator in contracted research. The person must be able to conduct independent research and development in materials characterization with an emphasis in ion beam technology and have the ability to interact with students and provide instruction in the use and interpretation of data. The individual must have extensive knowledge and experience in the operation and maintenance of materials characterization equipment. AMPAC is particularly interested in individuals desiring an academic setting to further their professional goals. The applications will be reviewed beginning April 17, 2000 and will continue to be reviewed until the position is filled.
Applicants should send vitae and three references to Dr. Vimal Desai, Director, 12424 Research Parkway, Suite 408, Orlando, FL 32826. UCF is an equal opportunity/affirmative action employer. As an agency of the State of Florida, all application materials and selection procedures are available for public review.
******************************************************************* Lucille A. Giannuzzi, Ph.D.
Associate Professor, Mechanical, Materials, and Aerospace Engineering Director, UCF/Cirent Materials Characterization Facility University of Central Florida 12443 Research Parkway, Suite 305 Orlando, FL 32826 email lag-at-mail.ucf.edu phone (407) 275-4354,5,6 fax (407) 275-4321 -------------------------------------------------------------------- "Good judgement comes from experience.
Experience comes from making bad judgement."
Mark Twain ********************************************************************
Gary, Are you concerned about rmp backstreaming, thereby contaminating your specimens with hydrocarbon. I assume you are using a filter with a baking element? -Ken
Hi Gary, How about a TEM film dessicator? Available from most EM suppliers, vacuum companies and some scrapped TEMs.
Ron
=========================================================================== Mr. Ron Doole e-mail ron.doole-at-materials.ox.ac.uk Department of Materials, phone +44 (0) 1865 273701 University of Oxford, fax +44 (0) 1865 283333 Parks Road. Oxford. OX1 3PH. UK. ============================================================================
you mention using a small vacuum cleaner to clean up resin dust. You don't specify whether it has an exhaust filter so I think I should just mention that the typical domestic vacuum cleaner exhausts fine particulates and may well make the problem worse because it may be pumping the very size of resin particles out that you want to avoid. We once used a small portable vacuum cleaner for cleaning up resin dust but stopped when I realized that there was a potential risk.
I know that there are now domestic and industrial vacuum cleaners which have very fine exhaust filters and there are also the small photocopier/toner vacuum cleaners. Has anyone investigated their use for resin dust?
Malcolm
Malcolm Haswell Electron Microscopy School of Sciences Fleming Building University of Sunderland SUNDERLAND SR1 3SD Tyne and Wear UK
Tel (0191) 515 2872 e-mail: malcolm.haswell-at-sunderland.ac.uk
greg erdos 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 solved the resin dust/ Dremel tool problem by rigging a small vacuum } cleaner with the intake very close to where we would be grinding, which was } usually under a disecting scope. I got the idea while watching a cast } being cut from a broken foot. The cast cutter had a vacuum attached. } Our rig takes care of the fine particles, while larger ones generally fall } to the earth and can be cleaned up later. } This more of an issue with epoxies, I suspect. LR White is a dental resin, } I believe, and the dentist will grind in your mouth or over you using your } chest as a table. I don't know if that means it is safe. } } Greg Erdos } University of Florida } Greg Erdos } 5410 SE 185th AVe } Micanopy, FL 32667 } 352-466-0843
In Australia we can readily purchase industrial dry N2, this obviates the drying agent and filtering of the gas. In WDS, gas proportional spectrometers are fed with about one bubble a second (holding the outlet into water). At that rate a cylinder routinely last for over a year and although an expensive gas is used for that purpose, cylinder rental invariably is the greater cost. A tight drying cabinet only requires a similar amount of gas to maintain a positive pressure, however slight. Specimens added to the cabinet should be dry and I think that a tray of desiccant is a good idea.
No additional cylinder would be required since Gary already has two. I used to deplete the N2 cylinder after use on the EMs, to nitrogen burst during film development. Since the EM duty cylinder does not go empty unexpectedly, the cylinder feeding the drying cabinet can be removed in time obtain a full cylinder. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Monday, March 27, 2000 1:59 AM, Dr. Gary Gaugler [SMTP:gary-at-gaugler.com] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } thanks to the many responders about this post. Let me add some } more info to help zero in on a solution--if there is one. } } The reason I am seeking a vacuum container is two-fold. First, } I have very limited N2 availability. I use industrial grade bottled N2 } and dry it with a molecular sieve for venting my SEM chamber } during specimen exchange. I vent at about 5psi. The bottles } are at 1900 psi and hold 130cu ft of N2. One bottle lasts about } two months. I have two bottles--on-line and standby. A "trickle" } of N2 through a dessicator would work but for how long per } bottle? I don't know. } } The other factor for a vacuum container is that if a specimen is } under a roughing pump vacuum, if it is at all defective, the body } of the specimen will implode. Thus saving the time of fooling with } it in the SEM. If it does not implode, then it will be dry and not } require very much pump down time before opening the column } isolation valve. Thus ensuring minimal effect on the column ion } pump. } } Steve D'Angelo showed a very nice metal dessicator unit, which } would be great if I had a lot of N2....I presume a lot of N2. If } I take one of my 130 cu ft cylinders and leak N2 into the dessicator, } about how long will it last? And what is the minimum leak pressure } needed to make the dessicator function as a drying unit? I could } get another cylinder of N2 and another sieve for this unit if the } gas volume was sufficient to run the box for two to three months. } } Appreciate your feedback. } } gary g. } }
Does anyone no of a good web source for info on light microscopy for undergraduates. Ideally I'm after reference material on brightfield, phase contrast and fluorescent microscopies. Some info on new areas would also be useful. Hope someone out there can help !
Thanks in Advance,
Barry Shaw
Barry Shaw Senior Imaging Technician Molecular & Cell Biology E Floor School Of Biomedical Sciences University of Nottingham Medical School NG7 2UH
I am looking to purchase an Zeiss INKO condenser with 4 Wollaston prisms. The turret position/Wollaston prism marked IIII was used in conjunction with the Plan Achromat 6.3x objective.
What was the difference between the INKO condenser (46 52 79) and the INKO slider III (47 44 44) and INKO slider II (47 44 31)? According to the literature, the type II INKO slider was for research microscopes and the type III for the the smaller STANDARD microscope.
Using a Photom I, I have tried the INKO condenser in position IIII with both type III and type II sliders and am not able to obtain DIC with a PLAN achromat x6.3 objective.
I am very familiar with the 3 position Wollaston prism condenser and am able to obtain great interference DIC with position I and the PLAN x16 objective. However, using the 4 position condenser on prism IIII yields no DIC using the x6.3 objective. As a side note, the prisms are in great shape and show no sign of separation.
I have noticed the dark fringe interference figures for IIII, III, and II are oriented at 11:00 and 5:00 when viewed through an inverted condenser, while in position I, the dark fringe is oriented at 1:00 and 7:00. (hand held with the condenser inverted, the polarizer is placed on top of the dove-tailed retainer ring to position and lock the condenser into the condenser holder, and the INKO slider is held at 45 degrees to the east -west oriented polarizer under the top condenser lens element, i.e., the entire system is inverted.) If this makes sense, great!
Any suggestions as to why the number IIII position does not yield DIC using the 6.3 objective?
Thanks Ken --------------- Ken Tiekotter, Adjunct Professor The University of Portland Dept. of Biology 5000 N Willamette Bldv. Portland, OR 97303
Hi, I am looking for carbon rods with spectrographic quality for Edwards vacuum evaporator. The rods should be about 1.5mm in diameter and 30mm in length. I have tried several suppliers without success. If any of you have information regarding who supplies this kind of carbon rods could you please let me know? Your help will be greatly appreciated. Thank you.
} Chesapeake Society for Microscopy } } Is pleased to announce } } The PhotoShop Workshop } } April 11th,2000 } } Morning Session } 9:00am-12: 00 noon } Introduction and Intermediate level } } Afternoon Session } 1:00pm- 4:00pm } More Advanced Issues and Discussion } } Location: NIH, Building 4 Room 433 } } } Seating is limited to 50 people } Preregistration for CSM members by March 31st No Charge } After March 31st Open Registration } CSM members No charge } Non-members - $20.00 per session } } To register Contact Andrea Weisberg } Phone: (301) 435-1977 } e-mail: aweisberg-at-nih.gov } } } Please register early, we can only seat 50 people. } Ask me about how to become a CSM member } Membership $10.00 per year } } Andrea S. Weisberg } NIH/NIAID/LVD } Bldg. 4/Room 210 } 4 Center Drive } Bethesda, MD 20892-0445 } phone: (301) 435-1977 } fax: (301) 480-1147 } e-mail: aweisberg-at-nih.gov }
I would be very concerned about vacuum pump oil contamination of your specimens if you us oil sealed roughing pumps on such a vacuum chamber. Oil backstreaming will occur unless you use a trap and are meticulous about trap maintenance.
Ronald Vane XEI Scientific
-----Original Message----- } From: Dr. Gary Gaugler {gary-at-gaugler.com} To: MSA listserver {Microscopy-at-sparc5.microscopy.com}
Hi Brian,
Service contract can be likened to "insurance policies'. We are in the service business & offer service contracts for less mostly because we can work on several manufacturers and save on travel time & costs. Still we allot about 20% of the contract costs to parts.
When you "go bare" (self insure) the equipment will probably be OK for about two years if the maintenance has been properly done. After that it is anyone's guess.
When we offer reduced service contract prices for reduced risk (customer buys the parts or limited number of service calls) the discount is about 20-30% to cover parts costs. The largest expenditure is labor and parts in that order.
Competition generally helps the equation but the question that I had is when "competition" is only done when faced with a start-up entrepreneurs. I would ask for a further discount of at least 20%.
Regards,
Earl Weltmer (714) 573-9158
Brian McIntyre 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 earl- } i've been considering the whole service contract routine lately since i } have an older sem, a newer fesem, and a vanilla tem under contract. the } sum of my contracts (from the vendors) is about $40K. i have a hard time } justifying this level of expenditure, but have been burned in the past with } replacement part costs under "limited" arrangements. in my role i have to } recover all lab costs, and contracts are a substantial portion of them. } } my main vendor (SEMs) has offered me a 5% "discount". this seems a bit } light (to me) because i am limited in terms of my cost recovery mechanisms. } a commercial or industrial lab would seem to have more flexibility than a } shared facility at a small university. currently i'm negotiating with them } to extend this discount. i'm not sure what will happen if i can't keep } these costs under control...perhaps this is an opportunity for a crafty } entrepreneur. at the very least it may be a signal to look for 3-rd party } service, in-house methods, and/or go bare on older equipment. } } brian } ******************** } } OK to rephase: How many have experienced an extreme reduction in service } } contract price (20% or more) when confronted with competition? } } } } Thank You, } } } } Earl Weltmer } } ---------------------------------------------- } Brian McIntyre } Electron Microscopy Lab- River Campus } Univ of Rochester } Rochester, NY 14627 } 716-275-3058/4875
A Cambridge S240 scanning electron microscope is available to any interested party for moving expenses and best offer. Please contact me directly. Thank you and kind regards, Diana
Diana L. Kittleson Pillsbury Technology East 737 Pelham Blvd. St. Paul, MN 55114 651-917-5859 651-917-5850 fax www.tpclabs.com
______________________________________________________________________ This e-mail and any attachment contains information which is private and confidential and is intended for the addressee only. If you are not an addressee, you are not authorized to read, copy or use the e-mail or any attachment. If you have received this e-mail in error, please notify the sender by return e-mail and then destroy it.
I have used a Snappy digitizer for the last 6 years with good results. I use it with a Javelin color camera. I,m curious, how did you get it to work with a monochrome camera?? The Snappy syncs on the color burst portion of the video waveform. I have never been able to get a oicture with a monochrome camera or electron microscope.
Dear Sir: I am looking for a PAM interface board/card which would plug into a slot in a Personal Computer. I have a Link Analytical EDX connected to a Hitachi S520LB SEM. The pulse processor for the EDX is a stand alone unit which connects via a 25 pin ribbon cable to a personal computer of older vintage i.e.. 80286 or 80386 technology. The connection is into a PAM Interface Card. The person I purchased this unit from was unable to locate the PAM Interface Board. I am looking for this board. Does anyone know where I can locate one? Thank You, David Browning dbrownng-at-stargate.net
Gary: Why not store samples in a standard vacuum desiccator (Fisher Scientific, VWR, etc). They come in glass (with ground glass or o-ring seals) and plastic (with o-ring seals). However, don't leave samples in a desiccator under active pumping. All pumps backstream. It's just a matter of how much contamination your samples can tolerate. Pump the desiccator to some nominal reduced pressure (a few seconds is fine) and close the valve. A silica gel or similar desiccant can be used concurrently in the dessicator and will more effectively pump residual water out of your samples at this reduced pressure.
Phil Swab Engineering Development Deposition Sciences Inc. Santa Rosa, CA 707-566-3718 phil.swab-at-depsci.com
-----Original Message----- } From: Dr. Gary Gaugler [SMTP:gary-at-gaugler.com] Sent: Sunday, March 26, 2000 7:59 AM To: MSA listserver Cc: Steve D'Angelo
thanks to the many responders about this post. Let me add some more info to help zero in on a solution--if there is one.
The reason I am seeking a vacuum container is two-fold. First, I have very limited N2 availability. I use industrial grade bottled N2 and dry it with a molecular sieve for venting my SEM chamber during specimen exchange. I vent at about 5psi. The bottles are at 1900 psi and hold 130cu ft of N2. One bottle lasts about two months. I have two bottles--on-line and standby. A "trickle" of N2 through a dessicator would work but for how long per bottle? I don't know.
The other factor for a vacuum container is that if a specimen is under a roughing pump vacuum, if it is at all defective, the body of the specimen will implode. Thus saving the time of fooling with it in the SEM. If it does not implode, then it will be dry and not require very much pump down time before opening the column isolation valve. Thus ensuring minimal effect on the column ion pump.
Steve D'Angelo showed a very nice metal dessicator unit, which would be great if I had a lot of N2....I presume a lot of N2. If I take one of my 130 cu ft cylinders and leak N2 into the dessicator, about how long will it last? And what is the minimum leak pressure needed to make the dessicator function as a drying unit? I could get another cylinder of N2 and another sieve for this unit if the gas volume was sufficient to run the box for two to three months.
Balzers Union renamed to BAL-TEC AG in 1992. But the product line was maintained and new developments are carried out. There are several coating systems of this older type in use. So of course we can deliver these targets furthermore. Article No. of this target: BU 007 129 -T
Our reprepresentative in USA / Canada:
Techotrade International 7 Perimeter Road Manchester, NH 03103-3343 Mr. Johnny Hagen T: +1 603 622-5011
Our representative will contact you directly to assist with further information.
Ulrike Zeile
BAL-TEC AG EM-Technology and Application FL-9496 Balzers Tel. +423 388 12 36 Fax +423 388 12 60
-----Ursprüngliche Nachricht----- Von: Mandayam V. Parthasarathy [mailto:mvp2-at-cornell.edu] Gesendet: Freitag, 24. März 2000 21:22 An: Microscopy-at-sparc5.microscopy.com Betreff: Balzers-Union putter coater
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Does anyone have for sale, or know of a supplier of Au/Pd target for the older model Balzers-Union Sputtering Device Type 07120-A, serial number 235? It is an early version of that coater series and uses targets for an SCD010 type coater with serial numbers 101-317. The target is ~5 cm diameter with a threaded mounting hole. Thanks!
Balzers Union renamed to BAL-TEC AG in 1992. But the product line was maintained and new developments are carried out. For additional information have a look at our webside www.bal-tec.com. There are several coating systems of this older type in use. So of course we can deliver these targets furthermore. Article No. of this target: BU 007 129 -T
Our reprepresentative in USA / Canada:
Techotrade International 7 Perimeter Road Manchester, NH 03103-3343 Mr. Johnny Hagen T: +1 603 622-5011
Our representative will contact you directly to assist with further information.
Ulrike Zeile
BAL-TEC AG EM-Technology and Application FL-9496 Balzers Tel. +423 388 12 36 Fax +423 388 12 60
-----Ursprüngliche Nachricht----- Von: Mandayam V. Parthasarathy [mailto:mvp2-at-cornell.edu] Gesendet: Freitag, 24. März 2000 21:22 An: Microscopy-at-sparc5.microscopy.com Betreff: Balzers-Union putter coater
of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html ------------------------------------------------------------- ----------. ++ ++ ++ Does anyone have for sale, or know of a supplier of Au/Pd ++ target for the ++ older model Balzers-Union Sputtering Device Type 07120-A, ++ serial number ++ 235? It is an early version of that coater series and uses ++ targets for an ++ SCD010 type coater with serial numbers 101-317. The target is ~5 cm ++ diameter with a threaded mounting hole. Thanks! ++ ++ ++ ******************************************************************* ++ ++ M.V. Parthasarathy ++ Prof. of Plant Biology, Adjunct Prof. of Biomedical Sciences (Vet), & ++ Director, Cornell Integrated Microscopy Center (CIMC) ++ Section of Plant Biology ++ 228 Plant Science Building ++ Cornell University, Ithaca, NY 14853 ++ E-Mail: mvp2-at-cornell.edu ++ Plant Biology Office: 268 Emerson; Telephone: 607-255-1734 ++ Plant Biology Fax: 607-255-5407 ++ CIMC Office: C1 054 Vet. Med. Center; Telephone: 607-253-3803 ++ CIMC Office Fax: 607-253-3803 ++ CIMC web site: http://www.cimc.cornell.edu ++ ++ ++
Like Malcolm I gave up on a hand held vacuum cleaner when sawing resin as I suspected it leaked fine particles. Now I saw up resin in a large bag in a fume cupboard. The bag has lasted 5 years. Anyone got a neater/safer method?
Dave
On Mon, 27 Mar 2000 10:35:33 +0100 Malcolm Haswell {malcolm.haswell-at-sunderland.ac.uk} 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 } -----------------------------------------------------------------------. } } } Greg } } you mention using a small vacuum cleaner to clean up resin dust. You } don't specify whether it has an exhaust filter so I think I should just } mention that the typical domestic vacuum cleaner exhausts fine } particulates and may well make the problem worse because it may be } pumping the very size of resin particles out that you want to avoid. We } once used a small portable vacuum cleaner for cleaning up resin dust but } stopped when I realized that there was a potential risk. } } I know that there are now domestic and industrial vacuum cleaners which } have very fine exhaust filters and there are also the small } photocopier/toner vacuum cleaners. Has anyone investigated their use for } resin dust? } } Malcolm } } Malcolm Haswell } Electron Microscopy } School of Sciences } Fleming Building } University of Sunderland } SUNDERLAND SR1 3SD } Tyne and Wear } UK } } Tel (0191) 515 2872 } e-mail: malcolm.haswell-at-sunderland.ac.uk } } greg erdos 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 solved the resin dust/ Dremel tool problem by rigging a small vacuum } } cleaner with the intake very close to where we would be grinding, which was } } usually under a disecting scope. I got the idea while watching a cast } } being cut from a broken foot. The cast cutter had a vacuum attached. } } Our rig takes care of the fine particles, while larger ones generally fall } } to the earth and can be cleaned up later. } } This more of an issue with epoxies, I suspect. LR White is a dental resin, } } I believe, and the dentist will grind in your mouth or over you using your } } chest as a table. I don't know if that means it is safe. } } } } Greg Erdos } } University of Florida } } Greg Erdos } } 5410 SE 185th AVe } } Micanopy, FL 32667 } } 352-466-0843 }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
I have a Hummer VII sputter coater, which has an automatic processing cycle. As the vacuum drops from 60 millitorr to 40 millitorr (during which time the high voltage should switch on), the entire process shuts down. I have tried running the system with the argon valve open and shut.
Any suggestions from users out there?
Thanks,
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
} Does anyone no of a good web source for info on light microscopy for } undergraduates. Ideally I'm after reference material on brightfield, } phase contrast and fluorescent microscopies. Some info on new areas } would also be useful.
Barry, Try looking at these web sites. The virtual microscopy is fun. You might want to filter out the stuff of interest to materials and geological sciences... or leave it in in the name of interdisciplinary microscopy !
A number of universities are now using "Optimizing Light Microscopy for Biological and Clinical Laboratories". It covers all the areas you mentioned and more. It also includes a number of small experiments which can be done as a group or on independent study. See our website for further info.
Caveat: MME does have financial interest in this book.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ******************************************************
At 01:46 PM 3/27/00 GMT0BST, BARRY SHAW wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Ping, The last time I went looking for spec-pure carbon rods, our Stores man found them at: ultra carbon, 900 Harrison St., Bay City, MI48708-8244. They weren't that exact size, but they are a good place to try. At 09:38 AM 3/27/00 -0400, you wrote:
} } Hi, I am looking for carbon rods with spectrographic quality for Edwards } vacuum evaporator. The rods should be about 1.5mm in diameter and 30mm } in length. I have tried several suppliers without success. If any of you } have information regarding who supplies this kind of carbon rods could } you please let me know? Your help will be greatly appreciated. Thank } you. } } Ping } 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
} Does anyone no of a good web source for info on light microscopy for } undergraduates. Ideally I'm after reference material on brightfield, } phase contrast and fluorescent microscopies. Some info on new areas } would also be useful. Hope someone out there can help ! } } Barry -
I can suggest a good CD-ROM: Pagliaro, l., et al., 1997 Microscopy-Tutor. You'll find a description and ordering information in the Project MICRO bibliography (URL below).
Caroline
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
} Here is a question for those of you who use sucrose to provide } cryoprotection for tissues: } } We are preparing perfusion fixed brain tissue by freeze substitution for } post-embedding immuno gold. Following fixation we prepare 500 micron } vibratome sections, which we then wish to infiltrate with 2 M sucrose in } buffer prior to freezing. We have been transferring the brain sections to } 1 M sucrose, then when they sink, transferring again to 2 M. The problem } is that the sections never sink in the 2 M sucrose. }
Dear Marie, The Handbook of Chemistry and Physics gives a specific gravety for 2 M sucrose of about 1.25. Is the sp. gr. of your tissue (exclusive of the exchangable fluids, of course) greater than this? If not, the sections will never sink. If it is marginally greater, then the sections will sink when hell freezes over, and you can do cryo easily. If you can determine the weight of a tissue section of known volume first with fixation fluid, then after infiltration with 1 M sucrose (and also measure the volume change), you could extrapolate to the situation for 2 M sucrose (or some- one with too much time on his/her hands could do this for many sucrose concentrations). Good luck. Yours, Bill Tivol
We need to localize zinc in prostate tissue for TEM. Has anyone used Sulfide-silver method or Zinc-dithizonate method? If so, is there a method better than the other or new methods that might work on such tissue?
I am also trying to locate a reference by Pihl E. Falkmer S: Trials to modify the sulfide-silver method for ultrastructural tissue localization of heavy metals. Acta Histochem 27:34-41, 1967. If you have a copy of this method please email me.
Thank you
Karen Kelley Senior Electron Microscopist UF Biotechnology Electron Microscopy Core Lab Box 118525 Gainesville Florida lab:352-392-1184 fax: 352-846-0251 email: klv-at-biotech.ufl.edu
} I had done an evapouration of cobalt on a quartz sample. Now I want to } etch the quartz sample to remove the substrate so that I can do } Transmission Microscopy for charactrization. My problem is that If I etch } the substrate with HF will it be harmful to carbon? If it is so what are } the other materials that can be used to etch quartz.
Dear Ramu, Quartz can be etched--slowly--in strong base. A 5-to-10 M solution of NaOH should do the trick, but if the quartz thickness is in the mm range, it will take a long time. I don't know whether the base will be harmful to either carbon or cobalt, but I don't think it will. Good luck. Yours, Bill Tivol
} Hi, I am looking for carbon rods with spectrographic quality for Edwards } vacuum evaporator. The rods should be about 1.5mm in diameter and 30mm } in length. I have tried several suppliers without success. If any of you } have information regarding who supplies this kind of carbon rods could } you please let me know? Your help will be greatly appreciated. Thank } you. }
Dear Ping, I got mine from Ted Pella, but I'm surprized you didn't find them in other suppliers' catalogues. Pella lists both carbon and graphite, both
spectroscopically pure and technical grade, in several diameters and lengths. I have no connection to Ted Pella, Inc. except that of customer. Yours, Bill Tivol
Hello everyone, I have a student who wants to prepare tissue culture cells for TEM in Agar blocks. I shall appreciate any suggestions. Thanks in advance. C.L.Singla
We are contemplating replacing our analytical TEM. A basic spec we envisage is STEM, EDS + GIF/PEELS. We are happy to choose the EDS, but we run into trouble with e-guns, i.e. to go LaB6 or FEG. In particular as we would like to bolt a GIF/PEELS on to the system.
We have several concerns: The safety of the FEG gun, considering we are a multi-user facility with users who have a frequent tendency to bend holders and press the wrong buttons. How problematic are FEGs, do you have much down time / running cost due to careless users?
GIF/PEELS: has anybody carried out a systematic comparison between a system with LaB6 or FEG. Is the extra cost of a FEG (over twice the price) really justified for GIF/PEELS or will it quite happily run on a LaB6.
The thoughts of fellow microscopy in the same quandary much a appreciated.
Keith.
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Dr. K. Moulding.
Materials Characterisation and Preparation Facility Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong.
Sucrose infiltration is an essential step in the preparation of cryosections for immunocytochemistry. Aldehyde-fixed biological tissue is infiltrated in 2.0M to 2.3M sucrose to protect the sample from freezing damage when it is subseuently frozen by immersion in liquid nitrogen. If the sample is fixed, the membranes become permiable to sucrose and the sample can be frozen into a vitreous state by immersion in liquid nitrogen. Although it is advised to infiltrate for 24 hr on a rotator, many years of experience has demonstrated that even after 15 min of exposure to sucrose, small pieces of aldehyde-fixed material are sufficiently cryoprotected to allow for successful vitrification. Take no heed of whether the sample has sunk to the bottom of the tube or not, check to see if it has been fixed and has spent sufficient time in the sucrose. If it has, it will be cryoprotected.
Useful reference: Griffiths, G, McDowall, A, Back, R, Dubochet, J. 1984. On the preparation of cryosections for immunocytochemistry. Ultrastruct. Res. 89:65-78 They showed by quantitative mass measurements that fixed cells are freely permeable to sucrose.
Best regards,
Paul Webster
Paul Webster, Ph.D. House Ear Institute 2100 West Third St. Los Angeles, CA 90057
Bill Tivoli wrote: The Handbook of Chemistry and Physics gives a specific gravety for 2 M sucrose of about 1.25. Is the sp. gr. of your tissue (exclusive of the exchangable fluids, of course) greater than this? If not, the sections will never sink. If it is marginally greater, then the sections will sink when hell freezes over, and you can do cryo easily. If you can determine the weight of a tissue section of known volume first with fixation fluid, then after infiltration with 1 M sucrose (and also measure the volume change), you could extrapolate to the situation for 2 M sucrose (or some- one with too much time on his/her hands could do this for many sucrose concentrations). Good luck.
Marie E. Cantino wrote: We are preparing perfusion fixed brain tissue by freeze substitution for post-embedding immuno gold. Following fixation we prepare 500 micron vibratome sections, which we then wish to infiltrate with 2 M sucrose in buffer prior to freezing. We have been transferring the brain sections to 1 M sucrose, then when they sink, transferring again to 2 M. The problem is that the sections never sink in the 2 M sucrose.
Hi, Our safety people are concerned about having our Polaron c.p.d pressure tested at regular intervals. Does any one out there know of anyone who does this sort of thing, preferably on site? Also I would be interested to hear how other users safety check this sort of equipment. thanks, Christine.
Dear Keith, If you are seriously worried about the experience of some of the users then you should consider the Philips/FEI Tecnai series of instruments. The person in charge of the microscope can determine the level of access to all the features for each user i.e. basic, medium and expert user profiles. The microcope, in a similar way to fly-by-wire aircraft, will simply prevent users from blowing the tip up or anything else detrimental to the 'scope. This should take out the worry about user/FEG problems.
The LaB6 and FEG work identically with the GIF/PEELS, they're only electrons after all. The FEG will give you a smaller energy width (good for EELS:ELNES .etc) and more current (good for Energy Filtered TEM, STEM:HAADF, EELS & EDS). Then there is the option of adding a biprism with the FEG if you want to try off-axis holography. The FEG machine will simply give you a wider tolerance range in terms of specimen thickness and experimental conditions i.e greater signal to noise ratios than with the LaB6, something that is a problem with STEM based methods. FEG alignment procedures will be slightly different too.
We have taken delivery of a 300kV Tecnai and I have to say that the machine is very impressive. The machine is very stable and the high brightness FEG is a major improvement over the LaB6. The GIF is also quite a God-send for zero-loss filtering and for mapping.
I hope this helps, Jon
P.S. I am not affiliated or have any commercial connection to Philips/FEI. ******************************************************** Dr Jonathan Barnard
Analytical Materials Physics The Angstrom Laboratory, Uppsala University P O Box 534, SE-751 21 Uppsala, Sweden Phone: +46-(0)18-4716838 Fax: +46-(0)18-500131 http://www.angstrom.uu.se/analytical/home.html
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Dear Friends,
The Pirani Gauge of the Oxford Hexland - CT1000 cryo unit mounted on SEM S120 is not working. Can anyone guide me in purchase of this? Could this be manufactured in-house? Please help me out.
Rajdeep Dongre Electron Microscopy Laboratory Agharkar Research Institute G.G. Agarkar Road Pune - 411 004, India Phone 91-20-5653680/5654357 e-mail : rajdeep-at-aripune.ernet.in
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We work with plant material and after ultra thin sectioning our sections show lines of compressions in cell wall areas. Of course, we try to stretch the Epon sections with a heat pen or with chloroform, but very often we are not successful. The sections just move on the water surface and nothing is happening. Any advice would be gratefully appreciated. Regards, Anne Heller -- Dr. Anne Heller, AG Elektronenmikroskopie, Institut fr Botanik (210), Universitt Hohenheim, Garbenstra§e 30 D-70593 Stuttgart
Hi Jo, Thanks for your answer. We use our GIF in a similar way, focusing around the energy loss we want to take the image at with a slit width covering the 3 energy windows if possible. This works best for rather thick samples (compared to our 5 nm particles)But as you say, maybe it's just the spatial resolution which is not good enough, our microscope was designed to enhance contrast for biological studies, unfortunately loosing some resolution in the process. We are not yet trying to get concentrations out of the pictures...rather get a qualitative mapping.
I tried to do EELS measurements a few times and got very few succes. The best results are obtained on beam resistant samples simply because I can then move the zero loss peak off the CCD and increase intensity (or acq time) at will. I start with a very low intensity and optimise it in turboview mode to get a good signal out. However I could seldom see well defined peaks. One point which I find strange is that we can often get reasonnably good elemental maps even when we don't see any peak in the EELS spectrum. The question is then if the map is really trustworthy...
Any comments welcome !
Olivier
Jo Verbeeck wrote:
} Hello, } } Regarding EFTEM on small particles, this should actually work quite well. } That is, elemental mapping works reasonably if you choose the right } objective apperture. You should simulate the spatial resolution to see } what can be attained under your conditions (HT, Cc, Cs etc). } However it will be very difficult to get real concentration information } out of these images. I'm trying EELS with nanoprobe but so far no succes } (everything gets destroyed before I take a spectrum, allignment is very } very difficult) } Still, focussing remains a problem. Comon practice is to focus at 100eV } and then suppose everything is OK for higher losses. I do not understand } the theory of this technique (blurring by finite slitwidth & Cc?) nor thus } it work well. Any comments on this are welcome. } } Jo } } ************************************************************* } * Jo Verbeeck * } * University of Antwerp * } * Dept. EMAT (Electron Microscopy for Materials Research) * } * e-mail: joverbee-at-ruca.ua.ac.be * } * tel: +32(0)3 218 02 49 * } * fax: +32(0)3 218 02 57 * } ************************************************************* } } On Tue, 21 Mar 2000, GuessWho wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Dear all, } } } } I was wondering if anybody was using a GIF filter. We have one mounted } } on a 120 kV TEM and are trying to image small particles (5-10 nm) and } } doing some energy filtering to image different elements in the particles } } (for example gold, silicon, copper, cerium...) . This is not really what } } I would call trivial work and we are still working out the set up } } (window size and positionning for example) in a rather empiric way. The } } main problem is often to get a decent signal in the interesting energy } } region and still keep the windows fairly narrow. } } I would add that we often have to work at low dose because of the beam } } sensitivity of our samples, but I guess our GIF setups can be improved. } } Any suggestion ? } } } } Any general comment on the GIF warmly welcome ! } } } } Thanks } } } } Olivier } } } } } }
} ------------------------------------------------------------------------ } 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 work with plant material and after ultra thin sectioning our sections } show lines of compressions in cell wall areas. Of course, we try to } stretch the Epon sections with a heat pen or with chloroform, but very } often we are not successful. The sections just move on the water } surface and nothing is happening. Any advice would be gratefully } appreciated. } Regards, } Anne Heller } -- } Dr. Anne Heller, AG Elektronenmikroskopie, } Institut fr Botanik (210), Universitt Hohenheim, } Garbenstra§e 30 } D-70593 Stuttgart } } Tel.: (0049)-711-459-2180 } Fax.: (0049)-711-459-3355
Hi Anne,
My first guess is that the mechanical properties of your Epon mix differs too much from that of the fixed walls. However, when I worked with plant material, I generally used Xylene to stretch floating sections (rarely in Epon). I'm not sure if was the aromatic character or the boiling point difference that made it better at swelling and relaxing the sections than CHCl3.
Other suggestions would be using a harder resin (my favorite was VCD and Quetol 651 (equiequivalent) hardened with NSA (or HSA), which I called Spurtol ;-)). I found it stains easier than Spurr's yet is low viscosity and about as hard as a medium Spurr's. Reducing the knife angle might help. Post curing the existing blocks at elevated temperature might harden them some more.
Cheers,
John
John Heckman MSM Department Michigan State University
I don't know if you have interest in this but it is good info. Wallace
-----Original Message----- } From: Parker, Jo Sent: Wednesday, March 29, 2000 8:45 AM To: All SOD Faculty & Staff
Dear Jonathan and Keith,
I just want to add a few comments to what Keith said about FEG's. One of the major advantages of a FEG over conventional souces for AEM is the spot size. The new FEG scopes have STEM resolution of 2 angstroms with great brightness. (I have only seen the Philips/FEI scope, but I think the other companies have similar specs.) When one is deciding whether or not to go for the FEG, I think it will depend on your applications. If a small intense beam and good energy resolution are necessary, go for the FEG. Of course, there is also the consideration of the cost of service contract ($$$$).
Christine Richardson wrote: =================================================== Hi, Our safety people are concerned about having our Polaron c.p.d pressure tested at regular intervals. Does any one out there know of anyone who does this sort of thing, preferably on site? Also I would be interested to hear how other users safety check this sort of equipment. =================================================== I might not be the last word on this, but I had described to me the kind of large liquid filled tank (I think it is water) that pressure vessels like this are tested in, and to me at least it did not sound like it would be the kind of thing that one would "carry around" in a portable kind of unit to do this kind of testing, on site. This description came, some years ago, from Dr. Wilf Gee (now deceased) and who was very much involved in the original manufacturing of this product and who had involvement for many years with the safety testing of the unit.
When your CPD unit was first delivered, it would have come with a copy of a report from the testing agency showing the results and conditions of the pressure testing. I might be off by a bit on this, but it was my recollection that the vessel is tested to a pressure level that is three or four times higher than what would be required to blow out the rupture disc. In other words the unit is tested to a level that, if the rupture disc did not blow exactly where it was supposed to blow, there is some very large margin of error so that the disc still would blow long before the vessel.
There are probably more CPD units of this design installed in the world than any other design. I have never heard of anyone having any kind of a problem with it (e.g. an explosion), except an occasional blowing of a rupture disc, but certainly not the pressure vessel itself. Just don't ever try to run the unit by replacing the rupture disc, if one is not handy, with a cut metal disc. That is very dangerous and should never be done. Believe it or not, we do uncover once in a while, people who do do that sort of thing, out of naivety of course and that is why I do not waste an opportunity to point out the danger in doing that sort of thing. That is why I always have recommended, for multi-user environments in particular, to always have a small supply of replacement rupture discs near by the unit just to avoid even the slightest temptation (such as by a student) to by-pass this important safety feature.
So I for one would be very interested in hearing the logic and rationale of your safety committee that is of the opinion that you should have your pressure vessel tested from time to time.
However, there is one kind of situation where one should have the vessel pressure tested and that is if one has attempted any mechanical modifications to the chamber itself. Then the system should be pressure tested. And since the manufacturer of this particular CPD unit (e.g. Polaron) has had on the order of thirty years of experience doing this kind of testing, on this particular design of vessel, I would recommend having them do the testing for you in the UK. Today the testing of the Polaron unit is done to the CE standard, without doubt the most stringent in the world.
If you do not have the original test certificate, if you send me your S/N I could try to get a copy of it for you. That alone might satisfy the questions being asked of your safety people.
Disclaimer: SPI Supplies has offered this particular CPD unit, especially to customers outside the USA, for nearly twenty five years and we are unaware of any requirement to have the vessel retested at periodic intervals for safety reasons.
Chuck
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I am passing this on for a fellow colleague. Thank you in advance.
Ginger (Baker) Hendricks EM Lab Manager Oklahoma State University College of Osteopathic Medicine
"I am looking for an immunogold-silver enhancement method for labelling a cytoplasmic antigen in cultured cells and viewing with Light Microscopy. We will be using Protein Aand a monoclonal antibody. Once we have determined that we have specific labelling, we will proceed to visualizing the gold with TEM."
} Hello everyone, } I have a student who wants to prepare tissue culture cells for TEM in Agar } blocks. I shall appreciate any suggestions. } Thanks in advance. } C.L.Singla } It would be helpful to know how the tissue culture cells are being grown...in plastic wells where they will be scraped off or on plastic or glass coverslips where the entire surface will be embedded. What is the ultimate goal of the TEM examination...morphology, interactions? Will the samples be ultimately embedded in plastic or expoxy?
Dr. Tina Schwach Microscopy Consulting Services, Inc.
I would appreciate it very much if someone can refer me to a good book or protocol for preparation of fungal spores for SEM and TEM. This is very foreign to me since I have been working with mammalian tissues exclusively for so many years but now that the lab is a core facility I am getting somel requests for other types of samples. Thank you,
Cora Bucana ******************************************************* Corazon D. Bucana, Ph.D. Department of Cancer Biology U.T. M.D. Anderson Cancer Center 1515 Holcombe Blvd. Box 173 Houston, Texas 77030 Phone: (713) 792-8106 FAX: (713) 792-8747 Email:bucana-at-audumla.mdacc.tmc.edu FAX: (713) 792-8747
} ------------------------------------------------------------------------ } 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, } Our safety people are concerned about having our Polaron c.p.d } pressure tested at regular intervals. } Does any one out there know of anyone who does this sort of thing, } preferably on site? } Also I would be interested to hear how other users safety check this } sort of equipment. } thanks, } Christine.
Hi Christine,
Over here in the US we have an Interstate Commerce Commission that checks pressure vessels via hydrostatic testing. This is required every 5 years and the tops of gas cylinders, at least, are stamped with the last test date. It's always amazed me just how long N2 cylinders last (reading hydrostatic test dates is something to do while your plates are processing). I've seen bottles with dates that were before W.W.I. All that age and 2250 psi. I've never heard of them requiring the testing of lab equipment, though. Might check with your bottled gas supplier to see if they'd do it.
} ------------------------------------------------------------------------ } 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 work with plant material and after ultra thin sectioning our sections } show lines of compressions in cell wall areas. Of course, we try to } stretch the Epon sections with a heat pen or with chloroform, but very } often we are not successful. The sections just move on the water } surface and nothing is happening. Any advice would be gratefully } appreciated. } Regards, } Anne Heller } -- } Dr. Anne Heller, AG Elektronenmikroskopie, } Institut für Botanik (210), Universität Hohenheim, } Garbenstraße 30 } D-70593 Stuttgart } } Tel.: (0049)-711-459-2180 } Fax.: (0049)-711-459-3355 } } } } Hi,
If I get any reaction in a section floating in a boat from xylene, heat pen, chloroform, etc., I know that something is wrong. My personal test of a good embedment is nonreactivity of floating sections. Why?
In industry, if it is important that there are no free monomers in the section, the material is exposed to the above solvents to soak them out.
If you have enough unpolymerized monomers in your section, solvents will affect that section. (Note: Embedments that are too soft in formulations are excluded from this discussion at this time).
So - keep your formulations well mixed at all times - do not allow it to sit in the hood. It must be in motion at all times. Push infiltration! More changes, longer times. If using epoxy of any sort, heat the infiltration medium to 37 deg C for 1 hour after every new change of resin while the sections are on the rotator. A plain 60W light bulb is ideal for this. Polymerize well. At least for 48 hours. If needed, repolymerize at 95 deg C for an hour especially if your formulation contains NMA.
Your formulation may be wrong for your material. Try going harder. Also with existing blocks, try reheating, then try a lower knife angle and a slower speed of cutting.
Most important: Keep records. Know what you are doing and why. Do not haphazardly try this, try that. You will get crazy! Embedments are very complex systems that belong into the realm of material science - that is why biologists have so much trouble with them. (I have had mega fights lasting years with epoxies)
Keith If you have a new FEG machine with a Schottky emitter, then these are relatively easy to operate and fairly robust, but they do, of course, cost a lot more and so you may not want to spend that money and have ham-fisted users operating it. For high-resolution analytical TEM/STEM there is no doubt that FEG is the preferred source. The small source size and small energy spread combined with high gun brightness gives great performance. A word of warning,though. When using a GIF with any TEM it is often necessary to operate at low mag because of the additional mag (18 - 20 times) introduced by the GIF. In this situation (depending on the mag), the LaB6 source may be preferable in certain situations because the total current is higher than the FEG source. I have used both types of machine-Schottky FEG plus GIF and LaB6 plus GIF and there do not appear to be any severe problems with the low mag mode operating with a Schottky source. The cold FEG source may not be so good from this point of view, since the source size is smaller than the Schottky FEG and so the cross-over probe size above which the current in the cold FEG probe is less than the LaB6 is small and probably in the range 10-100 nm. I have not used a cold FEG TEM with a GIF such as the Hitachi and so I can't really make an informed comment, but Profs Dravid and Marks at Northwestern University are very skilled users of the Hitachi machine and they can surely advise you. Best of luck.
Alan Fox Center for Materials Science and Engineering Naval Postgraduate School Monterey California 93940
Keith Moulding 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 } -----------------------------------------------------------------------. } } Greetings, } } We are contemplating replacing our analytical TEM. A basic spec we envisage } is STEM, EDS + GIF/PEELS. We are happy to choose the EDS, but we run into } trouble with e-guns, i.e. to go LaB6 or FEG. In particular as we would like } to bolt a GIF/PEELS on to the system. } } We have several concerns: } The safety of the FEG gun, considering we are a multi-user facility with } users who have a frequent tendency to bend holders and press the wrong } buttons. How problematic are FEGs, do you have much down time / running } cost due to careless users? } } GIF/PEELS: has anybody carried out a systematic comparison between a system } with LaB6 or FEG. Is the extra cost of a FEG (over twice the price) really } justified for GIF/PEELS or will it quite happily run on a LaB6. } } The thoughts of fellow microscopy in the same quandary much a appreciated. } } Keith. } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } Dr. K. Moulding. } } Materials Characterisation and Preparation Facility } Hong Kong University of Science and Technology, } Clear Water Bay, } Kowloon, } Hong Kong. } } FAX: (852) 2358 2451 } TEL: (852) 2358 8724 } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Phone a divers shop and ask who is testing their cylinders. Those people can also test the CPD. It would be a hydrostatic test and they would need to have a fitting to connect to the back of the CPD. To have any meaning the applied pressure would need to be higher than normally applied, I suggest by 50%.
I think its a bad idea to do such a test. The metal housing of those units is vastly over-designed and the window is quartz which ultimately would crack but not shatter. All the test would achieve is to burn up some of your funds and time. Stress the system and if your unlucky crack the window - would the "safety people" pay for that?
It would be interesting to learn how many systems have failed and if there were any special circumstances. I don't know of any instance of a CPD failure. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
Hi, Our safety people are concerned about having our Polaron c.p.d pressure tested at regular intervals. Does any one out there know of anyone who does this sort of thing, preferably on site? Also I would be interested to hear how other users safety check this sort of equipment. thanks, Christine.
ASSOCIATE DEAN, COLLEGE OF ENGINEERING AND APPLIED SCIENCES
Western Michigan University seeks applications and nominations for the position of Associate Dean for Undergraduate Programs and Assessment of the College of Engineering and Applied Sciences.
Western Michigan University is a Carnegie Doctoral I university with 750 FTE tenure-track faculty and an enrollment of about 27,000 students, 25% at the graduate level. It is advancing to Carnegie Research II classification and plans a substantial investment in the College of Engineering and Applied Sciences, including a new physical facility, to accomplish this goal. In addition to its Graduate College and Lee Honors College, Western supports seven degree-granting colleges: Arts and Sciences, Haworth College of Business, Education, Engineering and Applied Sciences, Aviation, Health and Human Services, and Fine Arts. These colleges offer 242 academic programs, including 60 at the masterâs level and 25 at the doctoral level.
The Associate Dean for Undergraduate Programs and Assessment reports to the Dean. This person provides leadership and oversees the development of college policy for undergraduate programs, accreditation, assessment, advising, recruitment, retention, student orientation, co-op, and intern programs. An earned terminal degree in one of the disciplines in the college or a related science is desirable. A doctoral degree, with at least one degree in an engineering or closely related science/engineering discipline, is required. University experience in academic programs and demonstrated excellence in teaching are required. Candidates should have excellent administrative and interpersonal skills and experience utilizing instructional and administrative technology. An excellent record of teaching and research and/or creative achievement that would warrant an appointment as associate or full professor with tenure in one of the academic units in the university is required.
The individual selected will assume academic and administrative responsibilities for undergraduate programs in a dynamic and growing college offering 16 baccalaureate, 11 masterâs and three doctoral programs. The collegeâs staff includes 70 full-time faculty, 10 administrators (8 with faculty rank), 31 funded staff positions, and dozens of contract staff and graduate student assistants, and teaches approximately 2300 students. The college has a strong commitment to education and research, which spans a broad spectrum of engineering and engineering technologies. The college offers courses and programs primarily on Western Michigan Universityâs main campus in Kalamazoo, but also serves as a major resource for off-campus instruction and economic growth at fives sites across West Michigan.
Candidates should submit 1) a curriculum vita, 2) three letters of recommendation, 3) an application letter stating their qualifications for the position, and 4) a statement outlining a personal vision for engineering education to:
Parviz Merati, Chair Associate Dean Search Committee Department of Mechanical and Aeronautical Engineering Western Michigan University Kalamazoo, MI 49008
Review of applications will begin on or about May 1, 2000. Applications will be accepted until the position is filled. Western Michigan University is an equal opportunity /affirmative action employer.
For additional information about WMU and the College of Engineering and Applied Sciences, refer to our Website: http://www.wmich.edu/. ===================================== Pnina Ari-Gur, D.Sc., Professor Materials Science and Engineering CMD Department Western Michigan University Kalamazoo, MI 49008 (616) 387-3372 FAX: (616) 387-6517 email: pnina.ari-gur-at-wmich.edu http://www.wmich.edu/cmd/arigur.htm =====================================
Not announced on ebay, and limited to MSA, I am offering an IBM Thinkpad 355 which is optionally available to make this camera truly portable. $350 extra gets the Thinkpad. You will still need a Slim SCSI PCMCIA card, but these are very low cost. I have one in a Sony VAIO and it works great with the Leaf.
I need information related to EMS software. There was a related posting (enclosed below) recently, replies to which I seem to have missed. My mail to the author of that posting also bounced back. Could somebody please help me get this info? Thanks very much. ---- Divakar R Physical Metallurgy Section, Indira Gandhi Centre for Atomic Research Kalpakkam 603102, India ----
-----Original Message----- } From: Divakar R [SMTP:divakar-at-igcar.ernet.in] Sent: Wednesday, March 29, 2000 8:23 PM To: 'Chengge Jiao'
Hi,
Does anybody know the price for following softwares for EM simulation.
1. Diffraction 2.0 (or the most latest version),
2. EMS.
Chengge Jiao
H.H.Wills Physics Laboratory University of Bristol, UK
In the matter of inexperienced users and FEG versus Lab6 I have no fears on our FEGTEM.
We have been running our JEOL3000F for over a year now without any trouble, it has a Schottky emitter so there is no flashing to be carried out. The emitter is the original one used for factory tests, shipped over, used for installation and still in use. We have experience of several LaB6 instruments over the past 15-20 years and are a materials science multi-user facility.
The FE gun runs 24 hours a day, the user just opens the valve to see the beam. The gun and emitter are run up by an experienced person whenever it needs it (every 2-3 months). We have a `Quick Emission Selector' that we have set up to give 3 more emission currents (higher for GIF, lower for less energy spread etc.) and the user cannot readjust these only select between them. There is no room for abuse (even for some of our users!).
The LaB6 guns have to be run up at the start of every session and every time a user changes specimen or films. After a period of time the LaB6 tips slowly deposit insulating LaB6 on the Wenhelt, this charges and the bias has to be adjusted to reset the emission correctly. When a user finishes the session this charge dissipates and the next user gets a high emission current which has to be reset using the bias. The bias has to be adjusted during the first part of the session as the charge builds up. This gives the user lots of room for abusing the tip, it is not possible to prevent access to the bias control as they do need to use it.
Regards, Ron
=========================================================================== Mr. Ron Doole e-mail ron.doole-at-materials.ox.ac.uk Department of Materials, phone +44 (0) 1865 273701 University of Oxford, fax +44 (0) 1865 283333 Parks Road. Oxford. OX1 3PH. UK. ============================================================================
Two days ago, the window in our CPD cracked. The run was in every respect normal, but the window cracked at 35 degrees/80 bar. This was a big surprise to us, as we have belived that this could never happen as long as temperature and pressure are within their normal ranges. We would be very interested to receive comments on this.
The week before, the equipment, which is more than 20y old, had been disassembled for cleaning and replacement of the window O-ring.
Best Regards, Gunnar Kopstad.
Vennlig Hilsen dr.ing Gunnar Kopstad overingeni¿r Avd f Patologi, Rit
For the record - and maybe this is a record - our Polaron E3000 CPDA was tested in Feb. 1974 to a proof load of 2,500 lbf/in2. Under "Remarks" on the test certificate it says "Proof pressure applied without any visual signs of distortion or leakage. Safety valve set at 1,900 lbf/in2."
I take "lbf/in2" to be "pounds per square inch".
The unit has been in quite regular use since 1974 with no problems other than failure of seals. I did once have to de-"fur" the waterways but that was a local water supply problem.
Keith Ryan Marine Biological Association Plymouth UK
} I tried to do EELS measurements a few times and got very few succes. The best } results are obtained on beam resistant samples simply because I can then move } the zero loss peak off the CCD and increase intensity (or acq time) at will. I } start with a very low intensity and optimise it in turboview mode to get a } good signal out. However I could seldom see well defined peaks. } One point which I find strange is that we can often get reasonnably good } elemental maps even when we don't see any peak in the EELS spectrum. The } question is then if the map is really trustworthy... } } Any comments welcome ! } } Olivier
With my experience I think it is rather normal than surprising that you can achive relatively good elemental maps in ESI mode but see no recognizable features in the EELS spectrum. With an elemental map you get energy loss information for each single image point. If there is a higher concentration of a certain element, the intensity of the corresponding pixel in the energy window on the edge will be higher than in neighbouring pixels not containing this element or only in less quantity. This does not necessarily mean that you would actually see a peak in the corresponding spectrum of this pixel, it can be just a change in the slope of the background. Since you are comparing the changes in intensity of every pixel this small differences become visible in an map. Furthermore, in spot mode or selecting a small area with an aperture, you will average usually over a larger area than a pixel in image mode. If the features of your specimen are smaller the jump ratio of the edge will be reduced due to the contribution of the surrounding material. If you thoroughly compare spectra acquired on a (large enough) area containing the element of interest and a spectrum of the matrix you will find differences. Furthermore you will see certainly more of the edges in your spectra of thick specimens if you go for a deconvolution. It is still surprising (at least for me) how much information you can gain from a spectrum which did not look much more than a steady decrease in intensity.
Petra
-------------------------------------------------------------- Dr. Petra Wahlbring Centre de Recherche Public - Gabriel Lippmann Laboratoire d'Analyse des Materiaux (LAM) 162a, av. de la Faiencerie L-1511 Luxembourg tel. +352-466644-402 fax +352-466644-400 e-mail: petra.wahlbring-at-crpgl.lu Visit our WWW site! http://www.crpgl.lu/~wahlbrin
} ------------------------------------------------------------------------ } 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 would appreciate it very much if someone can refer me to a good book or } protocol for preparation of fungal spores for SEM and TEM. This is very } foreign to me since I have been working with mammalian tissues exclusively } for so many years but now that the lab is a core facility I am getting somel } requests for other types of samples. Thank you, } } Cora Bucana } ******************************************************* } Corazon D. Bucana, Ph.D. } Department of Cancer Biology } U.T. M.D. Anderson Cancer Center } 1515 Holcombe Blvd. Box 173 } Houston, Texas 77030 } Phone: (713) 792-8106 } FAX: (713) 792-8747 } Email:bucana-at-audumla.mdacc.tmc.edu } FAX: (713) 792-8747
Dear Cora Bucana,
it is depending very much on the type of spore, e.g. mildew with high water content and "thin" walls is easy to prepare with standard protokolls for plant material. Dormant, thick walled spores with less water and high lipid content are quite tricky to prepare for TEM, but easy for SEM. I can send you a detailed protocol if you tell more about your fungal spores.
-- Dr. Anne Heller, AG Elektronenmikroskopie, Institut fr Botanik (210), Universitt Hohenheim, Garbenstra§e 30 D-70593 Stuttgart
} ---------------------------------------------------------------------- } -- 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: Garber, Charles A. * EMC.Ver #3.1 ] -- } } Christine Richardson wrote: } =================================================== } Hi, } Our safety people are concerned about having our Polaron c.p.d } pressure tested at regular intervals. } Does any one out there know of anyone who does this sort of thing, } preferably on site? Also I would be interested to hear how other users } safety check this sort of equipment. } =================================================== I might not be the } last word on this, but I had described to me the kind of large liquid } filled tank (I think it is water) that pressure vessels like this are } tested in, and to me at least it did not sound like it would be the } kind of thing that one would "carry around" in a portable kind of unit } to do this kind of testing, on site. This description came, some } years ago, from Dr. Wilf Gee (now deceased) and who was very much } involved in the original manufacturing of this product and who had } involvement for many years with the safety testing of the unit. } } When your CPD unit was first delivered, it would have come with a copy } of a report from the testing agency showing the results and conditions } of the pressure testing. I might be off by a bit on this, but it was } my recollection that the vessel is tested to a pressure level that is } three or four times higher than what would be required to blow out the } rupture disc. In other words the unit is tested to a level that, if } the rupture disc did not blow exactly where it was supposed to blow, } there is some very large margin of error so that the disc still would } blow long before the vessel. } } There are probably more CPD units of this design installed in the } world than any other design. I have never heard of anyone having any } kind of a problem with it (e.g. an explosion), except an occasional } blowing of a rupture disc, but certainly not the pressure vessel } itself. Just don't ever try to run the unit by replacing the rupture } disc, if one is not handy, with a cut metal disc. That is very } dangerous and should never be done. Believe it or not, we do uncover } once in a while, people who do do that sort of thing, out of naivety } of course and that is why I do not waste an opportunity to point out } the danger in doing that sort of thing. That is why I always have } recommended, for multi-user environments in particular, to always have } a small supply of replacement rupture discs near by the unit just to } avoid even the slightest temptation (such as by a student) to by-pass } this important safety feature. } } So I for one would be very interested in hearing the logic and } rationale of your safety committee that is of the opinion that you } should have your pressure vessel tested from time to time. } } However, there is one kind of situation where one should have the } vessel pressure tested and that is if one has attempted any mechanical } modifications to the chamber itself. Then the system should be } pressure tested. And since the manufacturer of this particular CPD } unit (e.g. Polaron) has had on the order of thirty years of } experience doing this kind of testing, on this particular design of } vessel, I would recommend having them do the testing for you in the } UK. Today the testing of the Polaron unit is done to the CE } standard, without doubt the most stringent in the world. } } If you do not have the original test certificate, if you send me your } S/N I could try to get a copy of it for you. That alone might satisfy } the questions being asked of your safety people. } } Disclaimer: SPI Supplies has offered this particular CPD unit, } especially to customers outside the USA, for nearly twenty five years } and we are unaware of any requirement to have the vessel retested at } periodic intervals for safety reasons. } } Chuck } } } ============================================ } } Charles A. Garber, Ph. D. Ph: 1-610-436-5400 } President 1-800-2424-SPI } SPI SUPPLIES FAX: 1-610-436-5755 } PO BOX 656 e-mail:cgarber-at-2spi.com } West Chester, PA 19381-0656 USA } Cust.Service: spi2spi-at-2spi.com } } Look for us! } ######################## } WWW: http://www.spi.cc } ######################## } ============================================ } } }
Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa R.Cross-at-ru.ac.za tel: +27 46 603 8168 - fax: +27 46 622 4377 http://www.ru.ac.za/emu/index.htm
** remember that ICEM-15 in 2002 is in Durban, South Africa **
Firstly, apologies for the earlier reply that was sent before I gave my comments!
} There are probably more CPD units of this design installed in the } world than any other design. I have never heard of anyone having any } kind of a problem with it
We have used two of these units for more than 25 years and the only problems we have experienced, other than seals which have had to be replaced from time to time, have been one ruptured disc and a leak which developed in the supply pipe from the CO2 cylinder. The latter was repaired (new one made up using existing connectors fitted to new high pressure piping) by our local compressed gas supplier.
Many years ago, having been told that pressure chambers should be checked from time to time, I sent one of our units back to be tested by Polaron in the UK. It came back with a certificate issued by Probe Technical Services saying that it had been tested to 2500 psi with the comment " Proof pressure applied for one minute without any visual sign of leakage or failure".
Regards
Rob
Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa R.Cross-at-ru.ac.za tel: +27 46 603 8168 - fax: +27 46 622 4377 http://www.ru.ac.za/emu/index.htm
** remember that ICEM-15 in 2002 is in Durban, South Africa **
Hi Christine, We have a Polaron E300 but no demand for routine testing from our 'safety people' presumably because they know that the burst disc will go long before the body starts to creak. After a major re-fit it was sent to testing specialists who gave it a hydrostatic test to 2500 psi for one minute for a safe working pressure of 2000 psi. It must be possible to do this on site, I cant imagine big autoclaves or air compressor reservoirs being posted off for their tests. It is not as exciting as it sounds. Holes are blanked off and then you pump it full of water so faults are revealed as leaks rather than the more entertaining earsplitting bang and shower of shrapnel which could result from the use of a compressible medium. If the safety people twist your arm try the Yellow Pages under Engineers, Inspection & Testing. ttfn, chris.smith-at-bbsrc.ac.uk Plant Path Dept., IACR-Rothamsted, Harpenden, Herts. UK.
Dear Gunnar It is most likely that there was at least one surface scratch or microcrack that was loaded in a way that resulted in major crack growth, whose result you observed. During glass removal and replacement, there were probably several actions that could have contributed to this -- scratching during cleaning or tool use, altered clamping of the glass, or turning a scratch on one side of the glass from inside to outside (or the reverse). Did anyone clamp the glass in a different way on replacing it, compared with before? (so that the applied stresses are less uniform or at least different, from before?) Moisture is well-known (scientifically and by all who specialize in cutting glass) to enhance crack growth when some component stress applied to the crack tip region helps to open the crack. All these could contribute to the failure. The timing of the failure indicates that one or more of these factors most likely resulted in cracking. Best wishes for avoiding the problem for another 20 years! Brian Robertson
------------------ Brian Robertson Assoc. Prof., Mechanical Engineering, University of Nebraska-Lincoln, on sabbatical at Department of Materials, University of Oxford, Parks Road, Oxford OX1 3PH, United Kingdom FAX 44+ 1865 273794 brian.robertson-at-materials.ox.ac.uk
* This e-mail message was sent with Execmail V5.0.x *
Olivier, I find it a bit bizarre that you cannot see our mapped features in your EELS. You're not mapping multiply scattered plasmons are you (a problem with lower lying edges)? How are you acquiring the EEL spectrum: Diffraction pattern (spot/CBED)? Or using the image? What sort of count rates are you getting in the pre-edge images and the maps? Are you sure it is not noise?
For EFTEM practice, I would if possible, try getting hold of some Al or Mg particles/powder (not smoke cubes), preferably mixed, and play with imaging the various plasmons i.e. see which energy windows are best and if possible see if you can image the oxide layers. It's fun because the signal is pretty high so easy to see.
Going to core-loss EFTEM the main problems are resolution: For low loss edges its spatial resolution (mainly delocalization) although there is evidence that this is not as restrictive as it looks on paper (see Z.L.Wang Ultramic Vol 67 (1997) pp105-111 for a demonstration in Al).
For higher edges the signal resolution (object resolution function as Berger & Kohl call it) tells you how good the statistics in a map are. A good paper is A.Berger & H.Kohl (Optik Vol:92 (1993), No4 pp175-193) which tells you about both spatial and signal resolution. The equations are quite rigorous, but it will show you how to get started measuring the signal-to-noise ratio (SNR). With a bit of practice you can then produce some SNR plots like those in Hofer, Grogger, Kothleitner & Warbichler (Ultramicroscopy 67 (1997)pp83-103) which tell you where the energy slit should be positioned to optimise SNR for a given slit width.
I suppose the best advice is find an easy material to practice with and work from thereon. Good luck and have fun.
Jonathan ******************************************************** Dr Jonathan Barnard
Analytical Materials Physics The Angstrom Laboratory, Uppsala University P O Box 534, SE-751 21 Uppsala, Sweden Phone: +46-(0)18-4716838 Fax: +46-(0)18-500131 http://www.angstrom.uu.se/analytical/home.html
Alan Fox, in a response to the Analytical TEM string, cautioned the question-raiser to beware of people who abuse instruments. Perhaps Nestor will forgive a little microscopy related humor and allow us to start a string on "Operator Horror Stories." Here's ours:
The "ham-fisted user" reference made us chuckle/cringe with the memory of a guest "microscopist" in our lab who hauled himself (220 pounds or so) out of his seat by pulling on the half-inserted specimen rod, bending it about 20 degrees or so!
Henceforth, when we saw him in the hall (he never came into the scope room again), we referred to him as "Conan the Microscopist"!
Ron Anderson, IBM, Hopewell Jct., New York, USA. anderron-at-us.ibm.com
IBM Analytical Services; http://www.chips.ibm.com/services/asg
Marie: You didn't mention the fixative and length of time fixed. Could it be possible that the tissue is "too well fixed" to allow easy exchange solutions? Also, have you tried thinner vibratome sections? We routinely use 40 micron thick brain sections for immuno labeling. Best regards, Henry *********************************************** Marie E. Cantino wrote: We are preparing perfusion fixed brain tissue by freeze substitution for post-embedding immuno gold. Following fixation we prepare 500 micron vibratome sections, which we then wish to infiltrate with 2 M sucrose in buffer prior to freezing. We have been transferring the brain sections to 1 M sucrose, then when they sink, transferring again to 2 M. The problem is that the sections never sink in the 2 M sucrose. ***************************** Henry Eichelberger, Manager Electron Microscopy Facility Department of Biological Sciences Binghamton University Binghamton, NY 13902-6000
Company Description: 30-year old designer and manufacturer of thin film optical interference filters used world-wide for laboratory and scientific equipment.
Position: Fluorescence Product Manager
Position Description: The Fluorescence Product Manager is responsible for any and all activities which help further the companyâs sales of fluorescence products. These activities include:
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Omega is seeking an individual who is excited about working with varied applications of fluorescence detection in the life sciences. The work involves discussing the best match between laboratory experiments and Omegaâs wide range of light filters for this science. Experience in fluorescence, microscopy, life sciences and instrumentation is essential.
Salary: Competitive with full benefit package.
Contact Information: Email response with attached Word file resume and salary history to: acoutermarsh-at-omegafilters.com or snailmail response to:
Andrew Coutermarsh, SPHR, HR Director Omega Optical, Inc. PO Box 573 Brattleboro, VT 05302
I am posting a message for a colleague who does not subscribe to this list.
He is looking for small embedding oven which can polymerize resins in the range of 40¡-70¡C, but which has a very precise temperature control (holds the temperature well, with very little fluctuation during polymerization). He is having problems with the oven he presently has because the temperature is not controlled well enough, and he is getting uneven polymerization.
Another condition is that he is in the middle of a project and needs the oven right away. He had an oven in mind, but it could not be delivered for 6 weeks!!!
Any suggestions from users or vendors are welcome. Please contact me offline, and I'll forward the replies to my colleague.
Thanks again for all your help.
Paula.
Paula Allan-Wojtas Research Scientist, Food Microstructure Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
Hallo friends, I have a Bomar SPC 1500 critical point dryer and I am looking for the rupture disc size or part number and a vendor. I looked in the Bomar manual but I can't find any information about it. I shall very much appreciate any information about it. Thanking you in advance. C.Singla
I agree with the disagreement that has been raised to my posting of my personal criteria for good embedment. (A section that stretches in the boat on exposure to solvent fumes is suboptimal and the embedment needs changing.)
I have not dealt much with plant material - mycobacterium is the closest I have come to dealing with cell walls which are tough as Michelin tires. I also have not used glass knives for more than a decade. Even my students start out on an old diamond knife. I am no longer familiar with the vagrancies of glass knives (every one of them seems to have a different cutting edge! Never twice the same!).
Certainly epoxies belong to the organic chemists. But they also belong to the material scientists. I have in front of me a thesis done under the mentoring of Dr. Giammara - A Material Science Evaluation of Epoxy Systems for TEM Applications. At one time I got into a huge fight with embedding filters of certain types. I had a computer program which calculated for me using WPE numbers and molecular weights 36 different formulations. I actually investigated 25 of them. Totally desperate (and mad) I started attending the material science symposia on polymers at the MSA meetings. I found this helpful.
My second favorite book is, HANDBOOK OF EPOXY RESINS. I adore it. (This has caused my friends to question my sanity and make peculiar remarks) This book has been of enormous help in my various battles (lots of them won) with epoxies. Certainly it deals mostly with organic chemistry.
On behalf of the many excellent microscopists I have known for many years I disagree violently with the statement that "and most EM biologists (AT LEAST THE GOOD ONES) understand a lot about organic chemistry." (Capitalizations are mine). One look at this most valuable of books makes it clear that most of the good EM biologists have no hope of knowing enough about polymer chemistry to understand or deal with the varied problems which can occur with epoxies.
Sections in our laboratory do not "stretch". Unless, of course, I want them to, for other special reasons. Our standard sections are the same size as the block face they come from. We deal with keratinized skin which is no picnic compared other biological tissues.
As usual, I always reserve the right to be wrong.
Hildy Crowley Sr. Electron Microscopist Dept of Biol Sciences University of Denver Denver, CO
P.S. My book is under lock and key at all times. If it is stolen, I will have to be hospitalized!
} This is on ebay now. Here is the URL: } } http://cgi.ebay.com/aw-cgi/eBayISAPI.dll?ViewItem&item=296173307 } } Not announced on ebay, and limited to MSA, I am offering } an IBM Thinkpad 355 which is optionally available to make this camera truly } portable. $350 extra gets the Thinkpad. You will still need a Slim SCSI } PCMCIA card, but these are very low cost. I have one in a Sony VAIO } and it works great with the Leaf.
It is on ebay and "Now announced on ebay, ..." It is NOT limited to MSA. Sorry about the poor eye-hand coordination.
Has anyone got a protocol for embedding LR white in a low-temperature chamber using UV light? I have a Leica AFS, so setting and maintaining a temp. is not a problem. I just tried it overnight at -20 C with UV (in filled gelatin capsules, but without activator) and did not get polymerization. Any tips would be appreciated.
Mary McKee Renal Unit MGH Charlestown, MA (617)726-5643
Dear UK CPD users, In the UK pressure systems which operate at pressures greater than 0.5 bar are subject to the Pressure Systems and Transportable Gas Container Regulations 1989. These rules require, among other things, regular inspection of the apparatus. You may also find that your institution's insurers impose a similar rule.
Regards, Eric
---------------------- Dr Eric Lachowski Department of Chemistry University of Aberdeen Meston Walk Old Aberdeen AB24 3UE Scotland +44 (0)1224 272934 fax 272921 e.lachowski-at-abdn.ac.uk
Originally Posted by: shens-at-focus.ruca.ua.ac.be
Hi,
How we can experimentally measure the collection angle when working with GIF? In other words, how to measure magnification between the plane of negatives and the plane of GIF entrance? Using a CCD camera? But normally a CCD camera magnifies an image 20 times of that on a negative, so pictures become uncomparible.
Originally posted by: Beverly_E_Maleeff-at-sbphrd.com
The Microscopy & Microanalysis 2000 Local Arrangements Committee is pleased to announce an additional social event at M&M2000---
TAKE ME OUT TO THE BALL GAME !!
Join us on Tuesday, August 15th as the PHILADELPHIA PHILLIES meet the ARIZONA DIAMONDBACKS at Veterans Stadium. Game time is 7:35 PM.
For $32.50, you'll get: - a ticket to the game (300 level, on the third base line) - a $10.00 coupon, good toward purchases at all concession and souvenir stands - round-trip transportation from the Convention Center to the stadium on a luxury coach (buses leave at 6:00 PM; return to the convention center immediately following the game)
A limited number of packages are available. First come, first served. Maximum 2 packages per person.
Checks, in US$ only, payable to "Microscopy & Microanalysis 2000" can be mailed to:
Bev Maleeff Treasurer, M&M2000 LAC c/o SmithKline Beecham Pharmaceuticals Mail Code UE 0462 709 Swedeland Road King of Prussia, PA 19406
Checks only, please. No credit cards accepted. Your cancelled check will serve as your receipt.
Tickets will be distributed at the M&M2000 Hospitality Booth on Monday and Tuesday, August 14th & 15th.
Further information about this and other M&M2000 events can be found on our web site: http://www.msa.microscopy.com/~mm2000/
Originally posted by: ron.doole-at-materials.ox.ac.uk
Hi All,
I enclose details of post doctoral positions currently available at Dept. of Materials here in Oxford. Please respond directly to the address given for each post. Good luck to any applicants,
Ron
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
********************************************************************** UNIVERSITY OF OXFORD
Department of Materials
POSTDOCTORAL RESEARCH POSITIONS - Salary range £16,286-£24,479p.a.
Applications are invited for the following positions:
Manufacture of Metal Matrix Composite Components
A position is available for a period of 20 months for a suitably qualified research assistant to join an inter-disciplinary research team in the Department of Materials and Department of Engineering Science researching into the manufacture of next generation aeroengine components from long fibre reinforced metal matrix composites. This post will study the fundamentals of fabrication mechanisms, and how these influence subsequent mechanical performance, using a combination of numerical modelling and experiments. Candidates should preferably have expertise and ability in one or more of the following areas: experimental solid mechanics, finite element analysis of plasticity or metal forming, microstructural characterisation of materials, composite materials. Please quote DJ00/6
Interface Modification in Organic LED
A position, sponsored by Opsys Limited, is available in the Department of Materials from April 2000 (or as soon as possible thereafter) for 18 months in the first instance. The project will explore routes to modifying the interfaces of the OLEDs with the objective to improve charge transport and injection. It will involve detailed investigation of the structural and physical properties of the modified interfaces between organo-lanthanide phosphors and other organic and inorganic materials. Techniques will include UPS, SIMS, AFM/STM and electro-optical characterisation of OLEDs. Expertise in one or more of these is highly desirable. Further details are available by email from: oleg.salata-at-materials.ox.ac.uk. Please quote DJ00/7.
Compositional inhomogeneities in information storage materials - effect on physical properties (Principal investigators: Amanda Petford-Long and Alfred Cerezo)
A 3-year position funded by the EPSRC, with support from Seagate Technology, is available in the Department of Materials from May 2000 (or as soon as possible thereafter). The aim of this project is to address how local inhomogeneities in the morphology and composition of thin layered films used for spin-valves and media in information storage technology are controlled by processing of the materials, and how this in turn determines their magnetic properties. The project will primarily involve the use of three-dimensional atom probe (3DAP) analysis to study the chemistry of the films at near-atomic resolution. Results from 3DAP analysis will be combined with measurements of magnetic properties to elucidate the role of the observed nanostructural features on the physical film properties. Some electron microscopy will also be required for comparative microstructural analysis. Expertise in materials characterisation is essential, and some knowledge of thin layered films or magnetic materials would also be helpful. Please quote ref. DJ00/8.
Applications including a full CV, list of publications and the names and addresses of three referees should be sent to The Administrator, Department of Materials, University of Oxford, Parks Road, Oxford, OX1 3PH, from whom further particulars are available. The closing date for applications is 20 April 2000.
It is not "horror" story, but a sort of... Many years ago the colleague from friendly Lab visited me with great project. The idea was simply and beautiful. She argues that because the image in the scope (TEM) is green (green fluorescence of the screen), she wants to modify the sample (I forgot what, some protein, I believe) by red-fluorescent dye to be able to see on the screen of the electron microscope the "double-staining": red on the green background. No comments...
Sergey
} Date: Thu, 30 Mar 2000 09:17:56 -0500 } From: "anderron-at-us.ibm.com"-at-sparc5.microscopy.com } Subject: Operator Horror Stories } To: microscopy-at-sparc5.microscopy.com } Importance: Normal } X-MIMETrack: Serialize by Router on D01ML065/01/M/IBM(Release 5.0.2b (Intl)|18 } January 2000) at 30/03/2000 09:18:01 } } ------------------------------------------------------------------------ } 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
As further comment on the pressure testing of the Polaron ® CPD units, I have been asked to submit the following statement from David Cheshire, a Manufacturing Manager at VG Microtech, the manufacturers of the Polaron brand of critical point dryers: ================================================= We use an independent test facility that has amongst its credentials the following international approvals : NAMAS, UKAS, NATA. The test certificates issued by ERA are for each individual chamber and we have copies of all these. The test itself is a sustained 50% overpressure (2000 psi) for 1 minute. Obviously any leakage would lead to failure and non certification. In the past units have been overpressured to 2500 and 3000 psi.
I think it is highly unlikely that a "local dive shop" would have the facilities and wherewithal to support the standards imposed by the various associations.
Incidentally the window is not quartz but a far denser and tougher material specifically manufactured for high pressure use.
Best regards Dave Cheshire
Polaron Range Development Manager VG Microtech The Birches Industrial Estate, Imberhorne Lane, East Grinstead, West Sussex. RH19 1UB. UK. =================================================== I believe there was some concern about having this kind of testing, if it was to be done, in any kind of facility that was not certified according to the bodies mentioned in David's message.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.spi.cc ######################## ============================================
In trying to adapt the SPAM filter last night to catch the latest porn site posting I accidently broke the software. Most messages got rejected after ~ 5 pm Chicago time. I will try to repost those messages , however, I may have missed a few. If you dont' see your posting by the end of the day, please resend it and accept my apology.
I would like to know why they use Argon in the metal coating devices. At our laboratory, we don't use Argon, we use air. What is the advantage of using Argon ? De Pauw Bart Faculty of Veterinary Medicine Morphology Salisburylaan 133 9820 Merelbeke Belgium Phone : 0032(0)9 264.77.19 Fax : 0032(0)9 264.77.90
Does anybody know where I can purchase Ni TEM grids with a holey carbon film ? I can find plenty of copper grids, but the Ni grids seem to be hard to find.
Thanks W. Erasmus
Willem Erasmus Snr. Scientist, Basic Catalysis Research Sasol Technology Tel : +27 +16 9603954 Fax : +27 +16 9602826 E-mail : willem.erasmus-at-sasol.com
Question: Many years ago the museum bought an AO microscope with an Expostar Shutter control. It appears that the control box (model 1190) has been lost. Is there anyway to buy a replacement? Does the company still exist? Is the microscope manufactored under another name?
Originally Posted by: george.bou-gharios-at-csc.mrc.ac.uk
I am writing to you in my capacity as secretary general of the XIth International Congress of Histochemistry and Cytochemistry which is held in YORK, UK 3-8 July 2000. The above mentioned Congress is Hosted by the Royal Microscopical Society (www.rms.org.uk) and n exciting programme has been put together which can be seen on
www.med.ic.ac.uk/external/ichc_2000/
Dr George Bou-Gharios Muscle Cell Biology Group MRC Clinical Sciences Centre Imperial College School of Medicine Hammersmith Hospital Du Cane Road London W12 0NN Tel: 0181-383 8261 Fax: 0181- 383 8264 email: george.bou-gharios-at-csc.mrc.ac.uk
Hi there folks, just a line to let you know that the schedule of presentations for the IUMAS 2000 meeting to be held in Kona, Hawaii, July 9th-14th 2000, is on the Web at:
http://www.microanalysis.org/iumas2000/
Thanks.
Jfm.
--
Dr. John Mansfield CPhys MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 763-2282 Cellular Phone: (734) 358-7555 (Leaving a phone message at 936-3352 is preferable to 358-7555) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42¡ 16' 48" Long. 83¡ 43' 48"
Hi there folks, just a line to let you know that the schedule of presentations for the IUMAS 2000 meeting to be held in Kona, Hawaii, July 9th-14th 2000, is on the Web at:
http://www.microanalysis.org/iumas2000/
Thanks.
Jfm.
--
Dr. John Mansfield CPhys MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 763-2282 Cellular Phone: (734) 358-7555 (Leaving a phone message at 936-3352 is preferable to 358-7555) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42¡ 16' 48" Long. 83¡ 43' 48"
I would like to know whether the ratio of plastidic to extraplastidic membranes changes in roots of Arabidopsis or any other plant in response to phosphate starvation. Does anyone know of a TEM ultrastructural morphometric analysis or any other type of analysis towards this end?
Thanks, Christoph Benning
Christoph Benning
Assistant Professor Dept. of Biochemistry Michigan State University East Lansing, MI 48824-1319 USA
The problem with Ni grids is that Ni is a magnetic metal, and so Ni grids can give all kinds of trouble. We used to use them back in the early days when they were about all that was available, and when resolution of EMs wasn't so great; now-a-days they have lost their popularity.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
He needs a source of plastic slides and/or film which can be machined and which do not fluoresce. Anybody have any ideas? Contact me offline.
Many thanks Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ******************************************************
} ---------- } From: Springett, Margaret J. } Sent: Friday, March 31, 2000 11:04 AM } To: 'springett.margaret-at-mayo.edu' } Subject: nickel grids } } I do an extensive amount of immunolabeling, and nickel grids are } economical, } but two factors need to be considered on a daily basis. The fact that } they } are magnetic can be an advantage as well as a detriment. Their magnetic } quality allows one to rinse grids floating on a bubble with a magnetic } stir } plate, this has been demonstrated in Dr. Bendyan's workshops on } goldlabeling } techniques. When trying to retreive a stubborn grid out of a grid box, a } magnetic tweezer is "heaven sent". Now about magnetic grids in the scope, } if you stigmate the scope after fine-focusing, your micrograph will be } focused. I have used these techniques to produce excellent micrographs of } caveolae at better than 100K magnification. For our use gold grids are } too } expensive, so "work around methods" with nickel grids are necessary. } Marge } } Margaret Springett IEM Specialist Electron Microscopy Core Facility Mayo Foundation and Clinic Rochester, MN 55905
There is an entire FTIR imaging industry out there. SpectraTech has been a leader in that industry and is a good place to start. Suggest you call Ed Manke (203-926-8998). Also, John Reffner has been one of the few people who has really bridged microscopy and spectroscopy. He is the real guru in this area. John is currently working with SensIR technologies and can be reached there at 203-207-9700.
The other companies who are in this area come from more of a spectroscopy background (Jobin Yvon/Instruments SA, Perkin Elmer, Hewlitt Packard, etc.) and would know less about the microscopy interface.
I taught a microscopy course to the apps specialists at Spectra Tech this January and had a chance to work with their Continuum, which has a combination of both glass and reflecting optics on its nosepiece. However, I think that the highest magnifications they had availalable were only about 32x objectives. The NAs were really great as I remember, so you could make use of electronics from the CCD to boost the actual mag to the screen.
Let me know how you make out.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ******************************************************
At 08:12 AM 3/31/00 -0600, Nestor J. Zaluzec wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
AO became part of Cambridge in 1986 then came under the Leica umbrella in 1990. I'd suggest that you start with Leica in Deerfield, IL. Wayne Buttermore is still there from the "old days" and may have a suggestion: 847-405-7044. The alternative would be to ask around in the used equipment market. Here are some of my contacts: MicroOptical Methods Dennis O'Leary Albany, NY 518-482-8200 Spectra Services Mike Specht Rochester, NY 716-654-9500 Vermont Optechs John Oren 802-425-2040 Martin Instruments Bob Martin Easley, SC 864-859-2688
Good luck. Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ******************************************************
At 08:13 AM 3/31/00 -0600, Nestor J. Zaluzec wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We once had a student twist the condenser control knob off a TEM. When asked to turn the knob clockwise, he just kept turning, until he handed me the knob.
William McManus Supervisor Electron Microscopy Facility Department of Biology Utah State University Logan UT 84322-5305
billEMac-at-cc.usu.edu 435-797-1920
-----Original Message----- } From: Nestor J. Zaluzec [mailto:zaluzec-at-sparc5.microscopy.com] Sent: Friday, March 31, 2000 7:02 AM To: Microscopy-at-sparc5.microscopy.com
Originally posted by: sryazant-at-ucla.edu
It is not "horror" story, but a sort of... Many years ago the colleague from friendly Lab visited me with great project. The idea was simply and beautiful. She argues that because the image in the scope (TEM) is green (green fluorescence of the screen), she wants to modify the sample (I forgot what, some protein, I believe) by red-fluorescent dye to be able to see on the screen of the electron microscope the "double-staining": red on the green background. No comments...
Sergey
} Date: Thu, 30 Mar 2000 09:17:56 -0500 } From: "anderron-at-us.ibm.com"-at-sparc5.microscopy.com } Subject: Operator Horror Stories } To: microscopy-at-sparc5.microscopy.com } Importance: Normal } X-MIMETrack: Serialize by Router on D01ML065/01/M/IBM(Release 5.0.2b (Intl)|18 } January 2000) at 30/03/2000 09:18:01 } } ------------------------------------------------------------------------ } 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 had one faculty operator several years ago who after a SINGLE lesson on the Phillips 200, decided to come back late at night and look some more. After taking some images, he couldn't get the door to the camera chamber off, since he had failed to vent it. After rummaging through the lab, he found a large screwdriver and commenced to pry the camera door off, unconcerned about the marks and gouges that he was putting in the brass. Needless to say, when the vacuum was finally broken, the mercury diffusion pump became very unhappy. Eventually we were able to replace the instrument and the user finally left after being denied tenure.
W. L. Steffens, Ph.D Dept. of Pathology University of Georgia
We get our Ni/holey carbon films from SPI, at http://www.2spi.com/. They will prepare excellent holey carbon films on just about any available grid material (for a price of course...).
Larry
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
Nestor J. Zaluzec 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 } -----------------------------------------------------------------------.} } Originally posted by: willem.erasmus-at-sasol.com } } Dear fellow microscopists } } Does anybody know where I can purchase Ni TEM grids with a holey carbon film } ? I can find plenty of copper grids, but the Ni grids seem to be hard to } find. } } Thanks } W. Erasmus } } Willem Erasmus } Snr. Scientist, Basic Catalysis Research } Sasol Technology } Tel : +27 +16 9603954 } Fax : +27 +16 9602826 } E-mail : willem.erasmus-at-sasol.com
We at Ladd Reseach can supply you with these, as would most of the other supply houses that make their own holey film. Please let us know what size mesh you want and a quantity and we can quote you.
Debbie Sicard Sales Manager
Disclaimer: Ladd Research is a vendor that sells EM supplies. --
LADD RESEARCH 131 Dorset Lane Williston, VT 05495 USA
TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net web site http://www.ladd.cc
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
The respected PhD I once worked with on our old JEOL JSM-U3 scanner, who was having trouble making out the details of the image on the monitor, so grabbed a flashlight and shined it at the screen so he could see the image a little better... :-).
Larry
} } } Originally posted by: sryazant-at-ucla.edu } } } } } It is not "horror" story, but a sort of... Many years ago the colleague } from friendly Lab visited me with great project. The idea was simply and } beautiful. She argues that because the image in the scope (TEM) is green } (green fluorescence of the screen), she wants to modify the sample (I } forgot what, some protein, I believe) by red-fluorescent dye to be able to } see on the screen of the electron microscope the "double-staining": red on } the green background. No comments... } } Sergey } } } Date: Thu, 30 Mar 2000 09:17:56 -0500 } } From: "anderron-at-us.ibm.com"-at-sparc5.microscopy.com } } Subject: Operator Horror Stories } } To: microscopy-at-sparc5.microscopy.com } } Importance: Normal } } X-MIMETrack: Serialize by Router on D01ML065/01/M/IBM(Release 5.0.2b } (Intl)|18 } } January 2000) at 30/03/2000 09:18:01 } } } } ------------------------------------------------------------------------ } } 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 } } -----------------------------------------------------------------------. } } } } } } } } Alan Fox, in a response to the Analytical TEM string, cautioned the } } question-raiser to beware of people who abuse instruments. Perhaps Nestor } } will forgive a little microscopy related humor and allow us to start a } } string on "Operator Horror Stories." Here's ours: } } } } The "ham-fisted user" reference made us chuckle/cringe with the memory of } } a guest "microscopist" in our lab who hauled himself (220 pounds or so) out } } of his seat by pulling on the half-inserted specimen rod, bending it about } } 20 degrees or so! } } } } Henceforth, when we saw him in the hall (he never came into the scope room } } again), we referred to him as "Conan the Microscopist"! } } } } } } } } Ron Anderson, IBM, Hopewell Jct., New York, USA. anderron-at-us.ibm.com } } } } IBM Analytical Services; http://www.chips.ibm.com/services/asg } } } } } } } } } } } _____________________________________ } } Sergey Ryazantsev Ph. D. } Electron Microscopy } UCLA School of Medicine } Department of Biological Chemistry } Box 951737 } Los Angeles, CA 90095-1737 } } Phone: (310) 825-1144 } Pager: (310) 845-0248 } FAX (departmental): (310) 206-5272 } mailto:sryazant-at-ucla.edu } http://www.bol.ucla.edu/~sryazant
Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
W. Erasmus asked: ================================================ Does anybody know where I can purchase Ni TEM grids with a holey carbon film ? I can find plenty of copper grids, but the Ni grids seem to be hard to find. ================================================= Some times the coating of Ni grids is a bit more difficult than the coating of copper grids. This translates to somewhat lower yields and sometimes higher prices. However, SPI Supplies regularly produces custom coated Ni grids on whatever mesh and grid the customer specifies and at only a slight price premium over copper.
Contact me off line for details or visit our webpage URL http://www.2spi.com/catalog/grids/cusctgrd.html
Some of the other major suppliers of consumables also will coat Ni grids, or at least that is my understanding.
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.spi.cc ######################## ============================================
One day a user of our TEM complained that she could see the beam with the specimen out, but as soon as she put in a grid the screen went black. After playing with it awhile and ruling out magnetism or something on the grid holder, I decided that there must be some small magnetic particle in the stage area that was being pushed around by the grid holder. Of course the service technician I got by phone wanted me to begin taking the column apart from the gun on down, but I grew impatient with this and soon went straight for the stage. I figured I would be looking for some small metal sliver. Imagine my surprise when I found an entire plastic pipettortip lying across the bore through the anticontamination plate in the column! It seems the previous user had been using these tips as forceps covers, and mysteriously lost one the day before. It had gone into the column through the airlock with her grid, not impossible with the Zeiss 10 top-loading system. Although not the only strange thing I have found in that microscope's column, at about an inch and a half long it is the largest.
Aloha, Tina **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
I am currently using the peroxidase/DAB pre-embedding method for localization of a novel extracellular matrix protein. I am using 40 micron vibratome sections. While my results give some extracellular labelling, most of the labelling is associated with dendritic microtubules. Has anyone experienced non-specific labelling of microtubules?
Microscpy Core Managers, Are there any EM/Confocal Microscope facilities within academia that are running in the black or at least breaking even without a subsidy. If so what are your rates and overhead, are your salaries included? Also are there any companies or facilities that do tele-microscopy? All info will be shared with the listserver, unless confidentiality is requested. Thank You.
Michael Delannoy Basic Science Microscopy Facility JHMI
Many, many years ago... I was asked to evaluate particle size of a powder. The powder was dried sludge from a radioactive waste storage tank. My downfall was permitting a co-worker to prepare the sample. When I received it, rather than a thin layer on carbon tape, it was caked on. Furthermore, the powder turned out to be sub-micron in size and almost pure Sr-90! Static and air currents began to spread contamination, even before I got it in the chamber. I quickly left the area before I became "hot" too.
I did finish the exam... In anti-contamination dress with a full face respirator at the console!
The job took a few hours. It took two weeks for me and my service engineer to disassemble the SEM, clean (and the room) it to reasonable levels and reassemble. The vacuum system was rather "warm" from that time on... Nevermore! Nevermore!
Moral: Always insist on making input and final approval if someone else wants to prepare your samples.
We had a cousin of Conan , who in his addled age, after inserting the injector tip into the stage of our EM400 could not find his grid in the microscope; he had dropped it on the floor. Of course the tip must have fallen off in the stage, so he promptly took a second tip and properly inserted the second tip on top of the first. Still no grid could be found. Ah ha, I will leave a polite note explaining the problem. It read as follows: " Please check the microscope, I had great difficulty inserting my specimens last evening." Philips kindly replaced the bulk of the stage just so they could keep the original for the museum of what not to do.
There once were two eager graduate students at Florida in the Materials Science and Engineering Department (one has the initials MK and now works for JEOL) that decided that they wanted learn how to align the Philips 300 TEM. Armed with the user manual, they proceeded to align the column, but couldn't do it. They had to call in the service engineer. The first thing that the service engineer did was to use a framing square to help to take the visible bow out of the column. It seems that the user manual they were working from was different from one for a microscope with a STEM attachment. (Not witnessed by me, but heard from reliable sources.)
A professor in the same department was escorting a visitor around the EM lab and the two were discussing possible experiments that might be performed in the Philips 300. They asked the EM technician to give them a hand opening up the microscope so they could look inside. The technician was busy and said that he would be with them in about 5 minutes. Well, busy professors can't wait for lowly technicians so they decided to open the chamber themselves. The professor cranked on the handle to open the gun chamber while it was under vacuum and the voltage was on. Bang, pop, whoosh, whistles, bells brought the technician running to see what had happened. After all, they only wanted to look inside. But, there was a happy ending to the story. The vacuum and diffusion pump were OK and the apertures were cleaned by the supersonic air. (I was around when this one happened.)
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
} -----Original Message----- } From: "anderron-at-us.ibm.com"-at-sparc5.microscopy.com } [mailto:"anderron-at-us.ibm.com"-at-sparc5.microscopy.com] } Sent: Thursday, March 30, 2000 9:18 AM } To: microscopy-at-sparc5.microscopy.com } Subject: Operator Horror Stories } } } -------------------------------------------------------------- } ---------- } 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 } } } -------------------------------------------------------------- } ---------. } } } } Alan Fox, in a response to the Analytical TEM string, cautioned the } question-raiser to beware of people who abuse instruments. } Perhaps Nestor } will forgive a little microscopy related humor and allow us to start a } string on "Operator Horror Stories." Here's ours: } } The "ham-fisted user" reference made us chuckle/cringe with } the memory of } a guest "microscopist" in our lab who hauled himself (220 } pounds or so) out } of his seat by pulling on the half-inserted specimen rod, } bending it about } 20 degrees or so! } } Henceforth, when we saw him in the hall (he never came into } the scope room } again), we referred to him as "Conan the Microscopist"! } } } } Ron Anderson, IBM, Hopewell Jct., New York, USA. } anderron-at-us.ibm.com } } IBM Analytical Services; http://www.chips.ibm.com/services/asg } } } }
This is my second go around with this, I got the Nestor porno bump this morning.
I had a student using my JEOL 100 CX II; loading and looking around went well. When she went to remove the holder she got half way out and yanked it toward her. This action put a 70 degree bend in the holder. I could never get the Z height to behave after that for some odd reason.
The other disaster was with my Philips 300, complete with Mercury Dif pump at that time. I needed to do some anode cleaning, so without deflating the air table I got on the console finished my work and got off. When the scope lurched to the right it dumped the contents of the mercury pump into the oil pump. This sent a cloud of mercury up the column creating an alloy wherever it went.
Believe it or not the scope is in fine working order with not a trace of mercury left in in the column. How did I do it? If you can figure it out I will send the winner a Lucent/Bell Labs brief case...at my cost of course.
John Grazul Lucent Technologies
"W. L. Steffens" 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 had one faculty operator several years ago who after a SINGLE lesson } on the Phillips 200, decided to come back late at night and look some } more. After taking some images, he couldn't get the door to the camera } chamber off, since he had failed to vent it. After rummaging through } the lab, he found a large screwdriver and commenced to pry the camera } door off, unconcerned about the marks and gouges that he was putting in } the brass. Needless to say, when the vacuum was finally broken, the } mercury diffusion pump became very unhappy. Eventually we were able to } replace the instrument and the user finally left after being denied } tenure. } } W. L. Steffens, Ph.D } Dept. of Pathology } University of Georgia
Once, over a decade back, I was working EM and Med Tech at the same time at a local hospital..
We had a CAP inspection, the officers were from another hospital in the state.
The question I was expected to reason and answer with a straight face was:
* What other protective measures besides standing behind a lead wall do you take when you put the glass slide into the TEM, to protect yourself from the flying electrons???
seriously!
I was rather stunned, and started looking from the wall socket to him and back, but he didn't catch the connection.
I then tried to tactfully explain that vacuum was required, and that we really didn't need that extra protection!
It was explained to me that he did too know all about EM because he had a one day workshop.
Web Pages: Lab Page http://treefrog.cvm.uiuc.edu Centeral States Microscopy Page http://treefrog.cvm.uiuc.edu/csmms Personal Home Page http://treefrog.cvm.uiuc.edu/lam
} We once had an operator on our JEOL 840A SEM complain about her } sample moving. Her sample was samples of vacuum grease she had gold } coated and was attempting to image in the SEM. When the electron } beam heated the grease it cracked up the gold coating and charged. } She was using the SEM to look for silicon in various greases. I } suggested she use a different technique so we did not end up with } vaporized grease inside the SEM.
David R. Hull NASA Glenn Research Center at Lewis Field Advanced Metallics Branch Mail Stop 49-1 21000 Brookpark Road Cleveland, OH 44135
It seems everyone has one of these. I had a person who one day complained of a burning smell while she was at the scope. I went to investigate and when I arrived in the room there was no smell... burning or otherwise. Several more times that session she came and got me to investigate this burning smell in the TEM room. This continued for a couple of sessions. I did go and investigate because after all, who wants their TEM on fire? I honestly didn't know what to do about it and was beginning to seriously question her sanity. Finally she convinced me to sit with her while she operated the scope and sure enough there was this faint smell of something burning! I jumped up and tried to find it, but it had gone away. So...she sat down again and began to work and the smell, like plastic kitchenware burning in a dishwasher, came back. Again I jump up and look around....to no avail! I was curious now. The burning smell only happens when she is sitting at the scope. Not being a big believer in spontaneous combustion, there had to be an answer! There was. As part of the TEM course I take the kick panel off the Zeiss 10 to show the students the guts of the vacuum system. The kick panel is right below the desk area where you sit up to the microscope. What she was doing was placing her tennis shoe on the heater of the diffusion pump and the plastic/rubber would melt and burn a bit -- just enough to give the room a burning smell. She never did notice her foot getting warm. When I pointed it out she did admit that her shoe was warm and had some ugly scars on it to boot! She never returned after that semester - I guess that she had enough of EM! Now I carefully point out the hazard and replace the cover quickly when we are finished with the class.
-- ================================================================== Greg Strout Electron Microscopist, University of Oklahoma WWW Virtual Library for Microscopy: http://www.ou.edu/research/electron/www-vl/ e-mail: gstrout-at-ou.edu Opinions expressed herein are mine and not necessarily those of the University of Oklahoma ==================================================================
And all of these cute blunders that happened way back when were caused by today's scientists/researchers.
Experience, good and bad, is the only way we all really learn.
Enjoying the stories.....
Harry Ekstrom Honeywell Materials Laboratory
-----Original Message----- } From: White, Woody N [mailto:Woody.N.White-at-mcdermott.com] Sent: Friday, March 31, 2000 2:46 PM To: Microscopy-at-sparc5.microscopy.com
Many, many years ago... I was asked to evaluate particle size of a powder. The powder was dried sludge from a radioactive waste storage tank. My downfall was permitting a co-worker to prepare the sample. When I received it, rather than a thin layer on carbon tape, it was caked on. Furthermore, the powder turned out to be sub-micron in size and almost pure Sr-90! Static and air currents began to spread contamination, even before I got it in the chamber. I quickly left the area before I became "hot" too.
I did finish the exam... In anti-contamination dress with a full face respirator at the console!
The job took a few hours. It took two weeks for me and my service engineer to disassemble the SEM, clean (and the room) it to reasonable levels and reassemble. The vacuum system was rather "warm" from that time on... Nevermore! Nevermore!
Moral: Always insist on making input and final approval if someone else wants to prepare your samples.
Purdue University Department of Biological Sciences
An opening is available immediately for a scientist specializing in macromolecular microscopy at Purdue University. The successful candidate would be responsible for overseeing the day-to-day operations of the high-resolution electron microscopy facility, training new users, and assisting outside visitors with data collection and analysis. He/she would also be expected to work with the cryoEM groups in the department at planning and implementing the future directions of the facility, including the development of new technologies for improved specimen preservation, data collection, and analysis. The ideal candidate should complement the existing strengths represented in the department and pursue independent research funding.
The Department of Biological Sciences at Purdue provides a rich intellectual environment as well as outstanding modern scientific facilities including Philips EM420, CM200-FEG, and CM300-FEG electron cryo-microscopes. Purdue University is home to over 50,000 students, faculty, and staff, and is located in West Lafayette, Indiana - approximately 1 hour northwest of Indianapolis and 2.5 hours southeast of Chicago. The area affords the advantages associated with a major university, while providing an affordable and attractive `small town' environment in which to live.
Requirements for this position include a Ph.D. in biophysics or one of the related sciences, a minimum of 5 years post-doctoral work experience in electron cryo-microscopy of biological molecules, and a proven ability to work well with others. Preference will be given to candidates who have successfully developed new technologies and methodologies for high-resolution electron cryo-microscopy. Salary will be commensurate with experience and includes full benefits.
Interested individuals should send their resume, relevant reprints, a statement of research interests and career goals, and provide contact information (name/phone/e-mail) for three individuals who have agreed to serve as references to:
Dr. Timothy S. Baker (tsb-at-bragg.bio.purdue.edu)
and/or
Dr. Amy McGough (amcgough-at-bcm.tmc.edu)
Department of Biological Sciences Lilly Hall of Life Science Purdue University West Lafayette, IN 47907-1392
Purdue is an equal access/equal opportunity university.
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Is there any truth in the story I was told about a lab in London that replaced the leaded glass on the microscope chamber with regular glass? Apparently the mistake was discovered when all the film on the shelf behind the operator become fogged!
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm
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Hello All, My two horror stories! I was working with an investigator who wanted to see matrix vessicles. Well, we were so happy that we had a nicely fixed sample with "lots" of vessicles. Nice whole round ones. Well, when the negatives were developed, my director and I discovered that what we really had were nicely fixed, beautiful bacteria!!!! No vessicles! Another time, in school, I was walking down the hallway later in the afternoon when I saw a scope room door slightly open. I popped my head in to see who was there and to ask if they wanted the door closed. When I looked inside, I found a student sound asleep, with hands still holding the knobs of the microscope!!!!!!! Funny huh? Jo
} ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } Alan Fox, in a response to the Analytical TEM string, cautioned the } question-raiser to beware of people who abuse instruments. Perhaps Nestor } will forgive a little microscopy related humor and allow us to start a } string on "Operator Horror Stories." Here's ours: } } The "ham-fisted user" reference made us chuckle/cringe with the memory of } a guest "microscopist" in our lab who hauled himself (220 pounds or so) out } of his seat by pulling on the half-inserted specimen rod, bending it about } 20 degrees or so! } } Henceforth, when we saw him in the hall (he never came into the scope room } again), we referred to him as "Conan the Microscopist"! } } Ron Anderson, IBM, Hopewell Jct., New York, USA. anderron-at-us.ibm.com } } IBM Analytical Services; http://www.chips.ibm.com/services/asg
Bart, I believe Argon is used because it is a larger molecule and sputters the metal more efficiently. I also use air with no problem. Possibly the amount of metal sputtered is slightly less per unit time. John Hunt
On Fri, 31 Mar 2000, Nestor J. Zaluzec 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 } -----------------------------------------------------------------------. } } } } Originally posted by: bart.depauw-at-rug.ac.be } } } Hello, } } I would like to know why they use Argon in the metal coating devices. At } our laboratory, we don't use Argon, we use air. What is the advantage of } using Argon ? } De Pauw Bart } Faculty of Veterinary Medicine } Morphology } Salisburylaan 133 } 9820 Merelbeke } Belgium } Phone : 0032(0)9 264.77.19 } Fax : 0032(0)9 264.77.90 } } }
Air contains water vapour, oxygen and carbon dioxide all of which can be broken down in the plasma generated in a sputter coater and gve rise to nasty active species which can damage delicate samples. Argon is used because of its relative high atomic number and this ensures there is multiple scattering of the sputtered metal. There is a Zenon sputter coater on the market which should sputter even better but I have no experience with it. Come to the Lehigh Short Course(contact {slc6-at-lehigh.edu) and we will tell you all about it.
Many years ago we had a Siemens EM-101. This was a relatively new microscope and beautifully machined with German precision. The camera chamber was a work of art. Each film plate (we used glass back then) was encased in it's own light-tight cassette. The camera would move a film cassette into position for exposure and pull off the cassette cover. The operator would then expose the film and move the casette into a drop box. The cassette cover would be pushed closed in the process. This camera worked almost flawlessly. On occassion, an operator, when loading the camera, would put the cassettes in on an angle. When you went to take the first image, the cassette would jam and not move into position. All that was necessary to unjam was to break vacuum, giggle the cassette with your hand to straighten it and then repump the camera chamber and get back to work...a 10 minute process as most. We had one operator who thought she knew it all. She was working on the microscope one evening and the camera jammed. Her solution was to take out the camera and take it apart...screw by screw and spring by spring. She ended up with an incredible heap of parts and of course had no clue as to how to put it back together. Neither did the Siemens service engineers. They had never seen one apart! They had to find another camera (not easy in those days with relatively few of these instruments around), take it apart..carefully marking where each piece came from...and then reassembled each camera simultaneously. It took hours!! Moral of the story...NO ONE fixed ANYTHING without permission of the lab personnel!!
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
I am preparing to purchase a sputter coater for Au/Pd SEM specimen preparation. I have narrowed the search to the VG-Scientific Polaron SC7620. My old Polaron E5100 is very tired.
I have located used Hummer VI, Fullam EMS-76M and Denton Desk II coaters for about $1000 less than new. Some of these include the pump. However, I have several dual stage mech pumps, so this is not an issue.
If anyone knows of a source for a good/recent used coater, I would appreciate knowing about it. Otherwise, I will go with a new one.
These are great stories and would make a wonderful collection, especially to hand out to students of EM. Yes, experience is a great way to learn but it's often best, and cheaper, to learn from some else's.
I heard that one years ago. Also one that supposedly happened at Univ. Wisconsin. Someone apparently cleaned a scope's internal parts in acetone and started pumping. Before it was fully pumped down, they turned on the filament. Bang! Nice story but could it possibly happen? I would think that any acetone residue would evaporate very quickly or is there some other phenomenon here relating to ratio of gases?
Damian Neuberger, Ph.D. Baxter Healthcare, Inc. WG3-2S P.O. Box 490 Round Lake, IL 60073 e-mail: damian_neuberger-at-baxter.co, Tel: 847-270-5888 Fax: 847-270-5897
} Is there any truth in the story I was told about a lab in London that } replaced the leaded glass on the microscope chamber with regular } glass? Apparently the mistake was discovered when all the film on the } shelf behind the operator become fogged! } } Paul Webster, Ph.D } House Ear Institute } 2100 West Third Street } Los Angeles, CA 90057 } phone:213 273 8026 } fax: 213 413 6739 } e-mail: pwebster-at-hei.org } http://www.hei.org/htm/aemi.htm
I am preparing to purchase a sputter coater for Au/Pd SEM specimen preparation. I have narrowed the search to the VG-Scientific Polaron SC7620. My old Polaron E5100 is very tired.
I have located used Hummer VI, Fullam EMS-76M and Denton Desk II coaters for about $1000 less than new. Some of these include the pump. However, I have several dual stage mech pumps, so this is not an issue.
If anyone knows of a source for a good/recent used coater, I would appreciate knowing about it. Otherwise, I will go with a new one.
These are great stories and would make a wonderful collection, especially to hand out to students of EM. Yes, experience is a great way to learn but it's often best, and cheaper, to learn from some else's.
I heard that one years ago. Also one that supposedly happened at Univ. Wisconsin. Someone apparently cleaned a scope's internal parts in acetone and started pumping. Before it was fully pumped down, they turned on the filament. Bang! Nice story but could it possibly happen? I would think that any acetone residue would evaporate very quickly or is there some other phenomenon here relating to ratio of gases?
Damian Neuberger, Ph.D. Baxter Healthcare, Inc. WG3-2S P.O. Box 490 Round Lake, IL 60073 e-mail: damian_neuberger-at-baxter.co, Tel: 847-270-5888 Fax: 847-270-5897
} Is there any truth in the story I was told about a lab in London that } replaced the leaded glass on the microscope chamber with regular } glass? Apparently the mistake was discovered when all the film on the } shelf behind the operator become fogged! } } Paul Webster, Ph.D } House Ear Institute } 2100 West Third Street } Los Angeles, CA 90057 } phone:213 273 8026 } fax: 213 413 6739 } e-mail: pwebster-at-hei.org } http://www.hei.org/htm/aemi.htm
My story is not as much of a horror as it is embarassing. Several years back I received a package of samples from a regular client without any paper work describing what the samples were. This is not unusual since we frequently recieve unknowns. I proceeded to open the package and was abruptly assaulted with an extremely strong odor of bannanas. Apparantly one of the samples was a vial of concentrated bannana flavoring. It was months (almost a year) before the odor completely disappeared from my office, and I was cajoled about it frequently. I by the way, hate bannanas.............
Lou Solebello -----Original Message----- } From: L R MELSEN {lmelsen-at-emory.edu} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
Here is a "just off the press" example (but this does come up at least once a year):
I fielded a phone call from a distraught SEM lab manager who told me that some months ago he put Dow Corning fluid into his diffusion pump "to save money". And now that he has had an accident, the silicone fluid of course, has contaminated his system, so he was asking us, "what organic solvent will easily remove it."
He was especially upset when I told him that his EDS detector will see Si everywhere also, because they do a lot of analyses for Si!
I won't repeat what he told me when I tried to explain the reality of his situation......
But it does go to show that there are a lot of new people entering our profession, some with less training and experience with vacuum than others, so such stories are very well worth repeating.
But just out of curiosity, is there some "recommended procedure" for removing silicone fluid from the internal parts of a column, and also removing it from the window of an EDS detector? I presume one can always call in an outside service provider with experienced people but a lot of users out there just don't have the budget for something like that. But they do have a good supply of student manpower.
While we are on the subject of silicone, a few weeks ago a well known TEM user got me on the phone to say "hello" and commented that he had just placed an order for silicone grease and yes, he said, he was going to be using it on the o rings of his column. I told him I thought that he should be using other greases and his response was "don't listen to dogma, I thought you read the listserver!". Am I correct, namely that one should not be using silicone grease on the o rings of a column instrument?
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.spi.cc ######################## ============================================
I am enjoying the "horror stories" (don't we all enjoy hearing about someone else's stupid mistake) and thought I would share one from the "software" category.
I had just joined Northern Scientific (later Tracor Northern, now Noran) and was writing EDS software for our first computerized analyzer (the venerable NS-880) -- the year was 1972, and my software, like any of the stuff being produced in those years where we wrote software on paper tape for machines with 4K memories, just wasn't all that "user friendly", so it was not unusual to have to walk people through difficult command sequences. Then too, computers were pretty new to everyone and people sometimes made silly-sounding mistakes out of pure inexperience (one highly intelligent customer, when instructed to 'type a space', dutifully typed "A SPACE" on the keyboard). One particular customer, however, defied all attempts at instruction and had passed well beyond "complaining" into the realm of "abusive" and "insulting". He insisted that he was following the instructions to the letter, but that our miserable software consistently malfunctioned. We were unable to duplicate the problem and finally several of us drove a considerable distance to his lab to get to the bottom of this. We went through the operations with him watching intently and the software performed flawlessly, which he acknowledged. We then asked him to operate the software himself and midway through, observed that he was responding to one of the dialog questions with a decidedly inappropriate response. We had him back out of this and repeat it with the correct response and the software then ran perfectly. To which his response was to draw himself up and state with great authority and indignation: "but when you run it THE WAY I DO, it doesn't work!"
Question: Many years ago the museum bought an AO microscope with an Expostar Shutter control. It appears that the control box (model 1190) has been lost. Is there anyway to buy a replacement? Does the company still exist? Is the microscope manufactored under another name?
It is not "horror" story, but a sort of... Many years ago the colleague from friendly Lab visited me with great project. The idea was simply and beautiful. She argues that because the image in the scope (TEM) is green (green fluorescence of the screen), she wants to modify the sample (I forgot what, some protein, I believe) by red-fluorescent dye to be able to see on the screen of the electron microscope the "double-staining": red on the green background. No comments...
Sergey
} Date: Thu, 30 Mar 2000 09:17:56 -0500 } From: "anderron-at-us.ibm.com"-at-sparc5.microscopy.com } Subject: Operator Horror Stories } To: microscopy-at-sparc5.microscopy.com } Importance: Normal } X-MIMETrack: Serialize by Router on D01ML065/01/M/IBM(Release 5.0.2b (Intl)|18 } January 2000) at 30/03/2000 09:18:01 } } ------------------------------------------------------------------------ } 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
Question: Many years ago the museum bought an AO microscope with an Expostar Shutter control. It appears that the control box (model 1190) has been lost. Is there anyway to buy a replacement? Does the company still exist? Is the microscope manufactored under another name?
Originally Posted by: george.bou-gharios-at-csc.mrc.ac.uk
I am writing to you in my capacity as secretary general of the XIth International Congress of Histochemistry and Cytochemistry which is held in YORK, UK 3-8 July 2000. The above mentioned Congress is Hosted by the Royal Microscopical Society (www.rms.org.uk) and n exciting programme has been put together which can be seen on
www.med.ic.ac.uk/external/ichc_2000/
Dr George Bou-Gharios Muscle Cell Biology Group MRC Clinical Sciences Centre Imperial College School of Medicine Hammersmith Hospital Du Cane Road London W12 0NN Tel: 0181-383 8261 Fax: 0181- 383 8264 email: george.bou-gharios-at-csc.mrc.ac.uk
Originally Posted by: shens-at-focus.ruca.ua.ac.be
Hi,
How we can experimentally measure the collection angle when working with GIF? In other words, how to measure magnification between the plane of negatives and the plane of GIF entrance? Using a CCD camera? But normally a CCD camera magnifies an image 20 times of that on a negative, so pictures become uncomparible.
Originally Posted by: george.bou-gharios-at-csc.mrc.ac.uk
I am writing to you in my capacity as secretary general of the XIth International Congress of Histochemistry and Cytochemistry which is held in YORK, UK 3-8 July 2000. The above mentioned Congress is Hosted by the Royal Microscopical Society (www.rms.org.uk) and n exciting programme has been put together which can be seen on
www.med.ic.ac.uk/external/ichc_2000/
Dr George Bou-Gharios Muscle Cell Biology Group MRC Clinical Sciences Centre Imperial College School of Medicine Hammersmith Hospital Du Cane Road London W12 0NN Tel: 0181-383 8261 Fax: 0181- 383 8264 email: george.bou-gharios-at-csc.mrc.ac.uk
In trying to adapt the SPAM filter last night to catch the latest porn site posting I accidently broke the software. Most messages got rejected after ~ 5 pm Chicago time. I will try to repost those messages , however, I may have missed a few. If you dont' see your posting by the end of the day, please resend it and accept my apology.
In trying to adapt the SPAM filter last night to catch the latest porn site posting I accidently broke the software. Most messages got rejected after ~ 5 pm Chicago time. I will try to repost those messages , however, I may have missed a few. If you dont' see your posting by the end of the day, please resend it and accept my apology.
Originally Posted by: shens-at-focus.ruca.ua.ac.be
Hi,
How we can experimentally measure the collection angle when working with GIF? In other words, how to measure magnification between the plane of negatives and the plane of GIF entrance? Using a CCD camera? But normally a CCD camera magnifies an image 20 times of that on a negative, so pictures become uncomparible.
It is not "horror" story, but a sort of... Many years ago the colleague from friendly Lab visited me with great project. The idea was simply and beautiful. She argues that because the image in the scope (TEM) is green (green fluorescence of the screen), she wants to modify the sample (I forgot what, some protein, I believe) by red-fluorescent dye to be able to see on the screen of the electron microscope the "double-staining": red on the green background. No comments...
Sergey
} Date: Thu, 30 Mar 2000 09:17:56 -0500 } From: "anderron-at-us.ibm.com"-at-sparc5.microscopy.com } Subject: Operator Horror Stories } To: microscopy-at-sparc5.microscopy.com } Importance: Normal } X-MIMETrack: Serialize by Router on D01ML065/01/M/IBM(Release 5.0.2b (Intl)|18 } January 2000) at 30/03/2000 09:18:01 } } ------------------------------------------------------------------------ } 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 would like to know why they use Argon in the metal coating devices. At our laboratory, we don't use Argon, we use air. What is the advantage of using Argon ? De Pauw Bart Faculty of Veterinary Medicine Morphology Salisburylaan 133 9820 Merelbeke Belgium Phone : 0032(0)9 264.77.19 Fax : 0032(0)9 264.77.90
I would like to know why they use Argon in the metal coating devices. At our laboratory, we don't use Argon, we use air. What is the advantage of using Argon ? De Pauw Bart Faculty of Veterinary Medicine Morphology Salisburylaan 133 9820 Merelbeke Belgium Phone : 0032(0)9 264.77.19 Fax : 0032(0)9 264.77.90
As further comment on the pressure testing of the Polaron ® CPD units, I have been asked to submit the following statement from David Cheshire, a Manufacturing Manager at VG Microtech, the manufacturers of the Polaron brand of critical point dryers: ================================================= We use an independent test facility that has amongst its credentials the following international approvals : NAMAS, UKAS, NATA. The test certificates issued by ERA are for each individual chamber and we have copies of all these. The test itself is a sustained 50% overpressure (2000 psi) for 1 minute. Obviously any leakage would lead to failure and non certification. In the past units have been overpressured to 2500 and 3000 psi.
I think it is highly unlikely that a "local dive shop" would have the facilities and wherewithal to support the standards imposed by the various associations.
Incidentally the window is not quartz but a far denser and tougher material specifically manufactured for high pressure use.
Best regards Dave Cheshire
Polaron Range Development Manager VG Microtech The Birches Industrial Estate, Imberhorne Lane, East Grinstead, West Sussex. RH19 1UB. UK. =================================================== I believe there was some concern about having this kind of testing, if it was to be done, in any kind of facility that was not certified according to the bodies mentioned in David's message.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.spi.cc ######################## ============================================
As further comment on the pressure testing of the Polaron ® CPD units, I have been asked to submit the following statement from David Cheshire, a Manufacturing Manager at VG Microtech, the manufacturers of the Polaron brand of critical point dryers: ================================================= We use an independent test facility that has amongst its credentials the following international approvals : NAMAS, UKAS, NATA. The test certificates issued by ERA are for each individual chamber and we have copies of all these. The test itself is a sustained 50% overpressure (2000 psi) for 1 minute. Obviously any leakage would lead to failure and non certification. In the past units have been overpressured to 2500 and 3000 psi.
I think it is highly unlikely that a "local dive shop" would have the facilities and wherewithal to support the standards imposed by the various associations.
Incidentally the window is not quartz but a far denser and tougher material specifically manufactured for high pressure use.
Best regards Dave Cheshire
Polaron Range Development Manager VG Microtech The Birches Industrial Estate, Imberhorne Lane, East Grinstead, West Sussex. RH19 1UB. UK. =================================================== I believe there was some concern about having this kind of testing, if it was to be done, in any kind of facility that was not certified according to the bodies mentioned in David's message.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.spi.cc ######################## ============================================
Hi there folks, just a line to let you know that the schedule of presentations for the IUMAS 2000 meeting to be held in Kona, Hawaii, July 9th-14th 2000, is on the Web at:
http://www.microanalysis.org/iumas2000/
Thanks.
Jfm.
--
Dr. John Mansfield CPhys MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 763-2282 Cellular Phone: (734) 358-7555 (Leaving a phone message at 936-3352 is preferable to 358-7555) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42¡ 16' 48" Long. 83¡ 43' 48"
Hi there folks, just a line to let you know that the schedule of presentations for the IUMAS 2000 meeting to be held in Kona, Hawaii, July 9th-14th 2000, is on the Web at:
http://www.microanalysis.org/iumas2000/
Thanks.
Jfm.
--
Dr. John Mansfield CPhys MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 763-2282 Cellular Phone: (734) 358-7555 (Leaving a phone message at 936-3352 is preferable to 358-7555) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42¡ 16' 48" Long. 83¡ 43' 48"
The problem with Ni grids is that Ni is a magnetic metal, and so Ni grids can give all kinds of trouble. We used to use them back in the early days when they were about all that was available, and when resolution of EMs wasn't so great; now-a-days they have lost their popularity.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
The problem with Ni grids is that Ni is a magnetic metal, and so Ni grids can give all kinds of trouble. We used to use them back in the early days when they were about all that was available, and when resolution of EMs wasn't so great; now-a-days they have lost their popularity.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
I would like to know whether the ratio of plastidic to extraplastidic membranes changes in roots of Arabidopsis or any other plant in response to phosphate starvation. Does anyone know of a TEM ultrastructural morphometric analysis or any other type of analysis towards this end?
Thanks, Christoph Benning
Christoph Benning
Assistant Professor Dept. of Biochemistry Michigan State University East Lansing, MI 48824-1319 USA
I would like to know whether the ratio of plastidic to extraplastidic membranes changes in roots of Arabidopsis or any other plant in response to phosphate starvation. Does anyone know of a TEM ultrastructural morphometric analysis or any other type of analysis towards this end?
Thanks, Christoph Benning
Christoph Benning
Assistant Professor Dept. of Biochemistry Michigan State University East Lansing, MI 48824-1319 USA
I had one faculty operator several years ago who after a SINGLE lesson on the Phillips 200, decided to come back late at night and look some more. After taking some images, he couldn't get the door to the camera chamber off, since he had failed to vent it. After rummaging through the lab, he found a large screwdriver and commenced to pry the camera door off, unconcerned about the marks and gouges that he was putting in the brass. Needless to say, when the vacuum was finally broken, the mercury diffusion pump became very unhappy. Eventually we were able to replace the instrument and the user finally left after being denied tenure.
W. L. Steffens, Ph.D Dept. of Pathology University of Georgia
He needs a source of plastic slides and/or film which can be machined and which do not fluoresce. Anybody have any ideas? Contact me offline.
Many thanks Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ******************************************************
He needs a source of plastic slides and/or film which can be machined and which do not fluoresce. Anybody have any ideas? Contact me offline.
Many thanks Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ******************************************************
I had one faculty operator several years ago who after a SINGLE lesson on the Phillips 200, decided to come back late at night and look some more. After taking some images, he couldn't get the door to the camera chamber off, since he had failed to vent it. After rummaging through the lab, he found a large screwdriver and commenced to pry the camera door off, unconcerned about the marks and gouges that he was putting in the brass. Needless to say, when the vacuum was finally broken, the mercury diffusion pump became very unhappy. Eventually we were able to replace the instrument and the user finally left after being denied tenure.
W. L. Steffens, Ph.D Dept. of Pathology University of Georgia
} ---------- } From: Springett, Margaret J. } Sent: Friday, March 31, 2000 11:04 AM } To: 'springett.margaret-at-mayo.edu' } Subject: nickel grids } } I do an extensive amount of immunolabeling, and nickel grids are } economical, } but two factors need to be considered on a daily basis. The fact that } they } are magnetic can be an advantage as well as a detriment. Their magnetic } quality allows one to rinse grids floating on a bubble with a magnetic } stir } plate, this has been demonstrated in Dr. Bendyan's workshops on } goldlabeling } techniques. When trying to retreive a stubborn grid out of a grid box, a } magnetic tweezer is "heaven sent". Now about magnetic grids in the scope, } if you stigmate the scope after fine-focusing, your micrograph will be } focused. I have used these techniques to produce excellent micrographs of } caveolae at better than 100K magnification. For our use gold grids are } too } expensive, so "work around methods" with nickel grids are necessary. } Marge } } Margaret Springett IEM Specialist Electron Microscopy Core Facility Mayo Foundation and Clinic Rochester, MN 55905
} ---------- } From: Springett, Margaret J. } Sent: Friday, March 31, 2000 11:04 AM } To: 'springett.margaret-at-mayo.edu' } Subject: nickel grids } } I do an extensive amount of immunolabeling, and nickel grids are } economical, } but two factors need to be considered on a daily basis. The fact that } they } are magnetic can be an advantage as well as a detriment. Their magnetic } quality allows one to rinse grids floating on a bubble with a magnetic } stir } plate, this has been demonstrated in Dr. Bendyan's workshops on } goldlabeling } techniques. When trying to retreive a stubborn grid out of a grid box, a } magnetic tweezer is "heaven sent". Now about magnetic grids in the scope, } if you stigmate the scope after fine-focusing, your micrograph will be } focused. I have used these techniques to produce excellent micrographs of } caveolae at better than 100K magnification. For our use gold grids are } too } expensive, so "work around methods" with nickel grids are necessary. } Marge } } Margaret Springett IEM Specialist Electron Microscopy Core Facility Mayo Foundation and Clinic Rochester, MN 55905
We once had a student twist the condenser control knob off a TEM. When asked to turn the knob clockwise, he just kept turning, until he handed me the knob.
William McManus Supervisor Electron Microscopy Facility Department of Biology Utah State University Logan UT 84322-5305
billEMac-at-cc.usu.edu 435-797-1920
-----Original Message----- } From: Nestor J. Zaluzec [mailto:zaluzec-at-sparc5.microscopy.com] Sent: Friday, March 31, 2000 7:02 AM To: Microscopy-at-sparc5.microscopy.com
Originally posted by: sryazant-at-ucla.edu
It is not "horror" story, but a sort of... Many years ago the colleague from friendly Lab visited me with great project. The idea was simply and beautiful. She argues that because the image in the scope (TEM) is green (green fluorescence of the screen), she wants to modify the sample (I forgot what, some protein, I believe) by red-fluorescent dye to be able to see on the screen of the electron microscope the "double-staining": red on the green background. No comments...
Sergey
} Date: Thu, 30 Mar 2000 09:17:56 -0500 } From: "anderron-at-us.ibm.com"-at-sparc5.microscopy.com } Subject: Operator Horror Stories } To: microscopy-at-sparc5.microscopy.com } Importance: Normal } X-MIMETrack: Serialize by Router on D01ML065/01/M/IBM(Release 5.0.2b (Intl)|18 } January 2000) at 30/03/2000 09:18:01 } } ------------------------------------------------------------------------ } 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
We once had a student twist the condenser control knob off a TEM. When asked to turn the knob clockwise, he just kept turning, until he handed me the knob.
William McManus Supervisor Electron Microscopy Facility Department of Biology Utah State University Logan UT 84322-5305
billEMac-at-cc.usu.edu 435-797-1920
-----Original Message----- } From: Nestor J. Zaluzec [mailto:zaluzec-at-sparc5.microscopy.com] Sent: Friday, March 31, 2000 7:02 AM To: Microscopy-at-sparc5.microscopy.com
Originally posted by: sryazant-at-ucla.edu
It is not "horror" story, but a sort of... Many years ago the colleague from friendly Lab visited me with great project. The idea was simply and beautiful. She argues that because the image in the scope (TEM) is green (green fluorescence of the screen), she wants to modify the sample (I forgot what, some protein, I believe) by red-fluorescent dye to be able to see on the screen of the electron microscope the "double-staining": red on the green background. No comments...
Sergey
} Date: Thu, 30 Mar 2000 09:17:56 -0500 } From: "anderron-at-us.ibm.com"-at-sparc5.microscopy.com } Subject: Operator Horror Stories } To: microscopy-at-sparc5.microscopy.com } Importance: Normal } X-MIMETrack: Serialize by Router on D01ML065/01/M/IBM(Release 5.0.2b (Intl)|18 } January 2000) at 30/03/2000 09:18:01 } } ------------------------------------------------------------------------ } 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
We get our Ni/holey carbon films from SPI, at http://www.2spi.com/. They will prepare excellent holey carbon films on just about any available grid material (for a price of course...).
Larry
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
We get our Ni/holey carbon films from SPI, at http://www.2spi.com/. They will prepare excellent holey carbon films on just about any available grid material (for a price of course...).
Larry
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
Nestor J. Zaluzec 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 } -----------------------------------------------------------------------.} } Originally posted by: willem.erasmus-at-sasol.com } } Dear fellow microscopists } } Does anybody know where I can purchase Ni TEM grids with a holey carbon film } ? I can find plenty of copper grids, but the Ni grids seem to be hard to } find. } } Thanks } W. Erasmus } } Willem Erasmus } Snr. Scientist, Basic Catalysis Research } Sasol Technology } Tel : +27 +16 9603954 } Fax : +27 +16 9602826 } E-mail : willem.erasmus-at-sasol.com
We at Ladd Reseach can supply you with these, as would most of the other supply houses that make their own holey film. Please let us know what size mesh you want and a quantity and we can quote you.
Debbie Sicard Sales Manager
Disclaimer: Ladd Research is a vendor that sells EM supplies. --
LADD RESEARCH 131 Dorset Lane Williston, VT 05495 USA
TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net web site http://www.ladd.cc
Nestor J. Zaluzec 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 } -----------------------------------------------------------------------.} } Originally posted by: willem.erasmus-at-sasol.com } } Dear fellow microscopists } } Does anybody know where I can purchase Ni TEM grids with a holey carbon film } ? I can find plenty of copper grids, but the Ni grids seem to be hard to } find. } } Thanks } W. Erasmus } } Willem Erasmus } Snr. Scientist, Basic Catalysis Research } Sasol Technology } Tel : +27 +16 9603954 } Fax : +27 +16 9602826 } E-mail : willem.erasmus-at-sasol.com
We at Ladd Reseach can supply you with these, as would most of the other supply houses that make their own holey film. Please let us know what size mesh you want and a quantity and we can quote you.
Debbie Sicard Sales Manager
Disclaimer: Ladd Research is a vendor that sells EM supplies. --
LADD RESEARCH 131 Dorset Lane Williston, VT 05495 USA
TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net web site http://www.ladd.cc
I am currently using the peroxidase/DAB pre-embedding method for localization of a novel extracellular matrix protein. I am using 40 micron vibratome sections. While my results give some extracellular labelling, most of the labelling is associated with dendritic microtubules. Has anyone experienced non-specific labelling of microtubules?
I am currently using the peroxidase/DAB pre-embedding method for localization of a novel extracellular matrix protein. I am using 40 micron vibratome sections. While my results give some extracellular labelling, most of the labelling is associated with dendritic microtubules. Has anyone experienced non-specific labelling of microtubules?
W. Erasmus asked: ================================================ Does anybody know where I can purchase Ni TEM grids with a holey carbon film ? I can find plenty of copper grids, but the Ni grids seem to be hard to find. ================================================= Some times the coating of Ni grids is a bit more difficult than the coating of copper grids. This translates to somewhat lower yields and sometimes higher prices. However, SPI Supplies regularly produces custom coated Ni grids on whatever mesh and grid the customer specifies and at only a slight price premium over copper.
Contact me off line for details or visit our webpage URL http://www.2spi.com/catalog/grids/cusctgrd.html
Some of the other major suppliers of consumables also will coat Ni grids, or at least that is my understanding.
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.spi.cc ######################## ============================================
W. Erasmus asked: ================================================ Does anybody know where I can purchase Ni TEM grids with a holey carbon film ? I can find plenty of copper grids, but the Ni grids seem to be hard to find. ================================================= Some times the coating of Ni grids is a bit more difficult than the coating of copper grids. This translates to somewhat lower yields and sometimes higher prices. However, SPI Supplies regularly produces custom coated Ni grids on whatever mesh and grid the customer specifies and at only a slight price premium over copper.
Contact me off line for details or visit our webpage URL http://www.2spi.com/catalog/grids/cusctgrd.html
Some of the other major suppliers of consumables also will coat Ni grids, or at least that is my understanding.
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.spi.cc ######################## ============================================
One day a user of our TEM complained that she could see the beam with the specimen out, but as soon as she put in a grid the screen went black. After playing with it awhile and ruling out magnetism or something on the grid holder, I decided that there must be some small magnetic particle in the stage area that was being pushed around by the grid holder. Of course the service technician I got by phone wanted me to begin taking the column apart from the gun on down, but I grew impatient with this and soon went straight for the stage. I figured I would be looking for some small metal sliver. Imagine my surprise when I found an entire plastic pipettortip lying across the bore through the anticontamination plate in the column! It seems the previous user had been using these tips as forceps covers, and mysteriously lost one the day before. It had gone into the column through the airlock with her grid, not impossible with the Zeiss 10 top-loading system. Although not the only strange thing I have found in that microscope's column, at about an inch and a half long it is the largest.
Aloha, Tina **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
One day a user of our TEM complained that she could see the beam with the specimen out, but as soon as she put in a grid the screen went black. After playing with it awhile and ruling out magnetism or something on the grid holder, I decided that there must be some small magnetic particle in the stage area that was being pushed around by the grid holder. Of course the service technician I got by phone wanted me to begin taking the column apart from the gun on down, but I grew impatient with this and soon went straight for the stage. I figured I would be looking for some small metal sliver. Imagine my surprise when I found an entire plastic pipettortip lying across the bore through the anticontamination plate in the column! It seems the previous user had been using these tips as forceps covers, and mysteriously lost one the day before. It had gone into the column through the airlock with her grid, not impossible with the Zeiss 10 top-loading system. Although not the only strange thing I have found in that microscope's column, at about an inch and a half long it is the largest.
Aloha, Tina **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Microscpy Core Managers, Are there any EM/Confocal Microscope facilities within academia that are running in the black or at least breaking even without a subsidy. If so what are your rates and overhead, are your salaries included? Also are there any companies or facilities that do tele-microscopy? All info will be shared with the listserver, unless confidentiality is requested. Thank You.
Michael Delannoy Basic Science Microscopy Facility JHMI
Microscpy Core Managers, Are there any EM/Confocal Microscope facilities within academia that are running in the black or at least breaking even without a subsidy. If so what are your rates and overhead, are your salaries included? Also are there any companies or facilities that do tele-microscopy? All info will be shared with the listserver, unless confidentiality is requested. Thank You.
Michael Delannoy Basic Science Microscopy Facility JHMI
Many, many years ago... I was asked to evaluate particle size of a powder. The powder was dried sludge from a radioactive waste storage tank. My downfall was permitting a co-worker to prepare the sample. When I received it, rather than a thin layer on carbon tape, it was caked on. Furthermore, the powder turned out to be sub-micron in size and almost pure Sr-90! Static and air currents began to spread contamination, even before I got it in the chamber. I quickly left the area before I became "hot" too.
I did finish the exam... In anti-contamination dress with a full face respirator at the console!
The job took a few hours. It took two weeks for me and my service engineer to disassemble the SEM, clean (and the room) it to reasonable levels and reassemble. The vacuum system was rather "warm" from that time on... Nevermore! Nevermore!
Moral: Always insist on making input and final approval if someone else wants to prepare your samples.
Many, many years ago... I was asked to evaluate particle size of a powder. The powder was dried sludge from a radioactive waste storage tank. My downfall was permitting a co-worker to prepare the sample. When I received it, rather than a thin layer on carbon tape, it was caked on. Furthermore, the powder turned out to be sub-micron in size and almost pure Sr-90! Static and air currents began to spread contamination, even before I got it in the chamber. I quickly left the area before I became "hot" too.
I did finish the exam... In anti-contamination dress with a full face respirator at the console!
The job took a few hours. It took two weeks for me and my service engineer to disassemble the SEM, clean (and the room) it to reasonable levels and reassemble. The vacuum system was rather "warm" from that time on... Nevermore! Nevermore!
Moral: Always insist on making input and final approval if someone else wants to prepare your samples.
This is my second go around with this, I got the Nestor porno bump this morning.
I had a student using my JEOL 100 CX II; loading and looking around went well. When she went to remove the holder she got half way out and yanked it toward her. This action put a 70 degree bend in the holder. I could never get the Z height to behave after that for some odd reason.
The other disaster was with my Philips 300, complete with Mercury Dif pump at that time. I needed to do some anode cleaning, so without deflating the air table I got on the console finished my work and got off. When the scope lurched to the right it dumped the contents of the mercury pump into the oil pump. This sent a cloud of mercury up the column creating an alloy wherever it went.
Believe it or not the scope is in fine working order with not a trace of mercury left in in the column. How did I do it? If you can figure it out I will send the winner a Lucent/Bell Labs brief case...at my cost of course.
John Grazul Lucent Technologies
"W. L. Steffens" 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 had one faculty operator several years ago who after a SINGLE lesson } on the Phillips 200, decided to come back late at night and look some } more. After taking some images, he couldn't get the door to the camera } chamber off, since he had failed to vent it. After rummaging through } the lab, he found a large screwdriver and commenced to pry the camera } door off, unconcerned about the marks and gouges that he was putting in } the brass. Needless to say, when the vacuum was finally broken, the } mercury diffusion pump became very unhappy. Eventually we were able to } replace the instrument and the user finally left after being denied } tenure. } } W. L. Steffens, Ph.D } Dept. of Pathology } University of Georgia
This is my second go around with this, I got the Nestor porno bump this morning.
I had a student using my JEOL 100 CX II; loading and looking around went well. When she went to remove the holder she got half way out and yanked it toward her. This action put a 70 degree bend in the holder. I could never get the Z height to behave after that for some odd reason.
The other disaster was with my Philips 300, complete with Mercury Dif pump at that time. I needed to do some anode cleaning, so without deflating the air table I got on the console finished my work and got off. When the scope lurched to the right it dumped the contents of the mercury pump into the oil pump. This sent a cloud of mercury up the column creating an alloy wherever it went.
Believe it or not the scope is in fine working order with not a trace of mercury left in in the column. How did I do it? If you can figure it out I will send the winner a Lucent/Bell Labs brief case...at my cost of course.
John Grazul Lucent Technologies
"W. L. Steffens" 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 had one faculty operator several years ago who after a SINGLE lesson } on the Phillips 200, decided to come back late at night and look some } more. After taking some images, he couldn't get the door to the camera } chamber off, since he had failed to vent it. After rummaging through } the lab, he found a large screwdriver and commenced to pry the camera } door off, unconcerned about the marks and gouges that he was putting in } the brass. Needless to say, when the vacuum was finally broken, the } mercury diffusion pump became very unhappy. Eventually we were able to } replace the instrument and the user finally left after being denied } tenure. } } W. L. Steffens, Ph.D } Dept. of Pathology } University of Georgia
Once, over a decade back, I was working EM and Med Tech at the same time at a local hospital..
We had a CAP inspection, the officers were from another hospital in the state.
The question I was expected to reason and answer with a straight face was:
* What other protective measures besides standing behind a lead wall do you take when you put the glass slide into the TEM, to protect yourself from the flying electrons???
seriously!
I was rather stunned, and started looking from the wall socket to him and back, but he didn't catch the connection.
I then tried to tactfully explain that vacuum was required, and that we really didn't need that extra protection!
It was explained to me that he did too know all about EM because he had a one day workshop.
Web Pages: Lab Page http://treefrog.cvm.uiuc.edu Centeral States Microscopy Page http://treefrog.cvm.uiuc.edu/csmms Personal Home Page http://treefrog.cvm.uiuc.edu/lam
Once, over a decade back, I was working EM and Med Tech at the same time at a local hospital..
We had a CAP inspection, the officers were from another hospital in the state.
The question I was expected to reason and answer with a straight face was:
* What other protective measures besides standing behind a lead wall do you take when you put the glass slide into the TEM, to protect yourself from the flying electrons???
seriously!
I was rather stunned, and started looking from the wall socket to him and back, but he didn't catch the connection.
I then tried to tactfully explain that vacuum was required, and that we really didn't need that extra protection!
It was explained to me that he did too know all about EM because he had a one day workshop.
Web Pages: Lab Page http://treefrog.cvm.uiuc.edu Centeral States Microscopy Page http://treefrog.cvm.uiuc.edu/csmms Personal Home Page http://treefrog.cvm.uiuc.edu/lam
} We once had an operator on our JEOL 840A SEM complain about her } sample moving. Her sample was samples of vacuum grease she had gold } coated and was attempting to image in the SEM. When the electron } beam heated the grease it cracked up the gold coating and charged. } She was using the SEM to look for silicon in various greases. I } suggested she use a different technique so we did not end up with } vaporized grease inside the SEM.
David R. Hull NASA Glenn Research Center at Lewis Field Advanced Metallics Branch Mail Stop 49-1 21000 Brookpark Road Cleveland, OH 44135
} We once had an operator on our JEOL 840A SEM complain about her } sample moving. Her sample was samples of vacuum grease she had gold } coated and was attempting to image in the SEM. When the electron } beam heated the grease it cracked up the gold coating and charged. } She was using the SEM to look for silicon in various greases. I } suggested she use a different technique so we did not end up with } vaporized grease inside the SEM.
David R. Hull NASA Glenn Research Center at Lewis Field Advanced Metallics Branch Mail Stop 49-1 21000 Brookpark Road Cleveland, OH 44135
It seems everyone has one of these. I had a person who one day complained of a burning smell while she was at the scope. I went to investigate and when I arrived in the room there was no smell... burning or otherwise. Several more times that session she came and got me to investigate this burning smell in the TEM room. This continued for a couple of sessions. I did go and investigate because after all, who wants their TEM on fire? I honestly didn't know what to do about it and was beginning to seriously question her sanity. Finally she convinced me to sit with her while she operated the scope and sure enough there was this faint smell of something burning! I jumped up and tried to find it, but it had gone away. So...she sat down again and began to work and the smell, like plastic kitchenware burning in a dishwasher, came back. Again I jump up and look around....to no avail! I was curious now. The burning smell only happens when she is sitting at the scope. Not being a big believer in spontaneous combustion, there had to be an answer! There was. As part of the TEM course I take the kick panel off the Zeiss 10 to show the students the guts of the vacuum system. The kick panel is right below the desk area where you sit up to the microscope. What she was doing was placing her tennis shoe on the heater of the diffusion pump and the plastic/rubber would melt and burn a bit -- just enough to give the room a burning smell. She never did notice her foot getting warm. When I pointed it out she did admit that her shoe was warm and had some ugly scars on it to boot! She never returned after that semester - I guess that she had enough of EM! Now I carefully point out the hazard and replace the cover quickly when we are finished with the class.
-- ================================================================== Greg Strout Electron Microscopist, University of Oklahoma WWW Virtual Library for Microscopy: http://www.ou.edu/research/electron/www-vl/ e-mail: gstrout-at-ou.edu Opinions expressed herein are mine and not necessarily those of the University of Oklahoma ==================================================================
It seems everyone has one of these. I had a person who one day complained of a burning smell while she was at the scope. I went to investigate and when I arrived in the room there was no smell... burning or otherwise. Several more times that session she came and got me to investigate this burning smell in the TEM room. This continued for a couple of sessions. I did go and investigate because after all, who wants their TEM on fire? I honestly didn't know what to do about it and was beginning to seriously question her sanity. Finally she convinced me to sit with her while she operated the scope and sure enough there was this faint smell of something burning! I jumped up and tried to find it, but it had gone away. So...she sat down again and began to work and the smell, like plastic kitchenware burning in a dishwasher, came back. Again I jump up and look around....to no avail! I was curious now. The burning smell only happens when she is sitting at the scope. Not being a big believer in spontaneous combustion, there had to be an answer! There was. As part of the TEM course I take the kick panel off the Zeiss 10 to show the students the guts of the vacuum system. The kick panel is right below the desk area where you sit up to the microscope. What she was doing was placing her tennis shoe on the heater of the diffusion pump and the plastic/rubber would melt and burn a bit -- just enough to give the room a burning smell. She never did notice her foot getting warm. When I pointed it out she did admit that her shoe was warm and had some ugly scars on it to boot! She never returned after that semester - I guess that she had enough of EM! Now I carefully point out the hazard and replace the cover quickly when we are finished with the class.
-- ================================================================== Greg Strout Electron Microscopist, University of Oklahoma WWW Virtual Library for Microscopy: http://www.ou.edu/research/electron/www-vl/ e-mail: gstrout-at-ou.edu Opinions expressed herein are mine and not necessarily those of the University of Oklahoma ==================================================================
Purdue University Department of Biological Sciences
An opening is available immediately for a scientist specializing in macromolecular microscopy at Purdue University. The successful candidate would be responsible for overseeing the day-to-day operations of the high-resolution electron microscopy facility, training new users, and assisting outside visitors with data collection and analysis. He/she would also be expected to work with the cryoEM groups in the department at planning and implementing the future directions of the facility, including the development of new technologies for improved specimen preservation, data collection, and analysis. The ideal candidate should complement the existing strengths represented in the department and pursue independent research funding.
The Department of Biological Sciences at Purdue provides a rich intellectual environment as well as outstanding modern scientific facilities including Philips EM420, CM200-FEG, and CM300-FEG electron cryo-microscopes. Purdue University is home to over 50,000 students, faculty, and staff, and is located in West Lafayette, Indiana - approximately 1 hour northwest of Indianapolis and 2.5 hours southeast of Chicago. The area affords the advantages associated with a major university, while providing an affordable and attractive `small town' environment in which to live.
Requirements for this position include a Ph.D. in biophysics or one of the related sciences, a minimum of 5 years post-doctoral work experience in electron cryo-microscopy of biological molecules, and a proven ability to work well with others. Preference will be given to candidates who have successfully developed new technologies and methodologies for high-resolution electron cryo-microscopy. Salary will be commensurate with experience and includes full benefits.
Interested individuals should send their resume, relevant reprints, a statement of research interests and career goals, and provide contact information (name/phone/e-mail) for three individuals who have agreed to serve as references to:
Dr. Timothy S. Baker (tsb-at-bragg.bio.purdue.edu)
and/or
Dr. Amy McGough (amcgough-at-bcm.tmc.edu)
Department of Biological Sciences Lilly Hall of Life Science Purdue University West Lafayette, IN 47907-1392
Purdue is an equal access/equal opportunity university.
Purdue University Department of Biological Sciences
An opening is available immediately for a scientist specializing in macromolecular microscopy at Purdue University. The successful candidate would be responsible for overseeing the day-to-day operations of the high-resolution electron microscopy facility, training new users, and assisting outside visitors with data collection and analysis. He/she would also be expected to work with the cryoEM groups in the department at planning and implementing the future directions of the facility, including the development of new technologies for improved specimen preservation, data collection, and analysis. The ideal candidate should complement the existing strengths represented in the department and pursue independent research funding.
The Department of Biological Sciences at Purdue provides a rich intellectual environment as well as outstanding modern scientific facilities including Philips EM420, CM200-FEG, and CM300-FEG electron cryo-microscopes. Purdue University is home to over 50,000 students, faculty, and staff, and is located in West Lafayette, Indiana - approximately 1 hour northwest of Indianapolis and 2.5 hours southeast of Chicago. The area affords the advantages associated with a major university, while providing an affordable and attractive `small town' environment in which to live.
Requirements for this position include a Ph.D. in biophysics or one of the related sciences, a minimum of 5 years post-doctoral work experience in electron cryo-microscopy of biological molecules, and a proven ability to work well with others. Preference will be given to candidates who have successfully developed new technologies and methodologies for high-resolution electron cryo-microscopy. Salary will be commensurate with experience and includes full benefits.
Interested individuals should send their resume, relevant reprints, a statement of research interests and career goals, and provide contact information (name/phone/e-mail) for three individuals who have agreed to serve as references to:
Dr. Timothy S. Baker (tsb-at-bragg.bio.purdue.edu)
and/or
Dr. Amy McGough (amcgough-at-bcm.tmc.edu)
Department of Biological Sciences Lilly Hall of Life Science Purdue University West Lafayette, IN 47907-1392
Purdue is an equal access/equal opportunity university.
Bart, I believe Argon is used because it is a larger molecule and sputters the metal more efficiently. I also use air with no problem. Possibly the amount of metal sputtered is slightly less per unit time. John Hunt
On Fri, 31 Mar 2000, Nestor J. Zaluzec 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 } -----------------------------------------------------------------------. } } } } Originally posted by: bart.depauw-at-rug.ac.be } } } Hello, } } I would like to know why they use Argon in the metal coating devices. At } our laboratory, we don't use Argon, we use air. What is the advantage of } using Argon ? } De Pauw Bart } Faculty of Veterinary Medicine } Morphology } Salisburylaan 133 } 9820 Merelbeke } Belgium } Phone : 0032(0)9 264.77.19 } Fax : 0032(0)9 264.77.90 } } }
Many years ago we had a Siemens EM-101. This was a relatively new microscope and beautifully machined with German precision. The camera chamber was a work of art. Each film plate (we used glass back then) was encased in it's own light-tight cassette. The camera would move a film cassette into position for exposure and pull off the cassette cover. The operator would then expose the film and move the casette into a drop box. The cassette cover would be pushed closed in the process. This camera worked almost flawlessly. On occassion, an operator, when loading the camera, would put the cassettes in on an angle. When you went to take the first image, the cassette would jam and not move into position. All that was necessary to unjam was to break vacuum, giggle the cassette with your hand to straighten it and then repump the camera chamber and get back to work...a 10 minute process as most. We had one operator who thought she knew it all. She was working on the microscope one evening and the camera jammed. Her solution was to take out the camera and take it apart...screw by screw and spring by spring. She ended up with an incredible heap of parts and of course had no clue as to how to put it back together. Neither did the Siemens service engineers. They had never seen one apart! They had to find another camera (not easy in those days with relatively few of these instruments around), take it apart..carefully marking where each piece came from...and then reassembled each camera simultaneously. It took hours!! Moral of the story...NO ONE fixed ANYTHING without permission of the lab personnel!!
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
Bart, I believe Argon is used because it is a larger molecule and sputters the metal more efficiently. I also use air with no problem. Possibly the amount of metal sputtered is slightly less per unit time. John Hunt
On Fri, 31 Mar 2000, Nestor J. Zaluzec 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 } -----------------------------------------------------------------------. } } } } Originally posted by: bart.depauw-at-rug.ac.be } } } Hello, } } I would like to know why they use Argon in the metal coating devices. At } our laboratory, we don't use Argon, we use air. What is the advantage of } using Argon ? } De Pauw Bart } Faculty of Veterinary Medicine } Morphology } Salisburylaan 133 } 9820 Merelbeke } Belgium } Phone : 0032(0)9 264.77.19 } Fax : 0032(0)9 264.77.90 } } }
Many years ago we had a Siemens EM-101. This was a relatively new microscope and beautifully machined with German precision. The camera chamber was a work of art. Each film plate (we used glass back then) was encased in it's own light-tight cassette. The camera would move a film cassette into position for exposure and pull off the cassette cover. The operator would then expose the film and move the casette into a drop box. The cassette cover would be pushed closed in the process. This camera worked almost flawlessly. On occassion, an operator, when loading the camera, would put the cassettes in on an angle. When you went to take the first image, the cassette would jam and not move into position. All that was necessary to unjam was to break vacuum, giggle the cassette with your hand to straighten it and then repump the camera chamber and get back to work...a 10 minute process as most. We had one operator who thought she knew it all. She was working on the microscope one evening and the camera jammed. Her solution was to take out the camera and take it apart...screw by screw and spring by spring. She ended up with an incredible heap of parts and of course had no clue as to how to put it back together. Neither did the Siemens service engineers. They had never seen one apart! They had to find another camera (not easy in those days with relatively few of these instruments around), take it apart..carefully marking where each piece came from...and then reassembled each camera simultaneously. It took hours!! Moral of the story...NO ONE fixed ANYTHING without permission of the lab personnel!!
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
Air contains water vapour, oxygen and carbon dioxide all of which can be broken down in the plasma generated in a sputter coater and gve rise to nasty active species which can damage delicate samples. Argon is used because of its relative high atomic number and this ensures there is multiple scattering of the sputtered metal. There is a Zenon sputter coater on the market which should sputter even better but I have no experience with it. Come to the Lehigh Short Course(contact {slc6-at-lehigh.edu) and we will tell you all about it.
Air contains water vapour, oxygen and carbon dioxide all of which can be broken down in the plasma generated in a sputter coater and gve rise to nasty active species which can damage delicate samples. Argon is used because of its relative high atomic number and this ensures there is multiple scattering of the sputtered metal. There is a Zenon sputter coater on the market which should sputter even better but I have no experience with it. Come to the Lehigh Short Course(contact {slc6-at-lehigh.edu) and we will tell you all about it.
I am preparing to purchase a sputter coater for Au/Pd SEM specimen preparation. I have narrowed the search to the VG-Scientific Polaron SC7620. My old Polaron E5100 is very tired.
I have located used Hummer VI, Fullam EMS-76M and Denton Desk II coaters for about $1000 less than new. Some of these include the pump. However, I have several dual stage mech pumps, so this is not an issue.
If anyone knows of a source for a good/recent used coater, I would appreciate knowing about it. Otherwise, I will go with a new one.
I am preparing to purchase a sputter coater for Au/Pd SEM specimen preparation. I have narrowed the search to the VG-Scientific Polaron SC7620. My old Polaron E5100 is very tired.
I have located used Hummer VI, Fullam EMS-76M and Denton Desk II coaters for about $1000 less than new. Some of these include the pump. However, I have several dual stage mech pumps, so this is not an issue.
If anyone knows of a source for a good/recent used coater, I would appreciate knowing about it. Otherwise, I will go with a new one.
I appreciate the comments by David Cheshire. Perhaps he could elaborate why a place (not necessarily a Dive Shop) certified for the testing of diving cylinders and regulators could not cope with a CPD? Is the implication that these testing places can not be trusted (sorry divers), or that a CPD is more complex than other equipment, or, horror, could these places be cheaper to get a CPD tested. and certified. Surely, hydrostatic testing is rather similar and certification and the relevant legislation would be for pressure vessels and not just for CPD.
I still believe that testing of CPD is counter-productive, but if required by legislation, then there is no option available.
I believe that CPD windows used to be quartz. This may well have changed and some manufacturers may now use other material e.g."bullet proof glass", which of course is a plastic. David Cheshire would have been more helpful to devulge which material is used. This would be of great interest because some people have used organic solvents as an intermediate fluid. In that case it would be important to know what solvents affect this "far denser and tougher material". Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
-----Original Message----- Sent: Saturday, April 01, 2000 12:03 AM To: MICROSCOPY BB
A couple of suggestions that may help your collogue and others in regards to ovens. Please note that PST sells ovens, but not to North America (wrong volts, costly shipping).
For resin curing most people purchase the cheapest oven type, which relies on convection currents to achieve reasonably even temperatures. Fan forced and jacketed ovens have greater temperature uniformity within the chamber. (See diagrams of oven types in our online: Home} Contents} E3} "click here" link) Its wise to place specimens always in a similar position within the chamber.
A thermostat control the heating elements and commonly they switch within 1.5 degree C. Metals absorb heat faster and conduct heat faster into specimens. It is quite possible to part-melt polyethylene capsules where they are in contact with metal. The simple solution is to place the specimens on a piece of wood or thick cardboard.
The offending oven may have a thermostat that switches to its own strange rhythm. More likely though, it has an unacceptably wide range when turning the element on and off. A possible solution to this problem is to enclose the specimen in an insulating box within the oven. This should cause the highs and lows to be leveled. That insulating box could be a small lid-less cardbox, upended over the specimen and its insulating support.
Uneven polymerisation is often due to varying times within the oven. An easy solution is a "lamplighter" timer. Plug the oven into this 24 hour timer and adjust the timer to come on at say 5 pm and switch off at 8 am. Specimen can be placed in the oven at any time and removed any time during the next day and cure for the same number of hours. Fewer fumes within the lab is another benefit. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Friday, March 31, 2000 4:02 AM, Paula Allan-Wojtas [SMTP:ALLANWOJTASP-at-EM.AGR.CA] wrote: } } } Hi, All, } } I am posting a message for a colleague who does not subscribe to this list. } } He is looking for small embedding oven which can polymerize resins in the } range of 40?-70?C, but which has a very precise temperature control (holds } the temperature well, with very little fluctuation during polymerization). He } is having problems with the oven he presently has because the temperature is } not controlled well enough, and he is getting uneven polymerization. } } Another condition is that he is in the middle of a project and needs the oven } right away. He had an oven in mind, but it could not be delivered for 6 } weeks!!! } } Any suggestions from users or vendors are welcome. Please contact me offline, } and I'll forward the replies to my colleague. } } Thanks again for all your help. } } Paula. } } Paula Allan-Wojtas } Research Scientist, Food Microstructure } Agriculture and Agri-Food Canada } Atlantic Food and Horticulture Research Centre } Kentville, Nova Scotia Canada B4N 1J5 } } Tel: (902) 679-5566 } FAX: (902) 679-2311 } } email: allanwojtasp-at-em.agr.ca }
These are great stories and would make a wonderful collection, especially to hand out to students of EM. Yes, experience is a great way to learn but it's often best, and cheaper, to learn from some else's.
Here is a "just off the press" example (but this does come up at least once a year):
I fielded a phone call from a distraught SEM lab manager who told me that some months ago he put Dow Corning fluid into his diffusion pump "to save money". And now that he has had an accident, the silicone fluid of course, has contaminated his system, so he was asking us, "what organic solvent will easily remove it."
He was especially upset when I told him that his EDS detector will see Si everywhere also, because they do a lot of analyses for Si!
I won't repeat what he told me when I tried to explain the reality of his situation......
But it does go to show that there are a lot of new people entering our profession, some with less training and experience with vacuum than others, so such stories are very well worth repeating.
But just out of curiosity, is there some "recommended procedure" for removing silicone fluid from the internal parts of a column, and also removing it from the window of an EDS detector? I presume one can always call in an outside service provider with experienced people but a lot of users out there just don't have the budget for something like that. But they do have a good supply of student manpower.
While we are on the subject of silicone, a few weeks ago a well known TEM user got me on the phone to say "hello" and commented that he had just placed an order for silicone grease and yes, he said, he was going to be using it on the o rings of his column. I told him I thought that he should be using other greases and his response was "don't listen to dogma, I thought you read the listserver!". Am I correct, namely that one should not be using silicone grease on the o rings of a column instrument?
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.spi.cc ######################## ============================================
My story is not as much of a horror as it is embarassing. Several years back I received a package of samples from a regular client without any paper work describing what the samples were. This is not unusual since we frequently recieve unknowns. I proceeded to open the package and was abruptly assaulted with an extremely strong odor of bannanas. Apparantly one of the samples was a vial of concentrated bannana flavoring. It was months (almost a year) before the odor completely disappeared from my office, and I was cajoled about it frequently. I by the way, hate bannanas.............
Lou Solebello -----Original Message----- } From: L R MELSEN {lmelsen-at-emory.edu} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
These are great stories and would make a wonderful collection, especially to hand out to students of EM. Yes, experience is a great way to learn but it's often best, and cheaper, to learn from some else's.
Here is a "just off the press" example (but this does come up at least once a year):
I fielded a phone call from a distraught SEM lab manager who told me that some months ago he put Dow Corning fluid into his diffusion pump "to save money". And now that he has had an accident, the silicone fluid of course, has contaminated his system, so he was asking us, "what organic solvent will easily remove it."
He was especially upset when I told him that his EDS detector will see Si everywhere also, because they do a lot of analyses for Si!
I won't repeat what he told me when I tried to explain the reality of his situation......
But it does go to show that there are a lot of new people entering our profession, some with less training and experience with vacuum than others, so such stories are very well worth repeating.
But just out of curiosity, is there some "recommended procedure" for removing silicone fluid from the internal parts of a column, and also removing it from the window of an EDS detector? I presume one can always call in an outside service provider with experienced people but a lot of users out there just don't have the budget for something like that. But they do have a good supply of student manpower.
While we are on the subject of silicone, a few weeks ago a well known TEM user got me on the phone to say "hello" and commented that he had just placed an order for silicone grease and yes, he said, he was going to be using it on the o rings of his column. I told him I thought that he should be using other greases and his response was "don't listen to dogma, I thought you read the listserver!". Am I correct, namely that one should not be using silicone grease on the o rings of a column instrument?
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.spi.cc ######################## ============================================
My story is not as much of a horror as it is embarassing. Several years back I received a package of samples from a regular client without any paper work describing what the samples were. This is not unusual since we frequently recieve unknowns. I proceeded to open the package and was abruptly assaulted with an extremely strong odor of bannanas. Apparantly one of the samples was a vial of concentrated bannana flavoring. It was months (almost a year) before the odor completely disappeared from my office, and I was cajoled about it frequently. I by the way, hate bannanas.............
Lou Solebello -----Original Message----- } From: L R MELSEN {lmelsen-at-emory.edu} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
I am enjoying the "horror stories" (don't we all enjoy hearing about someone else's stupid mistake) and thought I would share one from the "software" category.
I had just joined Northern Scientific (later Tracor Northern, now Noran) and was writing EDS software for our first computerized analyzer (the venerable NS-880) -- the year was 1972, and my software, like any of the stuff being produced in those years where we wrote software on paper tape for machines with 4K memories, just wasn't all that "user friendly", so it was not unusual to have to walk people through difficult command sequences. Then too, computers were pretty new to everyone and people sometimes made silly-sounding mistakes out of pure inexperience (one highly intelligent customer, when instructed to 'type a space', dutifully typed "A SPACE" on the keyboard). One particular customer, however, defied all attempts at instruction and had passed well beyond "complaining" into the realm of "abusive" and "insulting". He insisted that he was following the instructions to the letter, but that our miserable software consistently malfunctioned. We were unable to duplicate the problem and finally several of us drove a considerable distance to his lab to get to the bottom of this. We went through the operations with him watching intently and the software performed flawlessly, which he acknowledged. We then asked him to operate the software himself and midway through, observed that he was responding to one of the dialog questions with a decidedly inappropriate response. We had him back out of this and repeat it with the correct response and the software then ran perfectly. To which his response was to draw himself up and state with great authority and indignation: "but when you run it THE WAY I DO, it doesn't work!"
I am enjoying the "horror stories" (don't we all enjoy hearing about someone else's stupid mistake) and thought I would share one from the "software" category.
I had just joined Northern Scientific (later Tracor Northern, now Noran) and was writing EDS software for our first computerized analyzer (the venerable NS-880) -- the year was 1972, and my software, like any of the stuff being produced in those years where we wrote software on paper tape for machines with 4K memories, just wasn't all that "user friendly", so it was not unusual to have to walk people through difficult command sequences. Then too, computers were pretty new to everyone and people sometimes made silly-sounding mistakes out of pure inexperience (one highly intelligent customer, when instructed to 'type a space', dutifully typed "A SPACE" on the keyboard). One particular customer, however, defied all attempts at instruction and had passed well beyond "complaining" into the realm of "abusive" and "insulting". He insisted that he was following the instructions to the letter, but that our miserable software consistently malfunctioned. We were unable to duplicate the problem and finally several of us drove a considerable distance to his lab to get to the bottom of this. We went through the operations with him watching intently and the software performed flawlessly, which he acknowledged. We then asked him to operate the software himself and midway through, observed that he was responding to one of the dialog questions with a decidedly inappropriate response. We had him back out of this and repeat it with the correct response and the software then ran perfectly. To which his response was to draw himself up and state with great authority and indignation: "but when you run it THE WAY I DO, it doesn't work!"
I have a similar story to this one! A bright fellow from a senior University (I would embarrass them) had a dim TEM image, so rigged up an Anglepoise lamp to try and throw some more light on it!
Keith Ryan Marine Biological Association Plymouth, UK
} } } Larry Allard {l2a-at-ornl.gov} 03/31/00 06:08pm } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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} -----------------------------------------------------------------------. } Humorous, but not quite horror:
The respected PhD I once worked with on our old JEOL JSM-U3 scanner,
who was having trouble making out the details of the image on the monitor, so grabbed a flashlight and shined it at the screen so he could see the image a little better... :-).
Larry
} } } Originally posted by: sryazant-at-ucla.edu } } } } } It is not "horror" story, but a sort of... Many years ago the colleague } from friendly Lab visited me with great project. The idea was simply and } beautiful. She argues that because the image in the scope (TEM) is green } (green fluorescence of the screen), she wants to modify the sample (I } forgot what, some protein, I believe) by red-fluorescent dye to be able to } see on the screen of the electron microscope the "double-staining": red on } the green background. No comments... } } Sergey } } } Date: Thu, 30 Mar 2000 09:17:56 -0500 } } From: "anderron-at-us.ibm.com"-at-sparc5.microscopy.com } } Subject: Operator Horror Stories } } To: microscopy-at-sparc5.microscopy.com } } Importance: Normal } } X-MIMETrack: Serialize by Router on D01ML065/01/M/IBM(Release 5.0.2b } (Intl)|18 } } January 2000) at 30/03/2000 09:18:01 } } } } ------------------------------------------------------------------------ } } 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
} } -----------------------------------------------------------------------. } } } } } } } } Alan Fox, in a response to the Analytical TEM string, cautioned the } } question-raiser to beware of people who abuse instruments. Perhaps Nestor } } will forgive a little microscopy related humor and allow us to start a } } string on "Operator Horror Stories." Here's ours: } } } } The "ham-fisted user" reference made us chuckle/cringe with the memory of } } a guest "microscopist" in our lab who hauled himself (220 pounds or so) out } } of his seat by pulling on the half-inserted specimen rod, bending it about } } 20 degrees or so! } } } } Henceforth, when we saw him in the hall (he never came into the scope room } } again), we referred to him as "Conan the Microscopist"! } } } } } } } } Ron Anderson, IBM, Hopewell Jct., New York, USA. anderron-at-us.ibm.com
} } } } IBM Analytical Services; http://www.chips.ibm.com/services/asg } } } } } } } } } } } _____________________________________ } } Sergey Ryazantsev Ph. D. } Electron Microscopy } UCLA School of Medicine } Department of Biological Chemistry } Box 951737 } Los Angeles, CA 90095-1737 } } Phone: (310) 825-1144 } Pager: (310) 845-0248 } FAX (departmental): (310) 206-5272 } mailto:sryazant-at-ucla.edu } http://www.bol.ucla.edu/~sryazant
Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
What in the world is an Anglepoise lamp? Is this the same as a "Gooseneck"?
Don Marshall
} From Microscopy-request-at-sparc5.microscopy.com Sun Apr 2 17:01:37 2000 } } -----------------------------------------------------------------------. } } } I have a similar story to this one! A bright fellow from a senior } University (I would embarrass them) had a dim TEM image, so rigged up } an Anglepoise lamp to try and throw some more light on it! } } Keith Ryan } Marine Biological Association } Plymouth, UK }
} -----------------------------------------------------------------------. } } } } ------------------------------------------------------------------------ } } 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 } } } -----------------------------------------------------------------------. } } Humorous, but not quite horror: } } } The respected PhD I once worked with on our old JEOL JSM-U3 scanner, } } who was having trouble making out the details of the image on the } monitor, so grabbed a flashlight and shined it at the screen so he } could see the image a little better... :-). } } Larry } } } } } } } Originally posted by: sryazant-at-ucla.edu } } } } } } } } } } It is not "horror" story, but a sort of... Many years ago the } colleague } } from friendly Lab visited me with great project. The idea was } simply and } } beautiful. She argues that because the image in the scope (TEM) is } green } } (green fluorescence of the screen), she wants to modify the sample } (I } } forgot what, some protein, I believe) by red-fluorescent dye to be } able to } } see on the screen of the electron microscope the "double-staining": } red on } } the green background. No comments... } } } } Sergey } } } } } Date: Thu, 30 Mar 2000 09:17:56 -0500 } } } From: "anderron-at-us.ibm.com"-at-sparc5.microscopy.com } } } Subject: Operator Horror Stories } } } To: microscopy-at-sparc5.microscopy.com } } } Importance: Normal } } } X-MIMETrack: Serialize by Router on D01ML065/01/M/IBM(Release } 5.0.2b } } (Intl)|18 } } } January 2000) at 30/03/2000 09:18:01 } } } } } } } ------------------------------------------------------------------------ } } } 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 } } } } } -----------------------------------------------------------------------. } } } } } } } } } } } } Alan Fox, in a response to the Analytical TEM string, cautioned } the } } } question-raiser to beware of people who abuse instruments. } Perhaps Nestor } } } will forgive a little microscopy related humor and allow us to } start a } } } string on "Operator Horror Stories." Here's ours: } } } } } } The "ham-fisted user" reference made us chuckle/cringe with the } memory of } } } a guest "microscopist" in our lab who hauled himself (220 pounds } or so) out } } } of his seat by pulling on the half-inserted specimen rod, bending } it about } } } 20 degrees or so! } } } } } } Henceforth, when we saw him in the hall (he never came into the } scope room } } } again), we referred to him as "Conan the Microscopist"! } } } } } } } } } } } } Ron Anderson, IBM, Hopewell Jct., New York, USA. } anderron-at-us.ibm.com } } } } } } } IBM Analytical Services; http://www.chips.ibm.com/services/asg } } } } } } } } } } } } } } } } } _____________________________________ } } } } Sergey Ryazantsev Ph. D. } } Electron Microscopy } } UCLA School of Medicine } } Department of Biological Chemistry } } Box 951737 } } Los Angeles, CA 90095-1737 } } } } Phone: (310) 825-1144 } } Pager: (310) 845-0248 } } FAX (departmental): (310) 206-5272 } } mailto:sryazant-at-ucla.edu } } http://www.bol.ucla.edu/~sryazant } } Dr. Lawrence F. Allard } Senior Research Staff Member } High Temperature Materials Laboratory } Oak Ridge National Laboratory } 1 Bethel Valley Road } Bldg. 4515, MS 6064 } PO Box 2008 } Oak Ridge, TN 37831-6064 } } 423-574-4981 } 423-574-4913 Fax } l2a-at-ornl.gov } }
Date sent: Sun, 2 Apr 2000 21:34:13 -0400 (EDT) } From: donald j marshall {dmrelion-at-world.std.com} To: KPR-at-wpo.nerc.ac.uk
I heard this one at a conference last week. A PhD student was caught sawing a knob off a FE SEM. Apparently it was pushing against his desk so decided to saw it off and was going to glue it on at a 45 degree angle!
} ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } My story is not as much of a horror as it is embarassing. Several years } back I received a package of samples from a regular client without any paper } work describing what the samples were. This is not unusual since we } frequently recieve unknowns. I proceeded to open the package and was } abruptly assaulted with an extremely strong odor of bannanas. Apparantly one } of the samples was a vial of concentrated bannana flavoring. It was months } (almost a year) before the odor completely disappeared from my office, and I } was cajoled about it frequently. I by the way, hate bannanas............. } } Lou Solebello } -----Original Message----- } } From: L R MELSEN {lmelsen-at-emory.edu} } To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com} } Date: Friday, March 31, 2000 9:26 PM } Subject: Storys } } } } ------------------------------------------------------------------------ } } 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 had a cousin of Conan , who in his addled age, after inserting the } } injector tip into the stage of our EM400 could not find his grid in the } } microscope; he had dropped it on the floor. Of course the tip must have } } fallen off in the stage, so he promptly took a second tip and properly } } inserted the second tip on top of the first. Still no grid could be } } found. Ah ha, I will leave a polite note explaining the problem. It read } } as follows: } } " Please check the microscope, I had great difficulty inserting my } } specimens last evening." } } Philips kindly replaced the bulk of the stage just so they could keep } } the original for the museum of what not to do. } } } } } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
It does't have oxygen in it--with air, 02- is produced and accelerated at your specimens, eroding them. When done intentionally, this is called "plasma ashing," i believe. If your specimens look very smooth, this may be why. Argon produces Ar2+, which, if your coater is set up right, is accelerated at the gold target. JSIII
} Hello, } } I would like to know why they use Argon in the metal coating devices. At } our laboratory, we don't use Argon, we use air. What is the advantage of } using Argon ? } De Pauw Bart } Faculty of Veterinary Medicine } Morphology } Salisburylaan 133 } 9820 Merelbeke } Belgium } Phone : 0032(0)9 264.77.19 } Fax : 0032(0)9 264.77.90
Julian P.S. Smith III Dept. of Biology Winthrop University Rock Hill, SC 29733 803-323-2111 x6427 (vox) 803-323-3448 (fax)
Has anyone foud a solution for the following problem, I am getting desperate. There doesn't seem to be a ready-made solution available ?
We want to link an Illumination Technologies light-source to a ZEISS microscope. We are trying to get the right information about the necessary parts, but this seems to be non-trivial.
The following problems need a solution:
ZEISS Axioskop upright microscope and Illumination Technologies CF1000 ZEISS Axiovert 100M inverted microscope and Illumination Technologies 3900
In both cases we need a light guide and a fiber coupler to the microscope. Both light sources are to be used for epifluorescence applications.
Sincerely yours,
Peter Van Osta, MD
Senior Scientist Medical Image Analysis Biological Imaging Laboratory Life Sciences Department I - 6065 Janssen Research Foundation Turnhoutseweg 30 B-2340 Beerse Belgium Europe
I heard this one from the Philip's engineers: A brand new Philips microscope was being installed in a goverment laboratory. It was to be in a state of the art, brand-new building. Everything was there, house nitrogen, chilled water, etc. Even the darkroom was state of the art. Well, when the in-house plumbers hooked up the water to the microscope they weren't being very careful in their reading of the blueprint designs and they had D-19 developer running through the EM instead of water. I'm not sure, but I think they got a new microscope out of that blunder!
Margaret E. Bisher
NEC Research Institute 4 Independence Way Princeton, NJ 08540. Tel.: (609) 951-2629 Fax: (609) 951-2496 e-mail: peggy-at-research.nj.nec.com
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An additional story which had the potential of being a real personnal horror story is as follows:
I worked for Humberto Fernandez-Moran at the University of Chicao many years ago. For those of you who are not familiar with the name, he was instrumental in developing the first diamond knives, producing the first pointed filaments for routine use and construction of first cryo-TEM using liquid helium cooled lenses. It was a very interesting place for a young budding microscopist at the time!
Dr. Moran had a large scar on his nose. He said it was from the removal of a cancerous skin area. He claimed to have gotten the malignancy in that location due to using electron microscopes in the late 40's without the benefit of lead glass windows. They used to press their noses against the window when concentrating on the relatively dim image projected by those early instruments. I wonder if other early EM researchers eventually developed cancer which might be related to similar research experiences.
As it turned out, Dr. Moran lived a long life, although not without controversy through the years. He had a very unusual life history and was also a brilliant but erratic person to interact with....somewhat akin to what Bobby Knight is to basketball!
Debby --------------------------------------
Many years ago we had a Siemens EM-101. This was a relatively new microscope and beautifully machined with German precision. The camera chamber was a work of art. Each film plate (we used glass back then) was encased in it's own light-tight cassette. The camera would move a film cassette into position for exposure and pull off the cassette cover. The operator would then expose the film and move the casette into a drop box. The cassette cover would be pushed closed in the process. This camera worked almost flawlessly. On occassion, an operator, when loading the camera, would put the cassettes in on an angle. When you went to take the first image, the cassette would jam and not move into position. All that was necessary to unjam was to break vacuum, giggle the cassette with your hand to straighten it and then repump the camera chamber and get back to work...a 10 minute process as most. We had one operator who thought she knew it all. She was working on the microscope one evening and the camera jammed. Her solution was to take out the camera and take it apart...screw by screw and spring by spring. She ended up with an incredible heap of parts and of course had no clue as to how to put it back together. Neither did the Siemens service engineers. They had never seen one apart! They had to find another camera (not easy in those days with relatively few of these instruments around), take it apart..carefully marking where each piece came from...and then reassembled each camera simultaneously. It took hours!! Moral of the story...NO ONE fixed ANYTHING without permission of the lab personnel!!
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
I have just been asked to do auger analysis on a sample mounted in phenolic mounting resin. Normally I require that these samples be demounted for UHV compatibility but this person doesn't wish to do so.
Does anyone have any experience with putting phenolic mounting material in a UHV environment? Will I be seeing carbon on all my samples for the next few years?
Thanks, Henk
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://web.ceof.ohio-state.edu Murphy's Law: "If anything can go wrong, it will." Commentary: "Murphy was an optimist."
There is a new system which sounds like it a good fit for your application. It is a liquid light guide with coupler, lamp (long lived HBO) and power supply from EFOS. Contact: Allan Firhoj PH: 905-812-4302 Email: AllanF-at-EFOS.com URL: www.efos.com
Caveat: MME has no financial interest in this product.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ******************************************************
At 03:50 PM 4/3/00 +0200, Van Osta, Peter [JanBe] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I love reading the "horror stories". Here's a personal one:
When I was a grad student I did a CPD run with a fellow grad student JT. I worked on plant pathogens and JT worked on chick embryo hearts so we had little pieces of leaf tissue and tiny chick hearts to dry. When the run was finished I couldn't get the lid off the CPD (it was a twist type). JT volunteered to muscle it off and when she did the lid shot passed her head with a boom and hit the ceiling. She had a grazing wound on her forehead but was otherwise ok. We both burst into fits of nervous laughter...we both knew she was so lucky not to be seriously injured. Then we looked in the CPD and saw that all the lids had blown off the little white sample containers. We howled with laughter when we got down on our hands and knees to search the floor for the tiny hearts and leaf pieces. The CPD was fine, the samples were fine, and surprisingly we both graduated.
} [Beth wrote:] } I love reading the "horror stories". Here's a personal one:
I do, also. Although I have not posted to this List often (last time was for advice in purchasing a confocal which was very helpful and we are very pleased with the purchase), I've wanted to chime in and say how enjoyable it is to read these 'stories'. I think there is a very relevant component to these stories in that they help us be more aware of how easy it is for some very unusual, silly and sometimes dangerous things to happen when you thought such a thing was impossible if it even crossed your mind at all.
Sorry, but thankfully, (very thankfully) I have no horror stories, yet... Having said that, uh oh...
Gerald Harrison ================ } [Beth continued] } When I was a grad student I did a CPD run with a fellow grad student JT. I } worked on plant pathogens and JT worked on chick embryo hearts so we had } little pieces of leaf tissue and tiny chick hearts to dry. When the run was } finished I couldn't get the lid off the CPD (it was a twist type). JT } volunteered to muscle it off and when she did the lid shot passed her head } with a boom and hit the ceiling. She had a grazing wound on her forehead } but was otherwise ok. We both burst into fits of nervous laughter...we both } knew she was so lucky not to be seriously injured. Then we looked in the } CPD and saw that all the lids had blown off the little white sample } containers. We howled with laughter when we got down on our hands and knees } to search the floor for the tiny hearts and leaf pieces. The CPD was fine, } the samples were fine, and surprisingly we both graduated. } } Beth }
I can't even begin to think of why you would have D-19 developer in a pressurized pipe system? Did they process 1000's of negatives a day?
At 10:22 AM -0400 4/3/00, Peggy Bisher wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
David R. Hull NASA Glenn Research Center at Lewis Field Advanced Metallics Branch Mail Stop 49-1 21000 Brookpark Road Cleveland, OH 44135
The Philip's guy's did not give me such a clear answer. I can only assume that something stupid happened, like D-19 was put in the closed loop system of the Haskris Chiller, instead of the normal water. I do know that this darkroom was to service a large EM suite so perhaps there was this big tank of D-19 made up and they (the plumbers) grabbed it and used it. It does sound incredibly stupid, that's for sure.
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Margaret E. Bisher
NEC Research Institute 4 Independence Way Princeton, NJ 08540. Tel.: (609) 951-2629 Fax: (609) 951-2496 e-mail: peggy-at-research.nj.nec.com
These stories are a giggle, and also food for thought!
I would like to collect them for a future Net Notes, part of the News and Commentary section of Microscopy and Microanalysis. So keep them coming! I may have to edit them down to a reasonable number, and it would be a few months before they appeared, but they are too good to pass up.
Aloha, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
} We had a CAP inspection, the officers were from another hospital in the state. } } } The question I was expected to reason and answer with a straight face was: } } } * What other protective measures besides standing behind a lead } wall do you take when you put the glass slide into the TEM, to } protect yourself from the flying electrons??? }
Dear Lou Ann, In addition to the obvious ludicrous aspects to the question, the use of a lead shield to protect oneself against "flying electrons" is one of the worst things to do. One should use a low-Z absorber for elec- trons to reduce brehmsstrahlung x-rays, then, if desired, put lead between oneself and the low-Z shield to absorb those x-rays which are produced. Yours, Bill Tivol
} I heard that one years ago. Also one that supposedly happened at Univ. } Wisconsin. Someone apparently cleaned a scope's internal parts in acetone } and started pumping. Before it was fully pumped down, they turned on the } filament. Bang! Nice story but could it possibly happen? I would think } that any acetone residue would evaporate very quickly or is there some } other phenomenon here relating to ratio of gases?
Dear Damian, There must have been a lot of residue in the scope to have caused
an explosion. Not only would one need both acetone and oxygen in a suitable ratio, there would have to be enough so that the heat of combustion from one acetone molecule oxidising would cause other molecules to react, otherwise there would just (!) be a slow burning of the vapors. In addition, the acetone/oxygen ratio would be determined by both the amount of residual acetone and the relative pumping speeds for acetone and oxygen. No doubt there are people on this list who would know the pump speed ratio, so one could calculate the expected behavior of the acetone in the column for various values of the relevant parameters. Yours, Bill Tivol
If no one has any objections I would also like to post these on our upcoming section on microscopy for our museum's web site when it is finished. Ed Sharpe archivist for SMECC
I remember somewhere in the past a discussion of software for creating extended focus images (I think that is the correct term), in which a series of transmitted light images in different focal planes are combined in order to remove the out of focus information and show all parts of the image in focus. Can someone point to software that carries out this function? Thanks- Dave --
************************************************************ "Home of the 2000 NCAA Women's Basketball Champions"
Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 75 N. Eagleville Rd. U-125 Storrs, CT 06269 Knecht-at-uconnvm.uconn.edu 860-486-2200 860-486-4331 (fax) home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html ************************************************************
I've just noticed behavior which is different for my 2 e-beam instruments. If I bias my microprobe's gun for less emission, the beam current measured in a specimen faraday cup also goes down. However, if I bias my SEM's gun for less emission, the beam current (measured similarly) goes up(???). This may mean simply a shallower optic angle for the SEM and the anode allowing more electrons to pass, but I thought I'd throw the observation out there.
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
Hi everyone! I'm new to this microscopy society and 'am having trouble already!!To make it worse everyone is sending me panic stories!! Just kidding! I'm working on some poly-propylene wires. I need to take some TEM images but I'm having trouble at the first stage itself...ultramicrotomy. You see I need to fuse two PP wires and image the interface. Now the problem is, when I'm cutting it with the microtome, the interface just breaks off!!Right now I'm using a epoxy-hardner ratio of 10:4. will changing the proportions help? I'm thinking of using a diamond knife also. Does anyone have any bright ideas?!!Help me out on this!! Praveena ______________________________________________________ Get Your Private, Free Email at http://www.hotmail.com
In a message dated 4/3/00 6:23:17 PM, knecht-at-uconnvm.uconn.edu writes:
} I remember somewhere in the past a discussion of software for } creating extended focus images (I think that is the correct term), in } which a series of transmitted light images in different focal planes } are combined in order to remove the out of focus information and show } all parts of the image in focus. Can someone point to software that } carries out this function? Thanks- Dave
That function is performed in The Image Processing Tool Kit (http://members.aol.com/ImagProcTK/) by examining each pixel location in the two (or more) images and calculating the local variance in a 5 pixel wide circle. The pixel value that has the largest variance is kept, the rationale being that it has the most abrupt local variations and is likely to be in the sharpest focus. The method works pretty well for most microscope images where the magnification remains constant, but not so well for macroscopic images where the out-of-focus portions of the image are also shifted due to changes in focal length. It also doesn't handle cases with specular reflections to well. But it should be OK for your transmitted light case. There are other approaches in some software packages that use a simple high pass filter (e.g. a Laplacian) but this seems from my experiments to be no faster and more noise sensitive.
Lighten up - the delete button is easy enough to use :-) I often feel the same way about all the bio postings and these are a lot more fun to read, besides having some educational value.
Jan Markham Motorola FPDD 7700 South River Parkway, FPD22 Tempe, AZ 85284 Ph: (480)755-5509 FAX: (480)755-5115 Email: afp042-at-email.mot.com
-----Original Message----- } From: Kriho, Virginia [mailto:Vkriho-at-psych.uic.edu] Sent: Monday, April 03, 2000 2:46 PM To: 'List Server'
I don't mean to be a drag, but enough of these "horror stories". My email is clogged with these stories. I only want the facts, Sir. Thanks.
Here's another one. A customer on the west coast had a bottle of N2 that was used to vent their TEM . They also used the same bottle to agitate their developer. One day someone left the N2 on in the developer and it ran out. You can guess what happened next, the TEM was vented with D-19.
Group, Just a note to advise that I intend to publish a summary of these "horror" stories in an upcoming issue of Microscopy Today. For those of you who do not receive our publication, including those overseas, I will provide a copy of the summary - at no charge. To the authors (past, current and future), should you not wish to have your comment included on our summary, kindly advise by return email and I will insure that it does not happen. Best to all, Don Grimes, Microscopy Today
Hi all - Ditto for the Australian EM Newsletter. (Somebody has volunteered to put together a choice selection.). and one from us... -the guy who asked how long to wash between dehydration steps
and from another place a long time ago- -the Professor who offered to pay the electron microscopist by giving him some oil immersion objectives for the TEM.
Dr Sally Stowe Facility Coordinator, ANU EMU Box 475 Canberra ACT 2601 AUSTRALIA stowe-at-rsbs.anu.edu.au fax 61 (0)2 6249 3218 or 6279 8525
} } } "Don Grimes" {microtoday-at-mindspring.com} 04/04/00 01:30pm } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Group, Just a note to advise that I intend to publish a summary of these "horror" stories in an upcoming issue of Microscopy Today. For those of you who do not receive our publication, including those overseas, I will provide a copy of the summary - at no charge. To the authors (past, current and future), should you not wish to have your comment included on our summary, kindly advise by return email and I will insure that it does not happen. Best to all, Don Grimes, Microscopy Today
I know that Soft Imaging's AnalySIS with EFI does this. It takes a bunch of images that are focused at various points and combines them into one image which is totally in focus.
gary g.
At 05:09 PM 4/3/00 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I remember somewhere in the past a discussion of software for creating extended focus images (I think that is the correct term), in which a series of transmitted light images in different focal planes are combined in order to remove the out of focus information and show all parts of the image in focus. Can someone point to software that carries out this function? Thanks- Dave } -- } } ************************************************************ } "Home of the 2000 NCAA Women's Basketball Champions" } } Dr. David Knecht } Department of Molecular and Cell Biology } University of Connecticut } 75 N. Eagleville Rd. U-125 } Storrs, CT 06269 } Knecht-at-uconnvm.uconn.edu } 860-486-2200 860-486-4331 (fax) } home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html } ************************************************************
I strongly believe that many of us are able to extract facts even from "horror stories". Fact, what is that? In my point of view, the "horror stories" are pure extract from people's experience showing to us how manage EM facilities properly. I was surprised to know, for instance, that somebody was able to bent sample-holder standing up from the chair. I will include special topic about that case in my instructions for users now. If you don't like the way people share their experience, you may set filter tn the word "horror" in the E.mail program and direct those messages into the trash-folder. A little bit humor, here at ListServer is not bad.
Best wishes, Sergey.
P.S. It is not bad tradition, again, at ListServer to sign the messages.
} Date: Mon, 03 Apr 2000 16:45:46 -0500 } From: "Kriho, Virginia" {Vkriho-at-psych.uic.edu} } Subject: no more horror stories } To: 'List Server' {microscopy-at-sparc5.microscopy.com} } X-Mailer: Internet Mail Service (5.5.2448.0) } } ------------------------------------------------------------------------ } 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 have been responsible for a multi-user facility since 1970.
It has always run on the basis that anyone can walk in and learn to use the equipment. So we never have fights about access or possession.
We now have around 300 users of which about 150 start fresh each year.
How is it our machines are not all wrecks?
The FIRST lesson teaches two golden rules:
1. NEVER use force on any control
2. ALWAYS ask for help as soon as you dont understand what is happening.
Our few bad incidents have occurred because these rules were neglected.
When a user who is in trouble calls me in to sort it out I try very hard to always be cheerful and positive no matter how stupid they have been. I think if I am cheerful they will call me in next time they have a problem. If I am furious with them they will maybe try to hide their blunder, or worse, try to fix it themselves.
Of course, I must apply the rule to myself. When I dont understand what is happening, it is time to call the service engineer!
Dr. Mel Dickson, Deputy Director, The Electron Microscope Unit, Adjunct Associate Professor, School of Microbiology & Immunology The University of New South Wales UNSW SYDNEY 2052 Australia. Phone +612 9385 6383 Fax +612 9385 6400
Well, after reading Peggy Bisher's story, I couldn't help but add another along similar lines.
About 20-odd years ago, a prestigous institution purchased a state of the art new TEM. The main body came in a very large wooden container and was unloaded onto the loading dock at the back of the building. It sat there for quite a while, because it was too heavy to move with the regular forklift, and I think the lab still needed a few final things to be finished off. Anyway, one day, someone decided that they were going to move the TEM in, and loaded it on the forklift. About half way to the lab, the TEM started oscillating back and forth on the forklift - it wasn't strapped on securely, and an eyewitness said he just stood there and watched this thing slowly crash to the floor on its side. Not much use rushing in and getting crushed by a few 100 kilos of metal.
I'm not sure it ever worked properly, the camera was smashed and a few other things too. The workshop had to retool all the smashed bits as best they could.
Amazing how many ways there are to destroy precision instruments.
cheers,
Rosemary White Microscopy Centre CSIRO Plant Industry GPO Box 1600 Canberra, ACT 2601 Australia
I agree with Mel Dickson whole heartedly. I learnt many years ago that to bawl out a user for being stupid results in silence. The only way to find out what really happens is to be as helpful as possible whatever the situation. Rant and rave to let off steam later, in the privacy of another room. To this end I welcome the horror stories. It is better to tell users stories of other incidents to get them to be careful and to think about the situation. They are more willing to ask a `stupid' question without embarrasment if they are aware of the mistakes that have been made. If this prevents them making furhter mistakes then I'm all for it.
Ron
=========================================================================== Mr. Ron Doole e-mail ron.doole-at-materials.ox.ac.uk Department of Materials, phone +44 (0) 1865 273701 University of Oxford, fax +44 (0) 1865 283333 Parks Road. Oxford. OX1 3PH. UK. ============================================================================
What Jan is saying is that `"bio" postings have no educational value ; )
Markham Jan-AFP042 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 } -----------------------------------------------------------------------. } } Lighten up - the delete button is easy enough to use :-) I often feel the } same way about all the bio postings and these are a lot more fun to read, } besides having some educational value. } } Jan Markham } Motorola FPDD } 7700 South River Parkway, FPD22 } Tempe, AZ 85284 } Ph: (480)755-5509 } FAX: (480)755-5115 } Email: afp042-at-email.mot.com } } -----Original Message----- } } From: Kriho, Virginia [mailto:Vkriho-at-psych.uic.edu] } Sent: Monday, April 03, 2000 2:46 PM } To: 'List Server' } Subject: no more horror stories } } ------------------------------------------------------------------------ } 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 don't mean to be a drag, but enough of these "horror stories". My email } is clogged with these stories. I only want the facts, Sir. Thanks.
-- C. John Runions, Ph. D. Department of Plant Sciences University of Cambridge Downing St. Cambridge UK
} } } I remember somewhere in the past a discussion of software for } creating extended focus images (I think that is the correct term), in } which a series of transmitted light images in different focal planes } are combined in order to remove the out of focus information and show } all parts of the image in focus. Can someone point to software that } carries out this function? Thanks- Dave } -- } } Greetings David,
If you contact them they will send you a demonstrtion CD of this product.
regards
Arnold
_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/ Arnold Richard Pizzey Department of Haematology Royal Free and University College London Medical School 98 Chenies Mews London WC1E 6HX U.K
There are a couple of options. Autoquant will do this, as will modules from Soft Imaging Systems. I think some Zeiss software can do this too, as long as there is no lateral shift in sequential images (as is found in some dissecting scopes). These are packages we've tried, there may well be more.
cheers, Rsoemary
Rosemary White Microscopy Centre CSIRO Plant Industry GPO Box 1600 Canberra, ACT 2601 Australia
Hi Dave, you may have a look on the SIS (Soft Imaging Software) homepage. They offer an EFI -modul.
http://www.soft-imaging.de/
In our metallographic labority, we use the EFI-module for one year and we are very satisfied in working with it.
Hope ths answer will help.
Best regards Bernd Schweisfurth Lufthansa Technik Hamburg / Germany } ---------- } Von: David Knecht[SMTP:knecht-at-uconnvm.uconn.edu] } Gesendet: Dienstag, 4. April 2000 00:09 } An: microscopy-at-sparc5.microscopy.com } Betreff: extended focus software } } ------------------------------------------------------------------------ } 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 remember somewhere in the past a discussion of software for } creating extended focus images (I think that is the correct term), in } which a series of transmitted light images in different focal planes } are combined in order to remove the out of focus information and show } all parts of the image in focus. Can someone point to software that } carries out this function? Thanks- Dave } -- } } ************************************************************ } "Home of the 2000 NCAA Women's Basketball Champions" } } Dr. David Knecht } Department of Molecular and Cell Biology } University of Connecticut } 75 N. Eagleville Rd. U-125 } Storrs, CT 06269 } Knecht-at-uconnvm.uconn.edu } 860-486-2200 860-486-4331 (fax) } home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html } ************************************************************ }
I am trying to localize a beta-1,3-glucanases gene expression after Russian wheat aphid infestation. However I'm getting heavy labeling in the chloroplasts. Unexpected because no previous localization studies have shown glucanases to be expressed in the chloroplasts. My question is: How can I conform that the labeling I am getting is not background or artifacts of some sort?? I am not using osmium only uranyl acetate and lead citrate for the staining
Thank you for any assistance Martin Wilding
Martin Wilding Department Botany & Genetics University of the Orange Free State P.O. Box 339 Bloemfontein 9300 South Africa
Tel +2751 4012818 Fax +2751 4488772 Email paam-at-rs.uovs.ac.za
David, I wrote a macro for Optimas that does this; however, you should be able to do this with just about any image processing software. You can see an article I wrote in Microscopy Today, February/March 1988 for specifics. I can email you a copy of the article if you want it.
Everett Ramer National Energy Technology Laboratory Pittsburgh, PA, USA 412-386-4920 ramer-at-netl.doe.gov
} } } David Knecht {knecht-at-uconnvm.uconn.edu} 04/03/00 06:09PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I remember somewhere in the past a discussion of software for creating extended focus images (I think that is the correct term), in which a series of transmitted light images in different focal planes are combined in order to remove the out of focus information and show all parts of the image in focus. Can someone point to software that carries out this function? Thanks- Dave --
************************************************************ "Home of the 2000 NCAA Women's Basketball Champions"
Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 75 N. Eagleville Rd. U-125 Storrs, CT 06269 Knecht-at-uconnvm.uconn.edu 860-486-2200 860-486-4331 (fax) home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html ************************************************************
Group, As suggested by several of you, and in addition to publishing an edited summary of the stories in Microscopy Today, I will put the summary on my web site where it can be downloaded by any with an interest. I will advise when it is done. And, should Nestor decide to discourage further contributions, send comments to me directly and I will see that they are included. Regards to all, Don Grimes, Microscopy Today
One of the problems with many interfaces like yours, Praveena, is that there is not a lot of adhesion across them. Thus mechanical sectioning of any form places a stress on the interface and decohesion can occur. (In many cases of sectioned thin films at this lab, we've ended up with two debonded layers adhering to the epoxy but not to each other after sectioning, thus I don't think playing with the hardener ratio will help much). A diamond knife makes a much smoother 'cut' and thus reduces these stresses, as will a lower knife angle (like 35 degrees). Sectioning parallel to an interface is less stressful than perpendicular to it. If you have an inherently weak interface, however, chances are pretty good that decohesion will occur. You may increase your probability of finding a portion that is still together by trimming to an oversized block face.
Good luck.
Tom Malis Group Leader - Characterization Materials Technology Laboratory Natural Resources Canada (Govt. of Canada) 568 Booth St., Ottawa, Canada ph. 613-992-2310 FAX 613-992-8735 email: malis-at-nrcan.gc.ca ---------- From: Praveena Bhaskara [SMTP:bubbyp-at-hotmail.com] Sent: April 03, 2000 7:10 PM To: Microscopy-at-sparc5.microscopy.com Subject: ultramicrotomy: PP wires
------------------------------------------------------------------------ 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! I'm new to this microscopy society and 'am having trouble already!!To make it worse everyone is sending me panic stories!! Just kidding! I'm working on some poly-propylene wires. I need to take some TEM images but I'm having trouble at the first stage itself...ultramicrotomy. You see I need to fuse two PP wires and image the interface. Now the problem is, when I'm cutting it with the microtome, the interface just breaks off!!Right now I'm using a epoxy-hardner ratio of 10:4. will changing the proportions help? I'm thinking of using a diamond knife also. Does anyone have any bright ideas?!!Help me out on this!! Praveena ______________________________________________________ Get Your Private, Free Email at http://www.hotmail.com
This is less a horror story than a sad tale. When I took over managing the EM facility here, I ordered a new cylinder of CO2 for the CPD. When it arrived, I pointed out to the delivery person that he made a mistake and had not supplied a tank with a siphon tube. He replied that he delivered what he had always delivered. I checked the shipping records, and, indeed, my predecessor had used C02 from a tank without a siphon tube--in other words, for a decade he never critically point dried a single specimen, since only C02 gas would have entered the chamber!
Perhaps more than a few people may want check their cylinders. (In the US, the proper cylinder typically has a red band painted at the top of the cylinder and the words "w/ dip tube" stenciled on the side.)
DL
______________________________________________________________________ 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
Hi all: Many years ago when comunists were in power in Eastern Europe, thanks to a great person who knew how to deal with them, we got a beautiful pice of equipment, with all the possible stages. One of them - the heating stage - was especially impressive. All the people were amazed that in situ TEM heating experiment might be performed up to 1000 centigrade. Among them was a young scientist who was investigating some processes in aluminum. Probably, impressed by 1000 centigrade he forgot about melting temperature. So, the new stage was required. At this time it was a real horror story since the price of the stage was almost equal the price of the small car - Fiat 126. Have a nice day, Witold Z.
%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%% Witold Zielinski, Ph.D. Warsaw University of Technology Department of Materials Science and Engineering 02-507 Warszawa, Woloska 141 POLAND
You didn't read my tale too carefully. I said the "in-house plumbers" did the error. The Philip's folks had nothing to do with this blunder. They just told me the story because it involved one of their microscopes. Maybe I should have just left out who told me the story. I was only trying to give them the credit and not take it myself. I am sure all of our EM service engineers have some horror stories that would top any we have told, but are trying to be polite for all of our sakes.
} } Peggy told one on the Philips folks: } } } ... new Philips microscope ... goverment laboratory. ... house } } nitrogen, chilled water, etc. Even the darkroom was state of the } } art. Well, when the in-house plumbers hooked up the water to the } } microscope they weren't being very careful in their reading of the } } blueprint designs and they had D-19 developer running through the } } EM instead of water. I'm not sure, but I think they got a new } } microscope out of that blunder! }
Margaret E. Bisher
NEC Research Institute 4 Independence Way Princeton, NJ 08540. Tel.: (609) 951-2629 Fax: (609) 951-2496 e-mail: peggy-at-research.nj.nec.com
We use the AnalySIS software from Soft Imaging Systems in our lab and it works great. It's very easy to use and the results are an image which is in focus from top to bottom.
_______________________________
Bill Carmichael Electron Microscopy Faculty
Madison Area Technical College 3550 Anderson St. Madison, WI 53704 608-243-4309
Hi everyone! I'm new to this microscopy society and 'am having trouble already!!To make it worse everyone is sending me panic stories!! Just kidding! I'm working on some poly-propylene wires. I need to take some TEM images but I'm having trouble at the first stage itself...ultramicrotomy. You see I need to fuse two PP wires and image the interface. Now the problem is, when I'm cutting it with the microtome, the interface just breaks off!!Right now I'm using a epoxy-hardner ratio of 10:4. will changing the proportions help? I'm thinking of using a diamond knife also. Does anyone have any bright ideas?!!Help me out on this!! Praveena
I am not sure I understand the "fusing" part of your question. However PP has a Tg of -19 so you need cryo-microtome, if you are not perhaps that is part of the problem. By the way, hopefully these "panic" stories will not go on much longer. They are not usual to the list. Steve Stephen McCartney Research Associate Virginia Tech Materials Institute 2108 Hahn Hall Blacksburg, VA 24061-0344 USA
To All, As a field engineer, I fully agree with both Mel and Ron and have always told my customers, "The only stupid question is the one that you don't ask."
I will also say that occassionally it can be very difficult to hold one's temper when someone has done something stupid on an instrument covered by one's own service contract and then they try to lie about it. Users, don't add insult to injury. Tell us what you did so that we can more rapidly repair the damage by looking in the right direction.
Field engineers have horror stories, to:
I became an expert on the ETEC Autospec WDS as an ETEC field engineer by being impatient in looking for vacuum leaks. The Autospec about doubles the volume of the system and therefore takes a lot longer to vent. I had put a 13-1/2 stopper in the port for the secondary detector to see if the SED was the source of the leak. When it was about half vented, I pulled the stopper. Without the WDS, this wouldn't have caused any problem, but I drove the columnator/electron trap into the 4 crystal turret, broke the tape drive and blew the thin window detector, along with destroying 2 crystals and loosening all 4. The process of fixing all this was a three week intensive course in WDS alignment and operation.
The moral: Don't rush. Take it easy and (God forbid) THINK before you act. It was only milliseconds to create 3 weeks of work and some $4k in damaged parts.
Moral #2: Learn from the mistakes of others, because you'll never live long enough to make them all yourself.
Ken Converse owner Quality Images Delta, PA 17314 717-456-5491
I have the original drawings for MAC instruments that ETEC dumped. I can't vouch for completeness, but there are 4 boxes of tubes with B size drawings and larger and at least a ream of A size drawings.
I need room. If you need these drawings, please contact me before May 1. After that date I will dump them. They may be had for the cost of shipping.
Ken Converse owner Quality Images Delta, PA 17314 717-456-5491
Are there any of you left out there? I am trying to decide what to do with all of the parts aquired from ETEC. If you are still using your Omniscan, please let me know as I don't want to leave people stranded, but I could really use the space if there is no need for these parts.
Ken Converse owner Quality Images Delta, PA 17314 717-456-5491
O.K. I guess I didn't word that as well as I should have :-) However, my point remains valid - not all the postings are of interest to everyone, no matter what the content, and these have been fun.
Jan Markham Motorola FPDD 7700 South River Parkway, FPD22 Tempe, AZ 85284 Ph: (480)755-5509 FAX: (480)755-5115 Email: afp042-at-email.mot.com
-----Original Message----- } From: John Runions [mailto:cjr41-at-cam.ac.uk] Sent: Tuesday, April 04, 2000 1:40 AM To: Markham Jan-AFP042 Cc: 'Kriho, Virginia'; 'List Server'
Garber, Charles A. 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: Garber, Charles A. * EMC.Ver #3.1 ] -- } } Here is a "just off the press" example (but this does come up at least once } a year): } } I fielded a phone call from a distraught SEM lab manager who told me that } some months ago he put Dow Corning fluid into his diffusion pump "to save } money". And now that he has had an accident, the silicone fluid of course, } has contaminated his system, so he was asking us, "what organic solvent will } easily remove it." } } He was especially upset when I told him that his EDS detector will see Si } everywhere also, because they do a lot of analyses for Si! } } I won't repeat what he told me when I tried to explain the reality of his } situation...... } } But it does go to show that there are a lot of new people entering our } profession, some with less training and experience with vacuum than others, } so such stories are very well worth repeating. } } But just out of curiosity, is there some "recommended procedure" for } removing silicone fluid from the internal parts of a column, and also } removing it from the window of an EDS detector? I presume one can always } call in an outside service provider with experienced people but a lot of } users out there just don't have the budget for something like that. But } they do have a good supply of student manpower. } } While we are on the subject of silicone, a few weeks ago a well known TEM } user got me on the phone to say "hello" and commented that he had just } placed an order for silicone grease and yes, he said, he was going to be } using it on the o rings of his column. I told him I thought that he should } be using other greases and his response was "don't listen to dogma, I } thought you read the listserver!". Am I correct, namely that one should not } be using silicone grease on the o rings of a column instrument? } } Chuck } } ============================================ } } Charles A. Garber, Ph. D. Ph: 1-610-436-5400 } President 1-800-2424-SPI } SPI SUPPLIES FAX: 1-610-436-5755 } PO BOX 656 e-mail:cgarber-at-2spi.com } West Chester, PA 19381-0656 USA } Cust.Service: spi2spi-at-2spi.com } } Look for us! } ######################## } WWW: http://www.spi.cc } ######################## } ============================================ Chuck, I agree that he shouldn't be using silicone grease on his o-rings. Braycote 803 and 602 are what I use, for static and dynamic seals, respectively.
My experience with DC705 (Transene Vacoil-S), which all ETECs were shipped with, is that if the system is properly trapped and the vacuum logic is well thought-out, it seems to work just fine. The only user that I'm aware of that had problems with stray Si readings was one who had a SIMS system attached. SIMS is apparently very sensitive to Si.
If you burp your DP, it is diffcult to clean, although ispropanol seems to work fairly well. I've seen many burped systems, but have never had any latent problems with silicates causing excessive charging. The biggest plus of DC705 is that you can take it to atmosphere hot and the oil is indestructible. It's performance figures are quite acceptable, and, lets face it, most people don't think twice about what they stick in their vacuum system. A bullet-proof oil is nice to have.
Perhaps it's more of a problem on systems that don't have a sealed, full-length liner tube. I'm certainly open to thoughts on why, in 23 years of servicing SEMs, I haven't seen this problem.
Ken Converse owner Quality Images Delta, PA 17314 717-456-5491
A colleague has recently requested analysis with a hot stage light microscope. As I do not have one, I am looking to retrofit a very old Reichert compound microscope with a hot stage. Is there anyone out there who might be able to help me?
At Martin Marietta Labs (1977), Harry O. and myself were responsible for training, use, and maintenance of an ISI Mini-SEM. One morning we found the SEM with a blown filament after late-night operation by user or users unknown. Further investigation revealed the remains of a house fly affixed with Aquadag to a specimen stub -- and dispersed throughout the microscope. The innards of the house fly in the microscope prevented the SEM from reaching operating vacuum. It took two days to adequately clean the column and the vacuum system.
Apparently the guilty party wanted to examine a housefly in the SEM, but did not think about it exploding in high vacuum. The fact the fly was not sputter coated suggested the likely guilty party. He later confessed and was denied further access without close supervision.
Steve Stokowski Stone Products Consultants 10 Clark St., Ste. A Ashland, Mass. 01721 508-881-6364 (ph. and fax) http://members.aol.com/crushstone/petro.htm
Oh oh here comes the fight between the "bios" and the "mats". Jan's comment didn't bother me, I'm a bio, but to each their own. I think most materials stuff is also uninteresting. (neat pictures but...)
I realize this thread is getting tired, but as this one happened to me just yesterday, perhaps you'll bear with me as I tell it.
For those of you that don't know the instrument, the XL30 ESEM, like most modern SEM's, displays its image as 256 intensity levels on a computer monitor. Presumably the colour could be set to whatever the user chooses, but we use the default black-and-white.
I had a young photorapher working for a prestigious magazine who needed an SEM photograph of a human hair. I had the microscope set up before he arrived, so there was an image on the screen as he walked in. After a few minutes, he asked "I thought you said this was one of your own hairs". "Yes", I replied. He looked puzzled, looking closely at my head, then said "Did you take one of the grey ones?"
Tony.
** Anthony J. Garratt-Reed ** MIT Room # 13-1027 ** 77 Massachusetts Avenue ** Cambridge, MA 02139-4307 ** USA ** ** Phone: (+) 1-617-253-4622 ** Fax: (+) 1-617-258-6479 **
clogged? I have a whole 13 since yesterday among another dozen of real junk.
I can't believe some of these things have happened! I have been trying to dismiss all the anti user ideas that the faculty had here. Don't let students use it, don't let researchers use it, don't teach them how -- they can't remember how to use it. I was once a student and learned how to tear down an old ISI SEM to clean after a filament change (with manual valving). So if taught and monitored I said, then they should enjoy the fun part of EM. It's worked so far. (cross my fingers and knock on wood)
I don't have any stories but heard at a meeting of a visiting researcher from the Far East that was pipeting Osmium by mouth. When corrected he shook his head yes but was found doing it again so they banned him from the lab.
Most stages have the holes necessary to accept any of the conventional hot stages.
By the way, do you know which model of Reichert do you have?
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ******************************************************
At 11:48 AM 4/4/00 -0400, Harrison, Gail wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
our software, the EFI module for analySIS, DOES take into account the lateral shift typical for the dissecting microscopes. It corrects for this shift first before attempting to reconstruct the final image.
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== 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: Rosemary White [mailto:Rosemary.White-at-pi.csiro.au] Sent: Tuesday, April 04, 2000 3:45 AM To: microscopy-at-sparc5.microscopy.com
Dear Dave,
There are a couple of options. Autoquant will do this, as will modules from Soft Imaging Systems. I think some Zeiss software can do this too, as long as there is no lateral shift in sequential images (as is found in some dissecting scopes). These are packages we've tried, there may well be more.
cheers, Rsoemary
Rosemary White Microscopy Centre CSIRO Plant Industry GPO Box 1600 Canberra, ACT 2601 Australia
My horror story is about 3 very competent service engineers, all from the same company, who each blew the window in the same EDX detector in their own turn. Engineer #1 was just unlucky, I think. He was installing a STEM unit and the window just popped. Engineer #2 had disassembled the TEM goniometer and for some unknown reason turned on the roughing pumps. Evidently the inrush of air increased pressure on the window enough to cause it to pop. Engineer #3 just didn't listen to me. He had insisted that my detector bellows was causing a very small leak. He had taken the detector off 3 times, sure he would finally demonstrate that without the detector the TEM would hold vacuum. Each time, though, the vacuum would drift and he'd go find and solve another leak. After the 3rd time, I told him to not mess with the detector anymore for fear he would break it. Two days later after I returned from giving an out-of-town lecture, the detector was off the scope and the window was indeed blown...and the vacuum leak was still not solved. I ended up paying for a percentage of the last repair because the engineer had gotten permission to remove the detector from my trainee technician (2 months experience). The bellows was proved to be tight and a new butterfly valve solved the vacuum leak.
The companies and engineers remain nameless.
Chuck Butterick Engineered Carbons, Inc. Borger, TX
Oh no..... I really didn't mean to start another bio vs materials war. I just meant to point out that we have a variety of interests represented here and not every item is going to be relevant to every reader at every time (as a pharmacist, I have done some pharmacognosy in the past and enjoyed it). If you don't like something, just use the delete button and let others make their own judgements.
Pax everyone!
Jan Markham Motorola FPDD 7700 South River Parkway, FPD22 Tempe, AZ 85284 Ph: (480)755-5509 FAX: (480)755-5115 Email: afp042-at-email.mot.com
-----Original Message----- } From: "rlvaughn-at-unmc.edu"-at-sparc5.microscopy.com [mailto:"rlvaughn-at-unmc.edu"-at-sparc5.microscopy.com] Sent: Tuesday, April 04, 2000 10:45 AM To: Microscopy-at-sparc5.microscopy.com
Oh oh here comes the fight between the "bios" and the "mats". Jan's comment didn't bother me, I'm a bio, but to each their own. I think most materials stuff is also uninteresting. (neat pictures but...)
Oh no..... I really didn't mean to start another bio vs materials war. I just meant to point out that we have a variety of interests represented here and not every item is going to be relevant to every reader at every time (as a pharmacist, I have done some pharmacognosy in the past and enjoyed it). If you don't like something, just use the delete button and let others make their own judgements.
Pax everyone!
Jan Markham Motorola FPDD 7700 South River Parkway, FPD22 Tempe, AZ 85284 Ph: (480)755-5509 FAX: (480)755-5115 Email: afp042-at-email.mot.com
-----Original Message----- } From: "rlvaughn-at-unmc.edu"-at-sparc5.microscopy.com [mailto:"rlvaughn-at-unmc.edu"-at-sparc5.microscopy.com] Sent: Tuesday, April 04, 2000 10:45 AM To: Microscopy-at-sparc5.microscopy.com
Oh oh here comes the fight between the "bios" and the "mats". Jan's comment didn't bother me, I'm a bio, but to each their own. I think most materials stuff is also uninteresting. (neat pictures but...)
Oh no..... I really didn't mean to start another bio vs materials war. I just meant to point out that we have a variety of interests represented here and not every item is going to be relevant to every reader at every time (as a pharmacist, I have done some pharmacognosy in the past and enjoyed it). If you don't like something, just use the delete button and let others make their own judgements :-)
Pax everyone!
Jan Markham Motorola FPDD 7700 South River Parkway, FPD22 Tempe, AZ 85284 Ph: (480)755-5509 FAX: (480)755-5115 Email: afp042-at-email.mot.com
-----Original Message----- } From: "rlvaughn-at-unmc.edu"-at-sparc5.microscopy.com [mailto:"rlvaughn-at-unmc.edu"-at-sparc5.microscopy.com] Sent: Tuesday, April 04, 2000 10:45 AM To: Microscopy-at-sparc5.microscopy.com
Oh oh here comes the fight between the "bios" and the "mats". Jan's comment didn't bother me, I'm a bio, but to each their own. I think most materials stuff is also uninteresting. (neat pictures but...)
This is to confirm my subscription to the listserver and to ask my first question.
I wish to purchase a PC based scanner with a transparency adaptor to scan in TEM negative plates for digital processing and output. I'd like to keep the cost of the hardware under $1000. What hardware is available out there? I have looked at specs for HP 6390C and UMAX Powerlook III. What minimum specs should I be looking for? Thanks.
You don't say what the diameter of the wires is. If possible, you might try reducing the size of the sample area that needs to be sectioned. I don't know if you are sectioning at room temperature or cryo. The Tg of PP is around -19C, so if you are not doing cryo you might want to give that a try. A diamond knife should work better. Also, depending on what it is you are trying to see, if you can live with slightly thicker sections then I would try that.
I have received a another message concerning CPD window material (in the earlier string a Polaron/ VG manager noted that their windows were not quartz, but some unspecified superior material.
Ted Pella has advised that they are now using -
"sapphire windows (2 of them, one under the other), which are in turn covered by the bullet proof shield. I think sapphire has about 50% greater strength compared to quartz (about 9,000 psi vs about 6,000) - that's why we use sapphire, for greater safety. We have never heard of any accident with our unit.
We added the bullet proof shield ("Lexan") to add a second safety measure for the viewing ports.
Three other safety measures are included: rupture disc, over-temperature switch and over-pressure switch."
I have no doubt that all manufacturers of CPD are most concerned and make these units safe; afterall one accident could cost a year's manufacturing cost. Its nice to know what technology is now in use, thank you Ted Pella. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
Regarding the anomalous variation in beam current with emission which you observe between your instruments:
This is a good demonstration of the fact that as far as emission is concerned, more isn't always better. I commonly compare the problem of getting the best performance from the gun to the idea of trying to squirt water through a knothole some distance away -- the critical parameter isn't the flow rate of the hose, but rather the ability to direct it into a focused stream. In reality, the vast majority of the emission never makes it into the column (compare the emission current to your maximum beam current) so the quality of the emission is more important than the quantity. The dynamic of the triode gun is that as you reduce the bias (thereby increasing the size of the emitting area of the filament and generating more emission) you also reduce the amount of "focusing" and thus the emission is less convergent and a smaller fraction passes through the anode. Whether there is a beam increase or decrease depends on the specifics of where the gun is operating. In theory, you should be able to observe this "reverse trend" ( beam current goes down as emission increases) by reducing the bias sufficiently far on any instrument -- though a particular instrument may not have the range of bias adjustment to permit this.
Hope this helps.
Fred Schamber RJ Lee Instruments Limited
=shAf= 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've just noticed behavior which is different for my 2 e-beam } instruments. If I bias my microprobe's gun for less emission, the } beam current measured in a specimen faraday cup also goes down. } However, if I bias my SEM's gun for less emission, the beam current } (measured similarly) goes up(???). This may mean simply a shallower } optic angle for the SEM and the anode allowing more electrons to pass, } but I thought I'd throw the observation out there. } } cheerios, shAf } } {} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} } Michael Shaffer, R.A. - ICQ 210524 } Geological Science's Electron Probe Facility - University of Oregon } mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
A former colleague spent approx. six hours ( over three separate days ) analysing metal wear particles on the SEM. He was doing the work with a French engineer who had prepared the samples. She told which particles to analyse and he dutifully analysed them. I happened to pass and glanced at the screen and asked why they were analysing paper fibres. They both insisted that I was wrong ( unfortunately they didn't take my bet ! ), until I suggested that they try using the BSE detector. Strangely they had lower contrast than the Al stub. The engineer is almost finished her PhD now and my colleague has moved on to become a forensic scientist. Just goes to show the customer is always right ?
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2, Rep. of Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 Email: creid-at-tcd.ie Home Page: http://www2.tcd.ie/Electron_Microscope/emu/home.htm
If You are interested please have a look at our High performance slow-scan CCD for TEM at http://www.proscan.de/pakete1.htm
Mark YEADON schrieb:
} ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } Mike, } } You could check out Soft Imaging Systems' MegaView II at: } } http://www.soft-imaging-web.de/ {http://www.soft-imaging-web.de/} } } I'd like to hear other recommendations too... } } Mark } } %%%%%%%%%%%%%%%%%% } Mark Yeadon } Senior Research Fellow } Institute of Materials Research and Engineering } 3 Research Link } Singapore 117602 } } Assistant Professor } Department of Materials Science } National University of Singapore } Singapore 119260 } } TEL: (+65) 874 8591 } FAX: (+65) 872 0785 } Email: m-yeadon-at-imre.org.sg {mailto:m-yeadon-at-imre.org.sg} } } -----Original Message----- } From: Michael Coviello [SMTP:coviello-at-mae.uta.edu] } Sent: Thursday, March 23, 2000 4:44 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: TEM-Digital camera recommendations } } } ------------------------------------------------------------------------ } 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 Ya'll: } We are looking for a CCD camera for our Philips 430 TEM. We would } like } to get the best camera for the best price, e.g., either buying a } used } camera or a new non-Gatan camera (Gatan seems to be twice as } expensive } as the others). We would be using the camera for bright field and } high } resolution TEM of materials (semiconductors) rather than for } biological } specimens. Does anyone have a camera they would like to sell/donate } or } does anyone have recommendations as to a less-expensive camera that } they } know can be used for materials applications. } Thanks, Mike Coviello } Lab Manager } University of Texas -at- Arlington }
-- Best regards / Mit freundlichen Gruessen Dr. Frank Jenichen Proscan elektronische Systeme GmbH Tel.: +49 8195 999 -511 Fax: -512 mailto:Jenichen-at-proscan.de ------------------------------------------------------------ More information concerning our products and services can be found on our website http://www.proscan.de
I am having some difficulties obtaining a good, clean polish on gold-copper wires (SRM 482). I have embedded the wires in a thermoset resin, backed the wires with epoxy and polished with successively smaller SiC grit. I have tried a variety of final polishes including .25 um diamond, vibratory polishers, etc. I can achieve an excellent metallographic finish with regard to smooth surfaces, but I cannot remove some small spots from all of the wires. These are iron red in polarized light and do not have the morphology of the polishing compounds. A qualitative analysis by EDS does not show any elements other than copper and gold. Does anyone have experience polishing these alloys? Is there something I'm missing?
Thanks to all Robert Carlton Aventis Pharmaceuticals robert.carlton-at-rp-rorer.com
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What a great basis for an SEM exam question. I'm going to tuck this one away until needed! Debby
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
--------------------------------------
shAF,
Regarding the anomalous variation in beam current with emission which you observe between your instruments:
This is a good demonstration of the fact that as far as emission is concerned, more isn't always better. I commonly compare the problem of getting the best performance from the gun to the idea of trying to squirt water through a knothole some distance away -- the critical parameter isn't the flow rate of the hose, but rather the ability to direct it into a focused stream. In reality, the vast majority of the emission never makes it into the column (compare the emission current to your maximum beam current) so the quality of the emission is more important than the quantity. The dynamic of the triode gun is that as you reduce the bias (thereby increasing the size of the emitting area of the filament and generating more emission) you also reduce the amount of "focusing" and thus the emission is less convergent and a smaller fraction passes through the anode. Whether there is a beam increase or decrease depends on the specifics of where the gun is operating. In theory, you should be able to observe this "reverse trend" ( beam current goes down as emission increases) by reducing the bias sufficiently far on any instrument -- though a particular instrument may not have the range of bias adjustment to permit this.
Hope this helps.
Fred Schamber RJ Lee Instruments Limited
=shAf= 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've just noticed behavior which is different for my 2 e-beam } instruments. If I bias my microprobe's gun for less emission, the } beam current measured in a specimen faraday cup also goes down. } However, if I bias my SEM's gun for less emission, the beam current } (measured similarly) goes up(???). This may mean simply a shallower } optic angle for the SEM and the anode allowing more electrons to pass, } but I thought I'd throw the observation out there. } } cheerios, shAf } } {} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} } Michael Shaffer, R.A. - ICQ 210524 } Geological Science's Electron Probe Facility - University of Oregon } mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
Position available at the Smithsonian Institution:
The National Museum of Natural History in Washington, DC is seeking an experienced electron microscopist to fill a vacancy for SEM laboratory operation and management. The SEM facility is designed to serve both the biological and geological research communities in the museum, and houses two recent model SEMs and one state-of-the-art environmental microscope (to be installed in mid-2000). The principal responsibilities include training staff members and visiting scientists in proper use of equipment and theory of electron generation and detection, maintenance and troubleshooting all instrumentation (in conjunction with full service contracts), evaluation of new developments in SEM technology, and supervision of a support staff member. The successful applicant will also have the opportunity to gain experience in Focused Ion Beam (FIB) Microscopy and high spatial resolution secondary ion mass spectrometry HR SIMS.
This position will fill a federal government vacancy and is offered at the GS11 ($42,724) or GS12 ($51,204) levels, and carries promotion potential to grade GS 13. U.S. citizenship is required for this federal position. To obtain information concerning this vacancy call our automated Jobline at (202) 287-3102 (24 hours, 7 days a week), press 9 and request vacancy announcement 00MQ-2069. Announcements will be available beginning April 18th and applications must be received by May 16th, 2000. If questions arise after receiving and reading through the vacancy announcement please contact: Dr. Edward Vicenzi (Chair, SEM Lab Manager Search Committee) at vicenzi-at-volcano.si.edu. The Smithsonian Institution is an equal opportunity employer.
PLEASE NOTE: this position is open from April 18th to May 16th, 2000.
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Edward P. Vicenzi Smithsonian Institution Department of Mineral Sciences Washington, DC 20560-0119
A couple decades + ago while provding TEM services for the NIH neurology group, the Neurosurgeons were impatiently awaiting the Biopsy results of their surgical patient. The frozen section was not conclusive, and so they needed a TEM result which would confirm their findings. Not being satisfied with my answer that the results will take a couple days, or one day at the very earliest, the surgeon sent his first resident up to my lab and he began rummaging through my supply cabinets so he can prepare the sample quickly himself. This is before microwave embedding and fixation. He told me that he was instructed (by my boss) to cut a very thin slice of tissue (use a very sharp razor), coat it with a lot of glutaraldehyde (straight out of the bottle) and stick it into the scope (a Joel 100 at that time). He would be waiting in the OR with the patient until he got the result. He declared that he did not need any training. ... I was speechless and could not believe this attempt...I watched the fate with amusement. I Volutarily moved on to safer grounds thereafter.
Thomas Baginski
Thomas A Baginski, Room G-230 Technical Coordinator for Microscopy Uniformed Services University of the Health Sciences 4301 Jones Bridge Road Bethesda, MD 20814-4799
What is the current folklore on avoiding folds/creases in semi-thin resin sections? I am cutting a worm, approx. 1 mm in diamter, for light microscopy and photography. It is typical Annelid, i.e. external cuticle, muscular body wall with inner coelomic space containing another hollow tubular structure (the gut). It is embedded in araldite for normal TEM. I have tried drying sections onto glass slides at 35, 60, 70 and 110 degrees. They all wrinkle prior to Toluidine Blue staining on hotplate. I have also stained sections by flotation overnight prior to drying on slides. Help!
Keith Ryan Marine Biological Association Plymouth UK
Dear All, We've been fortunate here that we've not had too many horror stories (at least funny enough to print here) even though we've had many users of greatly differing abilities. Of the mild catastrophes we've had, all have been invaluable for teaching purposes. They teach new users what can go wrong and how you can become inadvertently famous by not thinking. They've also taught me how to write instructions to prevent accidents, and how creative people can be in coming up with new ways to run the microscope. I routinely tell stories of past users (without revealing identities) to novices to lighten the long hours of instruction. I've also gotten into the habit of attaching the price tag (figuratively) to fixing different components of the scope. There are the famous $18 plastic knobs, } $300 hexrings, and the $50k if you hit your head on the specimen holder while it's in the scope (actually happened!). This "scared straight" tactic is always tempered with humor and encouragement so not to paralyze the faint of heart. And most of all, we don't heap blame on users for errors. We encourage honesty, lest we get mysterious cases that take much longer to decipher (such as the case of the missing holder tip). Ciao for now, Ken
} } Regarding the anomalous variation in beam current with } emission which you observe between your instruments: } } ... } ... The dynamic of the triode gun is that as you reduce } the bias (thereby increasing the size of the emitting area of the } filament and generating more emission) you also reduce the } amount of "focusing" and thus the emission is less convergent } and a smaller fraction passes through the anode. } Whether there is a beam increase or decrease depends } on the specifics of where the gun is operating. } In theory, you should be able to observe } this "reverse trend" ( beam current goes down as emission } increases) by reducing the bias sufficiently far on any } instrument -- though a particular instrument may not have } the range of bias adjustment to permit this. } ...
Are you implying any "optimization" at the 1st cross-over by exploring the relationship of emission versus the number of electrons which get past the anode?? (... as if, the point at which decreasing the emission also decreases the beam current implies less than optimized (?) ...). What are your thoughts for this relationship regarding the "brightest crossover" versus "extended life of the electron emitter"??
I'm interested in localizing fluorescein-tagged gene probes for microscopy (in situ hybridization, light and EM) and blotting applications. Does anyone have any recommendations for the best (strongest labeling) anti-fluorescein antibody, or any experience comparing different clones, polyclonal vs. monoclonal, or different suppliers?
For those TEM operators who have not seen the effect of changing bias on the electron beam there is a neat experiment.
1. At "gun saturation" bring the condenser to crossover and magnify this to fill the screen.
2. Change the bias or emission control (not the filament heating)
Either a) The gun de saturates, if so turn the bias control in the other direction to increase the bias field Or b) The intensity increases slightly (the subject of current mail) Or c) The intensity decreases
If you see (b) the filament is in an position within the cathode that allows the full optimisation of the emission system; in my experience few people run under these conditions.
If you see (c) the filament is not in the optimised position within the cathode.
For SEM operators
1. Set up in the wave form or graph mode
2. Watch the trace as you increase the bias field (emission reduces)
Either a) With the filament position optimised for efficiency the trace should rise slightly Or b) With a filament positioned away from this point the trace will fall.
For those who do not understand the bias or emission control relationships-
A BIAS control will reduce the emission current when turned clockwise, higher bias.
An EMISSION control will reduce the emission current when turned anticlockwise, higher bias, or of you like turn up an emission control and the reduction in the bias field allows an increase in emission current.
They are acting on exactly the same area of the high voltage circuit but are simply wired in a different way.
Have fun
Steve Chapman Senior Consultant Protrain - for professional training in EM world wide e-mail protrain-at-emcourses.com web site www.emcourses.com Tel 44+ 1280 814774 Fax 44+ 1280814007
Hello folks, I was traveling and really wanted to zip in three short "horror stories" before the thread got yanked or burned. [1] Eons ago (Actually maybe 25 years) I was a service engineer with Philips and went to perform routine maintenance on a very early EM-300. I always cleaned the windows and in this instance, I found the smaller, left, projection window to be Plexiglas! No one would tell me how long the ersatz window had been in place but I really made my point when I demonstrated, with a Geiger Counter, that lots of x-rays were getting sprayed into the room. They quickly got as new window. I also had a similar experience when a customer stuck the Aluminum shipping plate in place of a broken window. They also bought a nice new leaded glass window. See, the window stories are true. [2] Some of the older microscopes used Mercury Diffusion Pumps to increase pumping speed. They were usually operated in tandem with the Oil Diffusion Pump. In one of the not terribly uncommon disasters where a blast of Mercury blasts up the column and turns all the beautifully Gold plated brass pieces into a amalgam covered mess, a service engineer who was particularly resourceful found out that Iodine readily combines with Mercury. He sprinkled Iodine crystals all over the contaminated parts of the column. Not long later, there was an explosion! No one was physically hurt, and I heard the microscope company cleaned up the mess. [3] Our laboratory was below the level of a creek so when there were sump pump failures, water would seep onto the floor, fairly quickly. A lovely and dedicated graduate student was working late a night when one of these pump failures occurred and she ended up with her feet under water while running our old JEOL 35C Scanner. I found her and said she would have to stop, as I yanked the wall switch. She was angry with me for interrupting her research until I was finally able to convince her she was very close to electrocuting herself.
Best regards,
Alex Greene SCIENTIFIC INSTRUMENTATION SERVICES, INC. PMB-499, 1807 West Slaughter Lane, Number 200 Austin, Texas 78748-6200 Phone: 512/282-5507 FAX 512/280-0702
QUALITY ELECTRON MICROSCOPE REPAIRS -----Original Message----- } From: "rlvaughn-at-unmc.edu"-at-sparc5.microscopy.com {"rlvaughn-at-unmc.edu"-at-sparc5.microscopy.com} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
The problem with using silicone compounds in electron microscopes is that if they get on parts where they are struck by the electron beam they break down to produce siliceous compounds which are non-conductive and so collect a static electric charge and deflect the electron beam in undesirable ways. These decomposition compounds are also very insoluble, and become difficult to remove. I recall our RCA EML microscope, which came charged with silicone DP oil, developed such deposits on the objective and condenser apertures. Fortunately, these apertures were made of platinum, and so we could clean them by heating them in a Bunsen burner to convert the deposits to silicon oxides and then treating them with concentrated hydrofluoric acid to dissolve these oxides - something that would be frowned on in most laboratories these days.
Because of the potential for generating this kind of a problem, I can see no reason whatsoever for using either a silicone grease or a silicone oil in a modern electron microscope. In fact, when I was managing electron microscope laboratories I refused to allow any of these materials in laboratories out of fear that some inexperienced person would use them on one of the instruments.
The reason for using a vacuum grease on an O-ring (or gasket) is to provide enough lubrication so that the O-ring will slide enough to fill the O-ring groove uniformly, without forming bumps or creases that can cause a leak. The grease should not be required to produce the basic vacuum seal - the groove should be smooth enough so that it could seal properly without the grease if the O-ring fitted into position properly. Thus, only enough grease should be applied to give a barely visible sheen to the O-ring - gobs and globs are not needed. While the silicone high vacuum grease is indeed a good lubricant for O-rings, it is no better than the Brayco and Krytox greases, which are based on polyphenylether compounds, and which do not introduce the possibility of having insoluble siliceous compounds formed on critical parts of the electron optical column. The function, use, and characteristics of vacuum greases are discussed in more detail in Chapter 10 of the book "Vacuum Methods in Electron Microscopy"
I also would not use a silicone fluid in a diffusion pump on an electron microscope, nor other equipment used in preparing electron microscopy specimens, for the same reason described above. Several diffusion pump fluids are now available that are nearly as stable to thermal and oxidative degradation as the silicone fluids, and which have vapor pressure characteristics that are comparably favorable. These include the Santovac-5 and Excello-54 polyphenyl ether fluids and the Lion-S and Alcatel-220 eicosyl naphthalene fluids, all of which will produce levels of vacuum as good as the DC 705 silicone fluid. These fluids are discussed in more detail in Section 5.4 of Vac. Meth. in EM.
The problem of removing silicone compounds from parts that have become contaminated with them is a very difficult one, Because these compounds are usually very viscous, and are not readily soluble. The Dow Corning Company, which manufactures them, recommends repeated wiping with cloth pads moistened with toluene, xylene, trichloroethylene, or perchloroethylene.
I have recently had fair success cleaning diffusion pump fluids off metal parts by first wiping the parts with dry paper towels to remove the bulk of the fluids, then spraying the surfaces with Tilex Soap Scum Remover and scrubbing them with a cloth pad, then rinsing them with hot running water. By repeating this process several times I have been able to get acceptable results in several instances. (I remove the water by rinsing with isopropyl alcohol, and then dry with a gas blaster.) It might be difficult to adapt this process to cleaning internal parts of an electron microscope, however. Other cleaning procedures are described on pp. 69 - 74 of Vac. Meth. in EM.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
One possible work-around is to cut your sections thinner than usual for light microscopy, down to 0.4 micro-meters, which then are more likely to dry wrinkle- free onto your slides. These thin sections then require an extreme stain to provide sufficient contrast. I refer you to two papers by del Cerro et al. on such a method, utilizing Stevenel's Blue as the stain. The articles are in Microscopa Acta Vol. 83, pp.117-121 and 217-220 (1980).
Hope this helps, Mike Nesson
Keith Ryan wrote:
} } What is the current folklore on avoiding folds/creases in semi-thin } resin sections? I am cutting a worm, approx. 1 mm in diamter, for } light microscopy and photography. It is typical Annelid, i.e. } external cuticle, muscular body wall with inner coelomic space } containing another hollow tubular structure (the gut). It is embedded } in araldite for normal TEM. I have tried drying sections onto glass } slides at 35, 60, 70 and 110 degrees. They all wrinkle prior to } Toluidine Blue staining on hotplate. I have also stained sections by } flotation overnight prior to drying on slides. Help! } } Keith Ryan } Marine Biological Association } Plymouth UK
-- _______________________________________________________________________ Michael Nesson, Ph.D. Department of Biochemistry & Biophysics 2011 Ag&LS, Oregon State University, Corvallis, OR 97331-7305 (541)737-5245 FAX:(541)737-0481 nessonm-at-ucs.orst.edu
I forgot about this recent one. We had a technician here who I taught how to develop film for our JEOL 2000FX. It had been awhile since he replaced the film, so after he left, I wanted to make sure that the film was loaded into the cassettes with the emulsion side up. When I opened the film cassette box, I couldn't take out the cassettes. He had put them in 180 degrees around and the slot in the film holders didn't match up with the alignment bar going up along the side of the box. The sides of the film box were actually bulging. I had to pry each cassette out of the box with a screwdriver. I left most of them for him to do the next day. Before showing him this, I innocently asked him if he had trouble putting the holders into the box. He said that the last couple were a little hard to put in. The good thing: he had loaded the cassettes with the emulsion side up. -Scott
Does anyone have a liquid nitrogen dewar in the 20 to 40 liter range with low evaporative losses that they would like to sell? Please contact me off-list.
A friend had just finished cleaning his TEM column parts with acetone, as he had been instructed. Being an impatient young man, he quickly reassembled everything and, as he was lowering the column back into place, noticed a few drops of acetone had fallen into the viewing chamber and onto the phosphorous screen. I guess he had the chamber open for cleaning as well, because he said he quickly grabbed the canned air and aimed it into the chamber to blow off the drops. I walked in just then to see a cloud of yellow dust settling all over the walls and floor of the EM lab... He cleaned out the viewing chamber as best he could, I suppose, but there was dust in the column and pumping system for months to come. Then there were the phosphorescent footprints and fingerprints that appeared all over the lab for weeks! Now every time I open my viewing chamber I hold my breath.
Aloha, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
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Many years ago, our lab had one "good" electron microscope-a JEOL 200. Every few years the massive high voltage cable would short out internally and require replacement. I watched the service technician unscrew the lock ring from the high voltage tank, which was about as large as a refrigerator to power many vacuum tubes. He used a chair to get on top of the unit and pull the cable out and hand the end to me. I heard a loud snapping sound and saw a frightened look on the fellow's face. In a hurry to get the job done, he had forgotten to use an insulated grounding rod kept in the area to bleed off the charge in the high voltage capacitors. Although he had a small burn on his hand, he and I finished the cable swap with a new one. A few years later, the instrument had a water hose leak which flooded all the control cable plugs at the rear of the console. The technician had to "rebuild" many of the plugs over a couple of days time. It ran well for a year or two and was shut down to make way for a newer instrument. It's always good to discuss major service procedures with someone to ensure that oversights don't occur unnecessarily. That person later rose to the higher eschelon's of his company.
Bernie Kestel Material's Science Division Argonne National Laboratory 9700 So. Cass Avenue Argonne, IL., 60439
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Back during the asbestos craze I was the last "little indian" doing TEM analysis, the other two left for bigger and better paying jobs elswhere. My business manangers (who knew the EM was big and needed electricity, but that was the extent of their knowledge) suggested cross-training some in-house employees to assist me with my work. The first guy walked into the scope room chewing gum (not like a normal human being, more like a horse). I started showing him the scope (JEOL 100sx) while he sat in the chair infront of the scope. The phone rang so I turned to answer it. Meanwhile that cute little handle on the camera door caught the new guy's attention. "what's this?" I heard, and then that all too familiar hiss followed by valves closing and pumps and power switches clicking off. He just reached out and turned the handle, venting the column. The scope handled the shock better than I did. I sent the guy to lunch, and locked the door behind him. Jon Ekman Associate Research Specialist University of Wisconsin Milwaukee 414-229-6471
A client of ours has a clever new way to present 3D images but needs some beautiful stereo pairs to generate a set of tests. These images need to be true stereo pairs, not anaglyphs. If you have anything you would like to share, please contact me directly. Also, there is a possibility that these materials may be used in future publications... of course, with credit, so please let me know if you would like the images used (a) just for the tests or (b) for both tests and publication, with your citation.
I look forward to hearing from you all!
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ******************************************************
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{blockquote TYPE=CITE} {pre} =shAf= wrote: Are you implying any "optimization" at the 1st cross-over by exploring the relationship of emission versus the number of electrons which get past the anode?? (... as if, the point at which decreasing the emission also decreases the beam current implies less than optimized (?) ...). What are your thoughts for this relationship regarding the "brightest crossover" versus "extended life of the electron emitter"?? {/pre} {/blockquote}
{p} {br} Yes, the whole mechanism relies on optimization of the electron trajectories so that they converge to a dense "crossover" in the region of the anode opening. Raw emission increases as bias is reduced but the amount of focusing is also reduced so that there is an optimum point where the maximum current is focused into the "virtual aperture" (entrance pupil) of solid angle where it will end up hitting the specimen. {p} Understanding the dynamics of the gun is really made much more complicated by the self-biasing (auto-bias) circuit which nearly every thermionic microscope uses (field emitters are, of course, a whole different story). If one had a direct-bias unit you could freely run the bias from low to high and clearly see the effects. At a very low bias, you get a flood of emission (usually limited only by the rating of the HV power unit) and at a sufficiently high bias you can cut the emission off entirely. However, because almost no one has a direct-bias setup, you can't adjust the bias directly -- you adjust the bias resistor -- which kind of does the same thing, but with one big difference -- you are now adjusting the operating point of the emission-stabilization circuit. A practical consequence is that you simply can't achieve cut-off -- since the actual bias is the voltage drop across the bias resistor due to the flow of emission current, it is clearly impossible to cut off the emission current completely (zero current would imply zero bias). Thus, the bias resistor acts like it is adjusting the bias directly (except in the opposite sense of the control) for low resistor values -- but at the other end of the range it behaves quite differently from a true bias control since it approaches "cutoff" asymptotically rather than abruptly. {p} Now look at the other part of the story -- the role of filament temperature. If one had a unit with an independent fixed bias, increasing the temperature would make the emission rise without limit (no "saturation" knee). But the autobias circuit means that at some point, as one increases the filament temperature, the increasing bias (due to the increased voltage drop across the bias resistor) starts trying to "cut off" the emission and you reach a stable operating point which microscopists like to call "saturation" which is, in fact, nothing more than the operating point of the autobias circuit. {p} So we have two effects we are trying to match up: (a) the need to optimize the projection of the crossover into the column's entrance pupil; and (b) an autobias circuit which will limit emission once a particular emitter temperature is reached. This is the purpose of adjusting the bias resistor. By setting it properly, you can make the optimal convergence condition occur at a desireable "saturation" temperature. This condition of optimal convergence is measured by a quantity we call "brightness", which is the product of the spatial and angular densities of the beam at the crossover (high brightness means the crossover has a small diameter and the beam doesn't diverge much -- just what you want to get the maximum beam down the column). {p} As to the life of the filament versus maximum brightness: There is a well-known equation due to Langmuir which gives the maximum brightness which can be achieved for a given filament temperature (for a given beam voltage and material work function). All other things being equal, the higher the temperature, the greater the attainable brightness. At the same time, the higher the temperature, the greater the rate of filament evaporation and the shorter its life. It should be possible on any microscope to set it up to approach the Langmuir limit over a range of operating temperatures (assuming appropriate wehnelt spacing and orifice size). So you have to make a choice, do you want high brightness or long filament life? You can't have both. It's too bad that our microscopes don't read filament temperature directly because then everyone could easily see what the REAL variable regulating filament life is. When you adjust the bias resistor, you adjust the emission stabilization operating point, and since microscopists are taught to "saturate" the filament, you are thus establishing the filament's operating temperature and thus its life. {p} Hope this somewhat long-winded answer addresses your question! {p} Fred {br} {blockquote TYPE=CITE} Fred writes ... {p} } {br} } Regarding the anomalous variation in beam current with {br} } emission which you observe between your instruments: {br} } {br} } ... {br} } ... The dynamic of the triode gun is that as you reduce {br} } the bias (thereby increasing the size of the emitting area of the {br} } filament and generating more emission) you also reduce the {br} } amount of "focusing" and thus the emission is less convergent {br} } and a smaller fraction passes through the anode. {br} } Whether there is a beam increase or decrease depends {br} } on the specifics of where the gun is operating. {br} } In theory, you should be able to observe {br} } this "reverse trend" ( beam current goes down as emission {br} } increases) by reducing the bias sufficiently far on any {br} } instrument -- though a particular instrument may not have {br} } the range of bias adjustment to permit this. {br} } ... {p} =shAf= :o) {/blockquote} {/html}
Dear Keith, My experience has been with parasitic helminths. Those of us who work to any degree of regularity on whole organisms find the varying consistency of tegument, musculature, and internal organs troublesome. Frequently the tegument expands differently than the circular, longitudinal, and tranverse musculature.
I have found the best solution to this situation is to adjust the hardness of the embedding media to match the most dense material in a given specimen. In addition to resin hardness, adequate infiltration is a must.
In previous list server responses to wrinkled sections, the techniques of infiltration and resin hardness, as well as the use of various solvents and heat to expand sections have been suggested.
Personally, I prefer using LR White methyl methacrylate, medium grade, or a harder version of the epon replacements with araldite. The difference in the appearance of the ultrastructure between these two resins will be obvious, but not necessary objectionable. I find LR White seems to 'relax' more than that of the epoxy resins, is easier to stain with multiple stains and requires less staining time with u.a. and pb.cit. However, LR White is the bane of many microscopists and therefore is not always the resin of choice.
This whole problem is interesting in that so many variables determine section quality, e.g., how do you pick up the sections, how clean are your microscope slides, how clean is your boat, how clean are the instruments you use to transfer sections to the microscope slide, how thick are your sections, on and on it goes!
My suggestion would be to make certain your slides, your glass knifes and boats, and the utensils you use to remove the sections are CLEAN! Next, try waiving a cotton swab dripped in cholorform very close to your sections and see if your sections expand enough to appear smooth. I prefer to do this step with the sections already on a droplet on a clean microscope slide. I watch the sections under the stereoscope to look for the smooth appearance. In some case, I actually use a heat wand in addition to the cholorform. If you do not have a heat wand, get a disposable heat pen (I believe most EM suppliers carry these). You can either replace the AA batteries when they go dead or, hook the heat wand to a variable DC transformer. Adjust the voltage to approximately 3 volts or until the 'filament' is slightly dull red. I usually rub a finger on my nose, GENTLY touch the filament and then turn on the voltage until the filament produces a wisp of smoke (strange technique eh?!)
One last comment: I usually turn my hot plate to almost 250 - 300 degree F. Watch the sections so no bubbles appear and try rocking the slide back and forth. I would also suggest you very the duration of staining time on the hot plate. Anything more than a minute and you will surely end up with some portion of the section lifting from the slide and consequently, get wrinkles.
Enough rambling... No horror stories with this one!
Cheers! -Ken ----------- Ken Tiekotter, Adjunct Professor The University of Portland Department of Biology 5000 N. Willamette Blvd. Portland, OR 97303
On Wed, 5 Apr 2000, Keith Ryan 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 is the current folklore on avoiding folds/creases in semi-thin } resin sections? I am cutting a worm, approx. 1 mm in diamter, for } light microscopy and photography. It is typical Annelid, i.e. } external cuticle, muscular body wall with inner coelomic space } containing another hollow tubular structure (the gut). It is embedded } in araldite for normal TEM. I have tried drying sections onto glass } slides at 35, 60, 70 and 110 degrees. They all wrinkle prior to } Toluidine Blue staining on hotplate. I have also stained sections by } flotation overnight prior to drying on slides. Help! } } Keith Ryan } Marine Biological Association } Plymouth UK } } }
Most mention using a heat pen or solvent vapour - I don't normally do this in the belief that the hotplate does the job anyway. It is set somewhere between 100-120 degrees.
My belief is that the problem is the specimen being a set of concentric tubes of slightly varying consistency?
Cleanliness has also been mentioned! I am normally perfect - of course (my wife told me so!).
The most interesting "wrinkle" (I had to get that pun in somewhere) is to vary the time on the hotplate. I will play with this today. I have been drying for a few minutes, with sections covered by a dish to prevent any effect from draughts (I go back a bit - cutting sections since 1969 - you'd think I have got it right by now!).
I was thinking also of removing the resin to maybe lessen the wrinkles' effect (sodium methoxide - made with saturated NaOH in methanol).
Keith
_______________________ Keith Ryan (Dr) Marine Biological Association Citadel Hill Plymouth Devon PL1 2PB England
To whom it may concern: I am a physics undergraduate and I am currently trying to work on an assignment in Microscopy. I am trying to find material on the theory and application of electron beam and x-ray methods of analysis with particular emphasis on SEM, TEM, EPMA, EDX, WDX, AES and XPS. I was given this website site as a possible source of information. Any assistance would be greatly appreciated.
Kindest Regards Chris MacWilliam. (ph95ccm-at-brunel.ac.uk)
{!doctype html public "-//w3c//dtd html 4.0 transitional//en"} {html}
{blockquote TYPE=CITE} {pre} =shAf= wrote: Are you implying any "optimization" at the 1st cross-over by exploring the relationship of emission versus the number of electrons which get past the anode?? (... as if, the point at which decreasing the emission also decreases the beam current implies less than optimized (?) ...). What are your thoughts for this relationship regarding the "brightest crossover" versus "extended life of the electron emitter"?? {/pre} {/blockquote}
{p} {br} Yes, the whole mechanism relies on optimization of the electron trajectories so that they converge to a dense "crossover" in the region of the anode opening. Raw emission increases as bias is reduced but the amount of focusing is also reduced so that there is an optimum point where the maximum current is focused into the "virtual aperture" (entrance pupil) of solid angle where it will end up hitting the specimen. {p} Understanding the dynamics of the gun is really made much more complicated by the self-biasing (auto-bias) circuit which nearly every thermionic microscope uses (field emitters are, of course, a whole different story). If one had a direct-bias unit you could freely run the bias from low to high and clearly see the effects. At a very low bias, you get a flood of emission (usually limited only by the rating of the HV power unit) and at a sufficiently high bias you can cut the emission off entirely. However, because almost no one has a direct-bias setup, you can't adjust the bias directly -- you adjust the bias resistor -- which kind of does the same thing, but with one big difference -- you are now adjusting the operating point of the emission-stabilization circuit. A practical consequence is that you simply can't achieve cut-off -- since the actual bias is the voltage drop across the bias resistor due to the flow of emission current, it is clearly impossible to cut off the emission current completely (zero current would imply zero bias). Thus, the bias resistor acts like it is adjusting the bias directly (except in the opposite sense of the control) for low resistor values -- but at the other end of the range it behaves quite differently from a true bias control since it approaches "cutoff" asymptotically rather than abruptly. {p} Now look at the other part of the story -- the role of filament temperature. If one had a unit with an independent fixed bias, increasing the temperature would make the emission rise without limit (no "saturation" knee). But the autobias circuit means that at some point, as one increases the filament temperature, the increasing bias (due to the increased voltage drop across the bias resistor) starts trying to "cut off" the emission and you reach a stable operating point which microscopists like to call "saturation" which is, in fact, nothing more than the operating point of the autobias circuit. {p} So we have two effects we are trying to match up: (a) the need to optimize the projection of the crossover into the column's entrance pupil; and (b) an autobias circuit which will limit emission once a particular emitter temperature is reached. This is the purpose of adjusting the bias resistor. By setting it properly, you can make the optimal convergence condition occur at a desireable "saturation" temperature. This condition of optimal convergence is measured by a quantity we call "brightness", which is the product of the spatial and angular densities of the beam at the crossover (high brightness means the crossover has a small diameter and the beam doesn't diverge much -- just what you want to get the maximum beam down the column). {p} As to the life of the filament versus maximum brightness: There is a well-known equation due to Langmuir which gives the maximum brightness which can be achieved for a given filament temperature (for a given beam voltage and material work function). All other things being equal, the higher the temperature, the greater the attainable brightness. At the same time, the higher the temperature, the greater the rate of filament evaporation and the shorter its life. It should be possible on any microscope to set it up to approach the Langmuir limit over a range of operating temperatures (assuming appropriate wehnelt spacing and orifice size). So you have to make a choice, do you want high brightness or long filament life? You can't have both. It's too bad that our microscopes don't read filament temperature directly because then everyone could easily see what the REAL variable regulating filament life is. When you adjust the bias resistor, you adjust the emission stabilization operating point, and since microscopists are taught to "saturate" the filament, you are thus establishing the filament's operating temperature and thus its life. {p} Hope this somewhat long-winded answer addresses your question! {p} Fred {br} {blockquote TYPE=CITE} Fred writes ... {p} } {br} } Regarding the anomalous variation in beam current with {br} } emission which you observe between your instruments: {br} } {br} } ... {br} } ... The dynamic of the triode gun is that as you reduce {br} } the bias (thereby increasing the size of the emitting area of the {br} } filament and generating more emission) you also reduce the {br} } amount of "focusing" and thus the emission is less convergent {br} } and a smaller fraction passes through the anode. {br} } Whether there is a beam increase or decrease depends {br} } on the specifics of where the gun is operating. {br} } In theory, you should be able to observe {br} } this "reverse trend" ( beam current goes down as emission {br} } increases) by reducing the bias sufficiently far on any {br} } instrument -- though a particular instrument may not have {br} } the range of bias adjustment to permit this. {br} } ... {p} =shAf= :o) {/blockquote} {/html}
Wil Bigelow 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 } -----------------------------------------------------------------------. } } The problem with using silicone compounds in electron microscopes is that } if they get on parts where they are struck by the electron beam they break } down to produce siliceous compounds which are non-conductive and so collect } a static electric charge and deflect the electron beam in undesirable ways. } These decomposition compounds are also very insoluble, and become difficult } to remove. I recall our RCA EML microscope, which came charged with } silicone DP oil, developed such deposits on the objective and condenser } apertures. Fortunately, these apertures were made of platinum, and so we } could clean them by heating them in a Bunsen burner to convert the deposits } to silicon oxides and then treating them with concentrated hydrofluoric } acid to dissolve these oxides - something that would be frowned on in most } laboratories these days. } } Because of the potential for generating this kind of a problem, I can see } no reason whatsoever for using either a silicone grease or a silicone oil } in a modern electron microscope. In fact, when I was managing electron } microscope laboratories I refused to allow any of these materials in } laboratories out of fear that some inexperienced person would use them on } one of the instruments. } } The reason for using a vacuum grease on an O-ring (or gasket) is to provide } enough lubrication so that the O-ring will slide enough to fill the O-ring } groove uniformly, without forming bumps or creases that can cause a leak. } The grease should not be required to produce the basic vacuum seal - the } groove should be smooth enough so that it could seal properly without the } grease if the O-ring fitted into position properly. Thus, only enough } grease should be applied to give a barely visible sheen to the O-ring - } gobs and globs are not needed. While the silicone high vacuum grease is } indeed a good lubricant for O-rings, it is no better than the Brayco and } Krytox greases, which are based on polyphenylether compounds, and which do } not introduce the possibility of having insoluble siliceous compounds } formed on critical parts of the electron optical column. The function, } use, and characteristics of vacuum greases are discussed in more detail in } Chapter 10 of the book "Vacuum Methods in Electron Microscopy" } } I also would not use a silicone fluid in a diffusion pump on an electron } microscope, nor other equipment used in preparing electron microscopy } specimens, for the same reason described above. Several diffusion pump } fluids are now available that are nearly as stable to thermal and oxidative } degradation as the silicone fluids, and which have vapor pressure } characteristics that are comparably favorable. These include the } Santovac-5 and Excello-54 polyphenyl ether fluids and the Lion-S and } Alcatel-220 eicosyl naphthalene fluids, all of which will produce levels of } vacuum as good as the DC 705 silicone fluid. These fluids are discussed in } more detail in Section 5.4 of Vac. Meth. in EM. } } The problem of removing silicone compounds from parts that have become } contaminated with them is a very difficult one, Because these compounds are } usually very viscous, and are not readily soluble. The Dow Corning } Company, which manufactures them, recommends repeated wiping with cloth } pads moistened with toluene, xylene, trichloroethylene, or } perchloroethylene. } } I have recently had fair success cleaning diffusion pump fluids off metal } parts by first wiping the parts with dry paper towels to remove the bulk of } the fluids, then spraying the surfaces with Tilex Soap Scum Remover and } scrubbing them with a cloth pad, then rinsing them with hot running water. } By repeating this process several times I have been able to get acceptable } results in several instances. (I remove the water by rinsing with } isopropyl alcohol, and then dry with a gas blaster.) It might be difficult } to adapt this process to cleaning internal parts of an electron microscope, } however. Other cleaning procedures are described on pp. 69 - 74 of Vac. } Meth. in EM. } } Wilbur C. Bigelow, Prof. Emeritus } Materials Sci. & Engr., University of Michigan } Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; } Fx:734-763-4788; Ph:734-662-5237
Wil, I have apparently always had a good handle on the problems of silicones and e-beams. However, it still begs the question of why I haven't seen these problems in 23 years of servicing ETECs. I've seen charging from actual droplets present in a contaminated system, but never after cleaning, and I KNOW I've never gotten a contaminated system really stripped of silicones. Perhaps it's because one characteristic of ETECs that customers have told me about is that they continue to image very well when incredibly contaminated compared to other systems. Just a thought.
Ken Converse owner Quality Images Delta, PA 17314 717-456-5491
The Electron Microcopy Analysis Center (EMAC) at West Chester University in Pennsylvania has an opening for a full-time electron microscopy technician in a multiple user facility. The successful applicant will have a minimum of a BachelorÕs degree and 3 years experience working with electron microscopes. Responsibilities will include daily operation and maintenance of the EMAC including training users on the instrumentation, specimen preparation and darkroom techniques. The successful applicant must demonstrate the necessary organizational, management and communication skills to efficiently operate the EMAC. Applicants must submit a cover letter describing their experience with different instrumentation, a resume and contact information for three references. Although not a requirement, a sample of electron micrographs showing the applicant's work would be useful for the selection committee. All materials are to be sent to the Department of Human Resources, 210 Carter Drive, West Chester University, West Chester, PA, 19383. Applications must be received by June 1 2000. Applicants must successfully complete the interview process to be considered a finalist. AA/EOE. Women and minorities are encouraged to apply.
The Electron Microcopy Analysis Center (EMAC) at West Chester University in Pennsylvania has an opening for a full-time electron microscopy technician in a multiple user facility. The successful applicant will have a minimum of a BachelorÕs degree and 3 years experience working with electron microscopes. Responsibilities will include daily operation and maintenance of the EMAC including training users on the instrumentation, specimen preparation and darkroom techniques. The successful applicant must demonstrate the necessary organizational, management and communication skills to efficiently operate the EMAC. Applicants must submit a cover letter describing their experience with different instrumentation, a resume and contact information for three references. Although not a requirement, a sample of electron micrographs showing the applicant's work would be useful for the selection committee. All materials are to be sent to the Department of Human Resources, 210 Carter Drive, West Chester University, West Chester, PA, 19383. Applications must be received by June 1 2000. Applicants must successfully complete the interview process to be considered a finalist. AA/EOE. Women and minorities are encouraged to apply.
I have a number of stereo pairs on my web site, some better than others...
http://woody.white.home.att.net
They are not intended for commercial use for profit, but may be used for test purposes. ...They were collected on my own time, but using the company's equipment.
Woody White McDermott Technology Inc. ----------------------------------------------- Hi,
A client of ours has a clever new way to present 3D images but needs some beautiful stereo pairs to generate a set of tests. These images need to Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
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Robert Carleton: Have ypu considered the possibility that the red spots are particles of Cu2O? This compound will show up as red partcles in polarized light.
Sam Purdy National Steel Technical Center Trenton, MI V 734-676-2682 F 734-676-2030 spurdy-at-nationalsteel.com
} ---------- } From: "Robert.Carlton-at-aventis.com"-at-sparc5.microscopy.com } Sent: 5, April 2000, 8:57 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Polishing SRM 482 Gold-Copper Wires } } ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } To all, } } I am having some difficulties obtaining a good, clean polish on } gold-copper } wires (SRM 482). I have embedded the wires in a thermoset resin, backed } the } wires with epoxy and polished with successively smaller SiC grit. I have } tried a variety of final polishes including .25 um diamond, vibratory } polishers, etc. I can achieve an excellent metallographic finish with } regard } to smooth surfaces, but I cannot remove some small spots from all of the } wires. These are iron red in polarized light and do not have the } morphology } of the polishing compounds. A qualitative analysis by EDS does not show } any } elements other than copper and gold. Does anyone have experience } polishing } these alloys? Is there something I'm missing? } } Thanks to all } Robert Carlton } Aventis Pharmaceuticals } robert.carlton-at-rp-rorer.com } } }
On Wed, 5 Apr 2000 17:47:17 -0500 "jekman-at-uwm.edu"-at-sparc5.microscopy.com wrote:
..... } Meanwhile that cute little handle on the camera door caught the } new guy's attention. "what's this?" I heard, and then that all too } familiar hiss followed by valves closing and pumps and power } switches clicking off. He just reached out and turned the handle, } venting the column.
I'd be willing to bet that almost everyone with a JEOL TEM can tell a similar story. I do wish they would do something about that handle...it sits there inviting itself to be turned. I've inadvertanly vented the scope myself several times by hitting it with the back of the chair.
WL Steffens, Ph.D Dept. of Pathology University of Georgia
Great thread. I think many an old timers could fill a book with those stories by themselves. Why though nothing autobiographical? There is no shame attached if the horror was a learning experience. We all know that education is expensive.
At only 23 years of age and only 18 months of intermittent "user experience" I found my self in charge of a small EM Lab with a Phillips 100C as the center piece. Interesting TEM, with the column in the near horizontal position and a transmission viewing screen.
There was some minor leak problem with the specimen holder. That TEM design prepumped around the holder on partial insertion, while a spring loaded ball sealed that space from the column. The holder was then pushed in completely and that pushed that ball aside. Simple and effective.
To fix the ball seal I had to open the column and so I also cleaned various bits and pieces and re-assembled. Pumped and realigned, I then pulled the specimen holder out, whereupon the TEM inhaled very deeply. I later found that that spring loaded ball assembly would fit 180 degrees reversed quite well, but in that position the ball would not roll back to cover the seal when the specimen holder was withdrawn.
The main damage was a clear center on that lovely transmission viewing screen, the coating had been vacuumed. It cost $300 then, which is more like $2000 today and I had another opportunity to hone my maintenance skills, cleaning column, oil and Hg pumps.
It was a very lonely job, with so little experience running that lab without any other assistance, but I never learned so much as during those 18 months in that lab. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Thursday, April 06, 2000 8:15 AM, Tina Carvalho [SMTP:tina-at-pbrc.hawaii.edu] wrote: } } Here's another oops. } } A friend had just finished cleaning his TEM column parts with acetone, as } he had been instructed. Being an impatient young man, he quickly } reassembled everything and, as he was lowering the column back into place, } noticed a few drops of acetone had fallen into the viewing chamber and } onto the phosphorous screen. I guess he had the chamber open for cleaning } as well, because he said he quickly grabbed the canned air and aimed it } into the chamber to blow off the drops. I walked in just then to see a } cloud of yellow dust settling all over the walls and floor of the EM } lab... He cleaned out the viewing chamber as best he could, I suppose, } but there was dust in the column and pumping system for months to } come. Then there were the phosphorescent footprints and fingerprints that } appeared all over the lab for weeks! Now every time I open my viewing } chamber I hold my breath. } } Aloha, } Tina } } **************************************************************************** } * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * } * Biological Electron Microscope Facility * (808) 956-6251 * } * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* } **************************************************************************** }
Tina (Weatherby) Carvalho wrote: ================================================= Here's another oops.
A friend had just finished cleaning his TEM column parts with acetone, as he had been instructed. Being an impatient young man, he quickly reassembled everything and, as he was lowering the column back into place, noticed a few drops of acetone had fallen into the viewing chamber and onto the phosphorous screen. I guess he had the chamber open for cleaning as well, because he said he quickly grabbed the canned air and aimed it into the chamber to blow off the drops. I walked in just then to see a cloud of yellow dust settling all over the walls and floor of the EM lab... He cleaned out the viewing chamber as best he could, I suppose, but there was dust in the column and pumping system for months to come. Then there were the phosphorescent footprints and fingerprints that appeared all over the lab for weeks! Now every time I open my viewing chamber I hold my breath. =============================================================== This might be a lot more serious, healthwise than just another "oops".
Until relatively recently, virtually all TEM viewing screens were coated with a Cd-containing phosphor. In recent years there has been an attempt to use non-Cd containing substitute materials (because of their much lower toxicity levels) but none really seem to be quite as good as the original Cd containing phosphors. That is why most of the bulk manufacturers of the Cd- containing phosphors stopped their production (because of liability concerns ) and that is also why you don't see Cd-containing phosphors readily available for sale any more.
But some of the people who do screen re-coating services, stockpiled some of the original Cd-containing phosphor because sometimes that is the level of performance demanded by the customer.
If the phosphor described by Tina was Cd-containing, I really don't think you want this to be around as an inhalation hazard. I don't know how common this kind of "oops" has been, but in the event it should happen in your lab, I would exercise great care to not inhale the particulates and to undertake a careful clean-up. And while we are on this topic, if you are doing your own screen coating and using a "lode" of phosphor purchased some years ago, the chances are very great that it contains Cd, and all precautions should be taken to avoid inhalation or other exposures when working with the powder
University of Pittsburgh Department of Materials Science and Engineering
Postdoctoral Research Position in Transmission Electron Microscopy
A POSTDOCTORAL POSITION has become available in the area of transmission electron microscopy of thin film data storage media as part of a collaborative research program between Professors J.M. Wiezorek and W.A. Soffa, Department of Materials Science and Engineering of the University of Pittsburgh, and the Seagate Corporation. Candidates should hold a Ph.D. in Materials Science, Physics or a related discipline and must have extensive hands-on experience in a broad range of imaging, diffraction and analytical electron microscopy characterization techniques, as well as with sample preparation methods. Demonstrated expertise in the preparation of thin film samples suitable for high-resolution TEM characterization in plan-view and cross-section is highly desirable. A basic knowledge of magnetism in materials and experience in instrument development and/or computer image processing/simulation would be beneficial. The appointment is for one year in the first instance and is available after June 01/00. Screening of applications will begin immediately and will continue until the post is filled. Applications from under-represented groups, including minorities, women and people with disabilities are encouraged.
Interested candidates should send a curriculum vitae, including publication list, and the names of at least three referees with postal addresses, telephone numbers and E-mail addresses to: Prof. J.M. Wiezorek, Department of Materials Science and Engineering, University of Pittsburgh, 848 Benedum Hall, Pittsburgh, PA 15261, USA. Email: wiezorek+-at-pitt.edu
________________________________ Jorg M. Wiezorek, Ph.D. Assistant Professor Director - Electron Microscopy Materials Science & Engineering University of Pittsburgh tel.: 412-624 0122 848 Benedum Hall fax.: 412-624 8069 Pittsburgh, PA 15261, USA
The MegaView II is a high-resolution side-mounted TEM camera. It provides large field of view and high frame rates as well as high-resolution images. For high-resolution TEM images, as in lattice images, a bottom-mount camera is more suitable. To that end we have the BioCam. Although the name seems to imply biological applications, the name is more a "historical" one and does not reflect the current applications for the camera.
Nestor, if this sounds like an advertisement, I apologize. Out MegaView II camera was mentioned in connection with high-resolution images in materials science, for which it may or may not apply, depending on the actual requirements.
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
Mark YEADON 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 } -----------------------------------------------------------------------. } } Mike, } } You could check out Soft Imaging Systems' MegaView II at: } } http://www.soft-imaging-web.de/ {http://www.soft-imaging-web.de/} } } I'd like to hear other recommendations too... } } Mark } } %%%%%%%%%%%%%%%%%% } Mark Yeadon } Senior Research Fellow } Institute of Materials Research and Engineering } 3 Research Link } Singapore 117602 } } Assistant Professor } Department of Materials Science } National University of Singapore } Singapore 119260 } } TEL: (+65) 874 8591 } FAX: (+65) 872 0785 } Email: m-yeadon-at-imre.org.sg {mailto:m-yeadon-at-imre.org.sg} } } -----Original Message----- } From: Michael Coviello [SMTP:coviello-at-mae.uta.edu] } Sent: Thursday, March 23, 2000 4:44 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: TEM-Digital camera recommendations } } } ------------------------------------------------------------------------ } 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 Ya'll: } We are looking for a CCD camera for our Philips 430 TEM. We would } like } to get the best camera for the best price, e.g., either buying a } used } camera or a new non-Gatan camera (Gatan seems to be twice as } expensive } as the others). We would be using the camera for bright field and } high } resolution TEM of materials (semiconductors) rather than for } biological } specimens. Does anyone have a camera they would like to sell/donate } or } does anyone have recommendations as to a less-expensive camera that } they } know can be used for materials applications. } Thanks, Mike Coviello } Lab Manager } University of Texas -at- Arlington }
A question for you immuno gurus out there. We have a client who is interested in looking again at some five-year old blocks, this time in order to label a protein. The specimens were fixed for standard TEM, i.e., in 2% glut / 2% paraformaldehyde, followed by osmium. They are embedded in Epon/Araldite.
I expect that this is going to be a tough one, if it can be done at all. My question is: are there any secret techniques for optimizing immunogold labeling in sub-optimal specimens like this? I'm aware of the silver enhancement methods (although I've never used them). Would that be a good route to go? Or is it hopeless?
Thanks.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
Alright, I've watched this string for days now and resisted the temptation, but it's just too much. So here's mine, before Nestor shuts the string down. I know several good ones, but I'll restrict this to one on me.
A year ago (recent history!), I was teaching a lab section in Electron Microscopy, and although overloaded with students, things were cruising along reasonably well. One day, however, in a fit of brainlessness, I instructed a student who was doing cell cultures in how to dehydrate samples for TEM. We were processing his samples in a culture plate. He was going to embed in Spurr's, so I took him through the ethanol dehydrations up to 100%, then moved on to, you guessed it, propylene oxide. The plastic cell culture plate he was using was not amused. Right in front of both of us, it melted and "embedded" his samples in a resin I was not prepared to risk a diamond knife on. His comment was something like, "Is it supposed to do that?"
He finished the semester with flying colors and presented a very respectable project. Even better, we're still friends.
Randy Tindall
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/
Anyone have any good sources for a rotatable stages for inverted scopes? I would like something with good x & y translation but that allowed the slide or tray on the stage to rotate while keeping centered. how do other confocal and deconvolution types solve this problem? thanks for any tips. 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)
Chris MacWilliam wrote: ========================================================= To whom it may concern: I am a physics undergraduate and I am currently trying to work on an assignment in Microscopy. I am trying to find material on the theory and application of electron beam and x-ray methods of analysis with particular emphasis on SEM, TEM, EPMA, EDX, WDX, AES and XPS. I was given this website site as a possible source of information. Any assistance would be greatly appreciated.
Kindest Regards Chris MacWilliam. (ph95ccm-at-brunel.ac.uk) ========================================================= I would suggest you visit the MICRO 2000 meeting and exhibition next week at the Novotel London West Hotel. You can get details from the Royal Microscopical Society website at www.rms.org.uk .
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.spi.cc ######################## ============================================
We have an IonTech (UK manufacture) dual ion thinner which is missing its rotating stage part; this is a round piece the size of a large washer, with cogs on the perimeter. It is an essential part. Does anyone know where we can obtain a replacement?
Patrick Deshaye Geomicrobiology Lab Portland State University deshayep-at-ch1.ch.pdx.edu
Let's face it... obtaining leads and reaching prospects is the life-blood for today's entrepreneur/small business owner.
The big corporations have traditionally had the advantage in this area. They have the marketing budget that allows them to flood the marketplace and get hundreds, if not thousands, of leads.
LEADS TURN TO GOLDEN PROSPECTS
Leads turn into prospects and the if the product or service is appealing, the result can be a substantial PROFIT!
How can you, the entrepreneur or home based small business person, level the playing field and compete for your share without taking out a second mortgage on your house?
THE INTERNET IS THE ANSWER
One area the savvy entrepreneur has turned to is the Internet.
But the Internet is huge. How do you stand out? How do you generate the leads that are so vital for successful sales?
================================================================ Here's a comparison of Internet/Traditional Advertising Methods:
A Direct Mailing to 250,000 prospects through the Postal Service will cost you about $150,000.
A Banner Ad that reaches 5 million Internet surfers will cost you in the neighborhood of $100,000.
Bulk E-Mail to 10 Million people can generate hundreds/thousands of leads for pennies. ================================================================
.Which Advertising Method is the most economical?
.Which Advertising Method almost guarantee's you'll receive more leads than you could ever wish for?
"Many business people are finding out that they can advertise in ways that they never could have afforded in the past. The cost of sending mass e-mail is extremely low, and the response rate is high and quick." - USA Today.
"E-Mail is an incredible lead generation tool." - Crains Magazine
E - M A I L M A R K E T I N G W O R K S ! !
It is the most cost effective, low budget, way to gain leads.
It's the tested way to reach thousands of people ... turn leads into prospects that result in orders that can build wealth for the entrepreneurial business person.
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HERE'S THE ANSWER:
M I L L I O N S V O L U M E 9
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We've sold mailing lists for over 5 years and we've never compromised on quality. We won't release any address list until it passes our "high standards" test.
This is not a rental list that is restricted to a one-time mailing. You are purchasing an e-mail address list for your own personal mailings and may use it over-and-over.
DON'T HESITATE or you will miss out on the least expensive, legal and most effective way to market... PERIOD!
Order the e-mail address list within 72 hours and we'll include the following FREE Bonuses...
1. To help you get started we include basic proven Professional Mailing Software. Not a demo, a full working version (SORRY, SINCE THE SOFTWARE IS FREE WE CANNOT OFFER TECHNICAL SUPPORT, however set-up instructions are included). Requirement: Windows 95/95/NT or a MAC with SoftWindows.
2. "HOW TO GENERATE LEADS WITH BULK E-MAIL" A manual/guide that addresses the Bulk E-Mail business. Especially useful for beginners. "HOW TO GENERATE LEADS WITH BULK E-MAIL" will answer most of your questions and concerns about Bulk E-Mail. An exclusive for our customers... INCLUDED FREE.
***SPECIAL BONUS*** Order within the next 72 hours and receive an additional 972,565 e-mail addresses as a prompt ordering bonus. Order Now!
O R D E R N O W . . . SAME DAY SERVICE (M-F) if your order is received before 2pm Pacific. If you have further questions or to place an order, call toll free: 1-800-375-6456 24hours a day.
For fastest service, you may fax your order to: 1-530-504-7707
To order, via credit card simply cut/paste and print out the EZ ORDER FORM below and fax to our office today.
***** MILLIONS CD - Volume 9 *****
***** NOW ONLY $247! *****
This "Special Price" is in effect for the next 72 hours, after that we go back to our regular price of $299.00 ... Don't delay... you can be in business tomorrow!
We accept Visa, Mastercard, Amex and Checks by Fax. Fax your order to: 1-530-504-7707
----------------------Cut & Paste---------------------- ---------------------EZ Order Form---------------------
_____Yes! I want everything! I am ordering within 72 hours. Include ALL bonuses along with my 10 Million E-Mail addresses (don't forget the 972,565 bonus addresses) at the special price of only $247.00 + shipping as I have indicated below.
_____Oop's I missed the 72 hour "special". I am ordering my 10 million E-Mail addresses at the regular price of $299.00 + shipping.
***PLEASE SELECT YOUR SHIPPING OPTION***
____I would like to receive my package FedEx OVERNIGHT. I am including $15 for shipping. (Hawaii & Alaska $20 - Canada $25, all other International add an *additional* $25 [$40 total] for shipping)
____I would like to receive my package FedEx 2nd Day delivery. I'm including $10 for shipping. (Sorry FedEx 2nd Day is NOT AVAILABLE for shipments to Alaska, Hawaii, Canada or any International destination - Continental U.S. shipping addresses only).
***Please Print Carefully***
NOTE: Orders cannot be shipped without complete information including your signature. No exceptions!
COMPANY NAME____________________________________________
ADDRESS_________________________________________________ (FedEx can only ship to street addresses - no P.O. boxes)
CITY, STATE, ZIP________________________________________
PHONE NUMBER____________________________________________ (required for shipping & tracking)
EMAIL ADDRESS___________________________________________ (Print Carefully - required in case we have a question and to send you a confirmation that your order has been shipped)
NAME ON CARD___________________________________________
TOTAL AMOUNT (Including Shipping): $___________________
DATE:x__________________
(Required) SIGNATURE:x_________________________________ I understand that I am purchasing the Millions Vol. 9 E-Mail Address List, the addresses are not rented, but are mine to use for my own mailing, over-and-over. Free bonuses are included, but cannot be considered part of the financial transaction. I understand that it is my responsibility to comply with any laws applicable to my local area. As with all software, once opened the CD may not be returned, however, if found defective it will be replaced with like product at no charge.
You may fax your order to us at: 1-530-504-7707 or 1-500-677-4016
CHECK BY FAX SERVICES!
Please Note: Sorry, we can only accept checks drawn on U.S. banks.
If you would like to fax a check, tape your check below and fax it to our office along with the EZ Order Form to: 1-530-504-7707
If You fax a check, there is no need for you to mail the original. We will prepare a one-time draft, with the exact information on your original check. All checks will be held for bank clearance. (7-10 days) Make payable to: "HW-Marketing"
******************************************************** Do not reply to this message - ******************************************************** To register your e-mail address for removal from unsolicited mailings, visit: http://www.OptList.com ********************************************************
Let's face it... obtaining leads and reaching prospects is the life-blood for today's entrepreneur/small business owner.
The big corporations have traditionally had the advantage in this area. They have the marketing budget that allows them to flood the marketplace and get hundreds, if not thousands, of leads.
LEADS TURN TO GOLDEN PROSPECTS
Leads turn into prospects and the if the product or service is appealing, the result can be a substantial PROFIT!
How can you, the entrepreneur or home based small business person, level the playing field and compete for your share without taking out a second mortgage on your house?
THE INTERNET IS THE ANSWER
One area the savvy entrepreneur has turned to is the Internet.
But the Internet is huge. How do you stand out? How do you generate the leads that are so vital for successful sales?
================================================================ Here's a comparison of Internet/Traditional Advertising Methods:
A Direct Mailing to 250,000 prospects through the Postal Service will cost you about $150,000.
A Banner Ad that reaches 5 million Internet surfers will cost you in the neighborhood of $100,000.
Bulk E-Mail to 10 Million people can generate hundreds/thousands of leads for pennies. ================================================================
.Which Advertising Method is the most economical?
.Which Advertising Method almost guarantee's you'll receive more leads than you could ever wish for?
"Many business people are finding out that they can advertise in ways that they never could have afforded in the past. The cost of sending mass e-mail is extremely low, and the response rate is high and quick." - USA Today.
"E-Mail is an incredible lead generation tool." - Crains Magazine
E - M A I L M A R K E T I N G W O R K S ! !
It is the most cost effective, low budget, way to gain leads.
It's the tested way to reach thousands of people ... turn leads into prospects that result in orders that can build wealth for the entrepreneurial business person.
The big question is:
Where do I get the e-mail addresses that will help me contact the millions of people on the Internet?
HERE'S THE ANSWER:
M I L L I O N S V O L U M E 9
***New - 10 Million addresses - Just Released***
The cleanest, most comprehensive e-mail address list in the world, BAR NONE!
We've sold mailing lists for over 5 years and we've never compromised on quality. We won't release any address list until it passes our "high standards" test.
This is not a rental list that is restricted to a one-time mailing. You are purchasing an e-mail address list for your own personal mailings and may use it over-and-over.
DON'T HESITATE or you will miss out on the least expensive, legal and most effective way to market... PERIOD!
Order the e-mail address list within 72 hours and we'll include the following FREE Bonuses...
1. To help you get started we include basic proven Professional Mailing Software. Not a demo, a full working version (SORRY, SINCE THE SOFTWARE IS FREE WE CANNOT OFFER TECHNICAL SUPPORT, however set-up instructions are included). Requirement: Windows 95/95/NT or a MAC with SoftWindows.
2. "HOW TO GENERATE LEADS WITH BULK E-MAIL" A manual/guide that addresses the Bulk E-Mail business. Especially useful for beginners. "HOW TO GENERATE LEADS WITH BULK E-MAIL" will answer most of your questions and concerns about Bulk E-Mail. An exclusive for our customers... INCLUDED FREE.
***SPECIAL BONUS*** Order within the next 72 hours and receive an additional 972,565 e-mail addresses as a prompt ordering bonus. Order Now!
O R D E R N O W . . . SAME DAY SERVICE (M-F) if your order is received before 2pm Pacific. If you have further questions or to place an order, call toll free: 1-800-375-6456 24hours a day.
For fastest service, you may fax your order to: 1-530-504-7707
To order, via credit card simply cut/paste and print out the EZ ORDER FORM below and fax to our office today.
***** MILLIONS CD - Volume 9 *****
***** NOW ONLY $247! *****
This "Special Price" is in effect for the next 72 hours, after that we go back to our regular price of $299.00 ... Don't delay... you can be in business tomorrow!
We accept Visa, Mastercard, Amex and Checks by Fax. Fax your order to: 1-530-504-7707
----------------------Cut & Paste---------------------- ---------------------EZ Order Form---------------------
_____Yes! I want everything! I am ordering within 72 hours. Include ALL bonuses along with my 10 Million E-Mail addresses (don't forget the 972,565 bonus addresses) at the special price of only $247.00 + shipping as I have indicated below.
_____Oop's I missed the 72 hour "special". I am ordering my 10 million E-Mail addresses at the regular price of $299.00 + shipping.
***PLEASE SELECT YOUR SHIPPING OPTION***
____I would like to receive my package FedEx OVERNIGHT. I am including $15 for shipping. (Hawaii & Alaska $20 - Canada $25, all other International add an *additional* $25 [$40 total] for shipping)
____I would like to receive my package FedEx 2nd Day delivery. I'm including $10 for shipping. (Sorry FedEx 2nd Day is NOT AVAILABLE for shipments to Alaska, Hawaii, Canada or any International destination - Continental U.S. shipping addresses only).
***Please Print Carefully***
NOTE: Orders cannot be shipped without complete information including your signature. No exceptions!
COMPANY NAME____________________________________________
ADDRESS_________________________________________________ (FedEx can only ship to street addresses - no P.O. boxes)
CITY, STATE, ZIP________________________________________
PHONE NUMBER____________________________________________ (required for shipping & tracking)
EMAIL ADDRESS___________________________________________ (Print Carefully - required in case we have a question and to send you a confirmation that your order has been shipped)
NAME ON CARD___________________________________________
TOTAL AMOUNT (Including Shipping): $___________________
DATE:x__________________
(Required) SIGNATURE:x_________________________________ I understand that I am purchasing the Millions Vol. 9 E-Mail Address List, the addresses are not rented, but are mine to use for my own mailing, over-and-over. Free bonuses are included, but cannot be considered part of the financial transaction. I understand that it is my responsibility to comply with any laws applicable to my local area. As with all software, once opened the CD may not be returned, however, if found defective it will be replaced with like product at no charge.
You may fax your order to us at: 1-530-504-7707 or 1-500-677-4016
CHECK BY FAX SERVICES!
Please Note: Sorry, we can only accept checks drawn on U.S. banks.
If you would like to fax a check, tape your check below and fax it to our office along with the EZ Order Form to: 1-530-504-7707
If You fax a check, there is no need for you to mail the original. We will prepare a one-time draft, with the exact information on your original check. All checks will be held for bank clearance. (7-10 days) Make payable to: "HW-Marketing"
******************************************************** Do not reply to this message - ******************************************************** To register your e-mail address for removal from unsolicited mailings, visit: http://www.OptList.com ********************************************************
David S. Phillips died suddenly of a heart attack on Saturday March 18, while hiking with his son's Boy Scout Group in Bandelier National Monument, near Los Alamos, New Mexico.
David was born on February 17, 1952, in Columbus, Ohio. Following graduation from Fairborn High School, Ohio, in 1970, he attended Case Western Reserve University in Cleveland, Ohio, where he received his B.S. degree in Metallurgy with highest honors in 1974. He continued on at Case for his M.S. and Ph.D. degrees researching in Ceramics with myself and Arthur Heuer. After a post-doctoral appointment with Jacques Castaing at the CNRS Laboratoire de Physique des Materiaux in Bellevue, France, David was on the faculty of the University of Illinois before joining Los Alamos National Laboratory in 1984.
David is survived by his wife Jane and by their sons Sam and Paul, all of Los Alamos. A memorial fund has been set up in the name of David Samuel Phillips at Los Alamos National Bank to benefit his sons.
David Phillips was a fine electron microscopist who applied his skills to a wide variety of problems in ceramics, superconductors, diamonds and nuclear materials. Amongst his many accomplishments were seminal papers on the use of high resolution and analytical electron microscopy to deduce the underlying cause for the star in star sapphire. He performed beautiful work in science and in everyday life and will be sorely missed.
xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx Terence E. Mitchell Laboratory Fellow Materials Science and Technology Division MS-K765 Los Alamos National Laboratory Los Alamos, NM 87545 voice mail: 505-667-0938 fax: 505-665-2992 e-mail: temitchell-at-lanl.gov xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx
There is still some space left for the North Carolina State University short course on Image Analysis and Stereology, but you should register soon to avoid disappointment.
There are two sessions of the workshop, May 11-13 and May 15-17, 2000, at N. C. State Univ. in Raleigh, NC. This course has been heavily attended and widely praised for the last 17 years. It offers hands-on training using computer-based image processing and analysis, couple with practical lectures by experts in the fields of digital imaging and stereology. Many attendees in the fields of materials, biology, medical applications, food science, industrial inspection, and other disciplines involving microscopy have found this course to be a very practical way to gain understanding and proficiency in the techniques for acquiring and interpreting structural information.
Full information, including course outlines, faculty information, and on-line registration, is available at http://members.aol.com/IPCourse/ You may also contact Cindy Allen at 919 515 8171.
VCR Group, Inc. sold Ion Tech products here in the U.S. many years ago. When we acquired VCR Group last year, we also acquired some old Ion Tech parts. I will talk to Vince Carlino and check our stock to see if we have what you need.
David Henriks Vice President 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
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by INTERNET:"DESHAYEP-at-ch1.ch.pdx.edu"-at-sparc5.microscopy.com } ----------------------------------------------------------------------- The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We have an IonTech (UK manufacture) dual ion thinner which is missing its rotating stage part; this is a round piece the size of a large washer, with cogs on the perimeter. It is an essential part. Does anyone know where we can obtain a replacement?
Patrick Deshaye Geomicrobiology Lab Portland State University deshayep-at-ch1.ch.pdx.edu
Haven't seen this one submitted to the list yet, but this is a nightmare we all have. Another department on campus was kind enough to give us a TEM, a JEOL 100CXII. It, of course, had minor vacuum leak which put it out of service, but besides that it was in mint condition. I had planned to move it myself but was overruled by my boss. There was concern for injury to myself or others in case it fell. I begged, whined, pleaded, and lost the discussion. A moving company would transport. I talked to the administrative person arranging the move and was told that the moving company said it had all the information it needed. Bells, whistles, lights, and red flags went off in my mind. I finally got hold of the moving company supervisor and was told he had done things like this before and knew what had to be done. I told him several times, NOT to use the liftgate on his moving truck, to use a forklift. The column was too heavy, and balanced wrong for liftgates to work well. He assured me HIS truck could easily lift the 3,000 lbs. More flags waved! I told him one last time to use a forklift to avoid problems. The day for moving came and this day, as well as the next two appointments, the movers gave the excuse the truck clutch was out, more red flags. When finally they did get the clutch repaired, I unfortunately (fortunately?) was not there to see the fireworks, they tried to use the lift gate, got the column several inches off the ground before both hydraulic cylinders for the truck lift gate sheared off, dropping the gate and giving the column a good bounce. As good luck would have it, the microscope didn't tip over, just wobbled a lot. Had it been higher, a lot of damage could have occurred as well as someone getting hurt. To top this story off, the moving company had the audacity to try to charge the University for the cost of repairing their truck. The damage to the microscope was limited to knocking the intermediate and projector lenses out of alignment and the instrument is working wonderfully in its new home. "Takes a lickin' and keeps on tickin'." Later; David L Bentley Imaging Facility The University of Arizona Tucson, AZ 85721-0036 (520)621-5097
Would this group be a good place to post micro-Raman questions or is there a better group?
We are just beginning to use this technique. Can anyone recommend a computer reference library for Raman? How do different wavelength lasers affect frequency shift and amplitudes?
Thanks, Ed
Ed Kurz Institute of Materials Science, U3136 97 North Eagleville Road University of Connecticut Storrs, CT 06269-3136 ekurz-at-mail.ims.uconn.edu (860) 486-4186 phone (860) 486-4745 fax
Email: vriesg-at-rpi.edu Name: Gwynne Vriesema School: Rensselaer Polytechnic Institute
Question: I am doing a project on the application of atomic force microscopy to making read/write devices that have a much higher data density than today's magnetic drives. Do you see any drawbacks to using AFM for this application? What aspect of AFM is most important to understand when figuring out how these drives would work? What are the advantages to using this method to store data? Thank you for your time!
I have a JEOL 2000FX and I am getting an "S" distortion in my images at all mags. The severity differs slightly with mag. I know that the distortion is there because I am looking at layered structures on Si. I put the samples with the interfaces parallel to the rod axis. As I move the sample along the rod axis, the "s" stays in the same place. I looked at the Kikuchi lines in an on-axis CBED pattern. They do not seem to suffer the same problem, but there is a little at low camera lengths at the outside parts of the pattern.
Does anyone know the source of this distortion and is there any cure for it?
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
I've operated a listserv for a couple of years called Irusers-l, which has been used by scientists who work in museums and laboratories that use FTIR or micro-FTIR to analyze historic and artistic works. Some members also use Raman. We've had brief discussions about expanding the group to any scientist who uses FTIR or Raman, regardless of application.
Would members of this list be interested in joining Irusers-l?
James Martin
Research Scientist in Chemistry Williams College james_martin.tripod.com/williams.htm
Director Orion Analytical, LLC
On Thu, 6 Apr 2000, Ed Kurz wrote:
} } Would this group be a good place to post micro-Raman questions or is there } a better group? } } We are just beginning to use this technique. Can anyone recommend a } computer reference library for Raman? How do different wavelength lasers } affect frequency shift and amplitudes? } } Thanks, } Ed } } } Ed Kurz } Institute of Materials Science, U3136 } 97 North Eagleville Road } University of Connecticut } Storrs, CT 06269-3136 } ekurz-at-mail.ims.uconn.edu } (860) 486-4186 phone } (860) 486-4745 fax } } } }
Waaay back, when I was an instructor at MIT in Cambridge, Mass., I was helping out in the physical metallurgy laboratory course when we were teaching the students how to develop glass metallographic plates (told you it was a long time ago). One of my students, the son of a famous metallurgist, placed his fully developed plate on the belt of a machine that he didn't notice was running. We never let him forget the day he tried to dry his plate in the cylindrical print drier.
George Langford, Sc.D., in PA trying to forget that everyone used mercury ice-point reference junctions in their thermocouples in the heat-treating lab in those days ... and how many got tipped over.
I have a colleague with advanced presbyopia who is doing consulting work and needs a stereo zoom to do botanical identification. He wants to stay below US $1,000.
He has found the following scopes on the web:
TT-5Z and TT-5B at the www.ken-a-vison.com web site
SMZ Bino zoom at www.microscope.org
Any comments on any of these or recommendations for other scopes in his price range.
Thank you for any good suggestions.
Don
(Direct replies from vendors are welcomed, but please identify your financial interest in your offers/recommendations.)
______________________________________________________________________ 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
They will supply replacement parts for most of their machines. (While you're about it, get more ceramic resistors, Al cathodes, cathode stops etc.) If you're rejuvenating one of these machines I'd be happy to describe the maintenance and alignment procedures - I have been taking these things to bits and reassembling them for a few years now.
Cheers,
Richard Beanland
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } We have an IonTech (UK manufacture) dual ion thinner which is missing its rotating stage part; this is a } round piece the size of a large washer, with cogs on the perimeter. It is an essential part. Does anyone } know where we can obtain a replacement? } } Patrick Deshaye } Geomicrobiology Lab } Portland State University } deshayep-at-ch1.ch.pdx.edu }
Shortly after our laboratory purchased our first analytical electron microscope, the microscope developed a problem in the electronics. The microscopist telephoned the vendor's service department who requested he measure some voltages to help diagnose the problem. The microscopist dutifully attached clip-leads to the appropriate test points and was attaching them to the voltmeter when one of the clip-leads slipped off, fell into the electronics chassis, and shorted out much of the electronics. The vendor service ingineers were in the lab for weeks repairing the problem. The rest of the lab teased him about this, giving the the microscopist the nickname, "clip-lead". A few years later when he left the group for a promotion to a staff job, he was presented with a pair of clip-leads where the metallic ends had been coated with liquid-rubber.
Best Regards,
John Minter Eastman Kodak Company Analytical Technology Division Rochester, NY 14652-3712 Phone: (716) 722-3407 FAX: (716) 477-3029 email: john.minter-at-kodak.com
Early one evening when the only people still in the building were us students, a guy from along the corridor came into our room saying that he could smell something downstairs. We all banded together and went down to investigate. After a while we noticed this sort of thick foggy smoke beginning to form below the ceiling along the length of the corridor and in the adjacent large teaching lab. We noticed it wafting down through some light fittings and gradually getting lower down and thicker. "Has to be electrical!" "Looks like the whole place could go up any minute!" It started looking pretty serious. When the fire brigade turned up we were told to get out as they proceeded to rip out chunks of the ceiling trying to locate the source. It was a very diffuse source and there was a very large area of ceiling to rip chunks out of. After a while their investigation moved upstairs -- where they found our carbon coater happily chugging away with it's bell jar off and the exhaust line feeding down into the ceiling cavity! You learn something new every day.
I think we remembered to say thanks....
Arthur Day, Electron Microscope Unit Phone: 61-2-9717-3457 Ansto Materials Division Fax: 61-2-9543-7179 PMB 1, Menai (Sydney), NSW, 2234 Email: ard-at-ansto.gov.au Australia www: http://www.ansto.gov.au/
{paraindent} {param} left {/param} {color} {param} 0100,0100,0100 {/param} {FontFamily} {param} Times New Roman {/param} {bigger} The Department of Biology, Acadia University, Wolfville, NS have two electron microscopes for disposal. The first is a Zeiss EM9A transmission electron microscope and the second a JEOL JSN-25S scanning electron microscope. Both microscopes are operable, but are surplus to the current needs of the department. The department is willing to donate these instruments to an institution, providing that institution is willing to assume the responsibility for removal and shipping to the new destination. For further information contact Dr. T.B. Herman (902) 585-1469. {/paraindent}
{nofill}
============================================================== Graham N. Cheeseman Biology Dept., Acadia University Wolfville, Nova Scotia B0P 1X0 TEL: (902) 585-1316 FAX: (902) 585-1059 INTERNET: Graham.Cheeseman-at-acadiau.ca ==============================================================
Hope this scary one helps some of you! We are a research and teaching lab (thankfully!) A very lucky (for us) student was in working on a weekend when she noticed white smoke pouring out of the lab into the lecture room, grabbed the extinguisher, and put out the electrical fire that started in the wall due to our specimen rotator motor shorting out (and not being properly wired). The fire was directly underneath the 1 gallon storage tanks for 100% ethanol, and pure Xylene..... {yipe} ! So bad, that it melted but did not break through the spigots... {big yipe} ! I also learned that when your filling a liquid nitrogen tank, and it gets full, and it begins to shoot up a little underneath a temperature sensor, you get to meet the fire chief! Life is good!
Tracey Pepper Supervisor Bessey Microscopy Facility Iowa State University ph: 515-294-3872 fax: 515.294.1337
Hi Randy One good "old" reference is Moise Bendayan and Max Zollinger. Ultrastructural Localization of Antigenic Sites on Osmicated-fixed Tissues Applying the Protein A-Gold Technique. J Histochem Cytochem 31 :101-109, 1983 Some times it works some times it doesn't. I would like to see some newer tricks if they are out there.
My horror story happened many years ago when I came into work one morning to find the Philips200 had been left running for a long period of time with no water coursing through its veins. The column was hot to touch so my immediate reaction was to turn on the water supply to try to cool it down. But hoses and o-rings had deteriorated from the heat, so instead of cooling the instrument down I now had water gushing from everywhere and filling up the columnâs viewing area so that it looked like a fish bowl. I called my Philips service engineer and he spent the next 3 days repairing and cleaning up the scope. He said the stage had come very close to a melt down and if that had happened I could have just dug a hole and buried the scope in the back yard. The scope survived to give the lab many years of service, thanks to Philips engineer, John Braunagel, whose expert service was given without a grumble or complaint.
I set up an lab in a hospital with a Siemens microscope-a wonderful instrument for resolution but a beast to align. The lenses were physically moved during alignment so that it did look odd to see different components of the column a few centimeters off. One morning I came in and a pathologist proudly told me that he had aligned the column for me. He had straightened it out very nicely and it took a full day for me to get it back in alignment. It did look nicer the way he did it but of course it was impossible to use. Joyce Craig Chicgo State University
How long should a gold target last? Our target looks gold except in the center. Problem is that we don't seem to get good coating. Joyce Craig Chicago State University
My horror story happened many years ago when I came into work one morning to find the Philips200 had been left running for a long period of time with no water coursing through its veins. The column was hot to touch so my immediate reaction was to turn on the water supply to try to cool it down. But hoses and o-rings had deteriorated from the heat, so instead of cooling the instrument down I now had water gushing from everywhere and filling up the column's viewing area so that it looked like a fish bowl. I called my Philips service engineer and he spent the next 3 days repairing and cleaning up the scope. He said the stage had come very close to a melt down and if that had happened I could have just dug a hole and buried the scope in the back yard. Surprisingly the scope survived to give the lab many years of service, thanks to Philips service engineer John B's expert service that he gave without a grumble or complaint.
This is a story that was told to me by Dr. Audrey Glauert of Cambridge University in the U.K. Even though it's not my own, it is so amusing that I can't help relaying it to you.
It seems that a number of years ago Dr. Glauert spent several months working in an electron microscopy laboratory in Africa. Every now and then the water supply to the laboratory would go off making it necessary to shut the electron microscope down for an extended period of time. Investigation eventually revealed that the problem arose because the town involved was getting it's water from a pond that was formed behind a dam that had been constructed across a nearby river. It seems that this pond was a favorite site for a herd of hippopotamuses to bathe, and every now and then one of them would manage to plug up the water inlet to the village water system, whereupon it was necessary for the villagers to go out and chase the hippos away and then open the inlet again. Since hippos are not very easy to chase, this could sometimes cause a rather prolonged period without water in the electron microscopy laboratory.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
With all these horror stories being shared, its surprising that nobody has related a "darkroom" tale yet. Surely everyone who has been in this field for several years has had the unpleasant experience of walking into the darkroom and finding everything coated with dried photographic fix residue? My darkroom experiences with users have been all in all much worse that microscope incidents.
WL Steffens University of Georgia Department of Pathology
{paraindent} {param} left {/param} {color} {param} 0100,0100,0100 {/param} {FontFamily} {param} Times New Roman {/param} {bigger} The Department of Biology, Acadia University, Wolfville, NS have two electron microscopes for disposal. The first is a Zeiss EM9A transmission electron microscope and the second a JEOL JSN-25S scanning electron microscope. Both microscopes are operable, but are surplus to the current needs of the department. The department is willing to donate these instruments to an institution, providing that institution is willing to assume the responsibility for removal and shipping to the new destination. For further information contact Dr. T.B. Herman (902) 585-1469. {/paraindent}
{nofill}
============================================================== Graham N. Cheeseman Biology Dept., Acadia University Wolfville, Nova Scotia B0P 1X0 TEL: (902) 585-1316 FAX: (902) 585-1059 INTERNET: Graham.Cheeseman-at-acadiau.ca ==============================================================
About a month after starting my post doc in the lab of Professor Ruhle in Stuttgart, Germany, I was working on the TEM late one Sunday night. I was using a double-tilt, analytical, double-specimen holder (read: expensive) and just putting it away. The holder was gripped in a Gatan holder stand and when I pushed the protective end sheath onto the tip the stage moved back so the front stand grip moved onto the narrower part of the holder where it doesn't grip. The motorized back end was heavy and the holder tipped backwards while I was still holding the protective end piece firm. What resulted is a holder that greatly resembled a Concord airplane coming in for a landing. It bent the holder at about a 30 degree angle right where the hinge was for the back specimen cup.
I sat and looked at it, beads of sweat forming on my forhead, contemplating the fact that I had an open ended return ticket and wondering if I could pack my stuff up and be gone back to the U.S. before anyone noticed. Of course I stayed and Professor Ruhle was very good about it, basically saying that mistakes happen but don't do it again! I didn't.
Lessons Learned: Be very careful when handling expensive specimen holders; think twice before you do anything.
I hope you all have a good weekend and don't mind my confessions!
Cheers, JSV
*************************** John Vetrano Pacific Northwest National Laboratory MSIN P8-16 P.O. Box 999 Richland, WA 99352 Phone: (509)372-0724 Fax: (509)376-6308 Email: mailto:john.vetrano-at-pnl.gov
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If anyone has any tips or a reference they could point me to for this, it would be much appreciated.
We have had several users with nanocrystalline ceramic samples from combustion or plasma syntheses who have had problems with the thin areas of their samples falling out during ion milling. We have tried a few schemes to correct this, without much success. Any help would be greatly appreciated.
The current user has 2 different nanocrystalline composites: TiB2/TiN and TiB2/TiC.
Valerie Leppert Dept. Chem. Eng. and Mat. Sci. University of California, Davis
That is a difficult question since it depends on so many variables (original target thickness, coating rates, time run, etc). I use my sputter coater often (but not as often as the carbon evaporator). Never kept accurate records of usage, but it is at least several years old.
In any event, the entire target should have a bright gold color. Some areas may have a matte vs shiny appearance, but should not be dark. If not all bright Au, it is either consumed or contaminated. Either case will cause sputtering to cease.
Woody White
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How long should a gold target last? Our target looks gold except in the center. Problem is that we don't seem to get good coating. Joyce Craig Chicago State University
My horror story happened many years ago when I came into work one morning to find the Philips200 had been left running for a long period of time with no water coursing through its veins. The column was hot to touch so my immediate reaction was to turn on the water supply to try to cool it down. But hoses and o-rings had deteriorated from the heat, so instead of cooling the instrument down I now had water gushing from everywhere and filling up the columnâs viewing area so that it looked like a fish bowl. I called my Philips service engineer and he spent the next 3 days repairing and cleaning up the scope. He said the stage had come very close to a melt down and if that had happened I could have just dug a hole and buried the scope in the back yard. The scope survived to give the lab many years of service, thanks to Philips engineer, John Braunagel, whose expert service was given without a grumble or complaint.
There was some discussion about scanners and SCSI bus problems. There are some recent developments that I'd like to share with those who are having SCSI-related problems.
If you have a PC which includes one or two USB ports, there is a new converter cable which should make your life much better. Microtech has a USB to SCSI-II converter cable (# USB-SCSI-HD50) that plugs into a USB socket and provides a high density 50 pin SCSI-II plug. This plug can be converted to SCSI-I wide Centronics ribbon-style connector if necessary. This converter cable works with PCs and Macs; and so far, all of my SCSI devices work with it (scanners, CD-R, CD-RW).
This item is available from various sources for about $80. I got mine from d-store.com but you might find other sources.
Does anyone have a functioning Zeiss 10c TEM that they would like to donate to a goverment office for a tax deduction or sell for cheap? I am the electron microscopist for the county veterinarian and we are trying to locate a Zeiss 10c to be used for virus identification work on animals and plants. Thank you. Cindy Shannon cshannon-at-nctimes.net
Suggest you visit the sites for both Digital Instruments and ThermoMicroscopes. Both have application notes which will be helpful. Also, suggest that you contact Sergei Magonov at DI and Jezz Leckenby at ThermoM. Both are extremely knowledgeable apps specialists. Sergei: 805-967-1400 Jezz: (408) 747-1600
Hope this is helpful.
Caveat: MME has no financial interest in either of these systems. Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ******************************************************
At 05:46 PM 4/6/00 -0500, wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
ELECTRON MICROSCOPIST Laboratory of Structural Biology Institute of Molecular Agrobiology, Singapore
A position is available for an experienced EM technician at the Institute of Molecular Agrobiology (IMA), Singapore, in the group of Dr. Terje Dokland, Laboratory of Structural Biology. The successful candidate will be responsible for the operation of the structural EM laboratory, and be involved in several research projects, in particular structural characterization of various plant and animal viruses of agricultural importance. Experience with cryo-EM of frozen-hydrated samples is a definite advantage, as is some experience with using computers, especially 3D reconstruction methods. IMA is a well funded and rapidly expanding research institute which was established in 1995 to conduct basic and applied research in agrobiology. It moved into a modern and spacious building at the National University of Singapore campus two years ago, and has state-of-the art equipment for structural and molecular biology, biochemistry, plant growth, tissue culture etc. There are currently two groups working in structural biology, including X-ray crystallography and electron microscopy, and there is extensive collaboration with other laboratories and institutes, both in Singapore and abroad. Further information about the institute can be found at http://www.ima.org.sg. Singapore is a cosmopolitan and multicultural society offering all the conveniences of a modern city, and is centrally located in Southeast Asia with easy access to other countries in the region. The position will be filled at the Research Officer or Assistant Research Officer level, depending on experience, and the contract will be initially offered for two years. IMA offers attractive salaries with bonus packages and benefits. Informal enquiries are welcome and can be directed to Terje Dokland at dokland-at-ima.org.sg. Applications, including a CV and names of two referees, should be sent to Terje Dokland Institute of Molecular Agrobiology 1 Research Link The National University of Singapore Singapore 117604
------------------------------------ Terje Dokland Senior Scientist Institute of Molecular Agrobiology 1 Research Link The National University of Singapore Singapore 117604 Phone: 65-872 7405 Fax: 65-872 7007 E-mail: dokland-at-ima.org.sg
"Convictions are greater enemies of truth than lies" F. Nietzsche
If anyone has any tips or a reference they could point me to for this, it would be much appreciated.
We have had several users with nanocrystalline ceramic samples from combustion or plasma syntheses who have had problems with the thin areas of their samples falling out during ion milling. We have tried a few schemes to correct this, without much success. Any help would be greatly appreciated.
The current user has 2 different nanocrystalline composites: TiB2/TiN and TiB2/TiC.
Valerie Leppert Dept. Chem. Eng. and Mat. Sci. University of California, Davis
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Maybe Judy Murphy's Delta College will diminish these "untrained" occurrences!
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Does anyone have a functioning Zeiss 10c TEM that they would like to donate to a goverment office for a tax deduction or sell for cheap? I am the electron microscopist for the county veterinarian and we are trying to locate a Zeiss 10c to be used for virus identification work on animals and plants. Thank you. Cindy Shannon cshannon-at-nctimes.net
This is actually a follow-on question to the thread on "Silicone Oils and Greases" which ran a few days ago.
In his typically thorough response to the discussion of why microscopists generally avoid silicone-based vacuum greases, Will Bigelow noted that
} While the silicone high vacuum grease is } indeed a good lubricant for O-rings, it is no better than the Brayco and } Krytox greases, which are based on polyphenylether compounds, and which do } not introduce the possibility of having insoluble siliceous compounds } formed on critical parts of the electron optical column. }
This mention of Krytox rang a bell and I checked Chapter 10 of Will's book "Vacuum Methods in Electron Microscopy" where he discusses oils and greases. In table 10.1 he lists both Braycote and Krytox as "Perfluorinated Polyether based" (not polyphenylether). In the subsequent discussion he states that they "probably should not be used ... in or near the electron gun because of the possibility that some of the perfluorinated base compound might get onto the high voltage insulator and cause microdischarges ... ". In chapter 5 he is more specific in his discussion of perfluorinated polyether diffusion pump oils where he again mentions Krytox and states that use of these pump oils was generally abandoned because it was found that "electron microscopes in which these fluids were used eventually developed high-voltage instabilities due to micro-discharges along the ceramic insulators in the electron guns." Will goes on to note that the problem seems to be more severe in TEMs (which operate at higher voltages) and these fluids "... have been used successfully for several years in scanning electron microscopes in some laboratories".
First question: I'm confused by Will's list-server statement that Krytox is a polyphenylether when it is listed in his book as a perfluorinated polyether. Not being an organic (or any other kind) of chemist, I might think that these are synonyms for the same family of materials except that Will also has a separate discussion of polyphenyl ether pump fluids (e.g., Convalex and Santovac) which is separate from that of the perfluorinated polyethers (Krytox and Fomblin). I think that Krytox actually is a perfluorinated polyether, isn't it?
Second question: The issue of whether Krytox and Braycote are perfluorinated polyethers is more than a trivial nomenclature issue because of the alleged problem of perfluorinated compounds contaminating high-voltage insulators. In addition to the somewhat equivocal cautionary statements in Will's book, I personally know of one lab which has banished these compounds from the premises because of an incident a number of years ago where the glassware used for cleaning of microscope parts got contaminated with Krytox -- this got transferred to several TEMs and resulted (so I'm told) in the need to replace multiple guns over a period of time -- and produced a lot of hair-pulling until the source of the problem was identified (actually, maybe this should be under the "horror stories" thread?). But this is the only direct report I have heard of such a problem and Will, though he notes the concerns, doesn't seem reluctant to recommend the stuff for EM use (and I do respect his depth of experience in such things). So my question: Is Krytox really the "bogeyman" I've been told it is? Or is this just another bit of microscopy folklore?
Given that: (1) there are lots of ways a vacuum grease could get transferred to a gun indirectly (i.e., via tools or contaminated gloves); (2) this stuff is reported to be essentially impossible to remove once it gets deposited on something; (3) the purported HV discharge behavior doesn't show up immediately but develops gradually over time; and (4) insulator cleanliness is enough of a problem without introducing this kind of sneaky contaminant -- IF true, it would seem that this class of compounds has no place in an EM lab. But if this is all a myth, I'm depriving myself of some otherwise great products. Insight anyone?
Most sputter coater manufacturers use one of two methods for mounting their targets.
1. Target material fixed by a clip or adhesive to an aluminium base (difficult to sputter without a special coater)
2. Target material fixed by adhesive to a brass base (this will sputter in relation to its composition e.g. 70%Cu 30%Zn)
You seem to have the type (2) target and are probably sputtering a mixture of gold (from the outside of the target) and brass (from the middle of the target). Check it out if you have x-ray analysis?
Sputter targets last a period in direct relationship to target thickness, coating thickness and degree of use, how long is a piece of string?
Good luck
Steve Chapman Senior Consultant Protrain - professional training in EM world wide Tel +44 1280 814774 Fax +44 1280914007 e-mail protrain-at-emcourses.com web site www.emcourses.com
"S" distortion or anisotropic distortion is produced through a balancing of aberrations. Take one imaging lens which is being used at low current and you will have pincushion distortion. Add another lens that is being used in the low to middle part of its current range and you will have pincushion distortion. Use the two lenses in a magnification system and the result is an image with anisotropic distortion.
When we design a TEM we try to balance the lenses so that the degree of distortion is minimised. Minimised does not mean totally removed it means NO distortion in the area that falls within the photographic frame. In early instruments, where the screen was very small, you hardly ever saw the problem, as screens have increased in size so the problem becomes more clear.
If the distortion has just started to become annoying this may be due to one or two reasons.
1) The high voltage has changed such that the lenses are not matched to the accelerating voltage as they were when the system was new?
2) A lens is not running at the correct level, its current is too low?
The most common reason for a change in lens or high voltage performance are their reference circuits, I am afraid I am not familiar with these circuits in the 2000FX.
Good luck hope this helps?
Steve Chapman Senior Consultant Protrain - professional training in EM world wide Tel +44 1280 814774 Fax +44 1280914007 e-mail protrain-at-emcourses.com web site www.emcourses.com
ELECTRON MICROSCOPIST Laboratory of Structural Biology Institute of Molecular Agrobiology, Singapore
A position is available for an experienced EM technician at the Institute of Molecular Agrobiology (IMA), Singapore, in the group of Dr. Terje Dokland, Laboratory of Structural Biology. The successful candidate will be responsible for the operation of the structural EM laboratory, and be involved in several research projects, in particular structural characterization of various plant and animal viruses of agricultural importance. Experience with cryo-EM of frozen-hydrated samples is a definite advantage, as is some experience with using computers, especially 3D reconstruction methods. IMA is a well funded and rapidly expanding research institute which was established in 1995 to conduct basic and applied research in agrobiology. It moved into a modern and spacious building at the National University of Singapore campus two years ago, and has state-of-the art equipment for structural and molecular biology, biochemistry, plant growth, tissue culture etc. There are currently two groups working in structural biology, including X-ray crystallography and electron microscopy, and there is extensive collaboration with other laboratories and institutes, both in Singapore and abroad. Further information about the institute can be found at http://www.ima.org.sg. Singapore is a cosmopolitan and multicultural society offering all the conveniences of a modern city, and is centrally located in Southeast Asia with easy access to other countries in the region. The position will be filled at the Research Officer or Assistant Research Officer level, depending on experience, and the contract will be initially offered for two years. IMA offers attractive salaries with bonus packages and benefits. Informal enquiries are welcome and can be directed to Terje Dokland at dokland-at-ima.org.sg. Applications, including a CV and names of two referees, should be sent to Terje Dokland Institute of Molecular Agrobiology 1 Research Link The National University of Singapore Singapore 117604
------------------------------------ Terje Dokland Senior Scientist Institute of Molecular Agrobiology 1 Research Link The National University of Singapore Singapore 117604 Phone: 65-872 7405 Fax: 65-872 7007 E-mail: dokland-at-ima.org.sg
"Convictions are greater enemies of truth than lies" F. Nietzsche
This is actually a follow-on question to the thread on "Silicone Oils and Greases" which ran a few days ago.
In his typically thorough response to the discussion of why microscopists generally avoid silicone-based vacuum greases, Will Bigelow noted that
} While the silicone high vacuum grease is } indeed a good lubricant for O-rings, it is no better than the Brayco and } Krytox greases, which are based on polyphenylether compounds, and which do } not introduce the possibility of having insoluble siliceous compounds } formed on critical parts of the electron optical column. }
This mention of Krytox rang a bell and I checked Chapter 10 of Will's book "Vacuum Methods in Electron Microscopy" where he discusses oils and greases. In table 10.1 he lists both Braycote and Krytox as "Perfluorinated Polyether based" (not polyphenylether). In the subsequent discussion he states that they "probably should not be used ... in or near the electron gun because of the possibility that some of the perfluorinated base compound might get onto the high voltage insulator and cause microdischarges ... ". In chapter 5 he is more specific in his discussion of perfluorinated polyether diffusion pump oils where he again mentions Krytox and states that use of these pump oils was generally abandoned because it was found that "electron microscopes in which these fluids were used eventually developed high-voltage instabilities due to micro-discharges along the ceramic insulators in the electron guns." Will goes on to note that the problem seems to be more severe in TEMs (which operate at higher voltages) and these fluids "... have been used successfully for several years in scanning electron microscopes in some laboratories".
First question: I'm confused by Will's list-server statement that Krytox is a polyphenylether when it is listed in his book as a perfluorinated polyether. Not being an organic (or any other kind) of chemist, I might think that these are synonyms for the same family of materials except that Will also has a separate discussion of polyphenyl ether pump fluids (e.g., Convalex and Santovac) which is separate from that of the perfluorinated polyethers (Krytox and Fomblin). I think that Krytox actually is a perfluorinated polyether, isn't it?
Second question: The issue of whether Krytox and Braycote are perfluorinated polyethers is more than a trivial nomenclature issue because of the alleged problem of perfluorinated compounds contaminating high-voltage insulators. In addition to the somewhat equivocal cautionary statements in Will's book, I personally know of one lab which has banished these compounds from the premises because of an incident a number of years ago where the glassware used for cleaning of microscope parts got contaminated with Krytox -- this got transferred to several TEMs and resulted (so I'm told) in the need to replace multiple guns over a period of time -- and produced a lot of hair-pulling until the source of the problem was identified (actually, maybe this should be under the "horror stories" thread?). But this is the only direct report I have heard of such a problem and Will, though he notes the concerns, doesn't seem reluctant to recommend the stuff for EM use (and I do respect his depth of experience in such things). So my question: Is Krytox really the "bogeyman" I've been told it is? Or is this just another bit of microscopy folklore?
Given that: (1) there are lots of ways a vacuum grease could get transferred to a gun indirectly (i.e., via tools or contaminated gloves); (2) this stuff is reported to be essentially impossible to remove once it gets deposited on something; (3) the purported HV discharge behavior doesn't show up immediately but develops gradually over time; and (4) insulator cleanliness is enough of a problem without introducing this kind of sneaky contaminant -- IF true, it would seem that this class of compounds has no place in an EM lab. But if this is all a myth, I'm depriving myself of some otherwise great products. Insight anyone?
Most sputter coater manufacturers use one of two methods for mounting their targets.
1. Target material fixed by a clip or adhesive to an aluminium base (difficult to sputter without a special coater)
2. Target material fixed by adhesive to a brass base (this will sputter in relation to its composition e.g. 70%Cu 30%Zn)
You seem to have the type (2) target and are probably sputtering a mixture of gold (from the outside of the target) and brass (from the middle of the target). Check it out if you have x-ray analysis?
Sputter targets last a period in direct relationship to target thickness, coating thickness and degree of use, how long is a piece of string?
Good luck
Steve Chapman Senior Consultant Protrain - professional training in EM world wide Tel +44 1280 814774 Fax +44 1280914007 e-mail protrain-at-emcourses.com web site www.emcourses.com
"S" distortion or anisotropic distortion is produced through a balancing of aberrations. Take one imaging lens which is being used at low current and you will have pincushion distortion. Add another lens that is being used in the low to middle part of its current range and you will have pincushion distortion. Use the two lenses in a magnification system and the result is an image with anisotropic distortion.
When we design a TEM we try to balance the lenses so that the degree of distortion is minimised. Minimised does not mean totally removed it means NO distortion in the area that falls within the photographic frame. In early instruments, where the screen was very small, you hardly ever saw the problem, as screens have increased in size so the problem becomes more clear.
If the distortion has just started to become annoying this may be due to one or two reasons.
1) The high voltage has changed such that the lenses are not matched to the accelerating voltage as they were when the system was new?
2) A lens is not running at the correct level, its current is too low?
The most common reason for a change in lens or high voltage performance are their reference circuits, I am afraid I am not familiar with these circuits in the 2000FX.
Good luck hope this helps?
Steve Chapman Senior Consultant Protrain - professional training in EM world wide Tel +44 1280 814774 Fax +44 1280914007 e-mail protrain-at-emcourses.com web site www.emcourses.com
Joyce Craig wrote: =================================== How long should a gold target last? Our target looks gold except in the center. Problem is that we don't seem to get good coating. =================================== This is not that uncommon of a question. The first rule of thumb is that if it does not look like gold, then it probably is not gold. There is one exception however, and that is when the sputtering process creates some kind of surface structure that leads to an optical effect, a non-gold looking gold color (more like grey). But that typically does not adversely effect the sputtering rate.
The other cases then would be either a) contamination from external sources (e.g. finger prints) or b) build up of contaminants from the use of gold with insufficient purity. If (a), then that "problem" is solved by a good solvent washing and scrubbing with something like acetone. If (b), then it will not rub off with solvent and it would be something building up in the way of impurities from within the gold foil itself.
There is a cost associated with taking gold from 0.98 to 0.99 and even more of a cost to 0.999 purity, then then to 0.9999 or 0.99995 still more cost. In other words, you really do want high purity gold in the cathodes, for this very reason, but the higher purities do cost more money. Putting it another way, a cathode of 0.9999 for example is a lot more expensive than one that is 0.98 or 0.99, even though the net gold content is about the same . I won't even begin to speculate on how many would see a difference between 0.99995 vs. 0.9999 or even 0.999. But there is a point where the impurity elements that don't sputter begin to build upon the surface, resulting in a mostly non-gold layer. It is my understanding that the original equipment manufacturers of sputter coaters and also the main firms offering replacement cathodes supply only cathodes at the higher end of the purity scale because of this reason. If you seek non-traditional EM sources as alternatives, you really have to see the documentation to know that you are getting high purity, but most of the alternative sources do not deal in such high purity gold.
If you feel you have followed some of the advice given out on this listserver about possibly saving money and you could be having a build up of alloying or contaminating elements, try polishing off this layer in a metallographic polishing table, in order to renew the original composition that apparently did work for some time. If you do this, I would be most appreciative if you could share your experience with the list, be it good or bad. It might create more of an awareness of the need for high purity gold when making cathodes.
Disclaimer: SPI Supplies offers replacement gold and other metal cathodes for sputter coaters used in the SEM laboratory so we would have a vested interest in customers purchasing their cathodes from SPI or our major competitors in the EM supplies business.
Chuck
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Frederick Schamber 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 actually a follow-on question to the thread on "Silicone Oils and } Greases" which ran a few days ago. } } In his typically thorough response to the discussion of why microscopists } generally avoid silicone-based vacuum greases, Will Bigelow noted that } } } While the silicone high vacuum grease is } } indeed a good lubricant for O-rings, it is no better than the Brayco and } } Krytox greases, which are based on polyphenylether compounds, and which do } } not introduce the possibility of having insoluble siliceous compounds } } formed on critical parts of the electron optical column. } } } } This mention of Krytox rang a bell and I checked Chapter 10 of Will's book } "Vacuum Methods in Electron Microscopy" where he discusses oils and greases. } In table 10.1 he lists both Braycote and Krytox as "Perfluorinated Polyether } based" (not polyphenylether). In the subsequent discussion he states that they } "probably should not be used ... in or near the electron gun because of the } possibility that some of the perfluorinated base compound might get onto the } high voltage insulator and cause microdischarges ... ". In chapter 5 he is } more specific in his discussion of perfluorinated polyether diffusion pump oils } where he again mentions Krytox and states that use of these pump oils was } generally abandoned because it was found that "electron microscopes in which } these fluids were used eventually developed high-voltage instabilities due to } micro-discharges along the ceramic insulators in the electron guns." Will goes } on to note that the problem seems to be more severe in TEMs (which operate at } higher voltages) and these fluids "... have been used successfully for several } years in scanning electron microscopes in some laboratories". } } First question: } I'm confused by Will's list-server statement that Krytox is a polyphenylether } when it is listed in his book as a perfluorinated polyether. Not being an } organic (or any other kind) of chemist, I might think that these are synonyms } for the same family of materials except that Will also has a separate } discussion of polyphenyl ether pump fluids (e.g., Convalex and Santovac) which } is separate from that of the perfluorinated polyethers (Krytox and Fomblin). I } think that Krytox actually is a perfluorinated polyether, isn't it? } } Second question: } The issue of whether Krytox and Braycote are perfluorinated polyethers is more } than a trivial nomenclature issue because of the alleged problem of } perfluorinated compounds contaminating high-voltage insulators. In addition to } the somewhat equivocal cautionary statements in Will's book, I personally know } of one lab which has banished these compounds from the premises because of an } incident a number of years ago where the glassware used for cleaning of } microscope parts got contaminated with Krytox -- this got transferred to } several TEMs and resulted (so I'm told) in the need to replace multiple guns } over a period of time -- and produced a lot of hair-pulling until the source of } the problem was identified (actually, maybe this should be under the "horror } stories" thread?). But this is the only direct report I have heard of such a } problem and Will, though he notes the concerns, doesn't seem reluctant to } recommend the stuff for EM use (and I do respect his depth of experience in } such things). So my question: Is Krytox really the "bogeyman" I've been told } it is? Or is this just another bit of microscopy folklore? } } Given that: (1) there are lots of ways a vacuum grease could get transferred to } a gun indirectly (i.e., via tools or contaminated gloves); (2) this stuff is } reported to be essentially impossible to remove once it gets deposited on } something; (3) the purported HV discharge behavior doesn't show up immediately } but develops gradually over time; and (4) insulator cleanliness is enough of a } problem without introducing this kind of sneaky contaminant -- IF true, it } would seem that this class of compounds has no place in an EM lab. But if this } is all a myth, I'm depriving myself of some otherwise great products. Insight } anyone? } } Fred Schamber
Fred, I've used Brayco 803 since about 1980 for static seals and have never had any problems. I used Apiezon L for dynamic seals until I was introdused to Brayco 602 at which point I noticed cleaner systems once the Apiezon was gone. Apiezon's vapor pressure numbers look pretty good until you elevate the temp a little (120F) and then you lose a couple of decades, if I recall correctly, and the stuff polymerizes like crazy in an e-beam, but it IS slippery. I've been using the Brayco 602 in gun translators for a number of years with no problems and much cleaner guns.
One other thing I've noticed: Old Apiezon oxidizes and turns to a sticky sludge but the Brayco greases don't seem to age at all.
Ken Converse owner Quality Images Delta, PA 17314 717-456-5491
I am an undergrad from Miami University of Ohio, and am beginning TEM research with Newt retina and lens. I have researched various sources to no avail. Luckily, I was introduced to your network of Microscopists. } } My main problem is finding a thorough protocol for sample preparation that includes such details as primary fixation, buffers, resin-type, and accurate times/temperatures. Using the limited knowledge I have aquired I was able to put together a experimental protocol, which is polymerizing at this moment. I would greatly appreciate any advice or leads on proper protocol in this area. } } } }
} Thanks, } } Thomas Litzinger
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Email: baddis-at-olypen Name: barnett addis Question: I am a retired Phd from another remote disipline in another era who has become interested im micromount minerals and wish to do some image grabbing with a mieji trinoc(on order) but have reached a quandry re cameras. Has anyone had any hands on experience with a pixeria or a kodak mds100.or other lower priced rigs. i am loking at these as affordable options that may provide a better image than the tried and true method of a color ( below $1,000) video camera and "snappy" as the grabber. Any comments, rumors, suggestions would be most appreciated
Hi Ian, the only really successful way to get rid of epoxy is to use fuming nitric or sulphuric. You can also try aggressive solvents like tetrahydrofuran or dimethylsulfoxide. What will work usually depends upon the composition of your sample - if it also etches in these acids, you may have difficulties. I can get you some recipes from the books on de-encapsulation of silicon chips we have here, if you like.
All these chemicals (particularly fuming nitric) are very unpleasant and need special precautions for use. Swelling of the epoxy is inevitable, and if the specimen is delicate you may need to go through several cycles of nitric/solvent to remove the epoxy without putting too much stress on your sample.
Richard
} Dear all, } I have made a cross section of something using M-Bond 610 resin and it } shifted in the clamp while curing so I now have a cross section that would } be almost impossible to polish to leave the interface vertical. I would } prefer not to just throw the piece in the bin and start again, due to lack } of material. Does anyone know a way to dissolve fully cured M-bond 610 so } that I could start again from scratch? } } Thanks } } Ian MacLaren } Beijing Laboratory of Electron Microscopy } Chinese Academy of Sciences } P.O. Box 2724 } 100080 Beijing } China } General Email: ian.maclaren-at-physics.org } Work (esp. large attachments): maclaren-at-image.blem.ac.cn } } ______________________________________________________ } Get Your Private, Free Email at http://www.hotmail.com
============================================================== Richard Beanland Caswell Technology, Caswell, Towcester, Northants NN12 8EQ
e-mail richard.beanland-at-gecm.com Tel. +44 1327 356363 Fax. +44 1327 356398 ============================================================== "The information contained in this message is legally privileged and confidential information intended for the eyes of the individual or entity named above. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution or copying of this message is strictly prohibited. If you have received this message in error, please notify us immediately by telephone. Caswell Technology is the trading name of Marconi Caswell Limited. Registered in London No. 3694360 Registered Office: One Bruton Street London W1X 8AQ. Holding Company: Marconi plc."
I had quite a few suggestions for getting rid of creases and wrinkles in semi-thin sections - as promised to some of you, here is a summary. At the end, i have appended some old references which I dug out of my store over the weekend. i don't know if they all/if any work! I am still trying. If I get good results, I may post another message! Meanwhile, on with the folk-lore!
And waiting for the moon to rise!
Keith Ryan Marine Biological association Plymouth UK
Folds in semi-thin sections
1. Use lower drying/staining temperatures - e.g. 60 C - when hotplate is still warming up 2. Dry flat at 50-55 C then go to hotter plate for 10-15 minutes 3. Longer on the hotplate 4. Dry sections on slide using a flame 5. Set hotplate to just below boiling the water 6. Vapour (6 messages) 7. Transfer to 10% acetone droplets 8. Harder resin (2 messages) 9. Softer resin 10. Longer infiltration times 11. Improper mixing 12. Heat pen (3 messages) 13. Chien grids (grid with 2 mm hole and tab for handling) to transfer thicks to water droplets 14. Wick away the water once section is spread 15. Use big drops, lots of water 16. Cut thinner sections 17. Cleanliness - of all items 18. Stain on drop of stain on hotplate, steam, transfer, water rinses, dry down 19. Put a nick in the corner of the cutting face so that the resin can expand differentially to the enclosed specimen 20. Etch the sections with ethoxide (ethanol + NaOH) 21. Remove resin to lessen apparent effect? Using saturated NaOH in methanol.
22. Coat the slide - One droplet of protein-glycerol is diluted in 1 ml dist. water, then the glass slide is dipped into this solution and dried in an oven at 30 C. Protein-glycerol: dissolve 1 gram ALBUMIN (MERCK) in 9 gram bidist. water at 37xC in an oven. Filter the solution and mix with the same amount of glycerol.
23. W.M. Harris (1978). Stain Technol. 53: 298-300. Transfer sections to a large drop of 10% acetone. Dry on hotplate set to 122 C (250 F). It says that! If the sections are known to be difficult to flatten, add more 10% acetone during the process. Remove the sections from the hotplate as soon as they are dried down. EXCESSIVE HEAT AFTER DRYING DOWN CAUSES WRINKLES. Finally, [place slides on a warming tray at 40-50 C and allow to dry down thoroughly, 30 minutes to overnight. Then stain.
24. M. Martins-Green (1978). Stain Technol. 53: 296-300. Sections spread with xylene vapour. Floated on toluidine blue stain (diluted, 1 ml of 1% stain plus 20 mls 2.5% sodium carbonate sol.) at 50 C on hotplate for 1 hour, then left in the dark at room temperature for 24 hours). Rinsing - not mentioned. Dried on 2-3 drops of double-distilled water for 10 minutes at 50 C. Cover-slipped. Method designed to allow collagen fibres to fully distend after cutting.
25. J.R. Sommer et al. (1979) Stain Technol. 54: 106-107 "wrinkles appear during staining rather than while the sections are being dried on the glass slides - eliminated when the stain is made up with 50% with respect to glycerol. Staining is not rapid - 12-24 hours in a covered dish at 50 C. Slides need not be albuminized" Imply staining by flotation at 60 C in a covered dish 12-24 hours, rinse briefly, dry on slides and view (without a coverslip).
26. J. Millonig (1980) Stain Technol. 55: 118-120. Paraphrasing: Wrinkles appear when sections are heated during staining, more so when there is a rim of resin around the tissue. During heating, the tissue expands more than the tissue. Transfer sections onto a drop of stain on the end of a slide. Heat in a flame. Place on a staining bridge (usually 2 minutes). After cooling, float sections off on water. Rinse in another beaker, transfer to acidified water. Transfer to slide, warm in a flame, wick away excess water. Do not need to albuminise the slide. 0.5 micron sections stain in abouit 2 minutes, thinner sections need a second heating and should be left floating on stain for longer.
27. N.B. Chandler & G.C. Schoenwlf (1983). Stain Technol. 58: 238-240. State that "wrinkling occurs only when mounted sections are *. heated". Sections dried overnight at 76 C on acid cleaned slides, prior to staining (76 C was the maximum temp. of their hotplate). Acid cleaning: soak slides for a minimum of 1 hour in 10% potassium dichromate plus 10% conc. sulphuric acid. Sections dried at 60 C often wrinkled, while those dried at 90 C often failed to stain properly. Sections dried less than 6 hours at 76 C often wrinkled during staining - hence dry overnight. Staining can be controlled more consistently in Coplin jars placed in a water bath rather than on a hotplate.
My horror story is pretty recent: Just last January. We had a local SEM service guy come in to do a semi-annual tune-up on our ESEM, and he did a great job. He did notice, though, that our ion pump wasn't giving us quite as good a vacuum as it should, and was kind of slow doing it. When he left he said "All you need to do is bake it off a bit with some heat for a few hours - that'll refresh the active ingredient (or parts) inside and it'll work better for you. I've got a heater that's designed for just such a job." True to his word, a couple days later he dropped off this device for me. Just a biggish aluminum box in two parts, with luggage clips to lock it together and a 1500 W heater element inside. Simplicity itself to use, just turn off your ion pump, clip this box over/around it, and plug it in for a while. The heater element rests right against the back of the ion pump. So I mount it on there, plug it in, wait a bit, and sure enough, things start heating up in there. OK, I think, I'll go see my colleague upstairs for a few minutes about those samples she was preparing..... I'm back down in the lab ten minutes later, and go back behind the scope to see how things are doing. There's a small hole in the back of the heater apparatus, and when I happen to glance in there, I see a small wall of blue flame. "This can't be good", thinks I....then "Now which type of fire extinguisher do you use on a burning ESEM? Water?......No, probably not. Powder?......uhhhhh.......no. CO2? Probably..." Meanwhile, I unplug the heater and mostly just stand there....thinking how close I've come to pensionable retirement age, only to lose it like this. But anyway, the flame started to die back a bit as soon as I unplugged the thing, and with some damp towels I was soon able to unclip it and remove it. It turns out our particular instrument had a clip mounted on the back of the ion pump to retain the HT cable in place, and this clip had been mounted on piece of black plastic, which I swear to God I thought was anodized aluminum. The plastic had melted with all that intense heat of course, and dripped right down onto the heater element, where it had burst into flame. I had our machine shop make me a metal replacement for the lost plastic bit, there was no other damage, by God, the ion pump now works great, and I figure local management doesn't need to hear about every little detail of life in the SEM lab, now, do they?
Frank Thomas MicroAnalysis Facility Geological Survey of Canada (Atlantic) Bedford Institute of Oceanography Dartmouth, Nova Scotia
G'day Cobbers! I could use some handy hints here. We've a student who wishes to analyse some hideous sludge - something to do with sewage me thinks. She is particularly interested in the state of the metals in this muck - whether it is present as sulphides, sulphates, nitrides, nitrates or bound to organic molecules. The stuff is rather fine grained, and I've suggested she filter and retain the } 10 micron fraction. She will then press it into a pellet with a flat, shiney surface if all goes well. Basically, I'm interested if there is anyone out there who has experience of analysing this type of material. What did you coat it with? How did you differentiate between the the various form of the metal compounds? Any other handy hints also gratefully received. Cheers, Malc.
-- Dr MP Roberts Phone: [+27](0)46 603 8313 Dept of Geology Fax: [+27](0)46 622 9715 Rhodes University Cell: 083 4060 262 (usually off) 6140 Grahamstown e-mail: malc-at-rock.ru.ac.za SOUTH AFRICA
I'm not sure what the Measurements Group uses to cross-link the M-Bond gage cement. The resin is listed as an "epoxy-phenolic adhesive". In several anhydride cross linked systems I've found that sodium ethoxide in ethanol worked well to de-resin samples. I made mine fresh from sodium and ethanol but fresh anhydrous AR grade should work as well. Something like 5 or 10% in dry ethanol should do it. This may be easier on your samples if they are acid sensitive. I'd be curious what works as I can see myself in a similar predicament someday. cheers, John
John Heckman MSM Department Michigan State University
} ------------------------------------------------------------------------ } 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 Ian, } the only really successful way to get rid of epoxy is to use fuming nitric or sulphuric. You can also try aggressive solvents like tetrahydrofuran or dimethylsulfoxide. What will work usually depends upon the composition of your sample - if it also etches in these acids, you may have difficulties. I can get you some recipes from the books on de-encapsulation of silicon chips we have here, if you like. } } All these chemicals (particularly fuming nitric) are very unpleasant and need special precautions for use. Swelling of the epoxy is inevitable, and if the specimen is delicate you may need to go through several cycles of nitric/solvent to remove the epoxy without putting too much stress on your sample. } } Richard } } } Dear all, } } I have made a cross section of something using M-Bond 610 resin and it } } shifted in the clamp while curing so I now have a cross section that would } } be almost impossible to polish to leave the interface vertical. I would } } prefer not to just throw the piece in the bin and start again, due to lack } } of material. Does anyone know a way to dissolve fully cured M-bond 610 so } } that I could start again from scratch? } } } } Thanks } } } } Ian MacLaren } } Beijing Laboratory of Electron Microscopy } } Chinese Academy of Sciences } } P.O. Box 2724 } } 100080 Beijing } } China } } General Email: ian.maclaren-at-physics.org } } Work (esp. large attachments): maclaren-at-image.blem.ac.cn } } } } ______________________________________________________ } } Get Your Private, Free Email at http://www.hotmail.com } } ============================================================== } Richard Beanland } Caswell Technology, } Caswell, } Towcester, } Northants NN12 8EQ } } e-mail richard.beanland-at-gecm.com } Tel. +44 1327 356363 } Fax. +44 1327 356398 } ============================================================== } "The information contained in this message is legally privileged and confidential information intended for the eyes of the individual or entity named above. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution or copying of this message is strictly prohibited. If you have received this message in error, please notify us immediately by telephone. } Caswell Technology is the trading name of Marconi Caswell Limited. Registered in London No. 3694360 Registered Office: One Bruton Street London W1X 8AQ. Holding Company: Marconi plc."
A microprobe wouldn't be my first choice for analyzing the sludge. Essentially, you'd wind up with a list of elements. Old fashioned x-ray diffractometry would yield a list of PHASES, which would be a much greater help. A microprobe's list of elements would, of course, be a great aid in the x-ray diffraction search/match operation. Check out the International Centre for Diffraction Data's web site for the latest computer search/matching things. www.icdd.com (or is it .org?)
Ron
Cobber?
(I have no financial interest in the ICDD)
Ron Anderson, IBM, Hopewell Jct., New York, USA. anderron-at-us.ibm.com
IBM Analytical Services; http://www.chips.ibm.com/services/asg
Dr Malcolm Roberts {malc-at-rock.ru.ac.za} on 04/10/2000 06:34:49 AM
To: Microprobe discussion group {Microscopy-at-sparc5.microscopy.com} cc:
G'day Cobbers! I could use some handy hints here. We've a student who wishes to analyse some hideous sludge - something to do with sewage me thinks. She is particularly interested in the state of the metals in this muck - whether it is present as sulphides, sulphates, nitrides, nitrates or bound to organic molecules. The stuff is rather fine grained, and I've suggested she filter and retain the } 10 micron fraction. She will then press it into a pellet with a flat, shiney surface if all goes well. Basically, I'm interested if there is anyone out there who has experience of analysing this type of material. What did you coat it with? How did you differentiate between the the various form of the metal compounds? Any other handy hints also gratefully received. Cheers, Malc.
-- Dr MP Roberts Phone: [+27](0)46 603 8313 Dept of Geology Fax: [+27](0)46 622 9715 Rhodes University Cell: 083 4060 262 (usually off) 6140 Grahamstown e-mail: malc-at-rock.ru.ac.za SOUTH AFRICA
We are attempting to critical point dry 100 micron thick Vibrotome cut cross sections of annelids (leeches). The end product looks fine in the SEM, except that the sections have curled/rolled up during the process, and we want them to remain flat. Attempts to flatten them after drying have not been very successful. Has anyone devised some sort of holder (sandwich?) that might overcome this problem by keeping them flat during their trip through the dryer?
This worked for a student a few months back working on nanoporous zirconia.
Ion mill the sample from one side long enough to remove the polishing damage from that side. Remove from the ion mill and coat the milled side with carbon. Return to the Ion mill and mill on the uncoated side. The carbon on the other side will help keep the thinned grains from falling out.
I got this trick from Scott Walck some time ago.
Ray
************************* Ray D. Twesten, PhD Center for Microanalysis of Materials University of Illinois (217) 244-6177 fax:(217) 244-2278
----- Original Message ----- } From: "Valerie Leppert" {vjleppert-at-ucdavis.edu} To: "'Microscopy Listserver'" {Microscopy-at-sparc5.microscopy.com} Sent: Friday, April 07, 2000 4:55 PM
Hi,
If you give me a worm and tell me to produce semi-thin sections that wrinkle, and semi-thin sections that do not wrinkle, I can do it. But I might need two worms!
Stretching, hot plate and water manipulations, cutting contortions, cleanliness, etc., are the band-aids. These methods do not address the basic problems of which there are at least two.
At the very base of the problem usually lies the fact that there is too high of a percentage of unbound monomers in the tissue. This can happen in at least several ways 1) The use of a good formulation which is not suitable for the tissue 2) the use of a formulation which has no basis in correct composition as far as molecular weights and WPE #s are concerned. These formulations are usually handed down from lab to lab and no one knows their origins. 3) inadequate infiltration procedures of a good and suitable formulation. All of the above will leave too many unbound monomers in the tissue.
So what? Unbound monomers are extremely active. They attract water, swell, pull, stretch the embedded tissue. Upon drying the water leaves since it is mechanically bound, and presto, nice wrinkles.
Let us look at 1) above. A formulation containing Araldite, and a hefty amount of DDSA is unsuitable for tough skin, collagen, worm architecture, and so on. Why? Because Araldite and DDSA are very long chains, and are extremely difficult to keep EVENLY mixed and EVENLY infiltrated into tough tissue. Note the capitalization of evenly! Some of the Araldite will stay outside of the worm gut, and there will be more there than inside the gut. Inside the gut there is an excess of DDSA. Crosslinkage and stability are reduced. Water will be absorbed in the boat by these loose monomers.
I cannot deal with 2) at all. I never use any formulation unless I understand it. No telling what the proportions are!
Let us look at 3). Inadequate infiltration procedures account for more trouble than than Monica Lewinsky! Formulations seperate, are allowed to infiltrate unevenly, or not thoroughly enough, or not slowly enough. Perhaps the dehydration was not adequate, and the formulation will not take up a space occupied by a polar substance. Again, you have loose monomers which should be bound or crosslinked.
What would I do with my worms? For wrinkles I would use Araldite-DDSA-Epon, infiltrate it poorly, underpolymerize it, and watch the pleats appear. For flat sections I would pick a formulation which would fit the material - perhaps the next to the hardest Luft's formulation, infiltrate it like crazy, never let it sit in the hood, heat every new change of new mixture to 37 degrees for one hour and a half with a light bulb while they tissue rotates, do many changes over perhaps 48 hours, at least. I would have really dehydrated well, gone to Propylene oxide, the infiltrate starting with a mixture of 3 parts propylene oxide and one part formulation, gradually working up to full epoxy mixture.
I would polymerize for 48 hours at least and cut the thinnest section I could tolerate for my study. I would use a sharp knife (of course).
Now, it might not work well the first time. So I would manipulate all the above until I was happy.
NOW. PLEASE NOTE: Suppose I have sections which are a little wrinkled, or some old blocks which yield severely wrinkled sections. I soak the slides in rt water for half an hour or less depending on how well they will stick to the slide. Then I would put the slides into a vaccum jar which has that strong, powdered dessicant which we used to use for drying EM film in a petri dish in the bottom. (Protect the slides from flying poweder by covering with hardened filter paper). Then evacuate the jar just short of implosion! Release the vaccum in about 24-48 hours. This will often save poor blocks or totally get rid of slight wrinkles.
Crowley, Hildegard Heinrich, and Ben H. Leichtling. Elimination or Reduction of Wrinkles in Semithin Epoxy Sections by Vaccum Drying. Stain Technology,Vol 64, No 5, September 1989.
Please do not ask me for reprints. I do not have any.
Have fun with the worms! Keep records! Know what you have done, and what you have not done!
Hildy
P.S. Very important! Formulations made from epon substitutes which contain Araldite or dilutants may not be suitable for use with difficult materials, since they may be too resilient or elastic. We have used Eponate 12 from Pella forever. Many years ago I had information regarding the spec readings of this substitute - it was very much like the old Epon. I am hazy on this point. Do not quote me. I do not have any monetary interests in Pella.
I'd suggest you also do so good old fashioned light microscopy on a "sludge smear". We've had the opportunity to use McCrone's Particle Atlas on CD ROM on several courses recently and have found it invaluable in identifying things like this. McCrone was running a special at PITTCON: $700 instead of the usual $900. The last time the PA came out in print, it was approximately 10 volumes in length. The CD has light, electron, and elemental analysis info. I would be surprised if there isn't a copy of either the print or CD version in your library.
Caveat: MME has no financial interest in this product.
Good hunting! Barbara Foster,President Microscopy/Microscopy Education 125 Paridon Street Suite 102 Springfield, MA 01118-2130 PH: 413-746-6931 FX: 413-746-9311 email: MME-at-map.com Website: www.MME-Microscopy.com/education
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At 12:34 PM 4/10/00 +0200, Dr Malcolm Roberts wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am writing this on behalf of an experienced SEM Technician, new to Canada who is looking for a part or full time position, (will even work as a volunteer for now) in the Greater Toronto Area.
She has 20 years of experience working on various materials on JEOL SEM's in Hungary but needs help in establishing contacts and learning the industry here.
Please contact me off-line if you are interested.
Karen Rethoret York University Microscopy Facility Toronto, Ontario 416-736-2100 x33289
} I could use some handy hints here. We've a student who wishes to } analyse some hideous sludge - something to do with sewage me thinks. She } is particularly interested in the state of the metals in this muck - } whether it is present as sulphides, sulphates, nitrides, nitrates or } bound to organic molecules.
I reported on using EDS to localize PCBs in sediment at the Microscopy and Microanalysis meeting in 1998 (page 498 in the proceedings). Fortunately for me, there is no Cl in sediments, so just locating Cl was indicative of PCBs. Your student could get chemical info from microanalysis with very high energy resolution by looking at the fine structure. Either EELS (on an instrument with a FEG) or WDS could have this resolution, and if the new technologies of superconducting tunnel junction diodes or micro- calorimeter detectors are available, they could provide sufficient resolution for EDS. Taking position-tagged spectra on, e.g., a Zeiss 912 could give her both the localization and chemical info she needs. I have no interest in either Zeiss or the new types of detector except as a would-be user and techno-geek.
} The stuff is rather fine grained, and I've } suggested she filter and retain the } 10 micron fraction. She will } then press it into a pellet with a flat, shiney surface if all goes well.
I just suspended the sediment, put a small amount on a grid, and took images and spectra. I used all but the mm-sized particles; most were in the micron to submicron size range. They were well- dispersed on the grid when the dilution of the suspension was ap- propriate, and I could get spectra from individual particles. This approach may be better than the pellet, since different particles can have different chemistries.
} Basically, I'm interested if there is anyone out there who has } experience of analysing this type of material. What did you coat it } with?
I didn't coat it; the high-voltage microscope is capable of getting good images from this material as is (no charging). The IVEM, likewise, was able to get good spectra.
} How did you differentiate between the the various form of the } metal compounds? Any other handy hints also gratefully received.
As I said, I didn't need to get chemical info, and as another poster said, using diffraction to characterize the material can add info about the form the metal is in. Reflection high-energy elec- tron diffraction (RHEED), low-energy electron diffraction (LEED), and photoelectron holography (PEH) can give info about surface structures (see, e.g., Leslie et al. (1999) Electron Crystallography in Surface Structure Analysis, Microscopy Research and Technique 46:160-177). If the metals are adsorbed to the sludge by adhering to a facet with a particular crystallographic orientation, either as occasional adatoms or, better still, as an epitaxial deposit, such surface techniques could be useful. Perhaps Larry Marks is the best person to talk to about this. Good luck. Yours, Bill Tivol
My profound apologies to everyone who read my comments on silicone oils and greases, in which I said that Brayco and Krytox greases are based on polyphenyl ether compounds. I was in too much of a hurry, and neglected to check my memory (which isn't getting any better as my maturity progresses).
Fred Schamber is right in calling attention to the fact that these greases are based on perfluorinated polyether compounds.
Thus, the information given on page 460 of my book 'Vacuum Methods in Electron Microscopy' is correct: the Brayco and Krytox greases are based on perfluorinated polyether compounds, NOT polyphenyl ether compounds.
Furthermore, Fred is correct in stating that great care should be exercised to keep the perfluoro-polyether compounds out of the electron guns of electron microscopes, because, as discussed on p. 187 of Vac. Meth. in EM, they have been known to cause microdischarges in electron guns causing undesirable high-voltage instabilities.
Although I have not bothered to try to keep up with all new developments since I finished writing my book, back in '94, I am not aware of any vacuum grease that is based on polyphenylether fluids. This is peculiar, because the polyphenyl ether diffusion pump fluids, Santovac-5, Excello-54, and Convalex-10, have all worked out so well. A grease is made by combining an oil with a gel-forming agent. Thus, to make an ordinary lubricating grease one might mix a soap such as sodium stearate with an ordinary lubricating oil. When heated to a proper temperature the oil and soap will interact to form a gel, which we call a grease. Thus, to make a grease based on the polyphenyl ether oils, all one would have to do is to find the appropriate gelling agent. This is not an entirely trivial task, however, because unless the correct agent is found the grease will 'bleed' (i.e. the oil will separate from the gelling agent), especially if the grease is heated or exposed to a vacuum. If a grease bleeds, you can end up with only the gelling agent remaining on the bearing or gasket that you wanted to lubricate, while the oil (which is the actual lubricating agent) spreads around and contaminates adjacent parts. The silicone- and perfluorinated polyether-based greases are remarkably stable and resistant to bleeding, whereas some other high vacuum greases are not.
Again, I apologize for my mistake.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
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I have also had an interesting, and aggravating problem with sputter coater targets recently. We have an older model Hexland (bought out by Oxford about 10 years ago) SEM cryo stage. Although not as fancy as current models, it usually does a reasonable job. Quite a number of years ago I purchased a large gold disc sputter coater target. I cut the 12mm discs needed for the sputter coater in the cryo prechamber from this larger disk with a simple stamping tool. This worked for years without problems. About a year or so ago, we started having problems getting good coatings...noticable because of excessive charging of coated samples. We immediately thought of contamination because the target was becoming discolored after only a couple of uses. If I removed it, cleaned it with metal polish, etc and reinstalled it, all was well for another couple of coatings. We tried new pumps, looking for oil leaks etc. but a good vacuum in the chamber ruled out large leaks and all other signs were negative. We also changed the argon tank thinking that perhaps this could be a source of contamination. The cryo stage is used in spurts and after a period of inactivity, we are again gearing up for heavier use. Recently the target was only giving a good coating on one sample before having to be cleaned....a major job when you have to warm to room temp from -170oC prior to opening up the chamber. I finally took the used target over to a Microprobe on campus and did a WDS analysis on it....turns out that the contamination was aluminum. The housing that holds the target in place is aluminum. Somehow the aluminum is being sputtered as well as the gold, producing a poor metal coating and contaminating the target as well. Oxford is trying to get replacement parts for the sputter head but we are at a loss to figure out why, after all these years, this is happening and what we can do about it if replacement parts cannot be found. By the way, a new crystage runs about $70,000 so we are not contemplating that move at the moment. Debby
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057 On Saturday, April 8, 2000, Garber, Charles A. {cgarber-at-2spi.com} wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id SAA04459 for dist-Microscopy; Mon, 10 Apr 2000 18:01:28 -0500 (CDT) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id SAA04454 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Mon, 10 Apr 2000 18:00:58 -0500 (CDT) Received: from bsd01.community.net.uk (bsd01.community.net.uk [195.72.164.132]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id SAA04447 for {Microscopy-at-sparc5.microscopy.com} ; Mon, 10 Apr 2000 18:00:44 -0500 (CDT) Received: from [195.72.167.200] ([195.72.167.200]) by bsd01.community.net.uk (8.9.3/8.9.3) with SMTP id XAA44780 for {Microscopy-at-MSA.Microscopy.com} ; Mon, 10 Apr 2000 23:55:21 +0100 (BST) Message-Id: {200004102255.XAA44780-at-bsd01.community.net.uk} To: MICROSCOPY BB {Microscopy-at-sparc5.microscopy.com}
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Ian MacLaren wrote: ======================================================= } Dear all, } I have made a cross section of something using M-Bond 610 resin and it } shifted in the clamp while curing so I now have a cross section that would
} be almost impossible to polish to leave the interface vertical. I would } prefer not to just throw the piece in the bin and start again, due to lack
} of material. Does anyone know a way to dissolve fully cured M-bond 610 so
} that I could start again from scratch? } } Thanks } } Ian MacLaren } Beijing Laboratory of Electron Microscopy } Chinese Academy of Sciences } P.O. Box 2724 } 100080 Beijing } China } General Email: ian.maclaren-at-physics.org } Work (esp. large attachments): maclaren-at-image.blem.ac.cn ============================================================= We have found over the years that the easiest way to remove not just M-Bond 610 but just about any epoxy or other organic polymer from a metal substrate is with the use of oxygen plasma etching. It was my recollection that one of our systems was purchased by the KYKY part of your facility and they would probably let you have access to it.
Disclaimer: SPI Supplies manufactures the Plasma Prep II plasma etcher which can be used to remove organics, including intractable cross-linked polymer systems, like M-Bond 610.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.spi.cc ######################## ============================================
rtch (another cobber from the Southern half of the world)
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Here is my guess about what has happened. I read someplace that alumina (aluminum oxide) does not sputter easily whereas pure uncoated aluminum metal sputter easily. Aluminum in air is quickly coated with alumina and it is protected for a while when placed in an area subject to ion bombardment for sputtering. The alumina layer on your target holder has been worn away by years of sputtering and cleaning, and in the cold vacuum of the cryo prechamber was not getting replaced by oxidation.
Cure: Take the holder out and have it anodized (a hard finish) or just heat it gently in air for a while to form a new layer of aluminum oxide.
Ronald Vane XEI Scientific
-----Original Message----- } From: Debby Sherman {sherman-at-btny.purdue.edu} To: message to: MSA list {microscopy-at-sparc5.microscopy.com}
Could the alumium have had some kind of coating to keep it from sputtering or could a short developed between the gold disk and the alumium housing? My guess is that there is a break down in the insulation between the target and the holder. If you have records of the current and voltage it sould show up when compaired with the old records when things were running right.
Could some other part of the system be contaminated by Al? If the rest of the system is not made of alumium you can use lye to clean it.
When a new machine costs $70 K you can afford to pay a good machinest for a lot of hours to make a part. With CAM mills they can make almost anything. And for what the computer controled mill can't make there are a few of us old guys that can finish the job with a file:) You porbably have a couple in one of the insterment shops on campus.
If you don't have a shop that can handle this kind of work I know a couple of guys out on the west coast that can. I don't have any connection with them except to admire their work.
Gordon
Gordon Couger gcouger-at-couger.com
Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
} } I have also had an interesting, and aggravating problem with sputter coater targets recently. We have an older model Hexland (bought out by Oxford about 10 years ago) SEM cryo stage. Although not as fancy as current models, it usually does a reasonable job. Quite a number of years ago I purchased a large gold disc sputter coater target. I cut the 12mm discs needed for the sputter coater in the cryo prechamber from this larger disk with a simple stamping tool. This worked for years without problems. } About a year or so ago, we started having problems getting good coatings...noticable because of excessive charging of coated samples. We immediately thought of contamination because the target was becoming discolored after only a couple of uses. If I removed it, cleaned it with metal polish, etc and reinstalled it, all was well for another couple of coatings. We tried new pumps, looking for oil leaks etc. but a good vacuum in the chamber ruled out large leaks and all other signs were negative. We also changed the argon tank thinking that perhaps this could be a source of contamination. } The cryo stage is used in spurts and after a period of inactivity, we are again gearing up for heavier use. Recently the target was only giving a good coating on one sample before having to be cleaned....a major job when you have to warm to room temp from -170oC prior to opening up the chamber. I finally took the used target over to a Microprobe on campus and did a WDS analysis on it....turns out that the contamination was aluminum. The housing that holds the target in place is aluminum. Somehow the aluminum is being sputtered as well as the gold, producing a poor metal coating and contaminating the target as well. } Oxford is trying to get replacement parts for the sputter head but we are at a loss to figure out why, after all these years, this is happening and what we can do about it if replacement parts cannot be found. } By the way, a new crystage runs about $70,000 so we are not contemplating that move at the moment. } Debby
The resin used was a TAAB Laboratories standard Hard pre-mix kit, mixed and kept in a freezer. Usually no problem but the resin mixture may have aged.
Infiltration may not have been sufficient, although it was overnight in the 50/50 mixture acetone/resin (our safety dept. is anti propylene oxide). We can usually get away with this procedure, but you may have some good points about this step. I am suddenly back about 20 years! Our library stopped taking Stain Technol. in the 1980's and I haven't searched it since!
The wrinkles seem to be most apparent in the area of the cuticle (which is really not very thick).
I hesitated a bit to post this message as I thought it would be too trivial, but my personal experience has shown me that people often do not know the basic concepts needed to acquire good quality images with a digital/video camera in light microscopy. I want to know if other people working in digital imaging agree on the very few concepts I think that are essential to acquire good quality digital images in microscopy.
I simply give an overview of the concepts for quantitative digital brightfield microscopy:
1) Understand the meaning of Numerical Aperture 2) Use white light, dim with neutral density filters if necessary, do not turn down the light to "reddish". 3) Set up Koehler illumination ! 4) Nyquist sampling (match magnification to CCD-array) 5) Understand the influence of the sampling density on the C.V. of your measurements Ian T. Young, Sampling density and quantitative microsocopy Analytical and Quantitative Cytology and Histology, vol. 10, 1988, pp. 269-275
6) Understand the dynamic range of your image acquisition system (camera + digitiser)
For a B/W camera: 1) Use a green filter for monochromatic light
For a color camera 1) Use a 3CCD camera, not a single CCD camera 2) Set the white balance
My personal opinion is that if you obey these basic rules you get good quality images, otherwise you don't. The quality of the images relates directly to the quality of your analysis and as such to the "quality" of your conclusions.
Regards,
Peter Van Osta, MD
Senior Scientist Medical Image Analysis Biological Imaging Laboratory Life Sciences Department I - 6065 Janssen Research Foundation Turnhoutseweg 30 B-2340 Beerse Belgium Europe
I am in the process of acquiring a Tissue Tek automatic tissue processor, second hand. I seem to have a choice between Tissue Tek II or Tissue Tek III, the later being more expensive of course. Since I have no experience with such machines, I would like to ask if in practice there are significant benifits from the more advanced model. Actually what I want is a reliable way of making lots of paraffin embeddings with a minimum of sofistication.
Thanks in advance for your answers
Dr. A.P. Alves de Matos Dental Medical School Lisbon
A grad student that I had recently trained to use our JEOL T-20 SEM was trying a little solo work. He had the specimen chamber door open, turned away from the scope, and looked back just in time to see one of our monster cockroaches disappearing into the scope. Rather than call me, or wait for the roach to reappear, he shut the door and pumped the scope down. He never did confess...
I will be more specific. I will need to process up to 60 specimens/week in two or three separated weeks during the year. The rest of the time hand processing is OK. Since the specimens are prone to dammage upon storage (immunohistochemical techniques on sponge tissues), I do not want to store them to facilitate processing. So, it is important to know if the machine can handle this amount of work. I have one technician that has many other things to do. Basic assistence should be available from our technical department. Anyhow I would not trust newer instruments just because they are more recent. I know of (and have) a few super-new machines (amaizingly expensive) that had to be thrown to garbage because no one (assistence included) was able to put them to regular work. It is important to know if the machine model is a well tested one that usually works for years with no problems or if it has regular problems. Since histology and histopathology labs of my knowledge use other tissue processors, I was not able to get specific advice locally about these machines.
Thanks A.P. Alves de Matos
----- Original Message ----- } From: Norm Granholm {granhona-at-email.uc.edu} To: A.P. Alves de Matos {apmatos-at-ip.pt} Sent: Tuesday, April 11, 2000 12:57 PM
Hello,
I`m looking for an EELS spectrum containing Oxygen K-edge and some other edge like Mn or Ti L-edge. Together with low loss spectrum and data about the angles. The spectra need to have a power of 2 (1024 for inst.) bins and low loss and high loss should have same size. I want to use this for comparison, i`m trying to do some quantitative EELS work but so far with limited success. I want to find out wether my program or my data is wrong;)
Thanks,
Jo
************************************************************* * Jo Verbeeck * * University of Antwerp * * Dept. EMAT (Electron Microscopy for Materials Research) * * e-mail: joverbee-at-ruca.ua.ac.be * * tel: +32(0)3 218 02 49 * * fax: +32(0)3 218 02 57 * *************************************************************
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Try sandwiching the sections between pieces of filter paper. We do this with lots of small samples which we are in danger of loosing. We use large washers as holders for the filter paper sandwiches. The pairs of washers can be secured using small binder clamps (an office product used like paper clips). Or you can knotch the washers so that you can use a piece of wire to hold them together. The knotches keep the wire from slipping off. If you cannot find washers with the correct internal and external dimensions, they are easily made in any machine shop. I would put the sandwiches together before or during dehydration and then carry the sections through the last ETOH changes as a unit. It takes a bit longer to CPD due to the absorption of the ETOH by the paper but you have a good chance of having flat sections at completion. Debby
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
On Monday, April 10, 2000, Dick Briggs {rbriggs-at-Science.Smith.edu} wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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Microscopist:
Our new lab is supposed to be built next to a four lane highway that has a lot of 18 wheeler truck traffic. I am concerned about vibration problems and would like to convince administration that the building should be built on the other side of the property as far away from the highway as possible. Any suggestions or known references covering this? Thanks in advance.
Phoebe J. Doss Manager/Adjunct Instructor Electron Microscope Lab Oklahoma State University
Looking for a few used B & L 7 to 30 power optical microscopes (or similar- A. O., etc.) to be given to local vo-tech school. I will pay shipping. Will trade something if required. Mike Urbanik www.crystalguru.com
This is a discussion archived on "Tips & Tricks" from a couple of years ago dealing with a similar problem. E-mail addresses of the poster and respondents are included so you might discuss things with them further.
Good luck
At 10:04 AM 4/11/2000 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall EM Technician Gainesville, FL 32610 University Of Florida ph 352-392-1184 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
They want to put your lab next to a 4-lane highway...this is a joke, right?
Larry ;-)
PS my suggested reference... 1. Common sense. Vol.1, No. 1, p. 1 another suggestion: make some vibration measurements and check it out...
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Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
Several years ago, Dow built a state-of-the-art analytical chemistry building (including a microscopy lab) in Midland, Michigan. One of the issues they addressed was nearby truck traffic, just like you have. They ended up closing the road. Bob Czeislinski (sp?), a member of this listservice, might be able to help you on some of the tech issues raised to get the change. (Sorry, I do not have his email address.)
} ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } Hildy } } Thanks for all the info. } } The resin used was a TAAB Laboratories standard Hard pre-mix kit, } mixed and kept in a freezer. Usually no problem but the resin mixture } may have aged. } } Infiltration may not have been sufficient, although it was overnight } in the 50/50 mixture acetone/resin (our safety dept. is anti propylene } oxide). We can usually get away with this procedure, but you may have } some good points about this step. I am suddenly back about 20 years! } Our library stopped taking Stain Technol. in the 1980's and I haven't } searched it since! } } The wrinkles seem to be most apparent in the area of the cuticle } (which is really not very thick). } } So long } } Keith Ryan } } Hi,
One should never leave tissue of any sort in a solution of solvent and epoxy overnight. It is deleterious to tissue structures and does not serve to infiltrate.
I do not have a worm. But if I suddenly had one to embed and I had no experience with worms, I would be very suspicious of it. I would immediately treat it as though it were really hard to handle. I would do all the dehydration steps for at least 30 min with a change of ethanol every 10 minutes. I would open a new bottle of 100% ethanol for the last three changes. I would use propylene oxide, and evaporate the waste from a bucket in the hood. I would use PO (dry acetone if you must) and epoxy in the ratios of 3 to 1 for 1/.2 hour, 2 to one for one hour, 1 to 1 for 2 hours. Then one hour in fresh expoxy. Then new, clean vials, with fresh epoxy. Perhaps 3 times for 2-3 hours each. Every time a new mixture is added I would heat with a light bulb directed at the rotator to 37 deg (not over 40) for about 1.5 hours. Then I would leave it overnight on the rotator, and start again in the morning. A pain in the neck! But then, science frequently sucks! In the evening I would embed in fresh material, and immediately polymerize at 60 deg. NOTE: All steps on rotator! Do not let material stand in the hood, not even for an hour.
I know nothing of your epoxy kit. I cannot comment on it. Since you have experience with it, I would not immediately throw it out. Just try pushing infiltration hard, and then try the vaccum trick. It saved a whole project for me once.
} ------------------------------------------------------------------------ } 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 Hildegard, } } To follow up a posting a few days ago, what epoxy are you using that does } not compress during sectioning? I am very curious! } } THanks, } } } Rosemary White } Microscopy Centre } CSIRO Plant Industry } GPO Box 1600 } Canberra, ACT 2601 } Australia } } phone 61-2-6246 5475 } fax 61-2-6246 5000 } email r.white-at-pi.csiro.au } } } } There is no formulation that can be said to not compress during sectioning. It depends on the way it is handled and the nature of the tissue being embedded. Please read my postings again which explain in detail what the basic problems are.
Someone asked me what I use in the laboratory for epoxies, and now I cannot find the person who asked.
Our general standard is the Luft' medium formulation. It is a requirement however for reembedding Vibratome or thick sections from glass slides, no matter what the original section was embedded in. We use Pella Eponate 12 and all other resins come from EMS. (I have no financial holdings in these companies)
If the end of the project goes only to thick sections, we use Mollenhauer's Araldite - Epon - DDSA - dibutyl, because this mixture sections like butter. However, we have to embed keratinized skin so the embedding procedure is really lengthy.
For immunocytochemistry we try to find out if we have huge quantities of antigen. We first try one embedment (this is for post-gold) with the above Araldite 502 mixture. If at first we don't succeed, we give up immediately! We then go to LR Gold which, in our case, reliably yields better ultrastructure than the LR White. (and of course, we get much more label)
All the above formulations are well known and can be found in any good textbook. Sometimes I construct other formulations (for embedding chocolate for paperweights for my co-workers) but those instances are applicable only to what I am about and not of general interest to anyone. ' Hope this is the answer you wanted.
This reminds me of a trick a Cambridge (yes, back then) service engineer pulled to demonstrate vibration in the SEM room at a former position. Fill a glass almost full and put it on the floor. Watch the pretty rings. Do a quick calculation on the height of the waves -- if they're 1mm high in the glass, at 10,000 times in the SEM they'd be 10 meters high. If any of the admin types have sail boats (or bass boats, this being Oklahoma), this might make an impression on them.
Phil
} Phoebe: } } They want to put your lab next to a 4-lane highway...this is a joke, right? } } Larry ;-) } } PS my suggested reference... 1. Common sense. Vol.1, No. 1, p. 1 } another suggestion: make some vibration measurements and } check it out... } } } Microscopist: } } } } Our new lab is supposed to be built next to a four lane highway that has a } } lot of 18 wheeler truck traffic. I am concerned about vibration problems } } and would like to convince administration that the building should be built } } on the other side of the property as far away from the highway as possible. } } Any suggestions or known references covering this? Thanks in advance. } } } } Phoebe J. Doss } } Manager/Adjunct Instructor } } Electron Microscope Lab } } Oklahoma State University } } Dr. Lawrence F. Allard } Senior Research Staff Member } High Temperature Materials Laboratory } Oak Ridge National Laboratory
}}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{ Philip Oshel Supervisor, AMFSC and BBPIC Dept. of Animal Health and Biomedical Sciences University of Wisconsin 1656 Linden Drive Madison, WI 53706-1581 voice: (608) 263-4162 fax: (608) 262-7420 (dept. fax)
I can also tell you first hand about the effect on light microscopy and microspectrometry equipment. I once had an assignment in a coking facility which used a switch engine to run coal cars around the yars. Forget about anything in the higher mag range!
Also, suggest that you talk to any of the EM apps people (Norm Burns, Tim Maitland?) from the old Cambridge/Leica SEM group (now part of LEO, Thornwood NY). Cambridge built that facility with a freight train running through the back yard.
There are companies which will do site evaluations, including some of the SEM groups. I don't know if there is a charge, but whatever it is, it would be less expensive than (1) not being able to do research in a brand new facility (administrators are traditionally very allergic to "egg on the face" syndrome") or (2) having to move the lab to another location once it is set up.
Hope this is helpful.
Best regards, Barbara Foster,President Microscopy/Microscopy Education 125 Paridon Street Suite 102 Springfield, MA 01118-2130 PH: 413-746-6931 FX: 413-746-9311 email: MME-at-map.com Website: www.MME-Microscopy.com/education
America's first national consortium of microscopy specialists offering customized on-site training in all areas of microscopy, image analysis, and sample preparation ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^
At 10:04 AM 4/11/00 -0500, Phoebe J Doss wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
In order to maximize usefulness, I need a dc coupled signal conditioner between my WDS analog rate output and the SEM line scan input.
Small and battery powered would be nice... Am open to suggestion, but what seems to be indicated is an amp with about +- 5 volt of INPUT offset with a gain of 1-25 (or more). Output swing should be capable of a minimum of +-5 volts.
(i.e. a couple of op-amps and 2 10- turn pots...)
I can build one, but would rather purchase if I can find one at a reasonable price. The MVA from my old ETEC would probably work, but is a NIM bin module and so would need to be repackaged and powered to stand alone.
I am currently taking measurement of SEM photos using IPP 4.1 (length measurements, angles, and thickness)and I am wondering if there is a way to "burn" these measurement into the photos for saving purposes. Has anyone done this or am I missing something. Thanks in advance.
Jim Arnold Senior Quality Technician / Failure Analysis Honeywell International, Inc. Aerospace Electronic Systems Microelectronics and Technology Center 9140 Old Annapolis Rd Columbia, MD 21045
This one is from grad school days. I was doing my thesis on the petrography and geochemistry of Miocene andesites as part of an international study on PreHellenic arc volcanism. Since I did all my own thin section preparations, I became the unpaid "volunteer" to keep the prep lab in good running condition, and to assist other students. Nothing compares to being an indentured servant. One day, a coastal sedimentary graduate student approached me. He was working on temporal barrier islands formed off the Gulf coast of Florida that formed as a result of Hurricane Helena in 1986. His major advisor thought it would be good for him make some impregnated thin sections from core samples, and stain them for carbonates and feldspars, map the distribution of them. I instructed him on all the staining procedures including safety procedures.......emphasizing safety procedured since he would be dealing with concentrated HF to etch the sections. I decided to stay in the lab to do some maintenance, and to keep an eye on him. Good thing I did, for what happened next could have turned into a real sad disaster. The student knocked over his slide drying rack into the HF bath. Before I could say or do anything, he immersed both of his hands into the concentrated HF to save his sections. Fortunately for me, I had been on a volunteer rescue squad in my teens, and was fully trained for all sorts of accidents. He was lucky........didnt lose his fingers, but did lose his finger nails, and his hands were scarred for life. Moral of this story? I should have done the procedure myself. I took on a potentially grave situation under my responsibility for a position I was not getting paid for, or properly insured for by the department. I am glad the fellow didnt suffer worse for his lack of thought, but I learned a valuable lesson, and held myself accountable and responsible for what happened. The student didnt.......... Lou Solebello
Has anyone devised some } sort of holder (sandwich?) that might overcome this problem by } keeping them flat during their trip through the dryer? } } We thank you in advance. } } Dick Briggs } Biology Department } Smith College } } } you can make a sandwich using a small reusable swinnex filter holder (holds 13 mm or larger polymer filters to filter liquids being expressed from a syringe). You saw off the connecting bits leaving the bits which screw together. Then you can sandwich things between two filters maybe with a spacer made by including a gasket between the filters. Then you process the whole assembly. Silver filters from Chuck Garber are good as they wont dissolve in liquid CO2..... Dr. Mel Dickson, Deputy Director, The Electron Microscope Unit, Adjunct Associate Professor, School of Microbiology & Immunology The University of New South Wales UNSW SYDNEY 2052 Australia. Phone +612 9385 6383 Fax +612 9385 6400
In a message dated 4/11/00 6:44:48 PM, jim.arnold6-at-honeywell.com writes:
} I am currently taking measurement of SEM photos using IPP 4.1 (length } measurements, angles, and thickness)and I am wondering if there is a way } to "burn" these measurement into the photos for saving purposes. Has anyone } done this or am I missing something. Thanks in advance.
With IPP you can label each feature with its measurement value for one measurement parameter.
Several years ago, one of my research students was using the Cambridge S250 SEM on an early solo session. The room was hushed, in almost in total darkness, and he was giving his total concentration to the screen while he adjusted the image. As he moved his hand to the specimen stage controls the turbo-molecular pump disintegrated without warning, making a crash that sounded like a metal tray full of spanners being dropped from a great height, followed immediately by the wailing of alarms. The poor chap was literally green with shock - he thought he had caused it!
When the column was opened up a glittering cloud of aluminium alloy powder drifted out. The turbo pump - a double-ended model - had its rotors and stators intertwined so forcibly that there was no free play. Presumably it had come to rest from 60,000 rpm in less than a single rotation. That works out at a damage rate of almost 1billion dollars per second!
Chris
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Dr Chris Jeffree University of Edinburgh Biological Sciences EM Facility Daniel Rutherford Building King's Buildings EDINBURGH EH9 3JH Tel: +44 (0) 131 650 5345 FAX: +44 (0) 131 650 6563
} } } From: Chris Jeffree {cjeffree-at-srv0.bio.ed.ac.uk} } } To: microscopy-at-sparc5.microscopy.com } } Subject: pre-final horror story? } } Send reply to: c.jeffree-at-ed.ac.uk } } Date sent: Tue, 11 Apr 2000 16:01:02 +0000 } } } } Several years ago, one of my research students was using the } } Cambridge S250 SEM on an early solo session. The room was } } hushed, in almost in total darkness, and he was giving his total } } concentration to the screen while he adjusted the image. As he } } moved his hand to the specimen stage controls the turbo-molecular } } pump disintegrated without warning, making a crash that sounded } } like a metal tray full of spanners being dropped from a great height, } } followed immediately by the wailing of alarms. The poor chap was } } literally green with shock - he thought he had caused it! } } } } When the column was opened up a glittering cloud of aluminium } } alloy powder drifted out. The turbo pump - a double-ended model - } } had its rotors and stators intertwined so forcibly that there was no } } free play. Presumably it had come to rest from 60,000 rpm in less } } than a single rotation. That works out at a damage rate of almost } } $1billion per second! } } } } Chris } } ------- End of forwarded message ------- } } ===================================================================== } } DR CHRIS JEFFREE } } BIOSEM - BIOLOGICAL SCIENCES EM FACILITY } } UNIVERSITY OF EDINBURGH } } Daniel Rutherford Building } } King's Buildings, Mayfield Road } } EDINBURGH, EH9 3JH, Scotland, UK } } Tel. #44 131 650 5345 } } FAX. #44 131 650 6563 } } Mobile 0410 585 401 } } email c.jeffree-at-ed.ac.uk } } SEM / TEM bookings sem-at-ed.ac.uk } } ===================================================================== } } }
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Dr Chris Jeffree University of Edinburgh Biological Sciences EM Facility Daniel Rutherford Building King's Buildings EDINBURGH EH9 3JH Tel: +44 (0) 131 650 5345 FAX: +44 (0) 131 650 6563
Maybe I should have headed this "funny horror stories". I used to run an EM Unit adjacent to a geology department. One of their more intellectual types decided that the only place to operate a rock-shaking machine was up against the outside wall of the darkroom and the SEM rooms. I never bothered to check the effects on the SEM, but observed that through the enlarger's focus magnifier the image was dancing. I told the grad student users that the shaker had to go, complete with reasons. Nothing happened, then I just turned the shaker off whenever it was on.
Eventually I was confronted by that intellectual quietly asking: "why was I against his shaker"? I figured years ago that anger was unprofessional and non-productive, but I lost it on that occasion. I asked rhetorically, how can you call yourself an intellectual, when you are unable to work out why a two bob (nickel and dime) shaker had no place next to an EM Unit. That shaker disappeared on next day.
Maybe I should have sent him an exam to sort out the problem: My instruments were there first Put the respective instrument cost into the equation Consider the difficulty involved in moving his shaker versus rehousing the EM Unit Also consider that a heavy shaker is almost designed to produce vibration. A microscope magnifies and not just objects but vibrations too. So one um of actually transmitted movement 50000x enlarged is 50mm - I trust that the movement was less than 1um.
In relation to the highway, it must be noted that you cannot switch that off. Also its difficult to foretell how much vibration would be transmitted. Its a question of risk: are those administrators willing to relocate the unit later or will they find the money to place several instruments on expensive antivibration devices. I don't believe its worth the risk; convince them. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
} } } Phoebe: } } They want to put your lab next to a 4-lane highway...this is a joke, right? } } Larry ;-) } } PS my suggested reference... 1. Common sense. Vol.1, No. 1, p. 1 } another suggestion: make some vibration measurements and check it } out... } } } } } Microscopist: } } } } Our new lab is supposed to be built next to a four lane highway that has a } } lot of 18 wheeler truck traffic. I am concerned about vibration problems } } and would like to convince administration that the building should be built } } on the other side of the property as far away from the highway as possible. } } Any suggestions or known references covering this? Thanks in advance. } } } } Phoebe J. Doss } } Manager/Adjunct Instructor } } Electron Microscope Lab } } Oklahoma State University } } Dr. Lawrence F. Allard } Senior Research Staff Member } High Temperature Materials Laboratory } Oak Ridge National Laboratory } 1 Bethel Valley Road } Bldg. 4515, MS 6064 } PO Box 2008 } Oak Ridge, TN 37831-6064 } } 865-574-4981 } 865-576-5413 Fax } allardlfjr-at-ornl.gov
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Reply to: Re: vibration due to truck traffic? Do this trick with a dish of water but reflect a beam of light off the water surface onto a wall and the vibrations are magnified before your eyes.
Paul Webster
Mssage from Phil Oshel:
This reminds me of a trick a Cambridge (yes, back then) service engineer pulled to demonstrate vibration in the SEM room at a former position. Fill a glass almost full and put it on the floor. Watch the pretty rings. Do a quick calculation on the height of the waves -- if they're 1mm high in the glass, at 10,000 times in the SEM they'd be 10 meters high. If any of the admin types have sail boats (or bass boats, this being Oklahoma), this might make an impression on them.
by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id CAA07737 for dist-Microscopy; Wed, 12 Apr 2000 02:44:54 -0500 (CDT) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id CAA07734 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Wed, 12 Apr 2000 02:44:24 -0500 (CDT) Received: from ulu.pbrc.hawaii.edu (ulu.pbrc.hawaii.edu [128.171.22.11]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id CAA07727 for {Microscopy-at-sparc5.microscopy.com} ; Wed, 12 Apr 2000 02:44:12 -0500 (CDT) Received: from halia.pbrc.hawaii.edu (halia [128.171.22.7]) by ulu.pbrc.hawaii.edu (8.8.8+Sun/8.8.8) with ESMTP id VAA24379 for {Microscopy-at-sparc5.microscopy.com} ; Tue, 11 Apr 2000 21:38:51 -1000 (HST) Received: from localhost (tina-at-localhost) by halia.pbrc.hawaii.edu (8.8.8+Sun/8.8.8) with ESMTP id VAA04910 for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 11 Apr 2000 21:38:48 -1000 (HST) X-Authentication-Warning: halia.pbrc.hawaii.edu: tina owned process doing -bs
If it ain't broke, don't fix it. This is the lesson I learned the hard way.
Several years back I was looking something up in the Balzers 400 freeze fracture manual, and noticed it recommended strongly that the turbo pump be sent in for reconditioning every 50,000 hours of use (or some such number). I had no previous experience with turbo pumps, and the consequences sounded pretty dire, so I was concerned. Since it had run 24/7 for several years, and then off and on for several more, it easily had whatever number of hours. The facility director was getting ready for a big project utilizing the instrument and, although he was reluctant, I convinced him that we should send the turbo pump in before he started.
It came back a few weeks later, and it was clearly not our pump, but another reconditioned one. As I lifted this large pump out of the box two small ball bearings bounced onto the floor and rolled away. Still holding the pump, I watched them go. Then I turned the pump all around in my arms, looking for any signs of damage or loose parts, but all looked fine. I considered calling the company, but it was Friday and with the time difference, it would be days before I got an answer. I figured what the heck, either it is going to work or not! So I installed it and turned it on, standing as far away as I could. It started up fine and achieved a reasonable vacuum, so I left it running over the weekend. Monday afternoon I decided it was OK, and turned it off.
My first thought was that a jet had crashed into the wall behind me and that I was going to die. And from the look on the faces of the others in the lab, they clearly thought they were going to, as well. The horrible screeching noise actually stops really suddenly as those pumps seize up, and then the quiet is deafening.
I opened the chamber to find it full of aluminum glitter.
The pump was replaced.
When I talked to some EM service people who had a lot more experience with turbo pumps, they all seemed to think that one should never overhaul a tp, but just wait until it crashes. Sure, it's a lot more expensive to repair it then, but apparently few do fail within the normal lifetime of the instruments they are on. Sigh.
Aloha, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
} In a message dated 4/11/00 6:44:48 PM, jim.arnold6-at-honeywell.com writes: } } } I am currently taking measurement of SEM photos using IPP 4.1 (length } } measurements, angles, and thickness)and I am wondering if there is a way } } to "burn" these measurement into the photos for saving purposes. Has anyone } } done this or am I missing something. Thanks in advance. } } With IPP you can label each feature with its measurement value for one } measurement parameter. } Years ago I used potassium iodide, calcium chloride and copper sulfate as a bleach. I don't remember the formula but it is not very critical. You could use this stuff to mark any silver photographic image. I think the silver ends up a silver iodide so you would need to refix and wash them. You would probably want to add a gelling compound to it to keep it from running starch or wall paper paste would be a good place to start.
A simpler method would be to scratch the film or use India Ink on it. Magic Marker felt tip pens should work as well. There might be some bleeding on the emulsion side.
For temporary marking there are opaquing paints to cover pin holes on litho negatives that is water based and should wash off the base side just fine. You nearest print shop or graphics art supply can fix you up with a life time supply for 5 bucks.
Good Luck Gordon
Gordon Couger gcouger-at-couger.com
Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
} } } Our new lab is supposed to be built next to a four lane highway that has a } } } lot of 18 wheeler truck traffic. I am concerned about vibration problems } } } and would like to convince administration that the building should be built } } } on the other side of the property as far away from the highway as possible. } } } Any suggestions or known references covering this? Thanks in advance. } } }
I take it you are going in on Hall of Fame. It is not as bad as you fear but it is bad.
Starting from scratch an air supported floor for the room would not be too expensive if ou designed it your self. Twenty Five cent pre pound steel and a decient air pump will handle low frequencie vibratin and active stuff does a great job from 10HZ up.
Now if you could just move a cross the road to the sheep fram most of your problems wold dissappear.
There are several minds on campus that have delt with simular situations and I will be glad to get you togeather.
Good luck Gordon
Gordon Couger gcouger-at-couger.com
Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
In semiconductor failure investigation various etchants and staining methods are used to obtain and or enhance selected features prior to FESEM investigation and image capture . As I now capture images digitally and have the increasing capabilities of image processing at my fingertips it's possible to ' modify' and possibly distort the initially accurate image obtained which could lead to misleading results .
I receive people's images and wonder if they are a result of different sem capabilities / preparation methods / etches or are they simply 'touched up '. They probably think the same about mine .
Therefore is there any way of controlling image processing to enable like for like comparison ? or any international standards ? way of indicating on an image it has been subject to processing or is it now a case of you cannot believe what you see ?
Was the cause ever determined? Or did I miss it somewhere in the text?
An excellent way to stop a turbo without going through the proper shut down sequence is to drop something into the turbines. An unwitting young prof at the grad school I went to inadvertantly dropped I can't remember what in a turbo pump in the UHV system that he was designing. This was with the pump running at several hundred thousand rmp. You can imagine the size of this pump. I am not sure that it stopped in less than one rotation, but it fried the turbo, non pun intended. He had to replace it more so because he borrowed the pump from another prof. than because he needed one for his experiments. You can image how green he was.
The moral of this story is to cover the inlet with a screen. The chances of anything falling into it is significantly reduced and you can save thousands of dollars.
Regards to you all, Andy
cjeffree-at-srv0.bio.ed.ac.uk on 04/11/2000 09:55:57 PM To: microscopy-at-sparc5.microscopy.com-at-INTERNET cc:
Several years ago, one of my research students was using the Cambridge S250 SEM on an early solo session. The room was hushed, in almost in total darkness, and he was giving his total concentration to the screen while he adjusted the image. As he moved his hand to the specimen stage controls the turbo-molecular pump disintegrated without warning, making a crash that sounded like a metal tray full of spanners being dropped from a great height, followed immediately by the wailing of alarms. The poor chap was literally green with shock - he thought he had caused it!
When the column was opened up a glittering cloud of aluminium alloy powder drifted out. The turbo pump - a double-ended model - had its rotors and stators intertwined so forcibly that there was no free play. Presumably it had come to rest from 60,000 rpm in less than a single rotation. That works out at a damage rate of almost 1billion dollars per second!
Chris
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Dr Chris Jeffree University of Edinburgh Biological Sciences EM Facility Daniel Rutherford Building King's Buildings EDINBURGH EH9 3JH Tel: +44 (0) 131 650 5345 FAX: +44 (0) 131 650 6563
Good trick Paul, but in my checkered past, we used a large (12" diameter) dish filled with mercury. That gave much better reflection of the light and very distinct vibration patterns on the wall.
Cheers,
Frank Shapiro.
Paul Webster wrote:
} } Do this trick with a dish of water but reflect a beam of light off the water surface onto a wall and the vibrations are magnified before your eyes. } } Paul Webster } } Mssage from Phil Oshel: } } This reminds me of a trick a Cambridge (yes, back then) service engineer pulled to demonstrate vibration in the SEM room at a former position. Fill a glass almost full and put it on the floor. Watch the pretty rings. Do a quick calculation on the height of the waves -- if they're 1mm high in the glass, at 10,000 times in the SEM they'd be 10 meters high. If any of the admin types have sail boats (or bass boats, this being Oklahoma), this might make an impression on them. } } Phil
You may want to take a few minutes and peruse the following site:
http://www.vibeng.com/index.htm
Vibration Engineering Consultants -VEC- has some useful information regarding site selection, vibration, EM and acoustic interferences, etc. However, it may help you more to talk with them personally. Craig Franklin came to our site in southern New Jersey a few years ago, to survey potential new locations for our SEM, and made some recommendations, as far as location, field cancellation and vibration insulation. We ended up buying the AC/DC field canceling system that he recommended, and a vibration table. These items have been a huge help in insuring the continued high resolution capabilities of our SEM. The people at VEC may be able to help make your case to your management, before the new facility is built.
Kristin Breen Staff Chemist, Marketing Technical Services Lab North American Region ExxonMobil Lubricants and Petroleum Specialties Co. Paulsboro, NJ (856) 224-2864
There is a difference between unwittingly taking pictures of artifacts and fraudulent digital manipulation. In the latter case there have been several strings on this listserver and elsewhere on digital manipulation, see the archives. Essentially, digital manipulation of image contrast, brightness, and gamma to produce a better image is the same as dodging, burning-in and choosing different contrast grade papers in an old-fashioned darkroom to enhance but not alter an image, this is OK. Beyond that, it is all to easy to alter an image digitally. "Alter," in the fraudulent sense. I don't see anything wrong with deleting specimen preparation artifacts--scratches, left-over polishing compound, etc.--as long as the true image content is unaffected. What is and what isn't in this category is a judgement call on my part. I assume that, unless proven otherwise, all of my colleagues out there are doing the "right" thing and clearly stating what manipulations of the image content were performed and why.
With all due respect, Martyn, if I were you I'd worry more about the former case. SEM imaging of chemically etched integrated circuit cross sections is horrendously prone to artifact production. We stopped this practice more than a decade ago, tuning instead to high-angle ion milling for short time periods. See our paper: Martinek, et al., 1989 MSA Proceedings, p. 720. We've been using GATAN Duo Mill ion millers for this from our TEM areas. GATAN has brought out a unit specifically to perform this function for SEM labs. It's the Model 682 (?) PECS or, the new PECS + RIBE system. (The question mark is mine--I'm not sure of the model number of the simple PECS system.) Good luck.
Ron
I have no business or financial relationship with GATAN other than being a long-time satisfied customer.
Ron Anderson, IBM, Hopewell Jct., New York, USA. anderron-at-us.ibm.com
IBM Analytical Services; http://www.chips.ibm.com/services/asg
"HARRISm/-at-esm-semi.co.uk" on 04/12/2000 06:33:00 AM
To: Microscopy-at-sparc5.microscopy.com cc:
A general enquiry regarding integated circuit X section images :
In semiconductor failure investigation various etchants and staining methods are used to obtain and or enhance selected features prior to FESEM investigation and image capture . As I now capture images digitally and have the increasing capabilities of image processing at my fingertips it's possible to ' modify' and possibly distort the initially accurate image obtained which could lead to misleading results .
I receive people's images and wonder if they are a result of different sem capabilities / preparation methods / etches or are they simply 'touched up '. They probably think the same about mine .
Therefore is there any way of controlling image processing to enable like for like comparison ? or any international standards ? way of indicating on an image it has been subject to processing or is it now a case of you cannot believe what you see ?
The TissueTek* brand of histology equipment has been around for many years, and in my opinion, is at the top of the quality, dependability, and reliability lists. The confusion here is what exact "TissueTek" models you are considering. Even using the suffixes "II" and "III" does not completely identify which TissueTek product you are talking about. It is best to acquire and use the individual Model numbers to prevent confusion.
What I call a "TissueTek II" processor is the [Model 4640]. This is a basic "dip and dunk" rotary processor and is considered an "open system" (no fume control). It is still being manufactured and it can be purchased as a new piece of equipment. No problem getting spare parts. You would have to provide some type of fume hood for it. The 4640 is compact enough that it would fit inside a standard chemical fume hood. If you would only be using it on a random basis and low volume, this would be a good choice. It can hold up to 120 specimens per run.
I consider the "TissueTek III" [Model 4660] as the first enclosed tissue processor with on-board fume control. It is also known as the "V.I.P." However, this Model was discontinued in 1982-83 and is totally OBSOLETE as far as spare parts and support from the manufacturer. There are a number of these units still in service around the world, however.
} From approximately 1983 to 1994 the next generation TissueTek V.I.P.s were the "K" series with [Model Numbers: 4617, 4618,4619]. They came in three volume sizes and and could be purchased in either a bench-top or floor configuration. They still supported by the manufacture, and spare parts are readily available. These, once again, are "closed systems" with fume control.
The current production models of the "V.I.P." is the "E" series [Models: 4890 & 4894] in the bench top configuration, and [Models 4892 & 4896] in the floor model configuration. They come in two volume sizes.
Of the above, the only processor that I would not recommend that you consider is the original TissueTek III VIP [Model 4660].
I hope this answers your questions.
* TissueTek, and TissueTek V.I.P. are registered trademarks of Sakura Finetek, Inc.
-- Ford M. Royer, MT(ASCP) Analytical Instruments, Ltd (Refurbished Histology, Cytology, & General Lab Equipment) 9921 13th Ave. N. Minneapolis, MN 55441-5004 phone: 800-565-1895, Ext. 17 fax: 612-929-1895 Email: froyer-at-bitstream.net web site: http://www.aibltd.com
"A.P. Alves de Matos" 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 } -----------------------------------------------------------------------. } } Thanks Norm for your suggestions } } I will be more specific. } I will need to process up to 60 specimens/week in two or three separated } weeks during the year. The rest of the time hand processing is OK. Since the } specimens are prone to dammage upon storage (immunohistochemical techniques } on sponge tissues), I do not want to store them to facilitate processing. } So, it is important to know if the machine can handle this amount of work. } I have one technician that has many other things to do. } Basic assistence should be available from our technical department. Anyhow I } would not trust newer instruments just because they are more recent. I know } of (and have) a few super-new machines (amaizingly expensive) that had to be } thrown to garbage because no one (assistence included) was able to put them } to regular work. It is important to know if the machine model is a well } tested one that usually works for years with no problems or if it has } regular problems. } Since histology and histopathology labs of my knowledge use other tissue } processors, I was not able to get specific advice locally about these } machines. } } Thanks } A.P. Alves de Matos } } ----- Original Message ----- } } From: Norm Granholm {granhona-at-email.uc.edu} } To: A.P. Alves de Matos {apmatos-at-ip.pt} } Sent: Tuesday, April 11, 2000 12:57 PM } Subject: Re: Tissue Tek machine evaluation } } } Dr. de Matos: } } } } I believe some general guidelines are appropriate for you to consider: } } 1. Get the newest machine you can afford. Older machines will have } more } } maintenance requirements and will become obsolete sooner. } } 2. Is instrument repair readily available? If not then any } automated } } system is potentially a problem. Automated systems do break. } } 4. Your note says "lots of paraffin embeddings". What is "lots"? } Hand } } processing is quite feasible for batches of small numbers and if personnel } help } } is not limiting (see next). No one likes doing it and an automated system } lets } } skilled individuals perform more appropriate tasks. All of this is, } however, an } } issue of resource utilization. You have considered these matters already, } } undoubtedly. } } 3. If personnel help is a limiting factor, the more automated the } better } } for you. If personnel help is not a limiting factor, then the less } automated the } } better for you. This may sound strange but I believe it is true. It all } depends } } upon who is paying the bills and whether the funds are available for other } uses. } } And there are always other uses for funds. } } } } Having raised all of those points, I'll tell you that I used a Tek II when } they } } first came out ( now some 20 years ago). And this for fewer than a dozen } samples } } per week. I wanted skilled individuals to do other tasks than hand dip } tissues. } } } } You might look, also, at Leitz/Leica tissue processors. My experiences } with } } Leica are that they provide outstanding equipment at reasonable prices. } Often } } they have trade in items available. Were I to have to chose I would head } this } } way. } } } } Best wishes, } } } } Norm } } (Norman.Granholm-at-uc.edu) } } Pathology, Univ. Cincinnati } } } } Voice 513 558 0182 } } Digital Pager 513 249 3889 } } ===============================
What's funny?Ê My lab is on the 4th floor of a semiconductor fab at the intersection of two 6-lane freeways.Ê We have 4 SEMs, 3 focused ion beams, and 2 200kV TEMs.Ê We routinely work over 150KX (SEM) with very few problems.Ê It's a well-designed building.Ê EMI is more of a problem.
Larry Allard wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } ToÊ Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Phoebe: } } They want to put your lab next to a 4-lane highway...this is a joke, right? } } LarryÊ ;-) } } PSÊ my suggested reference... 1. Common sense. Vol.1, No. 1, p. 1 } ÊÊÊÊÊÊ another suggestion:Ê make some vibration measurements and check it out... } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } ToÊ Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Microscopist: } } } } Our new lab is supposed to be built next to a four lane highway that has a } } lot of 18 wheeler truck traffic.Ê I am concerned about vibration problems } } and would like to convince administration that the building should be built } } on the other side of the property as far away from the highway as possible. } } Any suggestions or known references covering this?Ê Thanks in advance. } } } } Phoebe J. Doss } } Manager/Adjunct Instructor } } Electron Microscope Lab } } Oklahoma State University } } Dr. Lawrence F. Allard } Senior Research Staff Member } High Temperature Materials Laboratory } Oak Ridge National Laboratory } 1 Bethel Valley Road } Bldg. 4515, MS 6064 } PO Box 2008 } Oak Ridge, TN 37831-6064 } } 865-574-4981 } 865-576-5413Ê Fax } allardlfjr-at-ornl.gov
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky HoldfordÊ (r-holdford-at-ti.com) 972-598-1291 (pager) KFAB Physical Analysis Lab--SEM/FIB Kilby Center West Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Ê
I'm trying to track down a paper presented in the early '70s at an EM meeting in Erice, Italy. The papers were published in a volume called Electron Microscopy in Materials Science Vol. II, Valdre and Ruedl, eds. The paper I'd like to get is one by A. Metherell on Bloch waves. My quest has stymied our interlibrary loan folks here (I did get the 3rd vol). Anyone got a Vol II or the Metherell paper?
Thanks
John
John Heckman MSM Department Michigan State University
I wouldn't expect much vibration either, if I worked in a Billion $ building...Phoebe, what was your budget again?
Larry ;-)
} What's funny? My lab is on the 4th floor of a semiconductor fab at } the intersection } of two 6-lane freeways. We have 4 SEMs, 3 focused ion beams, and 2 } 200kV TEMs. We } routinely work over 150KX (SEM) with very few problems. It's a well-designed } building. EMI is more of a problem. } } Larry Allard 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 } } -----------------------------------------------------------------------. } } } } Phoebe: } } } } They want to put your lab next to a 4-lane highway...this is a joke, right? } } } } Larry ;-) } } } } PS my suggested reference... 1. Common sense. Vol.1, No. 1, p. 1 } } another suggestion: make some vibration measurements and } check it out... } } } } } ------------------------------------------------------------------------ } } } 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 } } } -----------------------------------------------------------------------. } } } } } } } } } Microscopist: } } } } } } Our new lab is supposed to be built next to a four lane highway that has a } } } lot of 18 wheeler truck traffic. I am concerned about vibration problems } } } and would like to convince administration that the building } should be built } } } on the other side of the property as far away from the highway } as possible. } } } Any suggestions or known references covering this? Thanks in advance. } } } } } } Phoebe J. Doss } } } Manager/Adjunct Instructor } } } Electron Microscope Lab } } } Oklahoma State University } } } } Dr. Lawrence F. Allard } } Senior Research Staff Member } } High Temperature Materials Laboratory } } Oak Ridge National Laboratory } } 1 Bethel Valley Road } } Bldg. 4515, MS 6064 } } PO Box 2008 } } Oak Ridge, TN 37831-6064 } } } } 865-574-4981 } } 865-576-5413 Fax } } allardlfjr-at-ornl.gov } } -- } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } Becky Holdford (r-holdford-at-ti.com) } 972-598-1291 (pager) } KFAB Physical Analysis Lab--SEM/FIB } Kilby Center West } Texas Instruments, Inc. } Dallas, TX } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ }
Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
} When I talked to some EM service people who had a lot more experience with } turbo pumps, they all seemed to think that one should never overhaul a tp, } but just wait until it crashes. Sure, it's a lot more expensive to repair } it then, but apparently few do fail within the normal lifetime of the } instruments they are on. Sigh.
Dear Tina, Yes, turbopumps canibalize themselves spectacularly--they're almost as much
"fun" as high-pressure Hg-vapor lamps. I'd like, however, to put in a good word for servicing them on a regular basis. The turbos that were on the HVEM when I got here were ~200 lbs, turned at ~10,000 rpm, and occasionally ate themselves. We soon replaced them with the TPU330 model pumps, which are ~30 lbs., turn at ~15,000 rpm, and have been humming along at "standby" speed for years. The vacuum is as good at standby as at full speed, so we have seen no need to go to the higher speed. We've changed the oil on a regular schedule and sent the pumps to Balzers every two years for bearing changes. There has been no deterioration in performance for ~15 years now. Yours, Bill
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Everyone's been discussing vibration in this thread (talk about slavish adherence to topic ...) but there's another consideration - magnetic disturbances. Those truck have steel frames, so they'll disturb the Earth's magnetic field as they pass by. I inadvertently made a magnetometer once, and I could see the inflence of vehicles passing by fifty feet away on my oscilloscope screen. At M.I.T. there was a lot of concern about the nearby elevator in one EM lab ...
Best regards, George Langford, Sc.D., who's got no vested interest in Earth's magnetic field except when out hiking. amenex-at-amenex.com
Several years ago (MSA in Cleveland if I recall correctly), there was some sort of roundtable discussing the ethics of image manipulation. I can't remember much more than that, but perhaps those with a better memory who may have participated in the discussion would have some good input.
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} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy. Message text written by INTERNET:"HARRISm-at-esm-semi.co.uk"-at-sparc5.microscopy.com } A general enquiry regarding integated circuit X section images :
In semiconductor failure investigation various etchants and staining methods are used to obtain and or enhance selected features prior to FESEM investigation and image capture . As I now capture images digitally and have the increasing capabilities
of image processing at my fingertips it's possible to ' modify' and possibly distort the initially accurate image obtained which could lead to misleading results .
I receive people's images and wonder if they are a result of different
sem capabilities / preparation methods / etches or are they simply 'touched up '. They probably think the same about mine .
Therefore is there any way of controlling image processing to enable like for like comparison ? or any international standards ? way of indicating on an image it has been subject to processing or is it now a case of you cannot believe what you see ?
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} ... } Question. } } In semiconductor failure investigation various } etchants and staining } methods are used to obtain and or enhance selected } features prior to } FESEM investigation and image capture . } As I now capture images digitally and have the } increasing capabilities } of image processing at my fingertips it's possible to } ' modify' and } possibly distort the initially accurate image obtained } which could } lead to misleading results . } } I receive people's images and wonder if they are a } result of different } sem capabilities / preparation methods / etches or are } they simply } 'touched up '. They probably think the same about mine . } } ...
Surely you are not implying similar distortions were never a possibility in the wet darkroom??
It is possible to put (for example) a fine checkerboard pattern in a small box which would imply this image has never been subjected to "blur", "sharpening", or many other kernal operations ... or you could install a standard grayscale to imply brightness, contrast and gamma are original ... BUT, you would need trust the author didn't install the markers after distorting.
} -----Original Message----- } From: "HARRISm-at-esm-semi.co.uk"-at-sparc5.microscopy.com } [mailto:"HARRISm-at-esm-semi.co.uk"-at-sparc5.microscopy.com] } Sent: Wednesday, April 12, 2000 5:33 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Subject : SEM + DIGITAL IMAGING } } } -------------------------------------------------------------- } ---------- } 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/MicroscopyListser} ver/FAQ.html } } } -------------------------------------------------------------- } ---------. } } } } A general enquiry regarding integated circuit X section images : } } Email . harrism-at-esm-semi.co.uk } Name . Martyn Harris Device Engineering - failure analysis } } } Question. } } In semiconductor failure investigation various etchants } and staining } methods are used to obtain and or enhance selected } features prior to } FESEM investigation and image capture . } As I now capture images digitally and have the } increasing capabilities } of image processing at my fingertips it's possible to ' } modify' and } possibly distort the initially accurate image obtained } which could } lead to misleading results . } } I receive people's images and wonder if they are a } result of different } sem capabilities / preparation methods / etches or are } they simply } 'touched up '. They probably think the same about mine . } } Therefore is there any way of controlling image } processing to enable } like for like comparison ? or } any international standards ? way of indicating on an } image it has } been subject to processing or is it now a case of you } cannot believe } what you see ? } } Regards. } } } }
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We have disconnected our old Philips EM-200 and will send it to salvage within the next week. If anyone wants parts from it or the others we have in storage (used for parts) please contact me immediately. All used parts (other than vacuum tubes) are free for the asking but you will need to pay shipping. We also have a Reichert OMU-3 ultramicrotome which can be used for parts. It is not functional at the moment. Debby
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
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Course Announcement:
FEBS Stereology and Immunocytochemistry course to be held at the University of Oslo in Norway. For details see: http://www.hei.org/htm/curs.htm
This is an intensive practical course covering all aspects of specimen preparation for immunocytochemistry. The practical part of the course is supported by in-depth lectures explaining preparation protocols and their theoretical background.
Subjects covered include stereology (quantitation), chemical fixation, rapid freezing, freeze substitution, cryoultramicrotomy, specimen contrasting, antibody labeling, pre-embedding labeling, antibody and colloidal gold preparation, and specimen evaluation.
Participants are encouraged to bring their own samples.
Due to the intensive practical nature of this course places are limited, so apply early. Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm
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Having gone through a similar problem with vibrations, I can fully appreciate what might happen. However, I cannot give you any formal information for you to support your cause, I can only offer you my story.
I moved to my insitute to set up an imaging laboratory but before I arrived, a vibration survey had been performed on the site chosen for the EM's. This site passed inspection.
When I arrived, I looked at the site and suggested it might not be suitable. I was instantly presented with the very professional survey file which said the site was suitable for electron microscopes, (as were about 10 other locations throughout the building).
When the EM supplier came in to survey the site, their standards were very different to the generic vibration expert, and the site failed (not surprizing considering it was in the middle of the 4th floor!). Every other site we looked at failed too, so we were stuck with attempting to make things work using an anti-vibration platform under the first microscope we installed (a TEM). Amazingly, the supplier of the anti-vibration platform did not even come to look at the site when I contacted them. They seemed to have no intention of doing a site survey, and were not even interested in obtaining information from the microscope supplier about the sort of problem we were attempting to correct.
We put the TEM on the very expensive air table, which made it 8' taller, and thus more difficult to operate, and tested it. The platform did not help at all in isolating the low frequency vibration that seemed to be running through the building. It seems that if the vibrating frequency is low enough, even the anti-vibration platform moves! It was also very difficult to operate a machine you couldn't touch when taking a picture. Eventually we found a stable site, renovated it and installed the microscopes there. All is now in order and the microscopes are performing to specification. Interestingly, the space is next to an in-door parking lot but the flow of traffic, although noticable to people in the lab, does not affect the microscopes. These are mostly slow moving cars with a few SUV's and pick-up trucks shaking the ground occassionally. My guess is that this is because the microscopes are installed between closely spaced support pillars on the side of the building. (Can't wait for the big earthquake to shake them up a bit!)
My advice would be to explain just how sensitive the EM's are to vibration, especially the low frequency shaking you can get from moving trucks and trains, and that the laboratory be located in as quiet a place as possible. Make sure that any vibration surveys are carried out by EM specialists, not by local heros, and keep away from floating platforms.
I am willing to re-tell ALL my experiences with sorting out our problem to anyone who wants or needs to listen. Correcting our mistakes was VERY costly.
Regards,
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm
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I thought about adding this story to the horror list but now it seems even more appropriate for the truck vibration problem. I quote directly from the book "Principles and Practise of Electron Microscope Operation" by Agar and Chescoe: The main sources of trouble, which can directly affect the resolution obtained with the instrument, are mechanical vibrations and electric fields. Several years ago we set up an EM facility in the basement of our new, state of the art, 5 story building here at Stanford. The engineers surveyed the room and proclaimed it fine for EM use. After using the microscope for a few weeks I noticed that I could never get a negative that was in perfect focus. I collected negatives from several other users and they all had the same problem. We contacted the engineers and one of them spent the entire day trying to get perfect image of fresnel fringes. By the time they turned of all the fans in our building (boy, did it get hot and stinky), he was able to focus. It turned out a giant electrical cable ran directly underneath the room to power the building ventilation. It was, of course, not in use yet when they surveyed the new building! Not to mention the nearby elevators that were not in use during survey time either. Sadly for us, rather than spend $10,000.00 on an H frame to eliminate the problem, they moved the old scope out and gave the space to a molecular biology lab instead. Here we are 7 years later without a decent EM facility in our building.
The service is important, no question about that. But I agree, also, with Tina, that it is not bad practice "do not disturb" working equipment. For instance, our TP on JEM1200-EX TEM is original pump coming with instrument 15+ years ago. It was never serviced at all. Our JEOL service engineer claimed that it is oldest working original TP on the JEM1200-EX series in US. From time to time they called me asking does TP still working? And it is work. I don't remember the exact model of TP. This is BALZERS for sure. I think that electronics life depends in many cases not from how long it operates but how many times you shut it ON and OFF. This kind of "law" works perfectly for major electronic components as well as for CRTs, computer components (HDs for instance). I guess, it may works for TPs too. During start/stop TP components may sense some stress: more stress, shorter life (we all know about that). I am experimented with magnetically levitated TP from SEIKO (no financial interest) now. It works great. It uses bearings only at low speed, at high speed the rotor is levitated. Combination of this TP and "scroll pump" (oil free) gives me absolutely oil-free vacuum system. I'll tell readers of this ListServer what happens with that TP later. I am not going to service it at all.
Sergey
} Date: Wed, 12 Apr 2000 10:34:14 -0400 } From: William Tivol {tivol-at-wadsworth.org} } Subject: Re: another turbo horror story } Sender: tivol-at-wadsworth.org } To: microscopy-at-sparc5.microscopy.com } X-Mailer: Mozilla 4.07C-SGI [en] (X11; I; IRIX64 6.5 IP21) } } ------------------------------------------------------------------------ } 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
As a new associate of Don G. of Microscopy Today I am interested located interesting techniques and Problem/Solution type of information on all types of microscopy.
In my dictionary I have found expression relevant to the situation: to go from one extreme to another. By my opinion to keep without service the mechanism rotated with 60,000 rpm is other extreme. By analogy it is possible to refuse to change oil in car motors...? Regards.
Victor Sidorenko, ANTRON Co.Ltd., scientific service, Moscow, Russia.
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} Sergey Ryazantsev Ph. D. } Electron Microscopy } UCLA School of Medicine } Department of Biological Chemistry } Box 951737 } Los Angeles, CA 90095-1737 } } Phone: (310) 825-1144 } Pager: (310) 845-0248 } FAX (departmental): (310) 206-5272 } mailto:sryazant-at-ucla.edu } http://www.bol.ucla.edu/~sryazant } } }
} From: Hall, Ernest L (CRD) Sent: Thursday, April 13, 2000 7:48 AM To: 'MSA Listserver Dist'
Please correct me if I am wrong, but I think I learned from this list that Coolwell is defunct? I have a model SE-075W CZ that I am having trouble with. No schematics and/or manuals to be found. Does anyone know where I might find these or is willing copy important info from manuals they have on this model? I would appreciate it.
Still in the fact finding phase but I suspect the thermostat is the main trouble with this unit. Do any of you know a supplier for these? Thank you in advance.
Responding to the message of {c5.40b6c0e.2626a950-at-aol.com} from "COFAB2-at-aol.com"-at-sparc5.microscopy.com: } } ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } As a new associate of Don G. of Microscopy Today I am interested located } interesting techniques and Problem/Solution type of information on all types } of microscopy. } }
Whooooooooo are you.......who-oo....oo-oo. } :o
(With appologies to Pete Townsend)
Gib
Gib Ahlstrand Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN. USA. 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.cdl.umn.edu http://biosci.umn.edu/MIC/consortium.html
I have a student that wants to look at Actinomycete colonies in the SEM. I have not looked at bacterial COLONIES before. How would one prepare such a sample?
TIA, Bill Chissoe
-- ============================================================= Bill Chissoe III Electron Microscopist University of Oklahoma 770 Van Vleet Oval Norman, Ok. 73019 E-mail: wchiss-at-ou.edu Ph. (405)325-4391 =============================================================
My apologies to everyone for not thoroughly reading the advertisement. I posted the message for our Human Resources Department. I didn't realize that Ft. Worth wasn't mentioned. Accordingly, to clarify the advertisement, the position would be at our R&D facility in Fort Worth, Texas, U.S.A.
Mitch ------------------- Mitchell D. McCartney, Ph.D. Associate Director Central Sciences
Scientist Retina Research - Degenerative Disease
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Seriously, George has brought up an interesting point as I myself have experienced the magnetic-interference-caused-by-powerline problem. The best thing to do in all these cases is have the vendor (in my case John D from JEOL) show up with the Earthquake Meter and Magnetic Treasure Finder and do a survey before the decision-makers decide anything. It's even better if you can work with the vendor BEFORE you let "them" plan anything.
This is one situation where asking forgiveness instead of permission doesn't bode well...
"They" are watching...
Laura
} } Hallo magnetic Microscopists ! } } Everyone's been discussing vibration in this thread } (talk about slavish adherence to topic ...) but there's } another consideration - magnetic disturbances. Those } truck have steel frames, so they'll disturb the Earth's } magnetic field as they pass by. I inadvertently made a } magnetometer once, and I could see the inflence of } vehicles passing by fifty feet away on my oscilloscope } screen. At M.I.T. there was a lot of concern about the } nearby elevator in one EM lab ... } } Best regards, } George Langford, Sc.D., who's got no vested interest in } Earth's magnetic field except when out hiking. } amenex-at-amenex.com }
Coolwell Co. (Out of Business 4/99) 26 Law Drive, Fairfield, NJ 07004 973 882-6611, 800 367-5665 Bought out by: Lytron, Inc. (They have schematics) 55 Dragon Court, Woburn, MA 01801 Application engineer, Greg Ducharm, 781 933-7300 East coast sales, Scott Martin Project engineer, Lonnie Fultz,4/00, Chillers Independent- Frank Haze 800 367-5665 (SEE PECO MANUFACTURING FOR TEMP CONTROL)
PECO Manufacturing Co., Inc. Portland, OR Sunn(?) Division 503 233-6401 Tempurature control for Coolwell chillers # TC103 025 C not available OEM only. Made for Coolwell in lots of 100 only (Coolwell list price was $170 each 1/99)
Switch used on temp control was made by: C & K (Unimax) Original part #WHB152-9-W C & K new part # HBS2KCB4SP011C (available from Newark Electronics #07F041) 22A, 125,277VAC, 15A 480VAC 1/4HP 125VAC, 1/2HP 250 270VAC
I have 3 Coolwell chillers (vintage 1993 and 1996), have replaced the temperature control on 2 of them in the past year and am out of spare controls. My units were over heating and shutting down. The temp control is no longer available (unless all of us Coolwell owners pitch in and buy 100 of them). [I suspect that the sensing bulb is ok but that the electrical switch has worn out. Test the bulb by immersing in hot water then cold water while watching for expansion and contraction near the switch actuator.] The switch could be replaced by drilling out it's securing rivots and replacing (part # above). Use appropriate screws to replace rivots as the switch must not slip. I have just ordered my new switches and will be testing this fix soon.
Please let me know what you find. Coolwells were used on many SEMs so I think we should have a few others interested in this problem. I have schematics for our SE series units with various options and Lytron has been very helpful. Good Luck. Jim
---------------------------
On 4/13/00 Joel McClintock wrote:
Please correct me if I am wrong, but I think I learned from this list that Coolwell is defunct? I have a model SE-075W CZ that I am having trouble with. No schematics and/or manuals to be found. Does anyone know where I might find these or is willing copy important info from manuals they have on this model? I would appreciate it.
Still in the fact finding phase but I suspect the thermostat is the main trouble with this unit. Do any of you know a supplier for these? Thank you in advance.
Joel McClintock U of Kentucky
James S. Romanow The University of Connecticut Physiology and Neurobiology Department Electron Microscopy Facility U-2131 Storrs, CT 06269-2131 bsgphy3-at-uconnvm.uconn.edu 860 486-2914 voice 860 486-1936 fax
As I recall, a few days ago someone asked about a source of an inexpensive abinocular optical microscope. Just yesterday I received an advertisment from the Cole Parmer Co. (800-323-4340; www.coleparmer.com) for a 20X binocular microscope for a base price of $159 (Cat. No.PP-03904-01), $220 when equipped with a top illuminator (PP-03904-02), and $240 equipped with both top and bottom illuminators (PP-03904-03).
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
It is possible that the reading you are getting from your vacuum gauge is not as good as you expect because the gauge itself became contaminated with oil. I presume the high vacuum gauge on your instrument is a Penning gauge, as is the case for most EMs these days. If so, it might be useful to take it off the instrument and clean it. We just had an incident where a Penning gauge got so badly fouled that it would not fire at all, and thus prevented the vacuum system from switching over to the high vacuum evacuation mode, and so the vacuum could never become good enough to allow the high voltage to be turned on.
It is usually not too difficult to clean a Penning gauge. The general construction of these gauges is discussed in Sect. 3.2.2, p. 99, of my book 'Vacuum Methods in Electron Microscopy" and illustrated in Fig. 3.12 on p. 101. Cleaning involves getting rid of the carbonaceous deposit that forms on the inside wall of the gauge tube, and on the insulator that supports the anode ring. Usually, depending on the design, these gauges can be disassembled, somewhat as shown in Fig. 3.12, whereupon various methods can be used to remove the carbonaceous deposits. One approach, of course, is to use some kind of an abrasive (try Revere Ware Stainless Steel Cleaner, or even a fine grade of metallographic polishing paper. Don't use steel wool, because pieces of it will get attracted by the strong magnet and become very troublesome to remove). The Hitachi service engineer was successful in cleaning our gauge by boiling the gauge tube (after removing the magnet from around it) and anode ring assembly in a STRONG detergent solution (try Alconox, or a similar Lab. detergent) for 30 to 60 minutes, periodically scrubbing with a toothbrush,
Good luck!
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
} Please let me know what you find. Coolwells were used on many SEMs so I } think we should have a few others interested in this problem. I have } schematics for our SE series units with various options and Lytron has been } very helpful.
We replaced the original thermostat on our Coolwell SE unit with an "ETC Single Stage Electronic Temperature Controller" and a "1309007-044 ETC Temperature Sensor" from Ranco (8115 U.S. 42 N., Plain City, Ohio, 43064). The unit bolts right to the front of the Coolwell chiller and works just fine. It works over a tempertature range of -30F to +220F and a differential of 1F to 30F. I seem to recall that the whole shebang was less than $100 and our campus refrigerator guy installed it with no problems. I can fax the spec sheets to anyone who is interested. I believe I got the idea from Maggy Piranian through this list.
Bob Dr. Robert R. Wise Associate Professor of Plant Physiology Department of Biology University of Wisconsin Oshkosh Oshkosh, WI 54901 (920) 424-3404 wise-at-uwosh.edu http://www.uwosh.edu/departments/biology/wise/wise.html
Dear Joel, I have had good luck getting a local HVAC (heating, ventilation and air conditioning) service firm to service my cooling water recirculators, which are Haskris. They have repaired all of them without problems. They may be able to fit replacement parts without having to resort to the manufacturer. At 09:50 PM 4/13/00 -0400, you wrote: } } Please correct me if I am wrong, but I think I learned from this list that } Coolwell is defunct? I have a model SE-075W CZ that I am having trouble } with. No schematics and/or manuals to be found. Does anyone know where I } might find these or is willing copy important info from manuals they have on } this model? I would appreciate it. } } Still in the fact finding phase but I suspect the thermostat is the main } trouble with this unit. Do any of you know a supplier for these? Thank you } in advance. } } Joel McClintock } U of Kentucky
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
Hi, I was wondering whether anyone out there can help me out on the forced modulation and phase contrast technoques. I'm familiar with the basics of the AFM ,ie., contact and the non-contact modes. Now I want to learn the advances methods. Can anyone suggest any good books that give the full techniques? Praveena
Praveena M Bhaskara MS Student Chemical Engineering Department University of Massachusetts, Lowell Lowell, MA 01854 Email:bubbyp-at-hotmail.com PH:978-459-0175 ______________________________________________________ Get Your Private, Free Email at http://www.hotmail.com
Email: deweese-at-fas.harvard.edu Name: Alex de Weese School: Harvard College Question: Hello, I just finished a lab where we did cell fractionation, and looked at nuclear and cytosolic fractions stained with toluidine blue under a zeiss compound light microscope. I was wondering why I couldn't see mitochondria. With 400 times magnification, I expected to be able to see them in the cytosolic fraction. Is it that the mitochondria are not staining? Is it a problem with contrast? Thanks for your help. Sincerely, Alex de Weese
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Reply to: Re: good ideas Well put.
Messages with only a name, or even less, I just delete now.
It is so easy to sign these things, and so interesting to know who is writing them.
Regards,
Paul Webster
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm
Gib Ahlstrand 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 } -----------------------------------------------------------------------. } } } Responding to the message of {c5.40b6c0e.2626a950-at-aol.com} } from "COFAB2-at-aol.com"-at-sparc5.microscopy.com: } } } } ------------------------------------------------------------------------ } } 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 } } -----------------------------------------------------------------------. } } } } } } As a new associate of Don G. of Microscopy Today I am interested located } } interesting techniques and Problem/Solution type of information on all types } } of microscopy. } } } } } } } Whooooooooo are you.......who-oo....oo-oo. } :o } } (With appologies to Pete Townsend) } } } Gib } } } } Gib Ahlstrand } Electron Optical Facility, University of Minnesota, Dept. Plant Pathology } 495 Borlaug Hall, St. Paul, MN. USA. 55108 (612)625-8249 } 612-625-9728 FAX, giba-at-puccini.cdl.umn.edu } http://biosci.umn.edu/MIC/consortium.html } } } } } RFC822 header } ----------------------------------- } } Received: from ultra5.microscopy.com [206.69.208.10] by mailhouse.hei.org } with ESMTP } (SMTPD32-5.05) id A88059603E4; Thu, 13 Apr 2000 17:38:24 -0700 } Received: (from daemon-at-localhost) } by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id JAA11014 } for dist-Microscopy; Thu, 13 Apr 2000 09:39:41 -0500 (CDT) } Received: from no_more_spam.com (sparc5 [206.69.208.10]) } by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id JAA11011 } for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Thu, 13 Apr 2000 } 09:39:11 -0500 (CDT) } Received: from mhub2.tc.umn.edu (mhub2.tc.umn.edu [128.101.131.42]) } by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id JAA11004 } for {Microscopy-at-sparc5.microscopy.com} ; Thu, 13 Apr 2000 09:38:59 -0500 (CDT) } Received: from puccini.cdl.umn.edu by mhub2.tc.umn.edu with ESMTP for } Microscopy-at-MSA.Microscopy.Com; Thu, 13 Apr 2000 09:33:42 -0500 } Received: from [160.94.81.129] (x94-81-129.ppath.umn.edu [160.94.81.129]) } by puccini.crl (8.9.1b+Sun/8.9.1) with SMTP id JAA22934 } for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 13 Apr 2000 09:41:50 -0500 (CDT) } Date: Thu, 13 Apr 2000 09:41:50 -0500 (CDT) } Message-Id: {200004131441.JAA22934-at-puccini.crl} } From: "Gib Ahlstrand" {giba-at-puccini.cdl.umn.edu} } Reply-To: "Gib Ahlstrand" {giba-at-puccini.cdl.umn.edu} } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: good ideas } Mime-Version: 1.0 } Content-Type: text/plain; charset="us-ascii" } X-Mailer: POPmail 2.3b7 } Errors-to: Microscopy-request-at-sparc5.microscopy.com } X-RCPT-TO: {pwebster-at-hei.org} } X-UIDL: 242869585 } Status: U } Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm
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Reply to: RE: Coolwell Manuals & info Yes Joel,
Frank Haze, who owned Coolwell (and what magnificent chillers they are) sold the company to Litron (phone: 781 933 7300). I do not know to what extent they will continue to support the chillers but the contact I was given at Litron was Lohny Fultz.
Our chillers are currently being maintained by our refrigeration service contractors and they appear to be doing a great job of it (Cascade Refrigeration, Irvine CA).
Regards,
Paul Webster
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm
Joel McClintock 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 } -----------------------------------------------------------------------. } } } Please correct me if I am wrong, but I think I learned from this list that } Coolwell is defunct? I have a model SE-075W CZ that I am having trouble } with. No schematics and/or manuals to be found. Does anyone know where I } might find these or is willing copy important info from manuals they have on } this model? I would appreciate it. } } Still in the fact finding phase but I suspect the thermostat is the main } trouble with this unit. Do any of you know a supplier for these? Thank you } in advance. } } Joel McClintock } U of Kentucky } } } } } RFC822 header } ----------------------------------- } } Received: from ultra5.microscopy.com [206.69.208.10] by mailhouse.hei.org } with ESMTP } (SMTPD32-5.05) id AF40260A03F8; Thu, 13 Apr 2000 16:58:56 -0700 } Received: (from daemon-at-localhost) } by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id IAA10912 } for dist-Microscopy; Thu, 13 Apr 2000 08:53:06 -0500 (CDT) } Received: from no_more_spam.com (sparc5 [206.69.208.10]) } by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id IAA10908 } for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Thu, 13 Apr 2000 } 08:52:35 -0500 (CDT) } Received: from smtp.uky.edu (smtp.uky.edu [128.163.2.17]) } by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id IAA10901 } for {Microscopy-at-sparc5.microscopy.com} ; Thu, 13 Apr 2000 08:52:24 -0500 (CDT) } Received: from pop.uky.edu (pop.uky.edu [128.163.2.16]) } by smtp.uky.edu (8.9.3/8.9.3) with ESMTP id JAA81247 } for {Microscopy-at-sparc5.microscopy.com} ; Thu, 13 Apr 2000 09:47:08 -0400 (EDT) } Received: from nc.gws.uky.edu ([128.163.193.169]) } by pop.uky.edu (8.9.3/8.9.3) with SMTP id JAA20017 } for {Microscopy-at-sparc5.microscopy.com} ; Thu, 13 Apr 2000 09:47:07 -0400 (EDT) } Message-Id: {2.2.32.20000414015029.009c4fa8-at-pop.uky.edu} } X-Sender: jmcclin-at-pop.uky.edu } X-Mailer: Windows Eudora Pro Version 2.2 (32) } Mime-Version: 1.0 } Content-Type: text/plain; charset="us-ascii" } Date: Thu, 13 Apr 2000 21:50:29 -0400 } To: Microscopy-at-sparc5.microscopy.com } From: Joel McClintock {jmcclin-at-pop.uky.edu} } Subject: Coolwell Manuals & info } Errors-to: Microscopy-request-at-sparc5.microscopy.com } X-RCPT-TO: {pwebster-at-hei.org} } X-UIDL: 242869584 } Status: U }
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Hello, We have in our lab a Durst Laborator 138S enlarger. After all those years of it's perfect working, there are some layers of dust on condenser lenses. Does anybody know, how to safely clean the surface of condenser lens? Thanking you in advance. O. Benada
+-----------------------------------+ Oldrich Benada Acad. Sci. CR Institute of Microbiology Laboratory of electron microscopy Videnska 1083 CZ - 142 20 Prague 4 - Krc Czech Republic +------------------------------------+ Phone: +420-2-4752399 Fax: +420-2-4715743
Praveena: One place to start is Digital Instruments web site--DI.com. They have many applications notes on the subjects you are interested in. I believe other manufacturers also have helpful web sites. Steve
Hi,I was wondering whether anyone out there can help me out on the forced modulation and phase contrast technoques. I'm familiar with the basics of the AFM ,ie., contact and the non-contact modes. Now I want to learn the advances methods. Can anyone suggest any good books that give the full techniques? Praveena
Stephen McCartney Research Associate Virginia Tech Materials Institute 2108 Hahn Hall Blacksburg, VA 24061-0344 USA
If it like most condenser enlargers, the condenser lenses are opposing each other and retained in a metal cylinder. Just remove the cylinder, lenses and clean them with a cloth towel, moistened in a solution of Simple Green or a store-bought lens cleaning solution. rinse the lenses, wipe them dry and use a duster to get all moisture and lint off of them. That ought to do it. Check your enlarger's lens while you are at it. And also see if the bellow and lens plate areas need blowing out. Lots of dust collects in those nooks and crannies.
gary g.
At 11:58 PM 4/13/00 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Can anyone tell me the typical magnetic field strength at the sample in an SEM and (if possible) how fast the field drops off with distance from the axis?. Any references would be gratefully received.
} I think that electronics life depends in many cases not from how } long it operates but how many times you shut it ON and OFF.
I agree that this is true, especially with computers and, in this case, pump controllers. BTW, we have had to fix the controllers on several occasions.
} This kind of } "law" works perfectly for major electronic components as well as for CRTs, } computer components (HDs for instance). I guess, it may works for TPs too.
Maybe not. There are two competing factors: There is wear on the bear- ings due to the inevitable friction of running. Changes due to acceleration of the pump during shutdown and startup cause an increase in wear. To get optimal performance, these factors must be balanced. Our pumps have mechanical bearings, which will wear during the normal operation of the pump, so we have chosen to get the bearings replaced every two years. Since we are replacing the part that wears, the pump lifetime should not be shortened by this procedure. We also change the bearing oil on a regular schedule; I think that this prevents any problems from oil breakdown and/or acidification. There is negligable bearing wear produced by the oil changes, and the bearing replacement will prevent the accumulation of even this wear.
} } During start/stop TP components may sense some stress: more stress, } shorter life (we all know about that). I am experimented with magnetically } levitated TP from SEIKO (no financial interest) now. It works great. It } uses bearings only at low speed, at high speed the rotor is levitated.
With this kind of pump stop/start cycles will cause the mechanical bearings to be used, so there is logic in leaving them running continually.
} } Combination of this TP and "scroll pump" (oil free) gives me absolutely } oil-free vacuum system. I'll tell readers of this ListServer what happens } with that TP later. I am not going to service it at all.
Sounds like a good system; I'm glad you plan to post how things go. Yours, Bill Tivol
We are currently in the process of replacing our aging Phillips 300 TEM with a brand new all digital instrument that is going to be installed in virtually the same position as the old scope. As supervisor of the surgical pathology EM facility I have to say we have never had to question the resolution of the 300 and given the often sub optimal specimens that we work with the photo's are crisp and sharp. Any out of focus photographs are usually the result of out of focus operators, the occasional helicopter landing on the roof and yes trucks. This is seldom an issue and certainly not the scope. Here's our problem. We just had a site visit to measure the electrical and magnetic forces in the room (standard for installation of any new microscope). And to my suprise the room has an electrical/magnetic problem that "May effect the resolution of the new instrument" and quite possibility "the new scope will not meet spec in this location". So what to do. I seriously doubt that the old scope is giving me better resolution than the new scope will and I can't change locations at this point. So for biology or really anyone embedding in epoxy resins is there a standard or a number in angstroms that we can say is the minimum we will accept. If I am pleased with my photo's taken at 60,000 X do really need to insist that the instrument be capable of taking a sharp photo at 300,000X.
this problem is not limited to image processing, of course. IP only makes it easier to use or abuse these things. If you think about how you get the signals from the SEM (or any other image source), there are many ways the signal can be distorted. For example, the detector may not be linear (as is the case for many old cameras and, for that matter, the human eye), signals may be distorted during transmission, noise is added, the film for photographing has certain characteristics, and the darkroom work can be more of an art than science. Forensic science has had to deal with this for some time. If an argument in a court of law can be made that a picture has been "doctored", it will lose it's effectiveness or be dismissed. In Forensics it is very important to keep a record of the whereabouts and the processing done to "evidence" at all times. Sometimes people burn a "gray scale" into the image before they do any processing. This gray scale will then show what happened to the image during processing. Of course, this is not foolproof as anybody can put on another gray scale after processing. But this is the same as inventing measurement data, which can be done, but is not a real problem in science as it just cannot be repeated independently. If you need to be absolutely sure, get a step-by-step recipe of the image processing and have someone else repeat it. If the results are different, you have reason to be concerned. If they are identical, you have your answer. You need to be familiar with the possibilities and shortfalls of image processing. You can start with an image that shows just noise, apply some Fourier-filters and other processing and end up with a periodic structure on the image. This, however, is not due to the image processing, it is due to using the wrong tools (or use the tools the wrong way).
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: David Henriks [mailto:Henriks-at-CompuServe.COM] Sent: Wednesday, April 12, 2000 11:14 AM To: Martyn Harris; Micro Listserver
Dear Martyn:
Several years ago (MSA in Cleveland if I recall correctly), there was some sort of roundtable discussing the ethics of image manipulation. I can't remember much more than that, but perhaps those with a better memory who may have participated in the discussion would have some good input.
David Henriks Vice President 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
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy. Message text written by INTERNET:"HARRISm-at-esm-semi.co.uk"-at-sparc5.microscopy.com } A general enquiry regarding integated circuit X section images :
In semiconductor failure investigation various etchants and staining methods are used to obtain and or enhance selected features prior to FESEM investigation and image capture . As I now capture images digitally and have the increasing capabilities
of image processing at my fingertips it's possible to ' modify' and
possibly distort the initially accurate image obtained which could lead to misleading results .
I receive people's images and wonder if they are a result of different
sem capabilities / preparation methods / etches or are they simply
'touched up '. They probably think the same about mine .
Therefore is there any way of controlling image processing to enable like for like comparison ? or any international standards ? way of indicating on an image it has been subject to processing or is it now a case of you cannot believe what you see ?
A student has come to me with the following problem. Can anyone assist us?
Ron L.
Please cc answers to:
saeeda-at-bu.edu
Does anybody know of any good fixing/dehydrating techniques for viewing this sample using the SEM? Alcohol and CPD dehydration techniques are not yielding good results and there is a lot of distortion of the gel. I am looking for something that will enable me to preserve the features of the gels, and analyze their morphology. These gels will be patterned with collagen and cells will be grown on them. I am interested in viewing these patterns of collagen, and the corresponding patterns of cell growth. Thanks.
If the lens is coated, which some Durst condensers are, I would suggest starting with a Blower Brush, available in most camera shops. Hopefully you can blow/brush most or all the dust away. If that fails, one of the new "microfiber" lens cloths work very well on most lenses. A high quality photographic lens cleaning solution should be safe, but I would try to clean without any chemicals first.
George Laing National Graphic Supply Albany, NY USA www.ngscorp.com
-----Original Message----- } From: Oldrich Benada [mailto:benada-at-biomed.cas.cz] Sent: Friday, April 14, 2000 2:58 AM To: Microscopy-at-sparc5.microscopy.com
Hello, We have in our lab a Durst Laborator 138S enlarger. After all those years of it's perfect working, there are some layers of dust on condenser lenses. Does anybody know, how to safely clean the surface of condenser lens? Thanking you in advance. O. Benada
+-----------------------------------+ Oldrich Benada Acad. Sci. CR Institute of Microbiology Laboratory of electron microscopy Videnska 1083 CZ - 142 20 Prague 4 - Krc Czech Republic +------------------------------------+ Phone: +420-2-4752399 Fax: +420-2-4715743
Does anyone know of a supplier of used ultramicrotomes? I'm looking for a Leica ultracut E, in good working order. Is there anyone who is dismantling a TEM lab who wants to sell one? Thanks.
Norman Michaud Director, Morphology Mass Eye and Ear Infirmary Ophthalmology-5th flr. 243 Charles St, Boston, MA 02114 norman_michaud-at-MEEI.Harvard.edu Tel:617-573-3316; Fax:617-573-4290
I can suggest two possibilities:1) atomic force microscopy of the hydrated gel; 2) high-pressure freezing of the gel, followed by either cryoSEM or cryo-ultramicrotomy and cryoTEM (cryoSEM would be simpler than cryo-ultramicrotomy).
Phil
} A student has come to me with the following problem. Can anyone assist us? } } Ron L. } } Please cc answers to: } } saeeda-at-bu.edu } } } Does anybody know of any good fixing/dehydrating techniques for viewing this } sample using the SEM? Alcohol and CPD dehydration techniques are not } yielding good results and there is a lot of distortion of the gel. I am } looking for something that will enable me to preserve the features of the } gels, and analyze their morphology. These gels will be patterned with } collagen and cells will be grown on them. I am interested in viewing these } patterns of collagen, and the corresponding patterns of cell growth. } Thanks. } } Saeeda
-- }}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{ Philip Oshel Supervisor, AMFSC and BBPIC Dept. of Animal Health and Biomedical Sciences University of Wisconsin 1656 Linden Drive Madison, WI 53706-1581 voice: (608) 263-4162 fax: (608) 262-7420 (dept. fax)
Hello world I think i have tried everything, but still!! Is there somebody, who nows how to detect Glycolipid/lipid on cryo ultrathin sections? And yes, i have read Dr. Wim's article, where he compare different metodes. I have been on a course in Austria, where they also told me, that this ( Dr. Wim) would be the way to decet lipid. But in the real world, it does not work. The lipid is floating all over the sections. Tanks for your help Sincerely Sus S¿rensen
Dear Bill, It was nice discussion there. Thanks for your nice reply. I totally agree with you. No question, the good service in time is a great deal! Unfortunately the service sometimes is not such great as you have with yours TPs. Again, sometimes people just do not have enough money to do service in time.
As for magnetically levitated TP, you right, it is good idea to keep it running continuously. Currently, I am working on my system in the way to be able to insert samples through air-lock, than it will be possible do not break vacuum to load samples. This is my plan. Wish me luck. Sergey
} Date: Fri, 14 Apr 2000 10:59:20 -0400 } From: William Tivol {tivol-at-wadsworth.org} } Subject: Re: TP service } Sender: tivol-at-wadsworth.org } To: microscopy-at-sparc5.microscopy.com } X-Mailer: Mozilla 4.07C-SGI [en] (X11; I; IRIX64 6.5 IP21) } } ------------------------------------------------------------------------ } 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
There are a LOT of suppliers of inexpensive light microscopes in the educational marketplace; many of the scopes are remarkably good. You'll find a list of suppliers on the Project MICRO web page (URL below). Get several catalogs; if you compare descriptions and photos, you'll realize that prices can vary almost 50% on the same item.
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/PCI/pci.html
I agree with Bill Tivol. It is less stressfull on electrical circuits and contacts to leave the power on. To do so prolongs the life of the part, as well as the reliability of voltage flow through an instrument. A good example is XRD and XRF tubes. Rule #1 I learned as a grad student was to always leave the power on, keep a current flowing through the tubes. Why? A power current "burns in" at a single point on the tube. If the power is shut off, the burn in point can drift. Drift actually weakens the tube, which is the opposite of what you would expect. The drift will also cause current fluctuations, creating another source of error reducing the precision and accuracy of the measurements. At $3000-$4000 per pop on a tube, it is prudent to do what ever you can to extend its life. Same applies to any electrical instrumentation or accessories.
Lou Solebello -----Original Message----- } From: William Tivol {tivol-at-wadsworth.org} To: microscopy-at-sparc5.microscopy.com {microscopy-at-sparc5.microscopy.com}
----- Original Message ----- } From: Michele Palmer {moonlite-at-csd.uwm.edu} To: {wchiss-at-ou.edu} Sent: Friday, April 14, 2000 7:53 PM
Hi
The question of much resolution do we really need and what resolution is obtainable often confronts the EM service engineer and the EM consultant?
As mentioned before we at Protrain buy instruments for clients or in conjunction with clients. On many occasions the proposed site does not meet the manufacturers requirements in their entirety but circumstances mean we must go ahead.
Whilst these situations are not ideal one must consider the clients application and the desired performance for that application. On many occasions checking an instrument in a clients laboratory, where the client does not see a problem in their results (5,000 to 30,000X), we see image instability at 200,000X. As a service engineer you judge the problem and decide if it is a simple fix or a cause for deep investigation. Often discussions with the client result in keeping going whilst constantly checking the magnitude of the fault. This route may be followed for many years before the engineer, as the client still has no visible micrograph problems, decides to get in and chase the fault.
So what am I saying? In the 70s high quality work was carried out on TEM that had a point to point resolution of around 1nm. We are all aware that 1nm at 100,000X is equal to 0.1mm on the micrograph and we need a hand lens to visualise this! In most routine medical observations, other than virus work, the typical max is about 30,000X, about 3nm limited by the section thickness (?) so would it not be sufficient to use a 1nm machine?
Many biologists still use very low kV, i.e. 80, so they are degrading their 0.3nm instrument still further. Running at 100 or 120kV will provide better quality images using dark room procedures to aid contrast. Higher accelerating voltage will help with fields and the modern antivibration systems, available as discussed earlier this month, will help overcome floor vibration.
So my advice after working with TEM for 36 years all over the world in all sorts of very poor environments is GO FOR IT! I will not take up space by relating the amazing stories of dreadful sites and superb microscopy, it certainly does happen!
Good luck
Steve Chapman Senior Consultant Protrain - for consultancy and training world wide Tel & Fax +44 01280 814 774 e-mail protrain-at-emcourses.com web www.emcourses.com
SUMMER I 2000 COURSE ANNOUNCEMENT - Transmission Electron Microscopy (BIO. 221-Section B)
NASSAU COMMUNITY COLLEGE, Garden City, Long Island, New York
A five week, Summer Session I 2000 semester, course in Biological Transmission 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 22 and end on June 22, 2000.
This is a "hands-on" course emphasizing biological specimen preparation, ultra-thin sectioning involving block trimming, glass knifemaking and operation of the ultramicrotomes (Sorvall MT-2B and LKB Ultrotome III), thick and ultra-thin section mounting and contrast staining (UA and Pb citrate), grid support films (formvar, carbon), student operation of the TEM (Hitachi HS-8, Philips EM 300) and production of electron micrographs through the process of black & white photography, and electron micrograph analysis. Students will work on a chosen sample(s) with the goal of producing a portfolio of high quality TEM photomicrographs of that sample(s).
The course is widely transferrable and the cost per credit is reasonable at $86 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 the beginnings of a 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/Sum00/index.html.
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 2000 COURSE ANNOUNCEMENT - Transmission Electron Microscopy (BIO. 221-Section B)
NASSAU COMMUNITY COLLEGE, Garden City, Long Island, New York
A five week, Summer Session I 2000 semester, course in Biological Transmission 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 22 and end on June 22, 2000.
This is a "hands-on" course emphasizing biological specimen preparation, ultra-thin sectioning involving block trimming, glass knifemaking and operation of the ultramicrotomes (Sorvall MT-2B and LKB Ultrotome III), thick and ultra-thin section mounting and contrast staining (UA and Pb citrate), grid support films (formvar, carbon), student operation of the TEM (Hitachi HS-8, Philips EM 300) and production of electron micrographs through the process of black & white photography, and electron micrograph analysis. Students will work on a chosen sample(s) with the goal of producing a portfolio of high quality TEM photomicrographs of that sample(s).
The course is widely transferrable and the cost per credit is reasonable at $86 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 the beginnings of a 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/Sum00/index.html.
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}
BX50 stand trinoc head 2ea 10X eyepieces 6-place nosepiece Phase condenser Phase inserts Rt stage UPlan FL Phase 4X, 20X, 40X, 60X, 100X, Plan 10X objectives 100 Watt lamphouse
Like new condition throughout.
Please contact me if you are interested in either of these system. Telecon is 916.791.8191
I also have two Olympus PM10AD computer control photo systems which include the control unit, shutter body, focusing telescope and connecting cable. PE eyepieces are also available.
Email: arthurmott-at-netzero.net Name: Arthur Mott
Question: I have a Bausch & Lomb 0.7X-3X which I wish to convert to gemological use(viewing gem stones). What type of lighting systems should I use , or where can I gather additional information.
} } } I agree with Bill Tivol. It is less stressfull on electrical circuits and } contacts to leave the power on. To do so prolongs the life of the part, as } well as the reliability of voltage flow through an instrument. A good } example is XRD and XRF tubes. Rule #1 I learned as a grad student was to } always leave the power on, keep a current flowing through the tubes. Why? } } A power current "burns in" at a single point on the tube. If the power is } shut off, the burn in point can drift. Drift actually weakens the tube, } which is the opposite of what you would expect. The drift will also cause } current fluctuations, creating another source of error reducing the } precision and accuracy of the measurements. } }
This is a pretty interesting way of looking at things, but it seems like an argument for leaving things eg X-Ray tubes running at their working conditions (kV and mA) fulltime. Can you expand on this "single point"? Is it some physical point on, for example, a filament, or is it kind of metaphorical? I can't quite understand the mechanism of the phenomenon you are describing. I've always just thought that the reason for leaving X-Ray tubes on, but at minimum power, was to avoid the current inrush through a cold (and therefore low-resistance) filament, plus the thermal stresses in repeatedly warming up and cooling down of the tube and all its glass-to-metal seals..
cheers
rtch
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
hi there Arthur, it is a dark field method of illumination and gemological assn. of America sells a base that the b and l pod will fit. there are also some third parties that do this base also but any of them can be pricey. check out gia on the net. also try search under key gemscope thanks Ed Sharpe archivist for SMECC
Whilst working in Australia we found a few labs using something called Shellite as a de-greaser to clean microscope parts. This works very well and seems to be very available. we then looked for the same product here in SA. We then found that this is a trade name only in Australia and is actually petroleum ether or spirit. We called the local Shell distributor to ask what the difference was between spirit and ether. Well, they were not sure on that one. Then we were told you must specify the temperature range of the petroleum Ether. 40 - 60, 60 - 80 or 80 - 100 deg Celsius. Again no explanation as to what the difference is an why. Can any one give us some more info on Petroleum ether / spirit and the advantages disadvantages on the different temperatures.
Thanks
Luc Harmsen Anaspec, South Africa Technical support on microscopy. Tel + 27 (0) 11 476 3455 Fax + 27 (0) 11 476 7290 anaspec-at-icon.co.za www.anaspec.co.za
Remember, ICEM 15 will be in 2002, Durban, South Africa. www.icem15.com
It is a real physical phenomonom. I dont think I understand it correctly myself, except if I think about like a spark plug in a car. You are right about the vacuum, but take a look at some of your old tubes if you have any. You should be able to see a gray smoky discoloration of the glass tubing at some area on the tube, usually on one side near the center. That is the burn in point.
That also brings up a curiosity question to me....Does it happen to the newer cermaic tubes? I dont know, and will have to ask. The newer ceramic tubes are supposed to have a longer life and reliability compared to the glass ones. -----Original Message----- } From: Ritchie Sims {r.sims-at-auckland.ac.nz} To: Lou Solebello {microls1297-at-mindspring.com} ; Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
Hi, Lou
} } } I agree with Bill Tivol. It is less stressfull on electrical circuits and } contacts to leave the power on. To do so prolongs the life of the part, as } well as the reliability of voltage flow through an instrument. A good } example is XRD and XRF tubes. Rule #1 I learned as a grad student was to } always leave the power on, keep a current flowing through the tubes. Why? } } A power current "burns in" at a single point on the tube. If the power is } shut off, the burn in point can drift. Drift actually weakens the tube, } which is the opposite of what you would expect. The drift will also cause } current fluctuations, creating another source of error reducing the } precision and accuracy of the measurements. } }
This is a pretty interesting way of looking at things, but it seems like an argument for leaving things eg X-Ray tubes running at their working conditions (kV and mA) fulltime. Can you expand on this "single point"? Is it some physical point on, for example, a filament, or is it kind of metaphorical? I can't quite understand the mechanism of the phenomenon you are describing. I've always just thought that the reason for leaving X-Ray tubes on, but at minimum power, was to avoid the current inrush through a cold (and therefore low-resistance) filament, plus the thermal stresses in repeatedly warming up and cooling down of the tube and all its glass-to-metal seals..
cheers
rtch
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
The difference between photobleaching and quenching is quite fundamental.
Basically when a molecule is photobleached it is converted to another molecule that is not fluorescent -i.e. a reaction takes place in which the fluorescing species is converted to another species.
Quenching on the other hand results from the excited fluorescent molecules having a non radiative route to release the energy it would typically release as fluorescence. The state of the molecule after the quenching however is the same as if it had fluoresced. ---------------------------------------------------------------------------- --------------------------- Dr. Giles Sanders Zeneca / SmithKline Beecham Centre for Analytical Sciences Chemistry Department Imperial College of Science, Technology and Medicine London SW7 2AY
(44) - 0171-594-5749
Never express yourself more clearly than you think. -- Niels Bohr (1885-1962) Danish physicist
---------------------------------------------------------------------------- ------------------------ ----- Original Message ----- } From: {"catchanangel-at-aol.com"-at-sparc5.microscopy.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: 15 April 2000 03:10
Luc, The terms "petroleum spirit" and "petroleum ether" are interchangeable. You may also encounter "light petroleum" and "pet ether". The numbers correspond to the boiling ranges in degrees celsius - they are just differnt fractional distillates. The 40-60 fraction is roughly hexane ( plus small amounts of larger and smaller hydrocarbon molecules). The higher boiling fraction will not evaporate as cleanly as the lighter ones, but all are good solvents for grease and oil. They are, of course, highly flammable.
Regards, Eric
---------------------- Dr Eric Lachowski Department of Chemistry University of Aberdeen Meston Walk Old Aberdeen AB24 3UE Scotland +44 (0)1224 272934 fax 272921 e.lachowski-at-abdn.ac.uk
Check the web site for the Gemmological Institute of America (GIA). I forget the exact URL, it's at home (www.gia.com? .org?). They sell a stereoscope designed for exactly this purpose, and looking at it will give you a good idea of what you need.
Briefly, it uses separate transmitted and reflected light sources. The transmitted light I believe is an "ordinary" tungsten microscope bulb, and the reflected source is a closely mounted fluorescent source. The stones are held by a "third-hand" type of gripper so that it can be examined from all angles. This is important not just in studying inclusions, but because the optical characteristics (including color) of some gems change with the crystal axis (the Usambara effect, if I've spelled that right). Different types of light are also needed, as color-change gems show different colors depending on the incident light. For best effect, I would also had an optical fiber source with dual goosenecks (not a ring-light, except in addition), and a *good* mirror to shine sunlight on the specimen. This is for judging stone color as well as color change.
The GIA site is a good source of information, but I'd check the Nat. Mus. Natural Hist./Smithsonian, and the Mus. Nat. Hist. in London also, to start.
Phil
} Can anyone answer this? } } Nestor } } -------------------------------------------------------------------------- } } Email: arthurmott-at-netzero.net } Name: Arthur Mott } } Question: I have a Bausch & Lomb 0.7X-3X which I wish to convert to } gemological use(viewing gem stones). What type of lighting systems should } I use , or where can I gather additional information. } } Arthur } } ---------------------------------------------------------------------------
-- }}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{ Philip Oshel Supervisor, AMFSC and BBPIC Dept. of Animal Health and Biomedical Sciences University of Wisconsin 1656 Linden Drive Madison, WI 53706-1581 voice: (608) 263-4162 fax: (608) 262-7420 (dept. fax)
Naturally I forgot the darkfield. (#*&$(&# Probably because it's important.
Phil
} hi there Arthur, } it is a dark field method of illumination and gemological assn. of America } sells a base that the b and l pod will fit. there are also some third parties } that do this base also but any of them can be pricey. } check out gia on the net. also try search under key gemscope } thanks Ed Sharpe archivist for SMECC
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
What do you mean by "convert"? Are you referring to a "dissecting" scope? If so, you multiply the number that you've given (presumabvly the power of the objective lens) times the power of the eyepiece, which is often 10x. That would mean that you may have 30x available, and that definitely requires good illumination. My son-in-law, who is a professional diamond cutter, really likes the flexibility and brightness of fiber optic illuminators - but they're expensive. The Edmund Optical catalog is a good place to begin. If you want specific gemological advice, contact the Gemological Institute of America (GIA).
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/PCI/pci.html
I don't think, there is an easy answer to your question. It depends on the microscope type (immersion lens or not), the lens configuration and design, and last but not least on the working distance, aside from the lens current, which depends on the acceleration voltage.
I have a few pointers for you, though. I remember faintly a book by Glaser, but that may be in German (anybody with a good reference for this book?).
Another book is: Ludwig Reimer (Scanning Electron Microscopy : Physics of Image Formation and Microanalysis (2nd Ed)(Springer Series in Optical Sciences, Vol 45) )
Here is a URL for the book at Amazon.com: http://www.amazon.com/exec/obidos/ASIN/3540639764/qid=955982926/sr=1-17/ 102-8036949-3232817
This book is a bit "theoretical", but has information about electron optics in the first two chapters.
You may want to search the net for "electron optics".
Hope this helps.
Michael
(disclaimer: I have no interest in Amazon.com. Check out other bookstores for prices)
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Chris Walker [mailto:chris.walker-at-physics.org] Sent: Friday, April 14, 2000 8:38 AM To: 'Microscopy-at-MSA.Microscopy.Com'
Can anyone tell me the typical magnetic field strength at the sample in an SEM and (if possible) how fast the field drops off with distance from the axis?. Any references would be gratefully received.
The pod, i.e., the top part that is the business end just comes out of the stand of a b&l zoom and drops in the gia base...
Of course for adventure one can build the illuminating base necessary by hand if so inclined.
Ed Sharpe
{ { Subj: Re: Bausch & Lomb Question: Date: 4/17/00 11:41:32 AM US Mountain Standard Time From: schooley-at-mcn.org (Caroline Schooley) To: arthurmott-at-netzero.net CC: Microscopy-at-sparc5.microscopy.com
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} ------------------------------------------------------------------------ } 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 } -------------------------------------------------------------------------- } } Email: arthurmott-at-netzero.net } Name: Arthur Mott } } Question: I have a Bausch & Lomb 0.7X-3X which I wish to convert to } gemological use(viewing gem stones). What type of lighting systems should } I use , or where can I gather additional information. } } Arthur -
What do you mean by "convert"? Are you referring to a "dissecting" scope? If so, you multiply the number that you've given (presumabvly the power of the objective lens) times the power of the eyepiece, which is often 10x. That would mean that you may have 30x available, and that definitely requires good illumination. My son-in-law, who is a professional diamond cutter, really likes the flexibility and brightness of fiber optic illuminators - but they're expensive. The Edmund Optical catalog is a good place to begin. If you want specific gemological advice, contact the Gemological Institute of America (GIA).
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.microsc } }
I am in need of a service contract for an ISI DS-130 SEM. Is there anyone who is particularly happy with a contract on their ISI (Topcon) instrument? ---------------------- Douglas R. Keene Associate Investigator Shriners Hospital Research Facilities 3101 S.W. Sam Jackson Park Road Portland, Oregon 97201 phone: 503-221-3434 FAX: 503-412-6894 (9-5 PST) e-mail: DRK-at-shcc.org
Try the following EELS database site. The have a number of O containing materials there with all of the acquisition parameters well documented. http://www.cemes.fr/eelsdb/
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
} -----Original Message----- } From: Jo Verbeeck [mailto:joverbee-at-ruca.ua.ac.be] } Sent: Tuesday, April 11, 2000 10:09 AM } To: Microscopy listserver message adress } Subject: EELS: need spectrum for comparing } } } -------------------------------------------------------------- } ---------- } 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`m looking for an EELS spectrum containing Oxygen K-edge and some } other edge like Mn or Ti L-edge. Together with low loss } spectrum and data } about the angles. } The spectra need to have a power of 2 (1024 for inst.) bins } and low loss } and high loss should have same size. } I want to use this for comparison, i`m trying to do some } quantitative EELS } work but so far with limited success. } I want to find out wether my program or my data is wrong;) } } Thanks, } } Jo } } ************************************************************* } * Jo Verbeeck * } * University of Antwerp * } * Dept. EMAT (Electron Microscopy for Materials Research) * } * e-mail: joverbee-at-ruca.ua.ac.be * } * tel: +32(0)3 218 02 49 * } * fax: +32(0)3 218 02 57 * } ************************************************************* } }
We have a JEOL 840A scanning scope with an Oxford Link X-ray analysis system. We would like to be able to get digital images out of it, for as few $$ as possible. I would be interested in comments on the following questions:
1) passive vs. active acquisition (my impression is that the resolution is better with active, but how much?)
2) relative merits of different commercially available systems (dPict, 4PI, GW Electronics, etc).
3) Are NIH Image and Photoshop sufficient for analysis and processing of images?
4) relative merits of various X-ray analysis software (such as FLAME, etc).
Thanks very much.
Kate L-P -- Kate Luby-Phelps Molecular & Cellular Imaging Facility UT Southwestern Medical Center 5323 Harry Hines Blvd. Dallas, TX 75390-9039 e-mail: lubyphel-at-utsw.swmed.edu Telephone: (214) 648-2190 Fax: (214) 648-6408 or -8885
Email: jeffc07-at-hotmail.com Name: Jeff Courter School: central michigan universty
Question: my problem is this: I have E. coli and P. mirabilis in 1% low temperature gelling agarose in spurr's. is there any way to visualize the specimen to trim it on a microtome other than taking sections individually and staining them until i find the agar and bacteria.
I understand that Shellite is a Shell Petroleum Co trade name, the generic name in Australia is white gas and this product is very similar to petrol (car fuel) - except it does not have the certain additives. ( I ran in desperation a Landcruiser for 50km on that stuff some years ago) It works well enough as a general solvent for a first degreasing and cleaning of EM parts, especially pumps. Its quiet cheap to purchase. Petroleum Ether much more volatile and a more powerful solvents. (more explosive too) They may be the same chemically, except that white gas would have a considerably higher boiling point than Pet. Ether. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Monday, April 17, 2000 8:47 PM, Anaspec [SMTP:anaspec-at-icon.co.za] wrote: } } } } Hi all } } Whilst working in Australia we found a few labs using something called } Shellite as a de-greaser to clean microscope parts. This works very well and } seems to be very available. we then looked for the same product here in SA. } We then found that this is a trade name only in Australia and is actually } petroleum ether or spirit. } We called the local Shell distributor to ask what the difference was between } spirit and ether. Well, they were not sure on that one. Then we were told } you must specify the temperature range of the petroleum Ether. 40 - 60, 60 - } 80 or 80 - 100 deg Celsius. Again no explanation as to what the difference } is an why. } Can any one give us some more info on Petroleum ether / spirit and the } advantages disadvantages on the different temperatures. } } } Thanks } } Luc Harmsen } Anaspec, South Africa } Technical support on microscopy. } Tel + 27 (0) 11 476 3455 } Fax + 27 (0) 11 476 7290 } anaspec-at-icon.co.za } www.anaspec.co.za } } Remember, ICEM 15 will be in } 2002, Durban, South Africa. } www.icem15.com
Kate, I saw your message on the Listserver. Here is my opinion on Digital Imaging.
Passive vs. Active - Passive resolution is limited to the maximum resolution the SEM can produce. If your scan generator has a maximum resolution of 2000 lines for the photo scan, then your passive resolution will also be 2000 lines. Passive just simply reads what's there already, including any character, text and markers. What you see on the Viewing CRT is "what you get" on the Passive System computer display. One neat thing that a Passive System can do is grab an image in split screen mode, one side a SE or BSE Image and the other half an x-ray dot map. An Active System, on the other hand, takes direct control of the scan coils(replacing the scan gen. in the sem), and digitizes the resulting video. Systems such as DIGISEM, can achieve images of 4K x 4K. I believe the 4Pi system goes even higher. I'm not sure about the rest. Active systems are typically suited for fast, high quality digital images, that are then viewed and modified with a program such as PhotoShop. Then, finally you have the standard TV Rate Frame Grabber. This only works in "TV" mode, and your resolution is typically limited to the resolution of the SEM TV scan rate(512 x 512?). Slow scan, either Active or Passive is the way to go. Do it yourself Active and Passive systems start out around $8,000.00, and of course, go up from there.
NIH IMAGE was originally written as a MAC application. However, it has been ported to the PC platform. In the process, the program has lost some of its functionality. PhotoShop, for both the PC and MAC is a superior program overall. There are some "shareware" programs out there worth looking at, such as Paint Shop Pro. It's cheap(less than $100) and can do just about anything PhotoShop can do(my opinion). Also, be aware that many imaging systems include image analysis function. This really drives the price up. If all you are interested in doing is simply grabbing and saving an image, I would stay away from the "high end" systems.
The 4Pi imaging system is available both in a MAC and PC version. I think everybody else is PC based.
EDS - What a can of worms. It seems everybody(including myself), offers a "PC based EDS upgrade". I have personally used the PGT Avalon(formerly American Nuclear Systems) and 4Pi Flame. The PGT Avalon eXcalibur software in its latest release is full 32 bit and can run on the Win95/98 and NT Workstation 4.0. PGT corrected many software problems after purchasing ANS. Very nice package. The Flame software uses "fuzzy logic" in its approach to quantitative analysis. The latest rendition of the Flame software seems pretty good, however, the MAC version is somewhat more stable. Also, an excellent product, along with 5***** customer support. Keep an eye out on 4Pi though, something new and exciting is in the works. Both are great systems, inexpensive and easy to operate. Again, the 4Pi system is available on both the PC and MAC platform. You will find prices on systems such as these are very close in price to each other. Prices for these "upgrades" typically start out around $15,000.00. Good luck in your quest for the right system.
Gary M. Easton Scanners Corporation www.scannerscorp.com
Note: I am not employed by any of the companies listed above(except for Scanners Corporation). However, I do have a commercial interest in these products, because I sell them and this is how I make a living(along with servicing SEM's). If I have slighted anybody, or any company, I apologize in advance. The opinions and facts I have stated here are a result of my 22+ years experience in the scanning electron microscopy field
----- Original Message ----- } From: Kate Luby-Phelps {lubyphel-at-SWVX12.SWMED.EDU} To: {Microscopy-at-sparc5.microscopy.com} Sent: Monday, April 17, 2000 4:37 PM
Douglas - We have Image Control, Inc., based in Orlando, FL, service our ISI-DS130C. We don't have a contract, just "as needed" service. I have been very happy with service and pricing. They do offer service contracts as well. I don't know what their geographical range is. Phone number is (407) 234-0676; fax (407) 292-7802.
Jill Jill Verlander Reed, D.V.M. Associate Scientist Director, College of Medicine Electron Microscopy Core Facility University of Florida P.O. Box 100215 Gainesville, FL 32610 verlaj-at-medicine.ufl.edu Phone: (352) 846-0820 Fax: (352) 846-3299
let me try to answer some of your questions. I am sending this cc to the listserver as some of your questions come up on a regular basis.
1) passive vs. active: Both types of acquisition will give you probably good images. The difference is, that in a passive system, the computer (or digitizer) simply digitizes the signals from the microscope. That means, that you are limited to whatever the microscope can supply in terms of resolution, dwell time, aspect ratio, etc. Most SEMs have a 1000 line or 2000 line option for taking images. That's what you get. A 1000x1200 image (for 1000 lines), or a 2000x2400 image (for 2000 lines). Plus of course the other modes of the microscope (slow scan, fast scan, etc). An active system actually takes control of the scanning coils, usually by replacing the scan generator during PC controlled acquisition. Thus the PC can select the resolution, dwell time, aspect ratio, etc. This gives you much more freedom in selecting an image format, plus it is much easier to acquire X-ray dot maps with an existing X-ray system. Typical max. resolutions are 4000x4000. These big images take time to acquire, though (seconds). Installation of a passive system may be easier in some circumstances, but on your 840 the installation of an active system is "plug and play".
2) different commercial systems. As we sell our own system (ADDA II), and since this goes to the listserver, I don't want to comment on this question. Call me if you need information about our system.
3) Software: Of the two programs you mention, I would prefer NIH. Photoshop is mainly for making photos look nicer and may not have all the tools you require (unless you buy other add-ons). Buy the software, however, with an eye on expansion and support. Our experience with other users is, that once they have added a digital acquisition system, they quickly find other things they can do with the system and sometimes need more software or hardware. For example, they realize that the old light microscope used for specimen preparation can be digitized too and be used much better than before. That, for example, would require a camera. So, if that is the case, you want to look for a system that also supports cameras. Another issue is the distribution of images. Many of our customers have predefined "reports" for their customers. In that case you want to look for software that supports the creation of custom templates, not just printing images. An embedded, network capable database with search capabilities is a big plus if you have several users.
4) X-ray software: This is outside my area of expertise, so I will refrain from making any comments.
I hope, this helps you decide about your SEM. If you have more detailed questions, please send me an email or call me.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
******************************************************* Disclaimer: Soft Imaging System produces and sells image acquisition and processing systems. We therefore have a vested interest in some of the items mentioned above. *******************************************************
-----Original Message----- } From: Kate Luby-Phelps [mailto:lubyphel-at-SWVX12.SWMED.EDU] Sent: Monday, April 17, 2000 2:37 PM To: Microscopy-at-sparc5.microscopy.com
We have a JEOL 840A scanning scope with an Oxford Link X-ray analysis system. We would like to be able to get digital images out of it, for as few $$ as possible. I would be interested in comments on the following questions:
1) passive vs. active acquisition (my impression is that the resolution is better with active, but how much?)
2) relative merits of different commercially available systems (dPict, 4PI, GW Electronics, etc).
3) Are NIH Image and Photoshop sufficient for analysis and processing of images?
4) relative merits of various X-ray analysis software (such as FLAME, etc).
Thanks very much.
Kate L-P -- Kate Luby-Phelps Molecular & Cellular Imaging Facility UT Southwestern Medical Center 5323 Harry Hines Blvd. Dallas, TX 75390-9039 e-mail: lubyphel-at-utsw.swmed.edu Telephone: (214) 648-2190 Fax: (214) 648-6408 or -8885
} the generic name } in Australia is white gas and this product is very similar to petrol (car fuel) } - except it does not have the certain additives.
White gas is the petroleum distillate fraction used to make petrol.
} They may be the same chemically, except that white gas would have a } considerably higher boiling point than Pet. Ether.
White gas contains hydrocarbons that have a boiling point roughly the same as that of octane. There are some chemical differences, since the higher-boiling-point fractions will contain more complex mixtures than the lower-boiling-point fractions. In particular, the lowest such fraction, which contains mostly pentanes, has no aromatics. Yours, Bill Tivol
Following the thread on processing tissue-culture cells for TEM, would C Singla reply back to me if they would like a protocol for embedding cell monolayers for TEM in the petri-dish.
Jeremy Sanderson
jb_sanderson-at-yahoo.com
__________________________________________________ Do You Yahoo!? Send online invitations with Yahoo! Invites. http://invites.yahoo.com
I'm currently in the process of setting up a Histology/Microbiology/EM facility. Being out of Histology for almost 30 years (been doing EM) I'm not sure of the best type of tissue processor to buy. I won't be doing that many specimens, so I'm leaning towards a rotary tissue processor. I have info on Shandon and Tissue-Tek. Are there others out there? Anyone having opinions on the 2 mentioned, I would appreciate any comments. I am looking for ease of use (changing solutions, fume safeguards), reliability on service and cost effectiveness. Thanks.
Peace be with you, Phil Rutledge (410)778-4136, 2120 prutledge-at-ars.usda.gov
I have been using folding grids for my sections for a long time (200 mesh, Cu). Lately however I have found it very difficult to fold the grids so that the two sides overlap properly and only a few of the holes are in good registry, the rest are partially blocked by the overlap. I am wondering if this is a manufacturing problem ( i.e. a bad batch ?) . Has anyone else experienced this problem with a batch of grids ? Suggestions are welcome.
There will be an electrical characterization by AFM seminar/workshop sponsored by Digital Instruments and Princeton University on April 25, 2000. For additional information and registration, please visit the DI web site at www.di.com and click on "workshops and seminars".
Regards, Jeff Doran Digital Instruments Veeco Metrology Group
There will be an electrical characterization by AFM seminar/workshop sponsored by Digital Instruments and Princeton University on April 25, 2000. For additional information and registration, please visit the DI web site at www.di.com and click on "workshops and seminars".
Regards, Jeff Doran Digital Instruments Veeco Metrology Group
} though (seconds). Installation of a passive system may be easier in some } circumstances, but on your 840 the installation of an active system is } "plug and play".
Actually, about 90% of the 840s have the internal scan relays soldered in place on the scan gen board. The other 10% will need the relays added. JEOL service is best for this as they have the relays in stock. To check, one must physically examine the scan gen board.
Scott ----------------------------------------------------------------------- Scott D. Davilla Phone: 919 489-1757 (tel) 4pi Analysis, Inc. Fax: 919 489-1487 (fax) 3500 Westgate Drive, Suite 403 email: davilla-at-4pi.com Durham, North Carolina 27707-2534 web: http://www.4pi.com
Anyone have an idea what the sale value might be for a Hitachi S-570 with a LaB6 gun & solid-state backscatter detector? No x-ray. Comes with a Gatan digital imaging system. Sorry, but I don't know the model of this system, but it uses a PowerMac with a NuBus card, so it's a bit hoary.
In good condition, recently PMed to give "to spec." performance in the upper stage at 200,000X.
Thanks.
Phil -- }}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{ Philip Oshel Supervisor, AMFSC and BBPIC Dept. of Animal Health and Biomedical Sciences University of Wisconsin 1656 Linden Drive Madison, WI 53706-1581 voice: (608) 263-4162 fax: (608) 262-7420 (dept. fax)
Hi, I'm looking for technical assistance or a service manual for a Leitz Orthomat E (7916) Photo System.The problem we are having is the unit will not allow a picture to be taken unless there is very little light. Once the system thinks it is in range, there is not enough light present to get the photograph. An adjustment of the light detector? Any assistance is appreciated. Lewis McCrigler Humboldt State University
G'day Luc, Shellite is a solvent manufactured by Shell, its used as stove fuel, lighter fuel, drycleaning solvent and as a rubber/adhesive solvent (also, as you have found, a great EM parts cleaner). Another name you may find Shellite listed under is Shell X55 solvent. Shellite is basically a highly volitile, low octane, unleaded petrol. According to Shell Australia it is not the same as white spirit, this is less volitile than shellite, therefore harder to evaporate from the metal surface. Below is part of the safety data sheet for Shellite. If anyone requires the complete 10 page safty data sheet please email me. Luc, see you next time you are in OZ. Regards JVN SHELL SHELlLITE
STATEMENT OF HAZARDOUS NATURE
HAZARDOUS ACCORDING TO WORKSAFE AUSTRALIA CRITERIA
SUPPLIER
Company: The Shell Company of Australia Limited Address: Shell House, 1 Spring Street (PO Box 872K) PO Box 2091 Melbourne Wellington VIC 3001 New Zealand Australia Telephone: (03)9666 5444 Telephone: 64 4 4720080 Fax: (03)96665008/64 44980100
HAZARD RATINGS
Flammability: 4
Toxicity: 2
Body Contact: 2
Reactivity: 0
Chronic effect: 2
Scale: Min / Nil = 0, Low = 1, Moderate = 2, High = 3 and Extreme = 4.
PERSONAL PROTECTIVE EQUIPEMENT FOR INDUSTRIAL/COMMERCIAL ENVIRONMENTS
Product Name: Shell Shellite Other Names: Product Code 00720
Used as rubber solvent, cleaning solvent, lighter fluid and as fast evaporating, highly volatile solvent in enamels, adhesives and lacquers. The use of a quantity of material in an unventilated or confined space may result in increased exposure and an irritating atmosphere developing Before starting consider control of exposure by mechanical ventilation.
PHYSICAL DESCRIPTION/PROPERTIES
APPEARANCE
Clear highly flammable liquid with a typical hydrocarbon liquid odour; floats on water. Classed as an aliphatic solvent; i.e has low aromatic content.
Molecular Weight: Not applicable. Boiling Point (deg C): 47-128 Melting Point (deg C): Not available. Vapour Pressure (kPa): 34.5 -at- 15 deg C Specific Gravity: 0.71 -at- 15 deg C Flash Point (deg C): {-30 Lower Explosive Limit (%): 1.0 Upper Explosive Limit (%): 7.5 Solubility in Water (g/L): Immiscible
INGREDIENTS
NAME CAS RN % paraffins, as liquid hydrocarbons Various } 60 naphthenes Notspec n-hexane 110-54-3 13 aromatic hydrocarbons total, including {5.0 toluene 108-88-3 3.5app ethylbenzene 100-41-4 benzene 71-43-2 {0.5 C8 and higher aromatics approx1
Anaspec 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 all } } Whilst working in Australia we found a few labs using something called } Shellite as a de-greaser to clean microscope parts. This works very well and } seems to be very available. we then looked for the same product here in SA. } We then found that this is a trade name only in Australia and is actually } petroleum ether or spirit. } We called the local Shell distributor to ask what the difference was between } spirit and ether. Well, they were not sure on that one. Then we were told } you must specify the temperature range of the petroleum Ether. 40 - 60, 60 - } 80 or 80 - 100 deg Celsius. Again no explanation as to what the difference } is an why. } Can any one give us some more info on Petroleum ether / spirit and the } advantages disadvantages on the different temperatures. } } Thanks } } Luc Harmsen } Anaspec, South Africa } Technical support on microscopy. } Tel + 27 (0) 11 476 3455 } Fax + 27 (0) 11 476 7290 } anaspec-at-icon.co.za } www.anaspec.co.za } } Remember, ICEM 15 will be in } 2002, Durban, South Africa. } www.icem15.com
-- **************************************************** John V Nailon Operations Manager Centre for Microscopy and Microanalysis The University of Queensland St. Lucia Queensland 4072 Phone: +61-7-3365-4214 Fax: +61-7-3365-4422 WWW: http://www.uq.edu.au/nanoworld/allstaff.html#Nailon ****************************************************
I carry a full line of refurbished histology equipment. Please contact me for price quotes and recommendations.
Ford Royer Analytical Instruments 9921 13th Ave. N. Minneapolis, MN 55441 (800) 565-1895, extension 17 fax: (612) 929-1895 email: froyer-at-bitstream.net
Phillip Rutledge 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'm currently in the process of setting up a Histology/Microbiology/EM facility. Being out of Histology for almost 30 years (been doing EM) I'm not sure of the best type of tissue processor to buy. I won't be doing that many specimens, so I'm leaning towards a rotary tissue processor. I have info on Shandon and Tissue-Tek. Are there others out there? Anyone having opinions on the 2 mentioned, I would appreciate any comments. I am looking for ease of use (changing solutions, fume safeguards), reliability on service and cost effectiveness. Thanks. } } Peace be with you, } Phil Rutledge (410)778-4136, 2120 } prutledge-at-ars.usda.gov
Just to throw in a further little complication of nomenclature:
In New Zealand, (and maybe in Australia, too), the general name we use for stuff like Shellite, is "white spirit".
In the US it's often called "unleaded gas(oline)", or "white gas(oline)", I think.
It's the stuff that Coleman camping stoves run on.
However, in the UK, "white spirits" is what I would call "mineral turpentine" ie the comparitively non-volatile solvent often used for thinning oil-based paints.
As I found a few years ago, shortly after my arrival in the UK to continue the camping holiday that I'd started in the US, it doesn't work in Coleman stoves.
rtch
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
we have recently started working on Transmission Electron Microscopy of several kinds of clusters in various martices. We frequently face the problem of analyzing particle size distribution and other features of the images.
We use several software programs for our particle analysis, but we always conclude that measuring the particle size by eye is the most reliable way to get accurate data.
We have both Digital Micrograph software and other programs (Scion image etc...), but we have never used them for this kind of application.
As we look both at bulk materials, and at thin films or powders, frequently our images show an uneven contrast, and defining a threshold is difficult.
Can anyone suggest (and eventually share) a script, a program or a set of user functions that allows to study particle size distributions?
Thank very much in advance for your help in this matter.
Massimo: Try to Fourier-filter (high-pass) the images before doing the particle analysis. In this way you might be able remove the influence of the uneven background. Sometimes this works nicely. You can easily do it in Digital Micrograph. Hope this helps,
Max Sidorov ---------------------------- Dr. Maxim V. Sidorov TEM Applications Specialist Philips Electron Optics, Applications Laboratory Building AAE, Achtseweg Noord 5 5600 MD Eindhoven, the Netherlands
} -----Original Message----- } From: Massimo Catalano [SMTP:massimo.catalano-at-ime.le.cnr.it] } Sent: Wednesday, April 19, 2000 09:17 } To: Microscopy-at-sparc5.microscopy.com } Subject: TEM of clusters and image analysis } } ------------------------------------------------------------------------ } 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 Listservers, } } we have recently started working on Transmission Electron Microscopy of } several kinds of clusters in various martices. } We frequently face the problem of analyzing particle size distribution and } } other features of the images. } } We use several software programs for our particle analysis, but we always } conclude that measuring the particle size by eye is the most reliable way } to get accurate data. } } We have both Digital Micrograph software and other programs (Scion image } etc...), but we have never used them for this kind of application. } } As we look both at bulk materials, and at thin films or powders, } frequently } our images show an uneven contrast, and defining a threshold is difficult. } } Can anyone suggest (and eventually share) a script, a program or a set of } } user functions that allows to study particle size distributions? } } Thank very much in advance for your help in this matter. } } Max } } Dr. Massimo Catalano } CNR-IME } Campus Universitario } Via Arnesano } 73100 Lecce - ITALY } tel: + 39 0832 322362 } fax: + 39 0832 325299 } email: massimo.catalano-at-ime.le.cnr.it } http://www.ime.le.cnr.it } http://www.ime.le.cnr.it/sime/sime.htm } } } }
On a recent business trip outside the US, I met some people who are using a LEO 440i SEM. They wanted to do cathodoluminescence (CL) with it and were not having too much success so I volunteered to post this message to the list to seek advice. The PMT is mounted in a housing/port that is located at about 2 O'clock (if you consider the door to be at 6 o'clock). The mounting of the PMT is horizontal and it looks like the distance to the PMT input is about 20 cm from the center of the chamber. There is a glass lens on a slider arrangement in front of the PMT input but they were unclear as to how to adjust this.
They do not know the type of PMT that was supplied and haven't opened up the system to determine this. It looks like it must be an end-on type.
One of their questions has to do with the adjustment, if any, for the PMT voltage and how they can do this and how they can monitor the value of the PMT voltage.
They believe that the PMT should have been mounted at an angle to better see the sample but the only other port where this could be done is not usable for the PMT when the EDS detector is in place.
The other work that they are doing with the instrument is completely satisfactory, so far as I could tell, and I suspect that someone on the list can provide the answers to these questions and help to get them in business doing monochromatic CL as well.
All suggestions from ohter users of this type of instrument, obviously, will be much appreciated.
Any vendors who provide CL accessories for this instrument should contact me directly.
Thank you.
Donald J. Marshall
Donald J. Marshall Relion Industries P.O. Box 12 Bedford, MA 01730 Ph: 781-275-4695 FAX: 781-271-0252 email dmrelion-at-world.std.com
Cathodoluminescence, mass spectroscopy, electron beam technology
"A weed is a flower out of place."
(Please note: Do not send email with attachments to this address. Instead, send it to donbarlen-at-aol.com. Thank you.)
Please note that the Seminar/Workshop at Princeton University on April 25, 2000 will be at PMI. For additional information and registration please see the DI web site at www.di.com and click on "Workshops and Seminars".
Regards, Jeff Doran Digital Instruments Veeco Metrology Group
Hi Richie & readers As I recall from the oil field days of the 80s, the term "white gasoline" referred to a condensate from natural gas production. People were known to collect (swipe) this free fuel at the well site & use it in their autos. I do not know what it's composition is but this was not really considered a good substitute for gasoline. Something about destroyed pistons & the such.
Bruce Brinson Rice U.
Ritchie Sims 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 } -----------------------------------------------------------------------. } } Just to throw in a further little complication of nomenclature: } } In New Zealand, (and maybe in Australia, too), the general name we } use for stuff like Shellite, is "white spirit". } } In the US it's often called "unleaded gas(oline)", or "white } gas(oline)", I think. } } It's the stuff that Coleman camping stoves run on. } } However, in the UK, "white spirits" is what I would call "mineral } turpentine" ie the comparitively non-volatile solvent often used for } thinning oil-based paints. } } As I found a few years ago, shortly after my arrival in the UK to } continue the camping holiday that I'd started in the US, it doesn't } work in Coleman stoves. } } rtch } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand
Yes, but Kate said, that they already have an EDX system attached. Normally these systems need access to the scan coils as well, that's why I mentioned "plug and play". I bet, that the EDX system is connected to connector JA2 on the back of the microscope, which is where we would also connect the digital acquisition system (of course with an additional connector for the existing X-ray system).
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== 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: Scott D. Davilla [mailto:davilla-at-4pi.com] Sent: Tuesday, April 18, 2000 3:45 PM To: Microscopy-at-sparc5.microscopy.com
} though (seconds). Installation of a passive system may be easier in some } circumstances, but on your 840 the installation of an active system is } "plug and play".
Actually, about 90% of the 840s have the internal scan relays soldered in place on the scan gen board. The other 10% will need the relays added. JEOL service is best for this as they have the relays in stock. To check, one must physically examine the scan gen board.
Scott ----------------------------------------------------------------------- Scott D. Davilla Phone: 919 489-1757 (tel) 4pi Analysis, Inc. Fax: 919 489-1487 (fax) 3500 Westgate Drive, Suite 403 email: davilla-at-4pi.com Durham, North Carolina 27707-2534 web: http://www.4pi.com
Has anyone had a demo for the new Zeiss CCD known as the "Axiocam"? Or better, has anyone ordered it and had it installed in their lab. I'd like to know the usual things regarding performance and overall satisfaction. Thanks.
Some of you asked what caused the turbo pump failure I related last week. I frankly don't know. There is some scope in the S250 for objects (samples, for example) to fall into the pump. Cambridge fitted a mesh screen to trap these, but it is possible that some small hard fragment got between the fan and stator blades somehow. In early Cambridge 250s there was no rough pumping sequence - the baffle valve was simply opened, dumping the specimen chamber air at 1 bar into the turbo pump while it was running at its top speed. The dramatic shreik of protest that results is a great party trick when demonstrating the machine to visitors, but slows the pump dramatically and undoubtedly causes great mechanical stress on the turbines and stator blades. Strangely, we never suffered pump failures during this stressful event, but only when the pumps were in apparently smooth running at top speed. I think we are now on about our fourth pump in twenty years, and this last one has survived unscathed for about 10 years with nothing more than an occasional oil-change, during which time it has mostly been run continuously, surviving endless air-dump cycles. Now that I have told you that I can probably expect our next failure almost immediately! Perhaps I'll order the skip now, just in case. ===================================================================== DR CHRIS JEFFREE BIOSEM - BIOLOGICAL SCIENCES EM FACILITY UNIVERSITY OF EDINBURGH Daniel Rutherford Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JH, Scotland, UK Tel. #44 131 650 5345 FAX. #44 131 650 6563 Mobile 0410 585 401 email c.jeffree-at-ed.ac.uk SEM / TEM bookings sem-at-ed.ac.uk =====================================================================
I saw the scope several weeks back but never saw through it, Zeiss could not get it to work. I know one lab on campus has one and they say it images beautifully, when it works...... they have been using our Optronics lately.
Thomas Moninger moninger-at-emiris.iaf.uiowa.edu University of Iowa Central Microscopy Research Facility http://www.uiowa.edu/~cemrf Views expressed are mine. On Wed, 19 Apr 2000, Chris Edwards 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 } -----------------------------------------------------------------------. } } } Has anyone had a demo for the new Zeiss CCD known as the "Axiocam"? Or } better, has anyone ordered it and had it installed in their lab. I'd like } to know the usual things regarding performance and overall satisfaction. } Thanks. } } }
Readers, At the upcoming Microscopy & Microanalysis Conference (13/17 August in Philadelphia) we will again hold a Just For Fun Image Contest. Concept is an image composed of one or more other images, one of which must be microscopical in nature. Prizes will be $300, $200 and $100 for first, second and third prizes respectively. One does not have to be present to win. Should you might be interested, kindly contact me direct and I will forward detail. Last year we had over 30 entries. Regards, Don Grimes, Microscopy Today
} Yes, but Kate said, that they already have an EDX system attached. } Normally these systems need access to the scan coils as well, that's why } I mentioned "plug and play". I bet, that the EDX system is connected to } connector JA2 on the back of the microscope, which is where we would } also connect the digital acquisition system (of course with an } additional connector for the existing X-ray system).
Maybe you misunderstand my intent. All I'm saying is don't assume a JEOL 840 has everything required for active scan control. Some don't, but it's not a show stopper provided the information is known and planned for in advance. Just because there is an EDX system attached does not mean that it is attached to the microscope beam control. There are more EDX systems sold without X-ray mapping than sold with X-ray mapping. In the same note, just because the JEOL 840 has a "plug and play" interface does not mean that the external scan relays are present. Both issues raise flags here and can mean the difference in a painless or painfull installation if the details are not addressed.
Scott ----------------------------------------------------------------------- Scott D. Davilla Phone: 919 489-1757 (tel) 4pi Analysis, Inc. Fax: 919 489-1487 (fax) 3500 Westgate Drive, Suite 403 email: davilla-at-4pi.com Durham, North Carolina 27707-2534 web: http://www.4pi.com
Many thanks for the various valuations sent me. We have enough now for our needs. For those interested, the valuations ranged from 15k$ to 45k$, with the average about 25 - 30 k$.
Phil -- }}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{ Philip Oshel Supervisor, AMFSC and BBPIC Dept. of Animal Health and Biomedical Sciences University of Wisconsin 1656 Linden Drive Madison, WI 53706-1581 voice: (608) 263-4162 fax: (608) 262-7420 (dept. fax)
We have a Kodak XLS8300 dye sublimation printer that needs repair. Kodak no longer supports this instrument, has no replacement parts we need and doesn't know where we can get service.
When we shipped it for repair, we got a report that said it needed a hard drive and possibly a saturn board (motherboard, I presume) and that they could not provide service.
I would appreciate getting suggestions of where to get this printer serviced.
Please reply directly to me, and not to the listserv.
Can someone help me with a US source to purchase, low fluorescence quartz coverlips for conventional microscope slides. Please respond directly at the following email address.
Steven Ridge Fryer Company Inc. 847-669-2000 Phone s.ridge-at-fryerco.com
I recently completed a research paper to complete my EE Degree which included a section on vacuum principles. I incorporated part of the in formation I got from an article by XEI scientific on contamination control. It spoke of oil deposits on cool windows or a loss of low energy xray. I thought I also read (on the web site) of how x-rays will turn ceramics yellow. I can not find the article again. Can you help me with this? What I am trying to decipher is, will concentrated xray emission through a ceramic cause it to yellow and could contamination deposits be the cause of this? Would appreciate any answers or leads as to where I might find the answer.
Once again there is a conference coming up and I have found myself with a few extra rooms. If anyone is need of hotel rooms at the San Francisco Marriott for the Materials Research Society meeting next week, please let me know. If nobody needs them, I will be cancelling them tomorrow night.
I have rooms for arrival on Saturday April 22 and departure on Friday April 28. Of course the dates could be changed if needed. They are rooms with 2 double beds and the confrence rate is $138 I think.
David Henriks Vice President 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
I did not say anything about yellowing of ceramics at the XEI Scientific web site. It is generally not a "contamination" problem. X-rays being an ionizing radiation are able to break bond and modify the crystalline structure of ceramics. When I did X-ray spectrometry I was very aware of the ability of high intensity, high energy x-rays to cause radiation damage and discoloration. However scanning electron microscope are generally operated at low energies ( {30KeV) and low beam currents. Xray damage is not a problem except with most sensitive materials.
Xray damage to oils could cause them to crosslink and yellow if oils are deposited on the surface of ceramic but I have not seen any complaints about this. Maybe a more specific description of your problem would help. Reply off list.
At 02:10 PM 4/19/00 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Rreally....can anyone identify a source of high quality #1 cover slips that are not made in China and don't have micro fractures in them?
Coverslip number 1 thickness is 0.13 to 0.17mm. Ours are 0.25 thick (and 25.4mm dia) I didn't know #1 quartz coverslips existed, made in China or outer space. Perhaps GG should not blame the Chinese manufacturer for flaws in #1 slips; maybe perfect #1 in quartz are impossible. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Thursday, April 20, 2000 2:33 PM, Dr. Gary Gaugler [SMTP:gary-at-gaugler.com] wrote: } } } At 02:10 PM 4/19/00 , you wrote: } } } Can someone help me with a US source to purchase, low fluorescence } } quartz coverlips for conventional microscope slides. Please respond } } directly at the following email address. } } } } Steven Ridge } } Fryer Company Inc. } } 847-669-2000 Phone } } s.ridge-at-fryerco.com } } } } -- } } BMv } } } Rreally....can anyone identify a source of high quality } #1 cover slips that are not made in China and don't have } micro fractures in them? } } gg }
Friends: I have always wondered about these solvents. One way to sort out intercontinental differences is to ask - what do they smell like? Gasoline has a distinctive odor, whether leaded or not. The Pet Ether I use in my lab is nearly odorless. The label describes it as having a "boiling range of 37.7 to 55.8 C/ 1 drop to dryness." Turpentine, or more correctly, oil of turpentine, has another distinctive odor. It is used to thin paints. Petroleum paint thinner, a good solvent, is much like pet ether, nearly odorless. All of the above are colorless, Kerosene, used in camp stoves and cabin heaters, another petroleum derivative, is yellow and possesses its own characteristic odor. All of these observations are from a US view point. How do these other solvents smell?
Sam Purdy National Steel Corp Tech Center Trenton, MI, USA spurdy-at-nationalsteel.com
} ---------- } From: Ritchie Sims } Sent: 19, April 2000, 12:10 PM } To: 'MSA listserver' } Subject: White Spirit } } ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } } } Just to throw in a further little complication of nomenclature: } } In New Zealand, (and maybe in Australia, too), the general name we } use for stuff like Shellite, is "white spirit". } } In the US it's often called "unleaded gas(oline)", or "white } gas(oline)", I think. } } It's the stuff that Coleman camping stoves run on. } } However, in the UK, "white spirits" is what I would call "mineral } turpentine" ie the comparitively non-volatile solvent often used for } thinning oil-based paints. } } As I found a few years ago, shortly after my arrival in the UK to } continue the camping holiday that I'd started in the US, it doesn't } work in Coleman stoves. } } rtch } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand }
OK, if not quartz, how about glass? The Chinese products come pre-made with micro cracks. Erie used to make slips but stopped. I can find round slips from Swiss glass but not square ones for 1" x 3" slides.
I think that 4 attempts with Chinese covers which all have the same problem is not a coincidence. These were bought from Ward's Scientific. They exchanged them without any hassle. The problem is that the slips are un useable. I'd be glad to send a few new ones to you so you can see the cracks for yourself.
gary g.
At 05:32 AM 4/20/00 , you wrote: } Coverslip number 1 thickness is 0.13 to 0.17mm. Ours are 0.25 thick (and } 25.4mm } dia) } I didn't know #1 quartz coverslips existed, made in China or outer space. } Perhaps GG should not blame the Chinese manufacturer for flaws in #1 slips; } maybe perfect #1 in quartz are impossible. } Cheers } Jim Darley } ProSciTech Microscopy PLUS } PO Box 111, Thuringowa QLD 4817 Australia } Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com } Great microscopy catalogue, 500 Links, MSDS, User Notes } www.proscitech.com } } On Thursday, April 20, 2000 2:33 PM, Dr. Gary Gaugler [SMTP:gary-at-gaugler.com] } wrote: } } } } } } At 02:10 PM 4/19/00 , you wrote: } } } } } Can someone help me with a US source to purchase, low fluorescence } } } quartz coverlips for conventional microscope slides. Please respond } } } directly at the following email address. } } } } } } Steven Ridge } } } Fryer Company Inc. } } } 847-669-2000 Phone } } } s.ridge-at-fryerco.com } } } } } } -- } } } BMv } } } } } } Rreally....can anyone identify a source of high quality } } #1 cover slips that are not made in China and don't have } } micro fractures in them? } } } } gg } }
I cannot believe that only Chinese coverslips are sold in the USA. We carry a comprehensive range of good German-made coverslips . Yes, and they don't have common flaws. If that sounds too much like an advertisement, I'll add that several suppliers in Australia offer a similar range. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Thursday, April 20, 2000 11:18 PM, Dr. Gary Gaugler [SMTP:gary-at-gaugler.com] wrote: } OK, if not quartz, how about glass? The Chinese products } come pre-made with micro cracks. Erie used to make slips } but stopped. I can find round slips from Swiss glass but not } square ones for 1" x 3" slides. } } I think that 4 attempts with Chinese covers which all have the } same problem is not a coincidence. These were bought from } Ward's Scientific. They exchanged them without any hassle. } The problem is that the slips are un useable. I'd be glad to } send a few new ones to you so you can see the cracks for } yourself. } } gary g. } } } At 05:32 AM 4/20/00 , you wrote: } } Coverslip number 1 thickness is 0.13 to 0.17mm. Ours are 0.25 thick (and } } 25.4mm } } dia) } } I didn't know #1 quartz coverslips existed, made in China or outer space. } } Perhaps GG should not blame the Chinese manufacturer for flaws in #1 slips; } } maybe perfect #1 in quartz are impossible. } } Cheers } } Jim Darley } } ProSciTech Microscopy PLUS } } PO Box 111, Thuringowa QLD 4817 Australia } } Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com } } Great microscopy catalogue, 500 Links, MSDS, User Notes } } www.proscitech.com } } } } On Thursday, April 20, 2000 2:33 PM, Dr. Gary Gaugler [SMTP:gary-at-gaugler.com] } } } } wrote: } } } } } } } } } At 02:10 PM 4/19/00 , you wrote: } } } } } } } Can someone help me with a US source to purchase, low fluorescence } } } } quartz coverlips for conventional microscope slides. Please respond } } } } directly at the following email address. } } } } } } } } Steven Ridge } } } } Fryer Company Inc. } } } } 847-669-2000 Phone } } } } s.ridge-at-fryerco.com } } } } } } } } -- } } } } BMv } } } } } } } } } Rreally....can anyone identify a source of high quality } } } #1 cover slips that are not made in China and don't have } } } micro fractures in them? } } } } } } gg } } }
} } } Can someone help me with a US source to purchase, low fluorescence } quartz coverlips for conventional microscope slides. Please respond } directly at the following email address. } } Steven Ridge } Fryer Company Inc. } 847-669-2000 Phone } s.ridge-at-fryerco.com } Steven, We can make anything you need. Email, fax or call with specifications. Mike Urbanik guru-at-crystalguru.com www.crystalguru.com Ph 941-645-5959 Fax 941-643-6058
I don't think we're in disagreement here. We've had our share of surprises (SEMs with external connectors for beam control, then nothing attached to the connectors, strange boards in the microscopes that should not have been there, etc.) We have had very few problems with the 840 (and 820, 6300, 6400 for that matter). But of course there is always a possibility that something is missing or not working, which must be ascertained in each case individually. As you said, it is usually not a show stopper, but requires some modifications of the microscope.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== 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: Scott D. Davilla [mailto:davilla-at-4pi.com] Sent: Wednesday, April 19, 2000 12:31 PM To: Microscopy-at-sparc5.microscopy.com
} Yes, but Kate said, that they already have an EDX system attached. } Normally these systems need access to the scan coils as well, that's why } I mentioned "plug and play". I bet, that the EDX system is connected to } connector JA2 on the back of the microscope, which is where we would } also connect the digital acquisition system (of course with an } additional connector for the existing X-ray system).
Maybe you misunderstand my intent. All I'm saying is don't assume a JEOL 840 has everything required for active scan control. Some don't, but it's not a show stopper provided the information is known and planned for in advance. Just because there is an EDX system attached does not mean that it is attached to the microscope beam control. There are more EDX systems sold without X-ray mapping than sold with X-ray mapping. In the same note, just because the JEOL 840 has a "plug and play" interface does not mean that the external scan relays are present. Both issues raise flags here and can mean the difference in a painless or painfull installation if the details are not addressed.
Scott ----------------------------------------------------------------------- Scott D. Davilla Phone: 919 489-1757 (tel) 4pi Analysis, Inc. Fax: 919 489-1487 (fax) 3500 Westgate Drive, Suite 403 email: davilla-at-4pi.com Durham, North Carolina 27707-2534 web: http://www.4pi.com
I have both a Spectra file in Link format and txt format saved as a MSA type file. Is there a software or an excel macro that could read these spectra ?
Steven Ridge wrote: ========================================================== Can someone help me with a US source to purchase, low fluorescence quartz coverlips for conventional microscope slides. Please respond directly at the following email address. =========================================================== SPI Supplies has offered quartz coverslips 0.2 mm thick for some time and the details can be found on URL http://www.2spi.com/catalog/ltmic/quartz.html
The optical characteristics of (fused) quartz are very very sensitive to the impurity levels present. Therefore it is important that one use coverslip that are of reproducible quality and of known impurity levels (also linked from above webpage). The particular quartz used for the manufacture of the SPI quartz coverslips is the same "electronic grade" used in many applications in the electronics industry. The coverslips and quartz are of USA manufacture (just in case one might wonder about origin).
Disclaimer: SPI Supplies has supplied quartz coverslips to users in the microscopy community for some number of years. Quartz coverslips are also offered by several other of the leading suppliers of consumables to the microscopy market.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.spi.cc ######################## ============================================
I take it you are referring to Link AN10000, QX2000 or eX/L spectra. There are a number of ways to analyze these spectra post facto. All depend on having them on a DOS disk, which presumably you do. If you have them on LINK formatted floppys, then you can read them on a DOS machine with a small program RDL2.EXE which I wrote a while ago, although that program will not read subdirectories on LINK floppys. It copies the files byte-by-byte to a DOS hard drive. Run the program to get three lines of useage instructions. Once they are in DOS, you can do a number of things:
1) Use my program LKSPCV.EXE which will convert them into a text file formatted for direct input into Excel or another spreadsheet or graphing program. LKSPCV knows about the differences between eX/L files and AN10/QX2000 files, and I believe (though I've never tested) that it will also handle the old LINK 860 files correctly, too. Usage: LKPSCV infilename.sp outfilename.txt
2) Use the program DTSA (Desktop Spectrum Analyser) which used to be sold by NIST. I know that the status of that program has changed, and I don't know if it is still available or not. However, it runs on a Mac, and can import both AN10/QX2000 and eX/L files, as well as MSA formatted files.
3) If you have access to a LINK ISIS analyser via a friend or colleague, these systems can import and process eX/L or AN10/QX2000 spectrum files.
LKSPCV.EXE and RDL2.EXE are available by FTP from prism.mit.edu:2101. Ther are a number of other files on that site, too, of which the useful ones are LKCONV.COM which formats LINK disks on a PC (which must be running plain DOS - not a DOS window) and LKCV3.EXE which extracts individual images and maps from LINK eX/L or AN10/QX2000 studies.
At 10:21 AM 04/20/2000 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
** Anthony J. Garratt-Reed ** MIT Room # 13-1027 ** 77 Massachusetts Avenue ** Cambridge, MA 02139-4307 ** USA ** ** Phone: (+) 1-617-253-4622 ** Fax: (+) 1-617-258-6479 **
I have a program that I wrote in Visual Basic 6 that will read the EMSA format EDS and EELS data, plot them, color, them, overlay them, print them, copies the graphical data to an RTF document, and can convert them to the EMSA format with two column X,Y pairs. I have been toying with the idea of selling the program through South Bay Technology inexpensively. I have no idea what the market would be for something like this. I have given a few copies of it out for evaluation and hoped to get feedback about what I should add to it, but no one has gotten back to me. The program is primarily geared for EELS spectra but works with EMIspec, Noran, and DTSA EMSA formatted EDS spectra. If you agree to give me feedback on the program (and a pitcher of beer at M&M 2000 if you go), I might be able to be convinced to send you a copy. I would like to know if it works with some of the other EDS systems.
The reason that I wrote it was that the Gatan EL/P program output the EMSA format in y-only format with columns of five. Gatan isn't the only company to do that, I believe that the Noran data comes out that way. That format is difficult to parse properly in Excel so that you can graph it. What I did was to open the ASCII data in a word processor, replace all the hard returns with a comma, then replace all the commas with a hard return, and then resave as an ASCII text file. Then you can open it in (or copy and paste the data) in Excel. Then you move the column over once and create the X values for plotting the data as X,Y pairs.
I originally wrote the program to just convert it to X,Y pairs so that I could use Excel but then it was so easy to graph it in VB6, that I just got carried away.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies." --
} -----Original Message----- } From: Filion, Christian [mailto:christian.filion-at-atlasstainless.com] } Sent: Thursday, April 20, 2000 10:21 AM } To: microscopy-at-sparc5.microscopy.com } Subject: EDS spectra on PC } } } -------------------------------------------------------------- } ---------- } 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 } } } -------------------------------------------------------------- } ---------. } } } Good morning, } } I have both a Spectra file in Link format and txt format } saved as a MSA } type file. Is there a software or an excel macro that could read these } spectra ? } } Thank you } } Christian Filion } Superviseur Laboratoires } Aciers Inoxydables Atlas } 1640 Marie-Victorin } Tracy, Qubec } J3R 5R5 } TelÊ: 450-746-5243 } faxÊ: 450-746-5241 }
I am trying to track down information on the short wavelength cut-off used in EDS microanalysis to determine the true electron accelerating voltage. I have a number of articles describing its determination, but little in the way of historical information. It is also known as the 'Duane-Hunt' limit. I cannot find any references to a Duane or a Hunt. Does anyone know the originator of the concept and why it is known as this?
Thanks Robert A. Carlton Aventis Pharmaceuticals Tel 610-454-3949 Robert.Carlton-at-aventis.com
We are looking to upgrade our EDS system with a 4pi based system or Evax. Does any one have any opinions about these systems or is there others you could recomend? The TEM is a CM20 with an EDAX detector.
The MSA text format is written in such a way so that the data should be directly importable into any standard spreadsheet program.
Just import it as ASCII text with a "comma" deliminator between columns.
I am presuming that you stored the data in 2 column (X,Y) format. If you did otherwise you will need to do a small amount of cleanup but it should not be difficult. The other obvious trick is to go back to the EDS system and make sure you store the data in X,Y single column format;
Nestor
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================================================================== Nestor J. Zaluzec Materials Science Division Building 212 Argonne National Lab 9700 S. Cass Ave Argonne, Illinois 60439 USA Tel: 630-252-7901, Fax: 630-252-4798 Email: Zaluzec-at-aaem.amc.anl.gov ================================================================== TPMLab: http://tpm.amc.anl.gov MMSite: http://www.amc.anl.gov ==================================================================
The box said "This program requires Win 95/98/NT or better..." so I bought a G3 Mac
The SEM lab manager position at the Smithsonian Institution is now open.
The National Museum of Natural History in Washington, DC is seeking an experienced electron microscopist to fill a vacancy for SEM laboratory operation and management. The SEM facility is designed to serve both the biological and geological research communities in the museum, and houses two recent model SEMs and one state-of-the-art environmental microscope (to be installed in mid-2000). The principal responsibilities include training staff members and visiting scientists in proper use of equipment and theory of electron generation and detection, maintenance and troubleshooting all instrumentation (in conjunction with full service contracts), evaluation of new developments in SEM technology, and supervision of a support staff member. The successful applicant will also have the opportunity to gain experience in Focused Ion Beam (FIB) Microscopy and high spatial resolution secondary ion mass spectrometry HR SIMS.
This position will fill a federal government vacancy and is offered at the GS11 ($42,724) or GS12 ($51,204) levels, and carries promotion potential to grade GS 13. U.S. citizenship is required for this federal position. To obtain information concerning this vacancy call our automated Jobline at (202) 287-3102 (24 hours, 7 days a week), press 9 and request vacancy announcement 00MQ-2069, then follow voice prompts to have information faxed or mailed to you. The application deadline is May 16th, 2000. If questions arise after receiving and reading through the vacancy announcement please contact: Dr. Edward Vicenzi (Chair, Search Committee) at vicenzi-at-volcano.si.edu. The Smithsonian Institution is an equal opportunity employer.
NOTE: Completed Smithsonian applications must be received by May 16th, 2000.
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Edward P. Vicenzi Smithsonian Institution Department of Mineral Sciences Washington, DC 20560-0119
The short wavelength limit for x-ray spectra produced by excitation with an electron beam is discussed and illustrated in the section on "The Continuous Spectrum" (I believe Sect. 1.3) in Chapter 1 of The book 'Elements of X-ray Diffraction', by B. D. Cullity, Addison Wesley.
Basically, you are dealing here with the Bremsstrahlung radiation which is generated by electrons in the incident electron beam which lose increments of their energy as they interact with the specimen. Each incremental energy loss gives rise to an x-ray photon having a photon energy equal to the energy loss increment, i.e. Energy loss of a beam electron = Energy of generated Bremsstrahlung photon. The maximum amount of energy a beam electron electron can give up occurs when it is stopped in a single collision, whereupon the energy loss equals the energy imparted to the electron by the applied accelerating voltage, and is numerically the same as the value of the accelerating voltage when expressed in keV. (i.e. an accelerating voltage of 20 kV gives electrons a kinetic energy of 20 keV) A maximum energy loss event of this kind produces a Bremsstrahlund photon with the highest possible energy. Photon wavelengths decrease as photon energies increase, and so this Bremsstrahlung photon of maximum energy also is the photon with the shortest wavelength. Ergo, if you measure the photon energy at the short wavelength limit, you know the value of the accelerating voltage.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
Very few people use quartz coverslips, but they should not be afraid. I understand that the only USA manufacturer is General Electric and almost certainly all pure quartz products made in USA are made from quartz supplied by GE. Our quartz slips and slides are made from GE quartz too. I think that it is important for endusers to know when supplies are in fact standard (however excellent) or "extra special". Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Friday, April 21, 2000 1:51 AM, Garber, Charles A. [SMTP:cgarber-at-2spi.com] wrote: } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } Steven Ridge wrote: } ========================================================== } Can someone help me with a US source to purchase, low fluorescence quartz } coverlips for conventional microscope slides. Please respond directly at } the following email address. } =========================================================== } } . . . .The optical characteristics of (fused) quartz are very very sensitive to the } impurity levels present. Therefore it is important that one use coverslip } that are of reproducible quality and of known impurity levels (also linked } from above webpage). The particular quartz used for the manufacture of the } SPI quartz coverslips is the same "electronic grade" used in many } applications in the electronics industry. The coverslips and quartz are of } USA manufacture (just in case one might wonder about origin). } } Disclaimer: SPI Supplies has supplied quartz coverslips to users in the } microscopy community for some number of years. Quartz coverslips are also } offered by several other of the leading suppliers of consumables to the } microscopy market. } } Chuck
Hi all, This is off the microscopy subject...sorry. Any recommendations on digital cameras for copy stand work and general photography? Has anyone tried the Nikon Coolpix 990? We're coming up on the end of the fiscal year and there might be funds available for a digital camera so any advice is greatly appreciated. thanks, Beth Richardson
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
Hi all, The status is now that DTSA is being given away free by NIST, go to http://www.cstl.nist.gov/div837/837.02/dtsa.html it is Macintosh only but imports many file types. Scott
} Anthony Garratt-Reed wrote: ..snip... } 2) Use the program DTSA (Desktop Spectrum Analyser) which used to be sold } by NIST. I know that the status of that program has changed, and I don't } know if it is still available or not. However, it runs on a Mac, and can } import both AN10/QX2000 and eX/L files, as well as MSA formatted files. ..snip...
------------------- note: new mailing address ------------------------ Scott Wight fax: 301-417-1321 NIST 222/A113 W voice: 301-975-3949 100 Bureau Dr STOP 8371 | email: scott.wight-at-nist.gov Gaithersburg, MD 20899-8371 \|/ disclaimer: Any opinion expressed is my own and does not represent those of my employer.
The 17th Annual New England Society for Microscopy (NESM) at Woods Hole, MA will be held Friday, May 12th and Saturday, May l3th.
Highlights of the meeting include: Invited Presentations in both the Biological and Physical Sciences arena, Commercial exhibits, Poster/Photomicrograph Award Contest, Banquet & Cocktail Get-Together, Discovery Cruise, Door Prizes, and much more!
For information re: registration, meeting agenda, poster/photomicrograph submission forms, accomodations, and directions, please contact Peggy Sherwood, Corresponding Secretary (NESM) at MESnesm-at-aol.com. A newsletter will be mailed to you. Please respond ASAP-registration deadline is Monday, May 1, 2000!
hi all.... we are considering getting a kodak MDS 120 digital camera for photomicroscopy (brightfield)...does anyone have any experience with this system...is it cost effective....? user friendly? Is there any other system that would be recommended? thanks for any feedback Ronald F. Mervis, Ph.D. RonMervis-at-aol.com ~~~~~~~~~~~~~~~~ Neuro-Cognitive Research Laboratories 2109 West Fifth Ave Columbus, OH 43212
Long time ago I had worked in a laboratory which had a linear electron accelerator. It was well known that regular glasses (made from glass, not plastic) left for a few days close to accelerator would turn in a pretty good sunglasses. Effect would last for a few month, and then glasses again would be clear. So, every spring a few glasses were sitting close to accelerator. Sure, energy was much higher than for SEM.
Vladimir Dusevich
} Dear Joe and list members: } } I did not say anything about yellowing of ceramics at the XEI } Scientific web } site. It is generally not a "contamination" problem. X-rays being an } ionizing radiation are able to break bond and modify the crystalline } structure of ceramics. When I did X-ray spectrometry I was } very aware of the } ability of high intensity, high energy x-rays to cause } radiation damage and } discoloration. However scanning electron microscope are } generally operated } at low energies ( {30KeV) and low beam currents. Xray damage } is not a problem } except with most sensitive materials. } } Xray damage to oils could cause them to crosslink and } yellow if oils are } deposited on the surface of ceramic but I have not seen any } complaints about } this. Maybe a more specific description of your problem would } help. Reply } off list. } } Notice: XEI Scientific sells anti-contamination systems. } } Ronald Vane } XEI Scientific } } -----Original Message----- } } From: Joe {jmetzger-at-suscom.net} } To: Microscopy-at-sparc5.microscopy.com } {Microscopy-at-sparc5.microscopy.com} } Date: Wednesday, April 19, 2000 5:59 PM } Subject: Xray } } } } ------------------------------------------------------------- } ----------- } } 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 recently completed a research paper to complete my EE } } Degree which included a section on vacuum principles. I } } incorporated part of the in formation I got from an article } } by XEI scientific on contamination control. It spoke of oil } } deposits on cool windows or a loss of low energy xray. I } } thought I also read (on the web site) of how x-rays will } } turn ceramics yellow. I can not find the article again. } } Can you help me with this? What I am trying to decipher is, } } will concentrated xray emission through a ceramic cause it } } to yellow and could contamination deposits be the cause of } } this? Would appreciate any answers or leads as to where I } } might find the answer. } } } } Sincerely } } Joe Metzger } } } } } } } } } }
I'll be attempting to do some post-embedding immunogold labeling of murine platelets. I'm soliciting advice on preferred fixatives and resins. I'd like to stay away from cryosectioning if possible. If anyone has a great technique or special tips, I'd like to hear about it.
Thanks in advance!
Lesley S. Bechtold Supervisor, Biological Imaging The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6191
Fuji just came out with a new 4.3million pixel camera (Fujifilm FinePix 4700) utilizing their new octagonal pixel technology (Super CCD) and it sells for {$1000. I saw it in a catalog from Publishing Perfection (http:/www.publishingperfection.com)
At 11:01 AM 4/24/00 -0500, Beth Richardson wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Beth: There is an excellent review of the Nikon 990 at http://www.dpreview.com/reviews/nikoncp990/. Maybe this addresses your question. I suspect that this camera would be somewhat limited for copy stand work. I do a lot of digital imaging in my lab and use a Nikon D1, but this is considerably more expensive then a Coolpix 990. A good alternative is the Leaf Lumina which is marketed by Electron Microscopy Sciences and others. I hope this helps!!!!!!
Ken Bart
Kenneth M. Bart Director, Electron Microscopy Facility Hamilton College Clinton, New York 13323 ---------------------------------------- Phone:315-859-4715 Fax: 315-859-4807 email: kbart-at-hamilton.edu
MME's Market Research survey has been extended one day in honor of Friday's holiday. If you have not returned your survey forms, please do so by end of business Tuesday.
If, for some reason, we missed you, please email me immediately with your fax number.
We already have nearly 10% return. Both a summary of our findings and the winner of the M&M '99 Master Report will be posted Wednesday on www.MicroscopyMarket.com
Thanks for your participation.
Barbara Foster,President Microscopy/Marketing & Education 125 Paridon Street Suite 102 Springfield, MA 01118-2130 PH: 413-746-6931 FX: 413-746-9311 email: MME-at-map.com Website: www.MME-Microscopy.com/marketing
MME is a full service technical marketing company specializing in microscopy and related imaging technologies. Catalytic information and more .... We help your company grow! %%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%
GE does sell quartz glass in the US. I don't recall seeing coverslips, but we have purchase quartz that was approximately 1 mm in thickness. Here is their contact information and a website.
4901 Campbell Road Willoughby, Ohio 44094 USA Phone: (216) 266-3590 or 1-800-438-2100 FAX: (216) 266-4043 or 1-800-258-3803
********************************************** Dr. Raj Lartius, CEO NovaScan Technologies Iowa State University Research Park 2501 North Loop Drive Ames, Iowa USA 50010
Email: rlartius-at-novascan.com Voice: 515-795-3164 Fax: 515-795-4414 Web: www.novascan.com ********************************************** "Innovative Tools to Explore the Microworld"
I was asked today if there exists a book that broadly covers all of the different types of Microscopy (LM, EM, Probe Microscopes, etc) and gives their pros & cons and perhaps examples of their use. The faculty member who made the request is not a trained microscopist, so they would prefer something more general.
If there is such a book, it sounds like it would be an interesting one to read.
Yours, Doug Cromey .................................................................... : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : Research Specialist, Principal University of Arizona : : (office: AHSC 4212A) P.O. Box 245044 : : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: doug-cromey-at-ns.arizona.edu): :...................................................................: http://www.pharmacy.arizona.edu/exp_path.html Home of: "Microscopy and Imaging Resources on the WWW"
I am looking for a stereomicroscope with Greenough optics that provides final magnification of ~10X to 64X or 80X. I want to use it for visualizing algal cells growing on agar petri dishes. The cells are about 5 microns in diameter and I want to push them around on the agar using fine glass needles. I understand that the old Zeiss DR-C works well for this application. Vermont Optech and Sciscope have been very helpful and may have usable, used systems. Any other suggestions/ideas are welcome.
William J. Snell, Ph.D. University of Texas Southwestern Medical Center T-214-648-2332 F-214-648-8694 email-William.Snell-at-email.swmed.edu
I am searching some device for holding (polycarbonate)filters during dehydration and CPD. I am trying to view mycoplasms with SEM. Has anyone experience with this ?
Greetings,
De Pauw Bart Ghent University Faculty of Veterinary Medicine Morphology Salisburylaan 133 9820 Merelbeke Belgium Phone : 0032(0)9 264.77.19 Fax : 0032(0)9 264.77.90
If you can help with the message below, please respond to BarbaraRBC-at-aol.com [mailto:BarbaraRBC-at-aol.com]
Please do NOT respond to the list
Susanne P Brandom MicroWorld
MESSAGE
Can you assist by providing a list of live cell blood testers in the California/Arizona area? If not, are you able to point me in the direction where I might find a list of microscopists who perform this type of test?
Thank you
Barbara Churchill (909) 624-2459 ( fax and voice mail)
} From: Bill Miller [mailto:microbill-at-mohawk.net] } Sent: Monday, April 24, 2000 2:57 PM } To: Beth Richardson; microscopy-at-sparc5.microscopy.com } Subject: Re: digital cameras for copy stand work?
} Fuji just came out with a new 4.3million pixel camera (Fujifilm FinePix } 4700) utilizing their new octagonal pixel technology (Super CCD) and it } sells for {$1000. I saw it in a catalog from Publishing Perfection }
Fuji has two new cameras coming out based on their new "Super CCD" technology, the Finepix 4700 and the S-1Pro SLR Camera. In scientific imaging, an important issue is image interpolation and you must be aware of how Fuji determines resolution of these new cameras. The Super CCD technology utilizes octagonal pixels which are aligned at an angle, as opposed to horizontal rows for a conventional CCD with square pixels. The advantages of this design include denser packing of pixels, and having pixels sensitive to R,G,B in each row. It is a very interesting technology to say the least. Look closely at any Fuji literature about these cameras. The Finepix 4700 has an IMAGE resolution of 4.3 million pixels, the S1 Pro has an IMAGE resolution of 6 million pixels. Note this is not CCD resolution. The 4700 has a CCD resolution of approximately 2.3 million pixels, the S1 Pro has a CCD resolution of 3.2 million pixels. Fuji's image size specs are based on their claim that the image quality with the S1 Pro will be equivalent to an image captured with a 6 million pixel "conventional" CCD camera, or a 4.3 million pixel conventional CCD camera with the 4700. I have no doubt that the new cameras will produce high quality images. I can also say for sure that the cost of the new cameras is very affordable when compared to similar products currently available. It remains to be seen what effects that Fuji's interpolation will have on image integrity. S-1 Cameras are expected in late May or early June, I will be happy to provide sample images to any who request them at that time. Finepix 4700 cameras are currently starting to ship from Fuji.
George Laing National Graphic Supply scisales-at-ngscorp.com (800) 223-7130 X3109
Our book, "Optimizing Light Microscopy" covers the light end of things and there is a short chapter covering EM, confocal, etc. Details are on our website: MME-Microscopy.com/education. A number of colleges and universities have started using it as a text, most recently, U Wash (Kip Hauch).
Caveat: MME does have a financial interest in this project.
Best regards Barbara Foster,President Microscopy/Microscopy Education 125 Paridon Street Suite 102 Springfield, MA 01118-2130 PH: 413-746-6931 FX: 413-746-9311 email: MME-at-map.com Website: www.MME-Microscopy.com/education
America's first national consortium of microscopy specialists offering customized on-site training in all areas of microscopy, image analysis, and sample preparation ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^
At 02:53 PM 4/24/00 -0700, Doug Cromey wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I was tempted to write a similiar response, but it would seem too biased. However, I just read a recent Wall Street Journal article that stated that Olympus Corporation has filed a law suit against Fuji. Appearantly, Fuji does not feel they have a strong defense as they are instructing Dealers to place a correction sticker over the resolution specifications on the boxes and make disclaimers in all advertisements.
Regards,
Lawrence Kordon Nikon, Inc. nikon-at-jagunet.com
George Laing 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: Bill Miller [mailto:microbill-at-mohawk.net] } } Sent: Monday, April 24, 2000 2:57 PM } } To: Beth Richardson; microscopy-at-sparc5.microscopy.com } } Subject: Re: digital cameras for copy stand work? } } } Fuji just came out with a new 4.3million pixel camera (Fujifilm FinePix } } 4700) utilizing their new octagonal pixel technology (Super CCD) and it } } sells for {$1000. I saw it in a catalog from Publishing Perfection } } } Fuji has two new cameras coming out based on their new "Super CCD" } technology, the Finepix 4700 and the S-1Pro SLR Camera. In scientific } imaging, an important issue is image interpolation and you must be aware of } how Fuji determines resolution of these new cameras. } The Super CCD technology utilizes octagonal pixels which are aligned } at an angle, as opposed to horizontal rows for a conventional CCD with } square } pixels. The advantages of this design include denser packing of pixels, and } having pixels sensitive to R,G,B in each row. It is a very interesting } technology } to say the least. } Look closely at any Fuji literature about these cameras. The Finepix 4700 } has an IMAGE resolution of 4.3 million pixels, the S1 Pro has an IMAGE } resolution } of 6 million pixels. Note this is not CCD resolution. The 4700 has a CCD } resolution } of approximately 2.3 million pixels, the S1 Pro has a CCD resolution of 3.2 } million } pixels. } Fuji's image size specs are based on their claim that the image quality } with the } S1 Pro will be equivalent to an image captured with a 6 million pixel } "conventional" CCD } camera, or a 4.3 million pixel conventional CCD camera with the 4700. } I have no doubt that the new cameras will produce high quality images. I } can } also say for sure that the cost of the new cameras is very affordable when } compared to } similar products currently available. It remains to be seen what effects } that Fuji's } interpolation will have on image integrity. } S-1 Cameras are expected in late May or early June, I will be happy to } provide } sample images to any who request them at that time. Finepix 4700 cameras are } currently } starting to ship from Fuji. } } George Laing } National Graphic Supply } scisales-at-ngscorp.com } (800) 223-7130 X3109
In a message dated 04/24/2000 10:32:32 AM US Mountain Standard Time, RonMervis-at-aol.com-at-sparc5.microscopy.com writes:
{ { we are considering getting a kodak MDS 120 digital camera for photomicroscopy (brightfield)...does anyone have any experience with this system...is it cost effective....? user friendly? } }
Hi Ron,
The Kodak MDS 120 seems to do a decent job. Part of the MDS (Microscopy Documentation System) with the camera is a c-mount adapter that you mount to the phototube of your microscope. It requires a 1.0X adapter in the phototube, and the adapter should have a c-mount on it. The Kodak adapter then mounts on the end of the 1.0X adapter.
You may encounter some vignetting with the camera, especially if you try to use the wide-angle setting on the camera's zoom lens. The vignetting seems to be a function of the photo adapters which are used, and Kodak has a disclaimer about this in the instruction manual. It may or may not be a problem with your scope/phototube/c-mount adapter.
Part of the package which we got is a 16 MB Flash Memory card for the camera, so you can either run the camera from your computer or store images on the memory card and then remove the card and take it to a computer to transfer the images to the computer. The package also had a PCMCIA card adapter in it. The memory card plugs into the end of the PCMCIA device, and you can then slip it into a PCMCIA slot on a laptop computer.
But if you need to use a desktop computer and don't have a PCMCIA slot, you need to get something like a SanDisk card reader for the computer. They're available at CompUSA, Fry's Electronics, OfficeMax, etc. for about $70.
The software with the camera is very nice. It lets you transfer images from the camera's memory directly or from from the memory card, preview images and download them directly from the camera, etc. as well as perform some basic image adjustments (flip, rotate, modify brightness, contrast, gamma, etc.)
Overall, I would say the Kodak system is a nice way to document general microscopy images in brightfield and phase contrast microscopy for under $2K. We haven't tried to use the camera for fluorescence imaging, so I have no history to contribute on this subject.
There is the Handbook of Microscopy: Applications in Materials Science, Solid State Physics and Chemistry /ed by S. Amelinckx et al., v.1-3, Weinheim, N.Y.: VCH (1997). It covers LM, EM (stationary and scanning beam methods), X-ray and acoustic microscopy, scanning probe techniques, magnetic and emission microscopies, image analysis, and microscopy applications to various classes of materials as well.
Vladimir
************************************ Vladimir P. Oleshko, Ph.D. Industrial Associates Program Center for Solid State Science Arizona State University Main Campus, PO Box 871704 Tempe, AZ 85287-1704 (480) 727-7666 Fax: (480) 965-9004 E-Mail:oleshko-at-imap3.asu.edu *************************************
-----Original Message----- } From: Doug Cromey [mailto:doug-cromey-at-ns.arizona.edu] Sent: Monday, April 24, 2000 2:54 PM To: microscopy-at-sparc5.microscopy.com
Colleagues,
I was asked today if there exists a book that broadly covers all of the different types of Microscopy (LM, EM, Probe Microscopes, etc) and gives their pros & cons and perhaps examples of their use. The faculty member who made the request is not a trained microscopist, so they would prefer something more general.
If there is such a book, it sounds like it would be an interesting one to read.
Yours, Doug Cromey ................................................................... : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : Research Specialist, Principal University of Arizona : : (office: AHSC 4212A) P.O. Box 245044 : : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: doug-cromey-at-ns.arizona.edu): :...................................................................: http://www.pharmacy.arizona.edu/exp_path.html Home of: "Microscopy and Imaging Resources on the WWW"
In addition to what George and Bill have pointed out, Fuji claims the imager (ccd) is roughly the size of APS film on the S1 Pro. What I cannot find in the literature is the pixel size specification whereby one can calculate or compare the field of view (on a microscope) of this camera with other mega pixel cameras. Shane
K. Shane Collins Scientific Instrument Company 805.444.4953 cell 310.568.9188 office 310.568.9189 fax
-----Original Message----- } From: George Laing [mailto:scisales-at-ngscorp.com] Sent: Tuesday, April 25, 2000 6:27 AM To: Microscopy-at-sparc5.microscopy.com
} From: Bill Miller [mailto:microbill-at-mohawk.net] } Sent: Monday, April 24, 2000 2:57 PM } To: Beth Richardson; microscopy-at-sparc5.microscopy.com } Subject: Re: digital cameras for copy stand work?
} Fuji just came out with a new 4.3million pixel camera (Fujifilm FinePix } 4700) utilizing their new octagonal pixel technology (Super CCD) and it } sells for {$1000. I saw it in a catalog from Publishing Perfection }
Fuji has two new cameras coming out based on their new "Super CCD" technology, the Finepix 4700 and the S-1Pro SLR Camera. In scientific imaging, an important issue is image interpolation and you must be aware of how Fuji determines resolution of these new cameras. The Super CCD technology utilizes octagonal pixels which are aligned at an angle, as opposed to horizontal rows for a conventional CCD with square pixels. The advantages of this design include denser packing of pixels, and having pixels sensitive to R,G,B in each row. It is a very interesting technology to say the least. Look closely at any Fuji literature about these cameras. The Finepix 4700 has an IMAGE resolution of 4.3 million pixels, the S1 Pro has an IMAGE resolution of 6 million pixels. Note this is not CCD resolution. The 4700 has a CCD resolution of approximately 2.3 million pixels, the S1 Pro has a CCD resolution of 3.2 million pixels. Fuji's image size specs are based on their claim that the image quality with the S1 Pro will be equivalent to an image captured with a 6 million pixel "conventional" CCD camera, or a 4.3 million pixel conventional CCD camera with the 4700. I have no doubt that the new cameras will produce high quality images. I can also say for sure that the cost of the new cameras is very affordable when compared to similar products currently available. It remains to be seen what effects that Fuji's interpolation will have on image integrity. S-1 Cameras are expected in late May or early June, I will be happy to provide sample images to any who request them at that time. Finepix 4700 cameras are currently starting to ship from Fuji.
George Laing National Graphic Supply scisales-at-ngscorp.com (800) 223-7130 X3109
I am forwarding this request in the hopes that someone would be able to assist these film producers to find some footage for a museum exhibit they are working on.
Thanks for our attention.
John Bozzola
+++++++++++++++++++++++++++++++++++++++++++++++++
Chedd-Angier is producing a series of exhibits for the Science Museum of Virginia in Richmond, VA.(. http://www.chedd-angier.com.) We currently produce the Scientific American Frontiers Program on PBS.(http://www.pbs.org/saf)
Our science museum project encompasses a wide variety of exhibits ranging from a voyage through a cell called cell watcher, to a life size journey through the human body called BODY PROBE. I am looking for a variety of footage pieces to use in these exhibits. They will all be used for North American non broadcast educational use only in a museum that DOES not have a separate admission policy for this exhibit. The electron micrograph footage that I am looking for is :
For cell watcher Mitochondria Golgi Bodies Cell membranes Nucleus Chromosomes Vesicles Endoplasmic Reticulum Lysosomes Fat cells expanding Mitosis White blood cells muscle cells contracting plant stem cells elongating cells from a fallopian tube amoeba sperm cells swimming This is a very extensive list, and if there are animation related footage sources I can use those as well. If you have any questions at all please contact me at 617-926-8300. Thank you
Andr Stark Producer Chedd-Angier 70 Coolidge Hill Rd Watertown, MA 02472 0101617-926-8300 617-926-2710(F)
As a new subscriber to the listserv I would like to ask your assistance in locating copies of any manuals or in-house guidelines for the basic care and maintenance of light microscopes.
I am currently assisting a group of microscopists in a humid / tropical environment to increase their ability to care for their microscopes. Care by the user is an important issue as is the ability to repair/maintain microscopes in-country. Whilst we can find quite a bit of information on how to use your microscope there is less on maintenance.
In addition does anyone have any idea on where we can purchase a "two-pin tool" for removing lenses?
Thankyou
Hazel Clothier
Regional Laboratory Scientist Pacific Regional Vector Borne Diseases Project Secretariat of the Pacific Community PMB Suva FIJI
Tel: (679) 321154 - direct (679) 320066 - ex 110 Fax:(679) 322714 hazelc-at-int
I am a beginning microscopist. I am preparing to take some TEM photos of granules isolated from mast cells. The technician I am working with would like to use phosphate buffer (+ glutaraldehyde) to fix the granules, but the literature I've read suggests using cacodylate buffer. What do you think about the benefits/disadvantages of phosphate vs cacodylate buffer for these purposes?
thanks, Aaron
__________________________________________________ Do You Yahoo!? Send online invitations with Yahoo! Invites. http://invites.yahoo.com
Question: when comparing phase contrast and interference contrast (Nomarski) the negative points a.o.(Ph) are halo. Interference contrast also (especially in minute organisms) has a drawback since depending on the position of the polarizer some part of f.i.bacteria do not show the relief image (the shadow) to its fullest. How do you call this phenomenon and how can it be explained?
if anyone has a used set of this avail let me know. also looking for encyclopedia of microscopy by gray (editor) thanks Ed Sharpe
{ { Subj: RE: General Microscopy book recommendation? Date: 4/25/00 2:29:52 PM US Mountain Standard Time From: Vladimir.Oleshko-at-asu.edu (Vladimir Oleshko) Reply-to: {A HREF="mailto:oleshko-at-imap3.asu.edu"} oleshko-at-imap3.asu.edu {/A} To: doug-cromey-at-ns.arizona.edu ('Doug Cromey') CC: microscopy-at-sparc5.microscopy.com
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There is the Handbook of Microscopy: Applications in Materials Science, Solid State Physics and Chemistry /ed by S. Amelinckx et al., v.1-3, Weinheim, N.Y.: VCH (1997). It covers LM, EM (stationary and scanning beam methods), X-ray and acoustic microscopy, scanning probe techniques, magnetic and emission microscopies, image analysis, and microscopy applications to various classes of materials as well.
Vladimir
************************************ Vladimir P. Oleshko, Ph.D. Industrial Associates Program Center for Solid State Science Arizona State University Main Campus, PO Box 871704 Tempe, AZ 85287-1704 (480) 727-7666 Fax: (480) 965-9004 E-Mail:oleshko-at-imap3.asu.edu *************************************
I am currently using a JEOL 5800LV SEM. My specimens are cells from cultures which we have grown in our labs. The low vacuum condition is 7Pa. The problem occurs when I increase the magnification, about 400 times and beyond. The picture I get will be enlarged and clear except for certain regions which displays some glaring effects. Playing around with the contrasts and brightness didn't help. I was told this was a charging problem, and increasing the vacuum might help, so I tried. It did help a little but the picture quality was compromised.
Are there other solutions to this problem of mine, such that the picture quality might remain the same.
Chris Lam BIOMAT Mechanical & Production Department National University of Singapore
Following the thread from Doug Cromey asking about books for Microscopy, I would like to suggest two manuals for light microscopy that are comprehensive in their coverage, *accurate in their educational content*, and easy to read and digest.
Title: Light Microscopy, An Illustrated Guide. Author: Ron OLDFIELD. Location: Sydney, Australia Published: 1994, Wolfe Publishing, imprint of Mosby-Year Book, Europe. 160 pages ISBN: 0-7234-1876-4 Available Amazon, no price given; ca. $20.
Title: Introduction to Light Microscopy Authors: Savile BRADBURY & Brian BRACEGIRDLE Location: Oxford, England Published: 1998, Bios Scientific Publishers Website: www.bios.co.uk Royal Microscopical Society Handbook No. 42, 122 pages. ISBN: 1-85996-121-5 Amazon Price: $32.95
Searching for, and finding, a suitable teaching and/or reference text is a personal thing. It depends on your requirements and preferences, but I suggest that these two books will help most people in most disciplines. I have used them for the last six years in teaching light microscopy to a wide spectrum of students from industry and academia.
For those intessted in digital imaging, although not a book specifically for microscopists, I have found the following text useful:
Title: Digital Imaging for Photographers, 3rd edn. Authors: Adrain DAVIES & Phil FENNESSY Published: 1998, Focal Press, Oxford, Boston.170 pages. website: www.bh.com/focalpress (Focal Press is an imprint of Butterworth-Heinemann) ISBN: 0-240-51538-2 Amazon Price: $31.96
I am both a professional and an amateur microscopist, please do not think I am denigrating the word amateur, but as an introductory text for amateur microscopists,or those starting out in a complex field, I would suggest:
Title: Exploring With the Microscope : A Book of Discovery & Learning Author: Werner NACHTIGALL Publisher: Stirling Publishing Corp. Inc., April 1997. 160 pages ISBN: 0-80690866-1 (check that this is 2nd edition) Amazon Price: $11.96
Finally, if you consider that getting school children interested in microscopy, and science generally, is an important investment for the future, then I would suggest the new Usborne Complete Book of the Microscope, which shows the relevance and the application of microscopy in many walks of life.
Title: The Usborne Complete Book of the Microscope Author: Kirsteen ROGERS Publisher: 1998, Usborne Publishing Ltd, 96 pages. website: www.usborne.com ISBN: 0-7460-3106-8 Amazon Price: $22.95
Whatever your field, light microscopy underpins, and is a necessary foundation for, electron microscopy, confocal and other techniques. I hope that these suggestions are useful. I would welcome anyone's views to me directly.
Jeremy Sanderson jb_sanderson-at-yahoo.com
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I have a few PE specimens on my desk that I want to prepare with the "permanganate etching method". Does anybody of you have some experience on this techique? Would you share some details or tips and tricks with me on how to obtain the best results? What is the best method to produce a replica from the etched specimen?
Petra -------------------------------------------------------------- Dr. Petra Wahlbring Centre de Recherche Public - Gabriel Lippmann Laboratoire d'Analyse des Materiaux (LAM) 162a, av. de la Faiencerie L-1511 Luxembourg tel. +352-466644-402 fax +352-466644-400 e-mail: petra.wahlbring-at-crpgl.lu Visit our WWW site! http://www.crpgl.lu/~wahlbrin
} ------------------------------------------------------------------------ } 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 a beginning microscopist. I am preparing to take } some TEM photos of granules isolated from mast cells. } The technician I am working with would like to use } phosphate buffer (+ glutaraldehyde) to fix the } granules, but the literature I've read suggests using } cacodylate buffer. What do you think about the } benefits/disadvantages of phosphate vs cacodylate } buffer for these purposes? } } thanks, } Aaron } } __________________________________________________ } Do You Yahoo!? } Send online invitations with Yahoo! Invites. } http://invites.yahoo.com } } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
Other EM websites around the world are invited to add this forum to their own sites - just follow the link on the Forum to customize your own version for your own web pages.
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-- ************************************************************ Duncan Waddell (BSc) Senior Scientific Officer Centre for Microscopy and Microanalysis The University of Queensland, St. Lucia, Qld, 4072 Australia Telephone: +61-7-3365-4216 Facsimile: +61-7-3365-4422 WWW: http://www.uq.edu.au/nanoworld/nanohome.html ************************************************************ Any opinion expressed is that of the writer, and not necessarily that of CMM or of the University. ************************************************************
Shane Collins - do you mean pixel size and numbers when you refer to field of view? Field of view, as I understand it has nothing to do with image quality, but is simply the physical size covered by a photographic system on the microscope. The crucial factors affecting field of view are objective lens mag. and the relation between photo eyepiece and the physical size of the film or CCD. 35mm film combined with a 3x photo eyepiece is about "normal", small CCD's require a much lower mag eyepiece, otherwise the system gives too greater magnifications, frequently beyond OM limits. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Wednesday, April 26, 2000 8:59 AM, Shane Collins [SMTP:kshanec-at-gte.net] wrote: } } } In addition to what George and Bill have pointed out, Fuji claims the imager } (ccd) is roughly the size of APS film on the S1 Pro. What I cannot find in } the literature is the pixel size specification whereby one can calculate or } compare the field of view (on a microscope) of this camera with other mega } pixel cameras. } Shane } } K. Shane Collins } Scientific Instrument Company } 805.444.4953 cell } 310.568.9188 office } 310.568.9189 fax } } } -----Original Message----- } } From: George Laing [mailto:scisales-at-ngscorp.com] } Sent: Tuesday, April 25, 2000 6:27 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: RE: digital cameras for copy stand work? } } } ------------------------------------------------------------------------ } 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: Bill Miller [mailto:microbill-at-mohawk.net] } } Sent: Monday, April 24, 2000 2:57 PM } } To: Beth Richardson; microscopy-at-sparc5.microscopy.com } } Subject: Re: digital cameras for copy stand work? } } } Fuji just came out with a new 4.3million pixel camera (Fujifilm FinePix } } 4700) utilizing their new octagonal pixel technology (Super CCD) and it } } sells for {$1000. I saw it in a catalog from Publishing Perfection } } } } Fuji has two new cameras coming out based on their new "Super CCD" } technology, the Finepix 4700 and the S-1Pro SLR Camera. In scientific } imaging, an important issue is image interpolation and you must be aware of } how Fuji determines resolution of these new cameras. } The Super CCD technology utilizes octagonal pixels which are aligned } at an angle, as opposed to horizontal rows for a conventional CCD with } square } pixels. The advantages of this design include denser packing of pixels, and } having pixels sensitive to R,G,B in each row. It is a very interesting } technology } to say the least. } Look closely at any Fuji literature about these cameras. The Finepix 4700 } has an IMAGE resolution of 4.3 million pixels, the S1 Pro has an IMAGE } resolution } of 6 million pixels. Note this is not CCD resolution. The 4700 has a CCD } resolution } of approximately 2.3 million pixels, the S1 Pro has a CCD resolution of 3.2 } million } pixels. } Fuji's image size specs are based on their claim that the image quality } with the } S1 Pro will be equivalent to an image captured with a 6 million pixel } "conventional" CCD } camera, or a 4.3 million pixel conventional CCD camera with the 4700. } I have no doubt that the new cameras will produce high quality images. I } can } also say for sure that the cost of the new cameras is very affordable when } compared to } similar products currently available. It remains to be seen what effects } that Fuji's } interpolation will have on image integrity. } S-1 Cameras are expected in late May or early June, I will be happy to } provide } sample images to any who request them at that time. Finepix 4700 cameras are } currently } starting to ship from Fuji. } } George Laing } National Graphic Supply } scisales-at-ngscorp.com } (800) 223-7130 X3109 } }
PO4 buffers can cause Calcium to precipitate out but the use of cacodylate is archaic. Microscopists tend to be the most refractory of all scientists to change. HEPES or PIPES are reasonable alternatives for almost all procedures in which cacodylate is used - certainly widely used for basic fixation such as you describe. they are non-toxic, less expensive, and better buffers. good luck
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The Fuji Super CCD is 23.3 x 15.6mm; 3040 x 2016 pixels. The S1 Pro uses Nikon mount lenses and has an equivalent 35mm frame size at 1.5X the lens' focal length.
What the equivalent FOV would be on a scope is still in question. If the S1 Pro works anywhere near how the Nikon E1 or E2 does, I fear the Fuji camera won't be applicable to microscopy at all. Without a lens, the camera must operate in aperture priority mode and center weighted exposure reading. Unless Fuji made some major changes to the basic Nikon (N-60) body and associated electronics, they may have just made a higher pixel count Nikon digicam.
gary g.
At 03:59 PM 4/25/00 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Refinement of Etching Techniques to Reveal Lamellar Profiles in Polyethylene Banded Spherulites Shahin,M.M., Olley,R.H., Blissett,M.J. J. Polym. Sci. Polym. Part B: Polym. Phys. 1999, vol.37, pp. 2279-2286
This is our latest development in the technique. If you don't have a copy, I can send you a reprint.
Generally, we use a two stage replication technique, making a first stage replica out of cellulose acetate, then putting Ta/W shadow and carbon on this and extracting the cellulose acetate.
If you can let me know a little bit more about your PE specimens, I can give you a few more details specifically adapted to your type of specimen. In particular, is it HDPE, LLDPE or LDPE?
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
Aron, Caco is more extractive which may (or may not) be a blessing if you are trying to clear out the cytoplasm a bit to get some more detail. Poshpate is more physiologic but may form artifactual granules with glutaraldehyde and osmium (in routine tandem use). I generally use caco for all non immuno work otherwise phosphate, pbs or hepes for IEM. Also remember caco contains arsenic. Good luck.
} ------------------------------------------------------------------------ } 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 a beginning microscopist. I am preparing to take } some TEM photos of granules isolated from mast cells. } The technician I am working with would like to use } phosphate buffer (+ glutaraldehyde) to fix the } granules, but the literature I've read suggests using } cacodylate buffer. What do you think about the } benefits/disadvantages of phosphate vs cacodylate } buffer for these purposes? } } thanks, } Aaron } } __________________________________________________ } Do You Yahoo!? } Send online invitations with Yahoo! Invites. } http://invites.yahoo.com } } } } Whenever possible (nearly all the time) use phosphate buffer. Cacodylate contains arsenic which is a potent carcinogen. One should never get into the habit of using arsenic buffer "just because". There must be a real reason for it! Once cacodylate is in permanent use in a laboratory, dust accumulates on the outside of bottles, on the inside of laboratory glassware waiting to be washed, and so on. The effects are cumulative. Arsenic is also a toxic to the kidneys, etc. Very dangerous stuff mostly because it is in use in many laboratories day in and day out.
} ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } Phosphate buffers can give rise to a precipitate with } uranyl acetate if it is used as an en block stain. } Cacodylate does not. } } Cacodylate contains arsenic and must be used with care. } } Dave } } } On Tue, 25 Apr 2000 22:56:46 -0500 Aaron Wheeler } {adog3050-at-yahoo.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 a beginning microscopist. I am preparing to take } } some TEM photos of granules isolated from mast cells. } } The technician I am working with would like to use } } phosphate buffer (+ glutaraldehyde) to fix the } } granules, but the literature I've read suggests using } } cacodylate buffer. What do you think about the } } benefits/disadvantages of phosphate vs cacodylate } } buffer for these purposes? } } } } thanks, } } Aaron } } } } __________________________________________________ } } Do You Yahoo!? } } Send online invitations with Yahoo! Invites. } } http://invites.yahoo.com } } } } } } } } ---------------------------------------- } Patton, David } Email: David.Patton-at-uwe.ac.uk } "University of the West of England" } } } If it is necessary to use UA with a buffer, use maleate systems. No precipitate!
Looking for the adjustable lucite bases made several years ago for adjusting a LM for improved ergonomic access/use. I had the information/supplier/manufacturer info around here someplace, but other than the somewhat random neuronal firing, I appear to have lost/misplace/misfiled/disposed of everything. I have a pathologist who needs to incline his Nikon Microphot FX-A about 5 degrees. We have begun the process of having a fixed wooden base constructed, but I much prefer the lucite devices. Any help/ideas? TIA.
Roger C Moretz, Ph.D.
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Looking for the adjustable lucite bases made several years ago for adjusting a LM for improved ergonomic access/use. I had the information/supplier/manufacturer info around here someplace, but other than the somewhat random neuronal firing, I appear to have lost/misplace/misfiled/disposed of everything. I have a pathologist who needs to incline his Nikon Microphot FX-A about 5 degrees. We have begun the process of having a fixed wooden base constructed, but I much prefer the lucite devices. Any help/ideas? TIA.
Roger C Moretz, Ph.D.
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Looking for the adjustable lucite stand marketed several years ago (at least within this fuzzy brain of mine it was that long ago) for improving the ergonomic interface to LMs. I have a pathologist who needs to incline his Nikon Microphot FX-A about 5 degrees. We had just about decided to have a fixed unit built by the carpentry shop, when I remembered the lucite devices. Can anyone help point me in the right directions? TIA.
Roger C Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc.
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Looking for the adjustable lucite bases made several years ago for adjusting a LM for improved ergonomic access/use. I had the information/supplier/manufacturer info around here someplace, but other than the somewhat random neuronal firing, I appear to have lost/misplace/misfiled/disposed of everything. I have a pathologist who needs to incline his Nikon Microphot FX-A about 5 degrees. We have begun the process of having a fixed wooden base constructed, but I much prefer the lucite devices. Any help/ideas? TIA.
Roger C Moretz, Ph.D.
_______________________________________________________ Get 100% FREE Internet Access powered by Excite Visit http://freelane.excite.com/freeisp
Looking for the adjustable lucite bases made several years ago for adjusting a LM for improved ergonomic access/use. I had the information/supplier/manufacturer info around here someplace, but other than the somewhat random neuronal firing, I appear to have lost/misplace/misfiled/disposed of everything. I have a pathologist who needs to incline his Nikon Microphot FX-A about 5 degrees. We have begun the process of having a fixed wooden base constructed, but I much prefer the lucite devices. Any help/ideas? TIA.
Roger C Moretz, Ph.D.
_______________________________________________________ Get 100% FREE Internet Access powered by Excite Visit http://freelane.excite.com/freeisp
On Wed, 26 Apr 2000 15:13:14 +0800 Lam xu Fu Christopher {eng81067-at-nus.edu.sg} 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 Sir, } } I am currently using a JEOL 5800LV SEM. } My specimens are cells from cultures which we have grown in our labs. } The low vacuum condition is 7Pa. } The problem occurs when I increase the magnification, about 400 times and } beyond. The picture I get will be enlarged and clear except for certain } regions which displays some glaring effects. Playing around with the } contrasts and brightness didn't help. } I was told this was a charging problem, and increasing the vacuum might } help, so I tried. It did help a little but the picture quality was } compromised. } } Are there other solutions to this problem of mine, such that the picture } quality might remain the same. } } Chris Lam } BIOMAT } Mechanical & Production Department } National University of Singapore } } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
Apologies for the multiple messages. The stupid excitemail server kept telling me the message could not be sent and then deleted the message from my inbox. So, I wrote another one, sent that, etc. Next time, I'll wait until I see if the listserver sends the one through before resending. :-( On Wed, 26 Apr 2000 12:32:40 -0700 (PDT), rmoretz-at-rdg.boehringer-ingelheim.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 } -----------------------------------------------------------------------. } } } } Looking for the adjustable lucite bases made several years ago for adjusting } a LM for improved ergonomic access/use. I had the } information/supplier/manufacturer info around here someplace, but other than } the somewhat random neuronal firing, I appear to have } lost/misplace/misfiled/disposed of everything. I have a pathologist who } needs to incline his Nikon Microphot FX-A about 5 degrees. We have begun } the process of having a fixed wooden base constructed, but I much prefer the } lucite devices. Any help/ideas? TIA. } } } Roger C Moretz, Ph.D. } } } } } } _______________________________________________________ } Get 100% FREE Internet Access powered by Excite } Visit http://freelane.excite.com/freeisp } }
Roger C Moretz, Ph.D.
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} Dear Sir,
} I am currently using a JEOL 5800LV SEM. } My specimens are cells from cultures which we have grown in our labs. } The low vacuum condition is 7Pa. } The problem occurs when I increase the magnification, about 400 times and } beyond. The picture I get will be enlarged and clear except for certain } regions which displays some glaring effects. Playing around with the } contrasts and brightness didn't help. } I was told this was a charging problem, and increasing the vacuum might } help, so I tried. It did help a little but the picture quality was } compromised.
} Are there other solutions to this problem of mine, such that the picture } quality might remain the same.
} Chris Lam } BIOMAT } Mechanical & Production Department } National University of Singapore
Chris,
I assume that your samples are not coated. 7 Pa may be too much of a vacuum for uncoated samples; enough gas molecules are needed to help neutralize the charge but too many tend to attenuate the signal. I generally operate the 5800 LV at 20-27 Pa without abundat charging artefacts. I would suggess 5 kV as maximum gun potential and a probe current of 10 or less.
Cheers,
Stephen M. Harmon Electron Microscopist United States Environmental Protection Agency M.S. 681 26 W. Martin Luther King Blvd. Cincinnati, OH 45268 513.569.7184 Harmon.Stephen-at-epa.gov
A beginning course on chemical microscopy covering the basic techniques of microchemical analysis using a microscope
The workshop will consist a weekend of lectures and hands on labs to cover theoretical and practical aspects of chemical microscopy. We expect to follow up with a second weekend course at a more advanced level for which this will be a prerequisite. The course instructors will be the well known and respected "Skip" Palenik of Microtrace, Inc. in Elgin, IL and N.Y.M.S. Instructor Don O'Leary.
WHEN: May 20 & 21, 2000 from 10 A.M. to 4 P.M.
WHERE: 1244 McBride Avenue, West Paterson, NJ. Phone (973) 812-8377 (Free parking, accessible by public transportation, Information on car pools and transportation will be provided.)
COST: $475 for N.Y.M.S. members, $495 for non-members (includes membership) Lunch and course materials are included. Checks made out to N.Y.M.S.
WHO: Those with some basic knowledge of chemistry and chemical analysis and use of the microscope.
HOW: Register using the form below. Limited to the first 12 registrants. Return form to Don O'Leary, 6 Chittenden Road, Fair Lawn, NJ 07410.
FURTHER INFORMATION: Call D. O'Leary (201) 797 -8849
PLEASE POST ---------------------------------------------------------------------------- ------------------------------------- Registration Form Chemical Microscopy
Are you sure it is a charging and not a burning? But anyway you can try some these steps: 1. If you are using slow scan, increase frequency of scanning and average frames. 2. Just decrease beam intencity and kV (may be you have too nice beam) until an image become too noisy. 3. Decrease beam intencity. If you are using BSE you can try to increase kV too. For SE (and I am not sure what type of it you can have) try increase pressure and kV, or decrease pressure and kV.
Good luck
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} -----Original Message----- } From: Lam xu Fu Christopher [mailto:eng81067-at-nus.edu.sg] } Sent: Wednesday, April 26, 2000 2:13 AM } To: 'Microscopy-at-MSA.Microscopy.Com' } Subject: SEM charging effects } } } -------------------------------------------------------------- } ---------- } 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 Sir, } } I am currently using a JEOL 5800LV SEM. } My specimens are cells from cultures which we have grown in our labs. } The low vacuum condition is 7Pa. } The problem occurs when I increase the magnification, about } 400 times and } beyond. The picture I get will be enlarged and clear except } for certain } regions which displays some glaring effects. Playing around with the } contrasts and brightness didn't help. } I was told this was a charging problem, and increasing the } vacuum might } help, so I tried. It did help a little but the picture quality was } compromised. } } Are there other solutions to this problem of mine, such that } the picture } quality might remain the same. } } Chris Lam } BIOMAT } Mechanical & Production Department } National University of Singapore } } }
I am a technical recruiter. Currently, I am working on a search for one of my Northeast clients for a Senior Metallographic Technician:
Will perform metallographic evaluation of coatings including work with SEM (Scanning Electron Microscopy) and X-ray. Will be responsible for the metallurgy laboratory, including organization of work, and operation and calibration of equipment. Will write technical and engineering reports. Will prepare projects for the metallurgy laboratory. Will maintain communication with customers.
AAS/BS plus fours years experience in a metallography laboratory.
Resumes should be sent by email, mail or fax. For best results, please send emails as attached Word.docs - I have Word 97.
Pearl Martin Image Associates Inc. 5254 Merrick Road Massapequa, NY 11758 Phone (516)798-3993 Fax (516)797-8703 Email: pearl-at-jobspot.com
Hi All, I am turning to all of you as a last ditch effort to see if my diamond knife is crazy or I am. I have been using this same knife for years (approximately 4 years) with the occasional difficulty. However, in the past few weeks I have been having a very hard time getting sections. The problem is this: water in the boat either wets the block or pulls away from the knife edge. There seems to be no in-between. Every once in a while, the stars seem to align, and the level achieves perfection for a few sections. I've tried several different cleaning methods, thinking that maybe the knife was dirty since I've been sectioning a lot of LR White. The only thing I've noticed (since I've been looking so very closely at the diamond) is that the seal (which was damaged when the knife arrived from the manufacturer) between the knife and the boat has detached. I would think that this would cause a leakage problem, which I haven't seen, not the problem I'm experiencing. Does anyone have any advice? Thanks for your help, Kristen Kristen A. Lennon Cell, Molecular & Developmental Biology Group Department of Botany & Plant Sciences University of California Riverside, CA 92521 kalen-at-citrus.ucr.edu
Tilting the sample toward the detector (up to about 45 degrees) helps eliminate some types of charging. It is easy enough and worth a try before more involved techniques are tried.
Mike Baxter Lehman College
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Petra: Are you using "permanganate etch" on a metallographic specimen? If so, Vander Voort's book "Meatallogrphy: Principals and Practice" covers the use of permanganate etches. See also Petzow's book on etching, in German or English.
Sam Purdy National Steel Technical Center Trenton, MI, USA } ---------- } From: Petra Wahlbring } Sent: 26, April 2000, 4:55 AM } To: microscopy-at-sparc5.microscopy.com } Subject: TEM: permanganate etching } } ------------------------------------------------------------------------ } 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 a few PE specimens on my desk that I want to prepare with the } "permanganate etching method". Does anybody of you have some experience on } this techique? Would you share some details or tips and tricks with me on } how to obtain the best results? What is the best method to produce a } replica from the etched specimen? } } Petra } -------------------------------------------------------------- } Dr. Petra Wahlbring } Centre de Recherche Public - Gabriel Lippmann } Laboratoire d'Analyse des Materiaux (LAM) } 162a, av. de la Faiencerie L-1511 Luxembourg } tel. +352-466644-402 fax +352-466644-400 } e-mail: petra.wahlbring-at-crpgl.lu } Visit our WWW site! http://www.crpgl.lu/~wahlbrin }
In a message dated 04/27/2000 5:27:39 PM US Mountain Standard Time, kalen-at-citrus.ucr.edu writes:
{ { water in the boat either wets the block or pulls away from the knife edge. There seems to be no in-between. Every once in a while, the stars seem to align, and the level achieves perfection for a few sections. I've tried several different cleaning methods, thinking that maybe the knife was dirty since I've been sectioning a lot of LR White. The only thing I've noticed (since I've been looking so very closely at the diamond) is that the seal (which was damaged when the knife arrived from the manufacturer) between the knife and the boat has detached. I would think that this would cause a leakage problem, which I haven't seen, not the problem I'm experiencing. } }
Kristen,
I had a knife that behaved similarly once upon a time. Let's assume the boat isn't leaking, since you aren't having problems with water collecting underneath the knife in the knife holder, correct? It would be difficult to imagine the knife has come loose in the boat, but that may be a very slight possibility. If it were loose in its mount it might not section with consistency. Either way, it would be a good idea to have the knife re-sealed properly.
The knife I had was almost impossible to get the proper amount of wetting on the diamond, but two things helped:
1. Try deliberately overfilling the boat to give a positive meniscus right at the knife edge. Leave the knife in this state for 15-20 minutes. Relax, go have a cup of coffee, whatever. Then try lowering the fluid to the proper level. This worked well for me.
2. For really difficult knives that refuse to wet, just a very little bit of saliva on the end of a dalmatian hair works great. A light brush against the tongue, then drag the hair in the boat fluid behind the knife edge. I know it sounds gross, but it works! Others have used Alconox crystals, detergents, etc. to break the surface tension, but good old saliva works every time.
IIn the following references, methods to adjust the best operating conditions (scan rate, Energy, current ) for SEM of uncoated insulators samples are proposed and discussed.
D. C. Joy, Scanning 11, 1 (1989). D. C Joy and C. S. Joy , Micron. 27, 247 (1996).
Good luck
***************************************************************** Mohamed Belhaj UFR SCIENCES Laboratoire d'Analyse des Solides Surfaces et Interfaces DTI/LASSI UMR CNRS BP 1039 Reims 51687 Cedex 2 Tel : (00 33) 03 26 91 33 27: (00 33) 03 26 91 33 14 Fax : (00 33) 03 26 91 33 12 ******************************************************************
Bob Chiovetti's ideas are right on the mark. It sounds crazy but saliva works wonders. The only thing I would change would be the tool to use to apply the saliva. It would probably be safer if you used the styrofoam stick provided by many diamond knife or EM supply folks. Clean the knife with the stick according to the instructions using a "bit of spit". Follow this up with DI water since saliva is surprisingly dirty.
Should restore any wetting properties your old knife has left.
Have fun, Joe Tabeling Delaware Diamond Knives, Inc. 800-222-5143
I am interested in getting information regarding processing images, specifically clay fabric using Adobe PhotoShop and Image Tool, (plug-ins and "The Image Processing Handbook"). Can anyone provide me with information on a short course or 3 to 5 day workshop on this subject?
} I am turning to all of you as a last ditch effort to see if my diamond } knife is crazy or I am. I have been using this same knife for years } (approximately 4 years) with the occasional difficulty. However, in the } past few weeks I have been having a very hard time getting sections. The } problem is this: water in the boat either wets the block or pulls away from } the knife edge. There seems to be no in-between. Every once in a while, the } stars seem to align, and the level achieves perfection for a few sections. } I've tried several different cleaning methods, thinking that maybe the } knife was dirty since I've been sectioning a lot of LR White. The only } thing I've noticed (since I've been looking so very closely at the diamond) } is that the seal (which was damaged when the knife arrived from the } manufacturer) between the knife and the boat has detached. I would think } that this would cause a leakage problem, which I haven't seen, not the } problem I'm experiencing.
} Kristen -
Sounds like you're describing a slow leak, which will lower the water level & wet the back of the knife. Try the "old-fashoned" knife sealing method: Melt a bit of dental wax on your slide-warmer, and warm the knife gently while the wax is melting. Use a toothpick to run a bit of wax into the cracked seal.
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/PCI/pci.html
This sounds crazy, but my new knife came from Micro Star Diamond knives, and they recommend only using dish detergent to wash their diamond knives. When I tried this, the first thing that I noticed is how easy it is to keep the knife wet. I first tried it with a problem knife, and it worked! Also, I don't have the block face wetting problem that I had had before. Try it, you'll be surprised. Jo Dee PS. I am not affiliated with Micro Star!
} ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } In a message dated 04/27/2000 5:27:39 PM US Mountain Standard Time, } kalen-at-citrus.ucr.edu writes: } } { { water in the boat either wets the block or pulls away from } the knife edge. There seems to be no in-between. Every once in a while, the } stars seem to align, and the level achieves perfection for a few sections. } I've tried several different cleaning methods, thinking that maybe the } knife was dirty since I've been sectioning a lot of LR White. The only } thing I've noticed (since I've been looking so very closely at the diamond) } is that the seal (which was damaged when the knife arrived from the } manufacturer) between the knife and the boat has detached. I would think } that this would cause a leakage problem, which I haven't seen, not the } problem I'm experiencing. } } } } Kristen, } } I had a knife that behaved similarly once upon a time. Let's assume the boat } isn't leaking, since you aren't having problems with water collecting } underneath the knife in the knife holder, correct? It would be difficult to } imagine the knife has come loose in the boat, but that may be a very slight } possibility. If it were loose in its mount it might not section with } consistency. Either way, it would be a good idea to have the knife re-sealed } properly. } } The knife I had was almost impossible to get the proper amount of wetting on } the diamond, but two things helped: } } 1. Try deliberately overfilling the boat to give a positive meniscus right } at the knife edge. Leave the knife in this state for 15-20 minutes. Relax, } go have a cup of coffee, whatever. Then try lowering the fluid to the proper } level. This worked well for me. } } 2. For really difficult knives that refuse to wet, just a very little bit of } saliva on the end of a dalmatian hair works great. A light brush against the } tongue, then drag the hair in the boat fluid behind the knife edge. I know } it sounds gross, but it works! Others have used Alconox crystals, } detergents, etc. to break the surface tension, but good old saliva works } every time. } } Hope this helps. } } Cheers, } } Bob Chiovetti } GTI Microsystems
Kristen, It may be that after 4 years your knife edge needs a good cleaning. I've found that when I start to have trouble wetting the knife edge I can treat/clean the knife with a dilute solution of household ammonia. However, remember the knife housing is made of anodized aluminum, prolonged exposure to ammonia will degrade the aluminum and the diamond bonding. After soaking your knife and cleaning it in your normal way, leave it wet, and clean it again in a dilute solution of ammonia, carefully cleaning the knife edge with a pithwood stick. Then rinse thoroughly in flowing water for several minutes.
Phil Swab Engineering Development Deposition Sciences Inc. Santa Rosa, CA 707-566-3718 phil.swab-at-depsci.com
-----Original Message----- } From: Kristen Lennon [SMTP:kalen-at-citrus.ucr.edu] Sent: Thursday, April 27, 2000 3:34 PM To: Microscopy-at-sparc5.microscopy.com
Hi All, I am turning to all of you as a last ditch effort to see if my diamond knife is crazy or I am. I have been using this same knife for years (approximately 4 years) with the occasional difficulty. However, in the past few weeks I have been having a very hard time getting sections. The problem is this: water in the boat either wets the block or pulls away from the knife edge. There seems to be no in-between. Every once in a while, the stars seem to align, and the level achieves perfection for a few sections. I've tried several different cleaning methods, thinking that maybe the knife was dirty since I've been sectioning a lot of LR White. The only thing I've noticed (since I've been looking so very closely at the diamond) is that the seal (which was damaged when the knife arrived from the manufacturer) between the knife and the boat has detached. I would think that this would cause a leakage problem, which I haven't seen, not the problem I'm experiencing. Does anyone have any advice? Thanks for your help, Kristen Kristen A. Lennon Cell, Molecular & Developmental Biology Group Department of Botany & Plant Sciences University of California Riverside, CA 92521 kalen-at-citrus.ucr.edu
I want to thank everyone who responded to my plea for help with my diamond knife problems. I am trying a variety of your suggestions. The strange, but overwhelming theme involved saliva as a wetting and even a cleaning agent. As I sit here (after trying a couple of suggestions), a beautiful ribbon of sections is forming in the boat. I hope that it holds up. Thanks, Kristen Kristen A. Lennon Cell, Molecular & Developmental Biology Group Department of Botany & Plant Sciences University of California Riverside, CA 92521 kalen-at-citrus.ucr.edu
} I wonder about TEM and radiation emitting. } is there some dangerous situation for old TEMs? } does anybody have an information about this. }
Dear Emrah, There could be x-rays generated by some old TEMs. We had a JEOL JEM-200 which emitted a stream of x-rays near the specimen airlock. Not only did this make it a bad idea to stand up at the micro- scope when the beam was on, it was also a danger for other people in the adjacent lab. The best way to see if there is a problem is to use a monitor (either a Geiger counter or an ionization chamber) and survey all around the instrument with the beam on, a heav- ily-stained specimen in the scope, and the settings at extreme positions. You want to survey at the worst possible conditions that a user could set up. Remember that x-rays can penetrtate the floor, so the area beneath the scope should be surveyed also (unless the scope is in the basement with no one able to work beneath it). It should take only a few hours to complete such a survey. Shielding for ~100 kV scopes is not too difficult in case there are radia- tion leaks. Resurvey after the shielding is in place. Good luck. Yours, Bill Tivol
K. Spencer wrote about Molecular Probes' Bodipy phalloidin not working or being excited by 488 nms, 568 and 647 laser lines. I would like to know which Bodipy you are referring to. I find it improper and misleading to talk about a reagent unless you are going to include your specifics. I have used several of these reagents in the past with excellent results. If you checked the probes catalogue you will note that there are 3 Bodipy phalloidins listed ( Bodipy 529/547, 558/569 and 584/592 ) along with several Bodipy phallicidins that also bind to filamentous actin. These are Bodipy 502 /512 and Bodipy-TR-X 589/617. The numbers refer to the excitation peak and the emission peak respectively. None of these probes should be excited with any efficiency by a 647 laser. You might see some if you are over stained with the 584/592, a common mistake with my users. This probe would be excited by a 568 laser but not by a 488 nm line. Secondly, before buying a probe, one is advised to find out what is excitation and emission spectra are ( not just peak numbers) so you know what possible cross talks one could possibly experience.
For these reagents I recommend the Bodipy 505/512 or Oregon Green for 488 excitation. AlexaFluor-568 phalloidin or rhodamine phalloidin for excitation with 543 or 568 laser. The Alexafluors would be my top choice because of superior spectral properties. I know of none that work with a 633 or 647 laser. Cy 5 is my preferred choice for the far red channel as a conjugated antibody and TOTO-3 as a DNA marker.
Recently someone complained of TOTO-3 bleaching as they imaged. The dye doesn't bleach, but rather gets displaced from the DNA when laser excites it. It only fluoresces when bound to DNA or RNA. The trick here is to use the lowest laser or excitation you can, and to include the dye at about 1-2 micromolar in your mounting medium so it can re-intercalate into the DNA.
Joe Goodhouse Confocal / EM Core Laboratory Department of Molecular Biology Princeton University
Visit us at http://www.molbio.princeton.edu/facility/confocal/index.html {http://www.molbio.princeton.edu/facility/confocal/index.html}
I concur with Bob Chiovetti's suggestions as possible solutions to aid in wetting of the diamond knife edge and eliminate block wetting. In particular, it seems that adding a bit of saliva to your eyelash tool, which is then wiped across the flat portion of the diamond, is very effective. In extreme cases we have subjected the knife to ionized air via glow discharge but this can cause water to be drawn to the back side of the diamond. Naturally, you should check the diamond for cleanliness. It may help to immerse the knife in soapy water after careful cleaning with ethanol dipped Styrofoam. After rinsing the soap away with purified water, we then rinse the boat in ethanol which also seems to help wetting of the edge.
Good luck,
Doug ---------------------- Douglas R. Keene Associate Investigator Shriners Hospital Research Facilities 3101 S.W. Sam Jackson Park Road Portland, Oregon 97201 phone: 503-221-3434 FAX: 503-412-6894 (9-5 PST) e-mail: DRK-at-shcc.org
} } I am searching some device for holding (polycarbonate)filters during } dehydration and CPD. I am trying to view mycoplasms with SEM. Has anyone } experience with this ? } We have a device from Tousimis that is designed to hold round glass coverslips, and I have used it to hold polycarbonate filters. It is a milled-out cylinder open along one side. Wavy washers are used to separate the stacked coverslips. There are two sizes, 13 mm and 22 mm, part numbers 8766 and 8767. You can contact Tousimis at 1-800-638-9558, fax 1-301-881-5374, http://www.tousimis.com
This device works for me! My only affiliation with Tousimis is as a satisfied customer.
Aloha, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
The TEM specimen preparation course advertised in the Lehigh "2000 Microscopy School" booklet, p. 20., has undergone revision. This is a completely new course that was not finalized when it was time to print the booklet. The material on p.20 was a place holder.
Scott Walck joined with me in organizing the course and we have secured commitments from a large number of specimen preparation tool vendors to provide staff and equipment for the course, listed below.
Title:
TEM Specimen Preparation 2000 With Emphasis on Recently Developed Tools and Methods
Course Description:
This course is designed to provide classroom instruction and laboratory demonstration of the newest methods for preparing SEM and TEM specimens. While an individual with no experience preparing specimens will benefit greatly from the course, the intended audience will consist of students with some degree of proficiency utilizing classical methods of specimen preparation and who wish to update their capabilities. Emphasis will be placed on the rapid preparation of specimens from very small pre-selected locations. The preparation of SEM samples will be treated as the first step in making TEM specimens. "Hands on" experience by the students will be available. Table-top exhibits and demonstrations of specimen preparation ancillary equipment, such as: saws, dimplers, disc cutting tools, etc., will be available during the lab periods. TEM examination of prepared specimens will be performed.
Instructors:
Ron Anderson, IBM Analytical Services and Scott Walck, PPG Industries, Inc.
Outline:
Thursday, June 22
10:00 am Registration (Whitaker Lobby) 1:00 pm Introduction Anderson/Walck 1:15 pm Specimen Preparation Flowchart, Initial Considerations, Recent Advances, Choice of Technique Anderson 2:15 pm Initial Thinning, Tools, Disk Cutting, Dimpling Walck 3:30 pm Mechanical Polishing Anderson 4:30 pm Automated Initial Preparation Methods Vendors Sawing: Sagitta Cleaving: SELA 7:45 pm Ion Milling Anderson Commercial Tool Overview Walck
Friday, June 23
8:30 am Lab: Tripod Polishing Lab: Sagitta Tool Lab: SELA Tool 11:00 am Focused Ion Beam (FIB) Methods Anderson/Walck 2:00 pm Lab: FIB Methods Lab: Ion Mill Tools
Saturday, June 24
8:30 am Mechanical Polishing Difficult Materials Anderson 9:00 am Ultramicrotomy Walck/Anderson 10:00 am Small Angle Cleavage Technique Video Igor 10:30 am Plasma Cleaning Specimens Walck 11:00 am Reactive Sample/Storage/Transporting Walck 11:15 am Lab: Student Use of Available Tools Lab: TEM Examination of Prepared Specimens. Discussion of Successes and Problems
Vendors providing instrumentation and staff: v Allied High Tech v Bal-Tec v FEI v EA Fischione v Gatan v Sagitta v SELA v South Bay Technology
The registration deadline is June 1st. Registration forms are in the "Lehigh Microscopy School" booklet, or they may be obtained, along with accommodation information etc., from :
Ms. Sharon Coe Dept. of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem, PA 18015-3195
On Fri, 28 Apr 2000 DDKJoe-at-aol.com-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 } -----------------------------------------------------------------------. } } } Karen, } } Bob Chiovetti's ideas are right on the mark. It sounds crazy but saliva } works wonders. The only thing I would change would be the tool to use to } apply the saliva. It would probably be safer if you used the styrofoam stick } provided by many diamond knife or EM supply folks. Clean the knife with the } stick according to the instructions using a "bit of spit". Follow this up } with DI water since saliva is surprisingly dirty. } } Should restore any wetting properties your old knife has left. } } Have fun, } Joe Tabeling } Delaware Diamond Knives, Inc. } 800-222-5143 } } Hi,
Saliva is dirty and full of debris and bacteria. Use a wetting agent instead as above and fill the boat with it or draw it across the edge whichever works best.
} ------------------------------------------------------------------------ } 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 All, } I am turning to all of you as a last ditch effort to see if my diamond } knife is crazy or I am. I have been using this same knife for years } (approximately 4 years) with the occasional difficulty. However, in the } past few weeks I have been having a very hard time getting sections. The } problem is this: water in the boat either wets the block or pulls away from } the knife edge. There seems to be no in-between. Every once in a while, the } stars seem to align, and the level achieves perfection for a few sections. } I've tried several different cleaning methods, thinking that maybe the } knife was dirty since I've been sectioning a lot of LR White. The only } thing I've noticed (since I've been looking so very closely at the diamond) } is that the seal (which was damaged when the knife arrived from the } manufacturer) between the knife and the boat has detached. I would think } that this would cause a leakage problem, which I haven't seen, not the } problem I'm experiencing. } Does anyone have any advice? } Thanks for your help, } Kristen } Kristen A. Lennon } Cell, Molecular & Developmental Biology Group } Department of Botany & Plant Sciences } University of California } Riverside, CA 92521 } kalen-at-citrus.ucr.edu } } The detachment of the knife from its mount may be the cause. It has happened in our laboratory. Try, however, to add one drop of Photo-flo to about 50ml of filtered or distilled water. Try that in the boat. Detachment tends to be a serious problem! Send it back and have it repaired and resharpened. After 4 years it probably needs it.
Spring cleaning and new equipment requires I dispose of a Mikros VE10 vacuum evaporator and a LKB 4800 ultramicrotome. Anyone needing parts from these old, non-functional units please contact me.
steve
Dr. Steven Barlow, Associate Director EM Facility/Biology Department San Diego State University 5500 Campanile Drive San Diego CA 92182-4614 phone: (619)594-4523 fax: (619) 594-5676 email: sbarlow-at-sunstroke.sdsu.edu website: http://www.sci.sdsu.edu/emfacility/
Chairman, Educational Outreach subcommittee promoting access to microscopes Microscopy Society of America http://www.msa.microscopy.com/
You may want to consider a digital camera that is a SLR camera. I have a Kodak DC 120 and it works reasonably well but when I want to take close up photos or copy stand shots, I have to take several exposures to get the subject critically centered. Of course, a SLR allows me to set up my photo exactly the way I want it; also getting the correct lighting is easier. For this kind of work I use a Kodak DCS 420, but at ~$12k it's a "bit expensive".
} Hi all, } This is off the microscopy subject...sorry. } Any recommendations on digital cameras for copy stand work and general } photography? Has anyone tried the Nikon Coolpix 990? } We're coming up on the end of the fiscal year and there might be funds } available for a digital camera so any advice is greatly appreciated. } thanks, } Beth Richardson
Damian Neuberger, Ph.D. Baxter Healthcare, Inc. WG3-2S P.O. Box 490 Round Lake, IL 60073 e-mail: damian_neuberger-at-baxter.co, Tel: 847-270-5888 Fax: 847-270-5897
The Kodak MDS 120 does a very good job for the price. My only recommendation is that you get the flash memory cards (I have a 64 MB card) and a card reader or adapter for a PCMCIA slot. Transferring the images via the built-in serial port is, of course, INCREDIBLY SLOW!!! The included software works well and you can easily get the images into other software such as Adobe Photoshop.
} Part of the package which we got is a 16 MB Flash Memory card for the camera, } so you can either run the camera from your computer or store images on the } memory card and then remove the card and take it to a computer to transfer } the images to the computer. The package also had a PCMCIA card adapter in } it. The memory card plugs into the end of the PCMCIA device, and you can } then slip it into a PCMCIA slot on a laptop computer. } } But if you need to use a desktop computer and don't have a PCMCIA slot, you } need to get something like a SanDisk card reader for the computer. They're } available at CompUSA, Fry's Electronics, OfficeMax, etc. for about $70. } } The software with the camera is very nice. It lets you transfer images from } the camera's memory directly or from from the memory card, preview images and } download them directly from the camera, etc. as well as perform some basic } image adjustments (flip, rotate, modify brightness, contrast, gamma, etc.)
Damian Neuberger, Ph.D. Baxter Healthcare, Inc. WG3-2S P.O. Box 490 Round Lake, IL 60073 e-mail: damian_neuberger-at-baxter.co, Tel: 847-270-5888 Fax: 847-270-5897
Just returned from a course on VP SEM (which was excellent) and just now can add to some of the digital camera discussion.
One thing not mentioned about using a SLR digital camera like the E1, N60 is vibration from the mirror and the shutter. If you are going to use a camera of this type you must be able to lift the mirror independently, like the old F1, whether it's film or CCD. I have a Kodak DCS420 that is based on the Nikon N90s camera body and it works ok at low magnifications but anything over about 200X just doesn't cut it.
At 08:33 AM 4/26/00 -0700, Dr. Gary Gaugler wrote: } The Fuji Super CCD is 23.3 x 15.6mm; 3040 x 2016 pixels. } The S1 Pro uses Nikon mount lenses and has an equivalent } 35mm frame size at 1.5X the lens' focal length. } } What the equivalent FOV would be on a scope is still in } question. If the S1 Pro works anywhere near how the } Nikon E1 or E2 does, I fear the Fuji camera won't be applicable } to microscopy at all. Without a lens, the camera must operate } in aperture priority mode and center weighted exposure } reading. Unless Fuji made some major changes to the basic } Nikon (N-60) body and associated electronics, they may have } just made a higher pixel count Nikon digicam.
Damian Neuberger, Ph.D. Baxter Healthcare, Inc. WG3-2S P.O. Box 490 Round Lake, IL 60073 e-mail: damian_neuberger-at-baxter.co, Tel: 847-270-5888 Fax: 847-270-5897
Neither the N60 or N80 bodies have mirror lock up. However, at equivalent ISO of 400-1600, it remains to be seen if lock up is necessary. And also, how ISO affects the resulting image. Based on how the E1 and E2 worked, I would not count on a miracle.
The Kodak 420 is obsolete and discontinued. These can be purchased used today for somewhere in the $2K-$4K range. But, with its tiny CCD, I would not have one again. The newer models like DCS460 have good features. But I have not been impressed by Kodak's tech support user friendliness. It would be a hard sell for me to buy any Kodak product these days over other manufacturer's offerings.
The mechanical shutter is a definite problem, both for image stability as well as camera reliability. The Leaf Lumina has a shutter but it operates only one time for each shot. It is very reliable. In contrast, the Polaroid DMC has a shutter that seems to always be "shuddering" the system. I would expect that the DMC would exhibit a poor reliability record. The advantage of the DMC is that it provides near-real time focusing through the camera's imager. The only other camera I have used that exceeds this is the Sony DKC-5000 (cat's eye) digital camera. But the DKC-5000 uses a very small CCD as well and suffers from tiny images. But focusing is true real time via a separate RGB color monitor. When a frame is shot, the main system box transfers the image to the computer via SCSI.
gg
At 08:45 PM 4/28/00 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
When using F-113, I preceed it with either or both methanol and acetone. The curious problem is that after either a lengthy immersion in acetone or methanol, the specimen (insect) floats when put in F-113. OK. So it is lighter than F-113. What is a good method of encompassing the specimen with F-113 to further dehydrate it prior to vacuum desiccation?
Any ideas?
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Modern surfers use PC boards. You can too at http://photoweb.net ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
I'm new to the list. I recently picked up a delightful Leitz Ortholux I, like at http://arsmachina.com/leitz_ortholux.htm and a non-working Zeiss OpMi-1 surgical scope, similar to the upper left quadrant of http://www.mednetlocator.com/opmi-1.gif with not much arm and no stand. I think it's missing a lense from the underside.
I'm temporarily illuminating the Ortholux with the 5V DC from a spare PC power supply. The lamp says 6 volts, I think. What's the voltage range from the original rheostat-style power supply?
Hi John! 5 volts is fine, the color temperature is a bit warmer but your bulb will last longer! I usually only run the bulb at full or over when I am taking a photograph. For normal observation I find the 5 volts fine. Congrats on the othrolux great classic scope! Ed Sharpe archivist for SMECC
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I'm new to the list. I recently picked up a delightful Leitz Ortholux I, like at http://arsmachina.com/leitz_ortholux.htm and a non-working Zeiss OpMi-1 surgical scope, similar to the upper left quadrant of http://www.mednetlocator.com/opmi-1.gif with not much arm and no stand. I think it's missing a lense from the underside.
I'm temporarily illuminating the Ortholux with the 5V DC from a spare PC power supply. The lamp says 6 volts, I think. What's the voltage range from the original rheostat-style power supply?
Ron: I've been using the Kodak DS120, which is the camera that is used in the system you asked about, and have had good results. There are a few things to keep in mind, however. The DS120 saves images to flash cards using a propriatary Kodak method that is not recognized by anything else. Therefore, some of the nifty print systems can't directly use the images. I use the Kodak-supplied software to copy the images from the flash card to hard disc via a card reader (SanDisk -- cheap and works real well). Once in my computer, I work on the images using PhotoShop. Final printing is performed on my Epson Photo 1200 using print paper. The results are very good.
I do note that when using the DS120 with the MDS adaptor on my Nikon microscope that there is a bright area in the center. I'm not sure who's fault that is.
The small viewing screen is a problem, but once the focus is properly set it is not a major disadvantage. With the system you are talking about (remember, I just have the camera and adaptor) it may be that the image can be displayed prior to collecting it onto the flash card.
Overall, I think it's a good value for the money.
If you'd like to talk about the camera, connect off line at edsworth-at-aol.com.
I'm trying to remember ... Freon 113: Peldri, yes? Or is this one that stays fluid? (And how did you get it, since Freons are mostly illegal anymore...) .
First, you might try vacuum, 1 atmosphere or so, maybe not that much, gently applied, and released.
For Peldri, I put specimens in small vials -- 4 mL Wheatons are my favorite -- and completely filled the vial, then capped, leaving no air. If there were enough alcohol:Freon intermediates, and enough changes in pure Freon, the specimens should be in pure Freon now.
Mind, I mostly did crustaceans and fish bits, not insects with their waxy epicuticles, but then some or all of that wax is lost in the alcohols anyway.
But I wouldn't use methanol or acetone. Ethanol works better for insects. Acetone is OK, but I've found ethanol better, and methanol is too extractive.
You don't have a CPD for this? Also, some insect parts can be dried from ethanol or even water. Mandibles of many species for instance, or the elytra of most beetles. Not whole insects, though.
I never got my specimens to sink in Freon 13, Back When, but they usually did sink in Peldri.
Phil
} When using F-113, I preceed it with either or both } methanol and acetone. The curious problem is that } after either a lengthy immersion in acetone or methanol, } the specimen (insect) floats when put in F-113. OK. } So it is lighter than F-113. What is a good method } of encompassing the specimen with F-113 to further } dehydrate it prior to vacuum desiccation? } } Any ideas?
}}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{ Philip Oshel Supervisor, AMFSC and BBPIC Dept. of Animal Health and Biomedical Sciences University of Wisconsin 1656 Linden Drive Madison, WI 53706-1581 Voice: (608) 263-4162 peoshel-at-facstaff.wisc.edu fax: (608) 262-7420 (dept. fax)
Last fall we actually fabricated one of those holders from some copper water pipe, and purchased the wavy washers from McMasters. I had seen the one Tina mentions in use and thought "heck, we could make one and save $100." Well we did just that. For less than $20, if you have a metal cutting saw, some solder and access to purchasing wavy washers, you can fabricate your own. One grad student has used it for the spring semester to dehydrate, and CPD, poly-l-lysine coated 12mm coverslips classifying the morphology and size of bacteria in the hind gut of Tipula abdomalis. I just reviewed his final work and the images looked great. He had no problems with debris or samples washing off. The advantage of making it yourself is that you can customize the size for the application for very little money. You would only be limited by the availibity of sizes of wavy washers.
-Geoff
} } I am searching some device for holding (polycarbonate)filters during } } dehydration and CPD. I am trying to view mycoplasms with SEM. Has anyone } } experience with this ? } } } We have a device from Tousimis that is designed to hold round glass } coverslips, and I have used it to hold polycarbonate filters. It is a } milled-out cylinder open along one side. Wavy washers are used to } separate the stacked coverslips. There are two sizes, 13 mm and 22 mm, } part numbers 8766 and 8767. You can contact Tousimis at 1-800-638-9558, } fax 1-301-881-5374, http://www.tousimis.com } } This device works for me! My only affiliation with Tousimis is as a } satisfied customer. } } Aloha, } Tina }
-- Geoff Williams
Electron Microscope Facility Supervisor Biology Department Brooks Hall Central Michigan University Mt. Pleasant, MI 48859
Dear Sir, I have been working for Istanbul University, Faculty of Fisheries, as a research Assistant on aquaculture. I am a doctorate student at the present. My studies are on Aquarium Fish ( ornamental ). The topic of my doctoral thesis is " A Study on the Artificial Propagation of Discus Fish ( Symphysodon spp. ) and Effective Factors on the Propagation ". Besides, I research too my thesis for discus 's embriological-larval developing and sperm developing with scanning elecron microscope ( SEM ). With regard to my doctoral thesis: Could you send me Scanning Electron Microscope SEM )' s use-book and example's processing ? I wish success your studying. Best regards,
Esra SAVAS I.U. Su Urunleri Fak., Ordu Cad., No: 200 34470 Vezneciler / Istanbul TURKEY
MAY 11TH SYMPOSIUM AT U.C. Davis TOPIC: Electron Backscatter Patterns (EBSP)
Featured Speakers: 1) David Dingley, TSL, "Automated Crystallography for the TEM: Recent Developments" 2) Adam Schwartz, LLNL, "Coupling Automated Electron Backscatter Diffraction with Transmission Electron and Atomic Force Microscopies". 3) Patrick Camus, Noran Instruments, "Phase Identification versus Phase Discrimination" 4) Pierre Rolland, Oxford Instruments, "Mapping Copper Interconnects for the Semiconductor Industry" 5) John Sutliff, HKL Technology, "Quantifying Polycrystalline Microstructures: The Automated-ESBP Technique and its Applications"
The symposium will be held in the AGR room in the Buehler Alumni & Visitors Center, UC Davis
Meeting starts at 3:00 No Host Social -at- 5:30 Dinner Buffet -at- 6:30 Meeting/Dinner will end by 9:00
Dinner Buffet- Rosemary & Orange Chicken Fettucine Alfredo Spring Mix Salad with Raspberry Vinaigrette Greek Salad topped with Crumbled Feta Cheese on a Bed of Greens Chef's Selection Vegetable Fresh Baked Rolls with Butter New York Cheesecake with Melba Sauce garnished with Mint Coffee, Decaf, Hot Tea & Iced Water
Please call Greg Lum at 415/338-1339 or Email: "glum-at-sfsu.edu" for dinner reservations by Monday May 8th. The symposium starts at 3:00 p.m. The dinner will start at 6:30 pm.
The Nikon D1 Digital SLR camera does not have a mirror lockup but does have an "Anti Vibration Mode" which causes the shutter opening to be delayed until mirror shock has subsided. The D1 has 2.7million pixels, ISO range 200-1600 and accepts most Nikon lenses, electronic releases,etc. It is excellent for both copystand and light microscopy applications.
George Laing National Graphic Supply 226 North Allen Street Albany, NY 12206 E-mail: scisales-at-ngscorp.com (800) 223-7130 X3109 (518) 438-8411 X3109
At 08:45 PM 4/28/00 , you wrote: } } Just returned from a course on VP SEM (which was excellent) and just now } can add to some of the digital camera discussion. } } One thing not mentioned about using a SLR digital camera like the E1, N60 } is vibration from the mirror and the shutter. If you are going to use a } camera of this type you must be able to lift the mirror independently, } like the old F1, whether it's film or CCD. I have a Kodak DCS420 that is } based on the Nikon N90s camera body and it works ok at low magnifications } but anything over about 200X just doesn't cut it. } } }
I have a Philips 501 SEM and 201 TEM that I am about to dismantle and discard. They were both operational until four months ago. If anyone needs them for spare parts please feel free to contact me.
Best Regards,
Kirk J. Czymmek, Ph.D. Director, Core Microscopy Facility Department of Biological Sciences University of Delaware Newark, DE 19716 kirk-at-udel.edu PH: (302) 831-1158
If you find them let me know I would like a copy. I will pay reasonable or slightly unreasonable charges for copying and mailing.
Gordon
G. C,. Couger 624 Cheyenne Stillwater, OK 74075-1411 405 624 2855 between 3:00 pm to 1:00 am CST 405 742 2758 cell ----- Original Message ----- } From: "John Foust" {jfoust-at-threedee.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Monday, May 01, 2000 7:28 AM
Meeting: "PASEM 2000" - THE PRACTICAL ASPECTS SERIES OF SHORT COURSES Dates: May 22 through June 2, 2000 Topic: The Practical Aspects Series consists of intensive 3.5 to 4.5 day short courses that provide participants with a thorough coverage of the basic theory and practice of scanning electron microscopy and associated techniques. Training is achieved through the use of easy to understand lectures coordinated with supervised laboratory sessions. The laboratory class sizes are purposely kept small so that participants will have an opportunity to gain extensive hands- on experience with available SEM and EDS instrumentation. Designed for the academic, government and industrial user, the courses are beneficial to microscopists and microanalysts at all levels from novice through advanced. Course titles and dates for our Year 2000 series follow:
1. SCANNING ELECTRON MICROSCOPY - May 22 through May 26, 2000 2. ADVANCED TOPICS IN SCANNING ELECTRON MICROSCOPY - May 30 through June 2, 2000 3. X-RAY MICROANALYSIS - May 30 through June 2, 2000
Sponsor: University of Maryland Location: College Park, Maryland, USA Interests: Both Physical & Biological Sciences Fields: SEM, EDS Contact: Tim Maugel, University of Maryland, Department of Biology, Building 144, Room 0240, College Park, Maryland, 20742, USA Tel: 301-405-6898 Fax: 301-314-9358 E-mail: tm11-at-umail.umd.edu WWW: http://www.life.umd.edu/pasem
If some of your spectra are only available in LINK AN/10000 format, then you could convert them to simple text files with a little program I wrote about a year ago. It's a 32 bit Windows program. You can select as many LINK spectrum files (in their original, not converted format) as you need. All the selected files will be converted in one run. I have tried it with more than 1700 spectra. The result of the conversion is a text file with the same name but with different file extension. An example from a converted spectrum file (from a series): -------------------------------LAK11S103.SP--------------------------- ak11s** 3
preset live time: 40 live time: 40 real time: 49 20.000000 eV/channel
Energy[keV], counts -0.200000, 0 -0.180000, 0 -0.160000, 0 -0.140000, 0 -0.120000, 3 -0.100000, 16 -0.080000, 42 -0.060000, 98 .. ---------------------------------------------------------------------- If you are still interested in a program like this, drop me a line and I'll send it to you.
With the best regards: Laszlo Varga
On 20 Apr 00, at 10:21, Filion, Christian 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 } -----------------------------------------------------------------------. } } } Good morning, } } I have both a Spectra file in Link format and txt format saved as a MSA } type file. Is there a software or an excel macro that could read these } spectra ? } } Thank you } } Christian Filion } Superviseur Laboratoires } Aciers Inoxydables Atlas } 1640 Marie-Victorin } Tracy, Québec } J3R 5R5 } Tel : 450-746-5243 } fax : 450-746-5241 } }
For those of you interested the Microscopy and Microanalysis 2000 Meeting Search Engine is now on line.
Using the Search engine you can find out dates/times/locations of any author, paper, subject or session at the upcoming meeting this August in Philadelphia.
We are looking for vendors of Peltier cooling stages for our Hitachi 2460N VPSEM. If you know of a company, please drop me a line. Also, if you have opinions on what features you would look for in such an item, I'd like to hear it. Thanks in advance, Randy -- Randy Nessler Views expressed are my own.
Never replaced a Pyramitome belt - but LKB microtome belts can be replaced by "regular" belt material - just match the width and springiness (did you keep any of the old belt fragments?). If your institution doesn't have a machine shop (those guys usually have extra belts floating around), try your local hardware store/superstore. I've had good luck replacing standard parts this way, and it is probably going to be light years cheaper than an "official" replacement part, if such a thing is available.
} From experience, giant rubber bands do not work very well.........
Tamara Howard CSHL
On Tue, 2 May 2000 mganger-at-aol.com-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 } -----------------------------------------------------------------------. } } } Date: Tue, 2 May 2000 08:08:07 EDT } Subject: Drive Belt for Pyramitome } To: Microscopy-at-sparc5.microscopy.com } MIME-Version: 1.0 } Content-Type: text/plain; charset="US-ASCII" } Content-Transfer-Encoding: 7bit } X-Mailer: AOL 5.0 for Windows sub 104 } } Greetings, } } I would like to know if anyone out there knows of a repair place that sells } parts for an LKB 11800 Pyramitome. The drive belt for the specimen arm has } literally fallen apart and I need to replace it. Any suggestions on where I } could get one would be greatly appreciated. } } Thanks in advance. } } Mike Ganger } Montclair State University } Montclair, New Jersey } mganger-at-aol.com } } } }
} I would like to know if anyone out there knows of a repair place that sells } parts for an LKB 11800 Pyramitome. The drive belt for the specimen arm has } literally fallen apart and I need to replace it. Any suggestions on where I } could get one would be greatly appreciated.
Be sure to look at vacuum cleaner drive belts. They come in many sizes and I've used them as drive belts for a lot of devices. They look like a giant O-ring, up to 1/4 inch thick and in many lengths.
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 530 754 7536 fax http://katie.ucdavis.edu raharris-at-ucdavis.edu
Ernest Orlando Berkeley National Laboratory One Cyclotron Road, Berkeley, California 94720 FOR IMMEDIATE RELEASE
Two places left for students to attend the NCEM Summer School on Computing in Electron Microscopy June 26-30, 2000 in Berkeley, California
(Berkeley, CA) The seventh annual Summer School on Computer-Interactive HRTEM Image Acquisition, Processing and Simulation will be held at the National Center for Electron Microscopy (NCEM), Lawrence Berkeley National Laboratory, University of California, Berkeley from June 26 through June 30, 1999.
The curriculum will focus on training participants in techniques of computer-assisted acquisition and interpretation of high-resolution electron microscope images, including HRTEM image processing and simulation, electron holography, focal-series reconstruction, and remote-control microscopy. Participants will learn general principles and apply them to specific cases. Instruction will focus on the use of computer assistance rather than microscope training, although participants will acquire images on NCEM microscopes as well as using specific application programs for image interpretation. Class size will be limited to 16. Deadline for applications, May 01, 2000, has been extended to May 04, 2000..
Participants who wish to apply newly acquired techniques to their own projects will be encouraged to extend their visit at NCEM into the next week. Please note: this type of arrangement requires advance submission of an NCEM microscope proposal (see: http://ncem.lbl.gov/frames/user.htm). Projects may involve prepared specimens for microscopy, images and diffraction patterns for processing, or crystal and defect data for simulations.
For more information and application materials, contact:
I have been reading a great deal on polymer microscopy lately and came across the statement that there was very little information to be gained by staining polymers and (as might be expected) that they did not take up stain well. Many years ago, I had heard of an application in which osmium tetroxide was used, I believe on polyethylene. Can anyone shed any light, either on the general comment or on the application using osmium tetroxide?
Many thanks, Barbara Foster,President Microscopy/Microscopy Education 125 Paridon Street Suite 102 Springfield, MA 01118-2130 PH: 413-746-6931 FX: 413-746-9311 email: MME-at-map.com Website: www.MME-Microscopy.com/education
America's first national consortium of microscopy specialists offering customized on-site training in all areas of microscopy, image analysis, and sample preparation ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^
Does anyone have a good, safe way of mounting Codonics dye sublimation prints onto poster boards? We would like to avoid the spray on "rubber cement" types of glues due to the mess.
Thank you.
#################################################################### John J. Bozzola, Ph.D., Director Micro-Imaging and Analysis Center 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 ####################################################################
Double sided tape or a product that picture framers use that just the glue without the tape. It is on a tape that releases the glue. It does an excellent job of mounting. I have pictures that were mounted with the Scotch brand 20 years ago that are just as good as the day I did them. It will also stick to the plastic use in resin coated photographic paper.
Any picture framing shop should have it.
Gordon
Gordon Couger gcouger-at-couger.com
Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00 ----- Original Message ----- } From: "John J. Bozzola" {bozzola-at-siu.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, May 02, 2000 6:58 PM
O.k., whereas not directly a microscopy question I was hoping some one would have a solution. Look for something that will function like aluminium foil but be non- reflective.
(1) shapable around objects (we're covering petri dishes), but not too massive
(2) be opaque (visble light 420-720nm)
(3) be non-reflective on at least one side. "Non-reflective": less than 15% reflectivity to visible spectrum 420-720nm.
Any suggestions? (Any good ideas for turning Al foil nice and black?)
Thanks.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
11th International Congress of histochemistry and Cytochemistry (ICHC 2000)
Cell Biology Tools for the New Century
ICHC 2000 is the premier meeting this year to address the very latest developments in visualisation techniques for Cell Biology. to be held in the magical medieval city of York, England between 3-8 September 2000.
We have a fabulous speaker listof over 100 including some of the biggest names in Cell Biology and Imaging such as Roger Tsien, Richard Haugland, Fred Bosman, Alan Boyde, Stefan Hell, Alan Fine, Jim Smith, Margaret Buckingham, Nick White, Angus Lamond, David Becker, Jeniffer Lipincott-Schwartz and many, many more.
Topics include -Tracking Molecules in Cells, Fluorescent markers of Gene Expression, Live Cell Imaging in plants and Fungi, Apoptosis, Cell Signalling, Environmental toxicology, Imaging Embryonic Development, Ion Imaging in Cells, Imaging in the Neurosciences etc., etc. etc.
See the science and have a chance to talk to the best in the field.
Come for a holiday as well as the science we can offer many great excursions.
For more information and details of how to register please visit our web-site at
www.med.ic.ac.uk/external/ichc_2000
Student bursaries are still available!
DON'T MISS OUT IT WILL BE ONE OF THE CONFERENCES OF THE YEAR
Gary Coulton
Dr. Gary Coulton Molecular Pathology Division of Biomedical Sciences Imperial College School of Medicine The Sir Alexander Fleming Building South Kensington London SW7 2AZ
tel 0044 (0)171 594 3190 fax 0044 (0)171 594 3022
PLEASE NOTE AS FROM APRIL 1ST 2000 THE FOLLOWING NUMBERS WILL REPLACE THOSE ABOVE AND SHOULD BE USED.
------------------------------------- Announcing the 11th International Congress of Histochemistry and Cytochemistry (ICHC 2000)
"Understanding Biocomplexity: The Post-Genome Challenge"
September 3-8, 2000, York, United Kingdom
ICHC 2000 will comprise 27 symposia addressing the latest developments and applications of histochemistry and cytochemistry in the life sciences including medicine.
Many leading experts to speak
On-line registration now open!!!!!!!!!!
For further details of the meeting and how to pre-register please visit our web-site at http://www.med.ic.ac.uk/external/ichc_2000
} O.k., whereas not directly a microscopy question I was hoping some one would } have a solution. Look for something that will function like aluminium foil but be non- } reflective. } } (1) shapable around objects (we're covering petri dishes), but not too massive } } (2) be opaque (visble light 420-720nm) } } (3) be non-reflective on at least one side. "Non-reflective": less than 15% } reflectivity to visible spectrum 420-720nm. } } Any suggestions? (Any good ideas for turning Al foil nice and black?) } } Thanks. }
I seem to remember that studio photographers use flat black aluminum foil. Try Porter's Camera, I think they are in Iowa. They are a large mail-order firm and probably have a website. Or try Calumet camera, 800 Supreme Drive, Bensenville, IL 1-800-225-8638.
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 **********************************************
The best stuff that I ever found for mounting pictures is 3M Positionable Mounting Adhesive. This is very good stuff and not messy at all. It comes in rolls and is a transfer type material. You roll it out, place your picture on it and cut it out with a sharp knife. You then press the adhesive with the backing sheet and peel off the sheet. The adhesive is transferred to every square inch of the print. You can position this stuff around and then you press it down hard and it sticks forever. The roll that I use is PMA 568. There is larger and smaller sizes available. I think that you have to order through a photographic supply house because I don't think any of the EM supply houses carry it.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
} -----Original Message----- } From: John J. Bozzola [mailto:bozzola-at-siu.edu] } Sent: Tuesday, May 02, 2000 7:59 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: dry mounting dye sub prints } } } -------------------------------------------------------------- } ---------- } 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/MicroscopyListse} rver/FAQ.html } } } -------------------------------------------------------------- } ---------. } } } Does anyone have a good, safe way of mounting Codonics dye } sublimation prints onto poster boards? We would like to avoid the } spray on "rubber cement" types of glues due to the mess. } } Thank you. } } #################################################################### } John J. Bozzola, Ph.D., Director } Micro-Imaging and Analysis Center } 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 } #################################################################### }
} } O.k., whereas not directly a microscopy question I was hoping some one would } have a solution. Look for something that will function like aluminium foil but be non- } reflective. } } (1) shapable around objects (we're covering petri dishes), but not too massive } } (2) be opaque (visble light 420-720nm) } } (3) be non-reflective on at least one side. "Non-reflective": less than 15% } reflectivity to visible spectrum 420-720nm. } } } Any suggestions? (Any good ideas for turning Al foil nice and black?) } } Thanks. } } Richard E. Edelmann, Ph.D. } Electron Microscopy Facility Supervisor } 352 Pearson Hall } Miami University, Oxford, OH 45056 } Ph: 513.529.5712 Fax: 513.529.4243 } E-mail: edelmare-at-muohio.edu } } "RAM disk is NOT an installation procedure." } }
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
As to pyramitomes, if anyone has one that is not in use and would like to sell it, please contact me. I used one years ago and wouldn't mind having one around. Debby
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
--------------------------------------
Greetings, Babara Foster asked about staining polymers. By chance, I recently looked at paper on staining polymers for electron microscopy. The paper compared osmium and ruthenium tetroxides and concluded that ruthenenium tet stained many more polymers than did the osmium. This paper has a kind of encylopedic feel to it because they checked a lot of polymers. The citation is Trent et al., 1983 Macromolecules 16: 589-598. I have no idea whether this would work with light microscopy or what info you could get from it exactly besides contrast, but perhaps you will find the paper interesting.
As ever, Tobias } } Hi, } } I have been reading a great deal on polymer microscopy lately and came } across the statement that there was very little information to be gained by } staining polymers and (as might be expected) that they did not take up } stain well. Many years ago, I had heard of an application in which osmium } tetroxide was used, I believe on polyethylene. Can anyone shed any light, } either on the general comment or on the application using osmium tetroxide? } } Many thanks, } Barbara Foster,President } Microscopy/Microscopy Education } 125 Paridon Street Suite 102 } Springfield, MA 01118-2130 } PH: 413-746-6931 FX: 413-746-9311 email: MME-at-map.com } Website: www.MME-Microscopy.com/education } } America's first national consortium of microscopy specialists offering } customized on-site training in all areas of microscopy, image analysis, and } sample preparation } ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^
Hi, We got a bolt of black cloth from a theatrical supply house. It is the kind of thing used to make curtains and other masking on the stage. I think it was called "covert black" but I could have that wrong. We were doing photobiological work and were forever having to limit the pesky wandering photons. The cloth is thick but not too thick to wrap around a dish. It is not perfectly opaque but if you need that I bet you could line the cloth with al foil, on the inside. Hope this helps, Tobias } } } O.k., whereas not directly a microscopy question I was hoping } some one would } have a solution. Look for something that will function like } aluminium foil but be non- } reflective. } } (1) shapable around objects (we're covering petri dishes), } but not too massive } } (2) be opaque (visble light 420-720nm) } } (3) be non-reflective on at least one side. } "Non-reflective": less than 15% } reflectivity to visible spectrum 420-720nm. } } } Any suggestions? (Any good ideas for turning Al foil nice and black?) } } Thanks. } } Richard E. Edelmann, Ph.D. } Electron Microscopy Facility Supervisor } 352 Pearson Hall } Miami University, Oxford, OH 45056 } Ph: 513.529.5712 Fax: 513.529.4243 } E-mail: edelmare-at-muohio.edu } } "RAM disk is NOT an installation procedure."
O.k., whereas not directly a microscopy question I was hoping some one would have a solution. Look for something that will function like aluminium foil but be non- reflective.
(1) shapable around objects (we're covering petri dishes), but not too massive
(2) be opaque (visble light 420-720nm)
(3) be non-reflective on at least one side. "Non-reflective": less than 15% reflectivity to visible spectrum 420-720nm.
Any suggestions? (Any good ideas for turning Al foil nice and black?)
Dear Richard,
How about depositing carbon black onto Al foil? It's messy, but fulfills
your requirements. Evaporating carbon seems like an inferior proceedure, but
that should also work. Just holding the foil above a low-temp flame (candle or
bunsen burner with the air turned down) seems the simplest. You can then
cover the carbon layer with something if necessary. Dipping the foil into laser-
printer toner could also work, and the toner could then be heated. Putting the
foil (cut to 8.5" x 11") through the printer is asking for trouble, but that could
also work if you can find a solid black character--WordPerfect 5.1 has such
a character, which can be accessed by ctrl-v, 3,3. Good luck.
I am looking for a protocol for Immunogold labelling membrane proteins on mitochondria / peroxisomes fractions..., I am hoping to use a formaldehyde type fixation..............to LR white resin, I would appreciate any advice, references, and if possible actual relevant protocols. Thank you in advance.
Dear John, Years ago I purchased "Mounting Film" from an EM supply house. This is double-sided sticky tape, two feet wide. You cut it to any size and press on. Mine is made by Lomacoll and is obviously German. At 06:58 PM 5/2/00 -0500, you wrote: } } Does anyone have a good, safe way of mounting Codonics dye } sublimation prints onto poster boards? We would like to avoid the } spray on "rubber cement" types of glues due to the mess. } } Thank you. } Best 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
We've used DryBond Adhesive Pads for mounting dye-sublimation prints. It's manufactured by Chartpak (their part number is DBS25 for 25 11" x 17" sheets). I order it from EMS (Cat. #77612-25), described on Page 311 of EMS Catalog XIII. Basically it consists of an array of tiny (well, actually quite large to electron microscopists) dots of rubber cement like stuff that you apply by placing the print on top, covering with a protective sheet then rubbing with a rubber brayer. Peel up the print, place on mounting board, cover with sheet and rub with brayer. We've used it on color dye-subs with and without the XtraLife coating with no problems. As far as longevity, I made a poster two years ago that is still adhered tight to the board, but that was with RC B&W paper. The oldest dye-sub has only been observed for about a year now.
I have no financial interest in Chartpak or EMS, the stuff is just nice to use.
Heather Owen
Heather A. Owen, Director Electron Microscope Laboratory Department of Biological Sciences University of Wisconsin - Milwaukee (414)229-6816
I use a product called Liquid Paper Dryline Permanent Adhesive made by the Gillette Company. It is supposedly acid free, glues instantly, no drying time, and comes as either permanent or temporary. I get it a office supply store.
} Does anyone have a good, safe way of mounting Codonics dye sublimation } prints onto poster boards? We would like to avoid the spray on "rubber } cement" types of glues due to the mess.
Damian Neuberger, Ph.D. Baxter Healthcare, Inc. WG3-2S P.O. Box 490 Round Lake, IL 60073 e-mail: damian_neuberger-at-baxter.co, Tel: 847-270-5888 Fax: 847-270-5897
I want to buy an inspection scope from which I can also acquire digital images.
I remembered that I had a flyer for the Kodak MDS 120 system ($1895).
I called Kodak, they still sell the same system. It seems a little "out dated" now w/ only 1.2 pixels and no USB connection.
Are there other product that offer similar simplicity and versatility (macroscopic off the scope capability), that also fall in the same price range, and have 2 million pixels w/ a USB connection?
Now that the program for the meeting is available on the website - Thanks Nestor - I would like to advertize the "LATE-BREAKING POSTER SESSION AT MICROSCOPY AND MICROANALYSIS 2000" for any remaining authors (or potential authors) who would like to contribute to the meeting.
Microscopy and Microanalysis 2000 will feature a poster session composed of presentations of newly acquired data or analyses which were unavailable for submission by the February 15 deadline. A short, half page abstract describing the studies is required. The abstract should include: Title, Authors, Authors affiliation, and a Brief Description of the studies. The description should include the Aim of the studies, a short characterization of the Methods, and a brief account of the Results and their Importance.
Abstracts should be e-mailed or faxed to the program chair, Stuart McKernan, at stuartm-at-tc.umn.edu (email) or 612-625-5368 (fax). Abstracts may be submitted immediately but must be received by June 23, 2000. Abstracts will be reviewed by members of the program committee. A limited number of poster boards are available and preference will be given to early submissions. Abstract authors will be notified of acceptance of their abstracts no later than July 1 (earlier for early submissions).
__________________ Stuart McKernan stuartm-at-tc.umn.edu Director Office:(612) 626-7594 IT Characterization Facility, University of Minnesota Desk:(612) 624-6009 100 Union Street S. E., Minneapolis, MN 55455 NEW-} Fax:(612) 625-5368
Can someone on the listserver offer any help, I am seeking the JEOL publication of the list of O Rings (part numbers, or size/type) that are used in the JEOL COATER JEE-400 and the JEE 4B/4C. If it is available could you please either email me a copy of Fax me a copy Fax Number is (in australia) +61 (2) 9385 6400. Thankyou for your help, Barry EM UNIT UNSW
My question is : I need to obtain a estimate of virus particle concentration in a sample of cell culture supernatant for an article to be submitted to a journal. I was told of a method called the 'latex particle reference method' for obtaining virus particle counts. Can anyone on the list supply information for this procedures ? Thanks for your time.
Sincerely,
Tom Doman, Project Associate Veterinary Science Department Penn State University
Dear Neal, A good fixative for immunogold work is still a combination of glut. + paraformaldehyde, but using a 'low' glutaraldehyde concentration. One you could try would be 0.25% glutaraldehyde + 4% paraformaldehyde in phosphate buffered saline pH 7.2. Formaldehyde alone results in poor structural preservation. From there on you could follow the instructions for dehydration, infiltration etc. from the LR White pamphlet which works well. For labelling: 1. Pre-treat with 0.02M glycine in PBS to block any free aldehyde groups (2x10mins), 2. Block with a suitable protein, e.g. BSA, ovalbuimin (1%- 1X20mins) 3. Place each grid on a 15-20ul drop of primary antibody, suitably diluted, and either leave overnight at 4 degrees C in fridge, or on lab bench for about 2 hours. 4. Rinse 6 times in drops of PBS with 1% protein block (BSA etc.) - (6X5mins). 5. Place each grid on a 20ul drop of Protein-A Gold complex, suitably diluted, for 1-1.5 hours. 6. Rinse in PBS (6X5mins). 7. Wash in distilled or millipore filtered water (in small beakers, 3 x several rapid dips(about 6), making sure the grids remain under the water at all times during dipping). 8. Stain with Uranyl Acetate and Lead Citrate.
** Blot (side-on) onto filter paper between steps 1+2, 2+3, 4+5, 7+8.
Good luck.
Cheers, Marilyn
Neal Leddy 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, } } I am looking for a protocol for Immunogold labelling membrane proteins on } mitochondria / peroxisomes fractions..., I am hoping to use a formaldehyde } type fixation..............to LR white resin, I would appreciate any advice, } references, and if possible actual relevant protocols. Thank you in } advance. } } Neal Leddy nleddy-at-tcd.ie
Please contact the person at the end of the post, not me, as I am posting at the request of someone without access. Thank you. ----- Original Message ----- Sent: Wednesday, May 03, 2000 8:59 PM
} JOB OPORTUNITY AT UNIVERSITY OF HOUSTON } } } Supervisor: Histology and Microscopy Laboratory; College of Optometry; } University of Houston, Texas } } Job Description: } } This position is in direct support of research and teaching faculty } where the subject of interest is typically ocular and neural tissue. The } duties range from tissue preparation, embedment, sectioning and } mounting, to operation of light and transmission electron microscopes, } and development and printing of photographic results. Knowledge and } competency in operation of ultramicrotomes and the transmission electron } microscope as well as current techniques in morphology, histology, and } immunocytochemistry is required. Experience with cryo-microscopy, in } situ hybridization, and computer image analysis is desirable. The } successful candidate also may participate as co-author of research } papers describing research performed in the laboratory. } } The supervisor is responsible for maintenance of stocks of laboratory } supplies and care of the equipment, and keeping the lab clean and } usable, including proper disposal procedures for hazardous wastes. Other } duties include instructing graduate students in common and specialized } anatomical techniques required for their various projects, and } supervision and consultation on their work as required. Some effort also } is applied to teaching of undergraduate optometry students including } preparation of teaching slides and technical instruction in anatomical } methods. } } This job is scheduled to begin on 1 May 1999. Salary will be negotiated } based upon qualifications and experience. For further information, you } may contact: } } Chris Kuether, Technical Services Manager } College Of Optometry, University Of Houston } 4901 Calhoun Blvd. Houston TX 77204-6052 } vox:(713)743-2049 fax:--2053; ckuether-at-uh.edu } } }
You can read up on Emitech cold stages on our site home} contents} K4 We are agents for Emitech for Australasia only (south of Singapore), just hope that you would find the info useful. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Tuesday, May 02, 2000 11:37 PM, nessler [SMTP:randy-nessler-at-uiowa.edu] wrote:
} We are looking for vendors of Peltier cooling stages for our Hitachi } 2460N VPSEM. If you know of a company, please drop me a line. Also, if } you have opinions on what features you would look for in such an item, } I'd like to hear it. } Thanks in advance, } Randy } -- } Randy Nessler } Views expressed are my own.
I have archived three discussions on this from the listserver. Go to:
http://www.biotech.ufl.edu/~emcl
and look in the TEM section of "Tips & Tricks" under virus. It also has a few other ideas. Good luck
At 06:48 PM 5/3/2000 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall EM Technician Gainesville, FL 32610 University Of Florida ph 352-392-1184 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
I think that I have used something that would work well for your application. It is an anodized Al foil that we use for laser safety purposes. The foil is very flat black and the black does not rub off easily. However, the foil is significantly stiffer than household Al foil. The brand we have used is BlackwrapTM and it can be purchased from The Great American Market, Hollywood CA 323-461-0200. They have a web site as well. I hope that this helps.
Matthew Ervin, Ph.D. U.S. Army Research Laboratory (301)394-0017 MErvin-at-ARL.mil
"Richard E. Edelmann" {edelmare-at-casmail.muohio.edu} on 05/03/2000 07:39:20 AM
Please respond to Edelmare-at-muohio.edu
To: microscopy-at-sparc5.microscopy.com cc:
O.k., whereas not directly a microscopy question I was hoping some one would have a solution. Look for something that will function like aluminium foil but be non- reflective.
(1) shapable around objects (we're covering petri dishes), but not too massive
(2) be opaque (visble light 420-720nm)
(3) be non-reflective on at least one side. "Non-reflective": less than 15% reflectivity to visible spectrum 420-720nm.
Any suggestions? (Any good ideas for turning Al foil nice and black?)
Thanks.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
Dear Listers, Does anyone currently use the Sony DKC-5000 CatsEye Digital Camera and if so would you recommend it for a shared technology lab. Rosemary Walsh, EM Facility for the Life Sciences Penn State University
Barbara, We use both osmium and ruthenium tetroxide to stain various polymers for EM observation. Ruthenium is less selective in its staining, and therefore does stain many more polymers. Both compounds stain unsaturated polymers pretty well. In olefins, like polyethylene, RuO4 is good for staining amorphous domains within the crystalline or semi-crystalline polymer fine structure. This is typically a TEM issue. With some rather coarse polymer blends (e.g. rubber inclusions in another polymer matrix) RuO4 or OsO4 can be used with SEM and FESEM imaging. I would expect to see very little staining contrast by LM due to the polymer domain sizes of typical co-polymers, polymer blends, or crystalline/amorphous polymer structure. Again the best reference I have is the book by L. Sawyer, Polymer Microscopy.
Brad Huggins BP Amoco, Naperville, IL
} ---------- } From: Barbara Foster[SMTP:mme-at-map.com] } Sent: Tuesday, May 02, 2000 4:57 PM } To: Confocal Microscopy List; microscopy-at-sparc5.microscopy.com } Subject: LM: Stains for polymers? } } ------------------------------------------------------------------------ } 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 have been reading a great deal on polymer microscopy lately and came } across the statement that there was very little information to be gained } by } staining polymers and (as might be expected) that they did not take up } stain well. Many years ago, I had heard of an application in which osmium } tetroxide was used, I believe on polyethylene. Can anyone shed any } light, } either on the general comment or on the application using osmium } tetroxide? } } Many thanks, } Barbara Foster,President } Microscopy/Microscopy Education } 125 Paridon Street Suite 102 } Springfield, MA 01118-2130 } PH: 413-746-6931 FX: 413-746-9311 email: MME-at-map.com } Website: www.MME-Microscopy.com/education } } America's first national consortium of microscopy specialists offering } customized on-site training in all areas of microscopy, image analysis, } and } sample preparation } ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ } ^^ }
We use routinely this method in our laboratory. Here is the used protocol of quantitation of virus particles by negative stain electron microscopy:
A selected volume (100 uL) of supernatant containing virus particles is mixed with a selected volume (100 uL) of polystyrene latex beads of known concentration (about 10e8 beads by mL) and a size between 100 - 200 nm in diameter. The mixture is placed in an Beckman Airfuge 240 µL-tube. A Formvar and carbon-coated grid is inserted into the bottom of the microtubes.* The tubes are placed in a Airfuge A-100 fixed angle rotor (30¡) and centrifuge at 20 psi (120 000 g) for 5 minutes. The grids are recovered with fine self-closing tweezers, dried with bibulous paper, stained 1 minute with phosphotungstic acid (PTA 3%, pH6.0) and dried again with bibulous paper. Samples are visualized under a transmission electron microscope with an approppriate magnification. On two different grids, 200-500 particles (latex beads or virus particles) are counted from at least five different areas on each grid. That way, from the ratio of the two types of particles in the suspension, the ratio of the volumes added and the known concentration of latex particles, the concentration of viral particles can be calculated.
Virus particle concentration (particles/mL) = (virus count / latex beads count) X (latex beads concentration) X (1/test article dilution)
The level of sensitivity of this procedure is between 10e6 and 10e10 particles per mL. Less than 10e6, there is not enough virus to get a realistic count and more than 10e10, there are too much virus to well differentiate them. This procedure is used principally to quantify Retrovirus type-A and -C particles in cells supernatant, but can be used for any virus particles.
*R. Alain et al, J.Virol. Meths, 16 (1987), 209-216 *Alain, R. Microscopy Today, May 1997, issue#97-4, D. Grimes Ed, p. 20
If you need more explanation you can contact us. If you are not the equipment to do this technique, we can offer this service at low price.
Robert Alain
********************************************************** Robert Alain, M.Sc. Microscopie lectronique
INRS-Institut Armand-Frappier Centre de Microbiologie et Biotechnologie 531 boul. des Prairies Laval, Qubec CANADA H7V 4B7 Tel: (450)687-5010 ext#4388 Fax: (450)686-5626 e-mail: Robert.alain-at-INRS-iaf.uquebec.ca or Robert_alain-at-hotmail.com Http://www.iaf.uquebec.ca/iaf/recherche/viro/me.html **********************************************************
----- Original Message ----- } From: Tom Doman {jtd1-at-psu.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, May 03, 2000 7:48 PM
{html} You may want to look at the Olympus DP-11 digital camera. You can find it on their web page at: {a href="http:///"} http://www.olympusamerica.com/product.asp?c=21&s=11&p=18&product=612 {/a} {br} It has very high resolution, live video output (which is quite useful at low magnification as a focusing and framing aid) and features a c-mount so that it can be used on any microscope, or with a macro lens. It uses SmartMedia cards as the recording media, and combined with a USB SmartMedia card reader, download to the computer is fast and easy. {br} {br} Configured with an AC adapter, 64MB SmartMedia card, USB card reader and a 9" Sony video monitor, it is priced at $4,349.00. This is more than you indicated you would like to spend, but I think that you will find that its features, resolution and ease of use may justify the additional expense. {br} {br} At 03:23 PM 5/3/00 -0400, you wrote: {br} >------------------------------------------------------------------------ {br} >The Microscopy ListServer -- Sponsor: The Microscopy Society of America {br}
Thanks to everyone who responded - I was thinking anodizing myself but Matt Ervin actually had an Off-the-Shelf source.
===============
Its exam time here and I wanted to share this answer from my EM Theory Final:
Ques: What does "EELS" stand for? What does it detect (Be specific!)?
Ansr: Energized Eucaliptic Leaf Shooter. If properly used it can be used to lure the better part of the world's Koala Bear population in a general area to get a more precise population density reading.
[The student recieved 1 point for creativity]
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
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Hello listers- I am hoping that someone might have a current address or phone for "Cambride Image Technologies Ltd." or "Nuclide" A colleague of mine is looking to purchase a cold cathode luminoscope and was given these two names. Or if someone knows of another supplier any information would be appreciated. Thank you, Sarah
Sarah A.W. Lundberg Lab (EPMA) (702) 895-2660 or Electron Microanalysis and (SEM) (702) 895-2462 Imaging Laboratory Office (702) 895-1134 Department of Geoscience, UNLV Fax (702) 895-4064 4505 S. Maryland Parkway Box 454010 Las Vegas, NV 89154-4010 Dept Office (702) 895-3262
I believe that what Geoff was referring to is called "Black Wrap." It is a heavy foil with a matte black coating. It is used by theater lighting people worldwide to create custom masks and light shields. It is made by several companies. My local lighting house sells black wrap made by Great American ... in 12 inch by 50 foot or 24 inch by 25 foot rolls for US$27.50. Since you're at a university you could probably go over to the theater department and get a little piece to try.
Larry
Larry D. Ackerman Lily & Yuh Nung Jan Laboratories Howard Hughes Medical Institute UCSF, Box 0725, Rm U226 533 Parnassus Ave. San Francisco, CA 94143
At 05:59 AM 5/4/00 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
After some time of getting used to it, the 5000 is OK. It uses a small CCD so the image files are small and not really high resolution. But for relatively modest final image output size, it is a good camera. It provides real time focusing via a separate RGB+sync color monitor. However, adjusting the monitor to match the captured image's exposure is tricky.
If you are going to be moving the camera around, remember that it uses a SCSI interface to download captured images. You won't be able to move it very far from the SCSI port unless you use a notebook or laptop computer.
Scotch 924 Transfer Tape 1/2 inch wide is the product I mentioned earlier. It cost about 6.95 a roll for a life time supply for many of us. http://productinfo.mmm.com/us/office/products/office.jhtml?powurl=4JLD1SMBbe ZCZYKRQ9geT1T4S9TCgvPDJGVQ33gl
The way I used it to mount photos was stick a band of it around the edge of the photo as close as I could to the edge. That's all I did for 5X7's for 8X10 I put a cross diagonally from corner to corner. I didn't mount prints larger than that but If I had I would have added some tape in the open areas. Some one made a small version that had an applicator but the refills cost as much as a full sized roll and it is not much if any easier to use.
I tried a lot of things spray glue, two sided tissue and a number of others. This was the easiest and most consistent other than Seal mount tissue and a hot press.
The hardware store where I drink coffee is will sell mail order. I have not interest in the hardware store other than a bunch of us drink their coffee. Their web page address is www.studyboards.com or 405 372 2644 ask of Sandy or A.J.
Gordon
Gordon Couger gcouger-at-couger.com
Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
You can find information on www.bal-tec.com --} products--} vct100
Best regards, Udo Graf BAL-TEC AG +423 388 12 26 +423 388 12 60 (Fax) udo.graf-at-bal-tec.com
+ -----Ursprüngliche Nachricht----- + Von: nessler [mailto:randy-nessler-at-uiowa.edu] + Gesendet: Dienstag, 2. Mai 2000 14:37 + An: miscroscopy listserver + Betreff: SEM Peltier stage vendors? + + + We are looking for vendors of Peltier cooling stages + for our Hitachi + 2460N VPSEM. If you know of a company, please drop me a + line. Also, if + you have opinions on what features you would look for in + such an item, + I'd like to hear it. + Thanks in advance, + Randy + -- + Randy Nessler + Views expressed are my own. +
I have customer here requiring some help. They are making replicas of fossil mandibles for SEM analysis. Their problem is how to localise the areas of interest when the specimen is in the SEM. I was sent the quotation below, and we are trying to find out whether we, or anyone else, can supply the sort of material they need. My background is mainly materials, so I'm farming this out to the list to see if anyone has any suggestions, or maybe even recognises the fragment below.
"Fiberglas screening material: fiber thickness (310 microns), hole width (1, 240 microns), and length (1,340 microns), wiped with Permabond 910 adhesive and pressed onto the surface so that contact was made everywhere".
Thanks in advance
Tim
***************************************************************** Tim E. Harper Managing Director CMP Cientifica s.l. Space & NanoTechnology Division Phone +34 91 640 71 85 Fax +34 91 640 71 86 http://www.cmp-cientifica.com/ {http://www.cmp-cientifica.com/}
That sounds a lot like fiberglass window screen to me, but the holes here are probably a bit smaller. There is also a fiberglass screening used for backing plaster repairs. It might have a pitch more similar to what you described.
At 05:09 PM 5/5/2000 +0200, you wrote: } Hi, } } I have customer here requiring some help. They are making replicas of fossil } mandibles for SEM analysis. Their problem is how to localise the areas of } interest when the specimen is in the SEM. I was sent the quotation below, } and we are trying to find out whether we, or anyone else, can supply the } sort of material they need. My background is mainly materials, so I'm } farming this out to the list to see if anyone has any suggestions, or maybe } even recognises the fragment below. } } } "Fiberglas screening material: } fiber thickness (310 microns), hole width (1, } 240 microns), and length (1,340 microns), wiped with Permabond 910 adhesive } and pressed onto the surface so that contact was made everywhere". } } } Thanks in advance } } Tim } } ***************************************************************** } Tim E. Harper Managing Director } CMP Cientifica s.l. } Space & NanoTechnology Division } Phone +34 91 640 71 85 Fax +34 91 640 71 86 } http://www.cmp-cientifica.com/ {http://www.cmp-cientifica.com/}
We have had the Sony DKC-5000 for quite a few years and hve never had a problem with the hardware despite many clumsy users. We have it running on a Mac computer taking the Sony RGB signal to SVHS via a Harmonic Research CV-233P video encoder. The SVHS signal is input to a ATI Xclaim video board in the Mac. So the live window and the Sony acquire plug in run on the same monitor. The density and color balance rarely coincide between the two displays. We have had many problems with users changing settings and adding software, etc. on the Mac resulting in problems with the video display. As Dr Gaugler said, the resolution is not very high. There should be better systems now but I continue to recommend that our scientists record their images on color slide film and then scan it with a $2000 slide scanner (Nikon, Polaroid etc.) much cheaper, higher resolution and bandwidth and easy convenient storage!
Larry D. Ackerman Lily & Yuh Nung Jan Laboratories Howard Hughes Medical Institute UCSF, Box 0725, Rm U226 533 Parnassus Ave. San Francisco, CA 94143
We are still not set-up well for Cryo-SEM, and although we have made some progress and learned much about these techniques from you all on this Listserver and others, I am still looking for a "cryomicroscopy facility" who has, for example, an E-SEM with Peltier Stage/controlled cooling rate capabilities and with cryostage (capable down to temperatures as low as approximately -75C); who would be interested in working together with one of my clients, in a large Chemical/Oil Company, to investigate/observe the low-temperature structures of various Synthetic Fluids.
Anyone interested, please contact: Brad Huggins at BPAmoco, Naperville, IL hugginbj-at-bp.com 630 420-3668
Hi All, Does anyone have a good protocol for silver enhancement of immunogold labeled sections for TEM? I have a paper in front of me, but they used a kit. We do light level silver enhancement without a kit all of the time, but I don't want to chance trying to adapt that method before consulting all of you. Thanks for your help! Have a wonderful weekend, Kristen Kristen A. Lennon Cell, Molecular & Developmental Biology Group Department of Botany & Plant Sciences University of California Riverside, CA 92521 kalen-at-citrus.ucr.edu 909-787-4525
If I understand correctly how you are using the DKC-5000, then you are actually obtaining half the image resolution that the system can provide.
The DKC-5000 has a 1/2" 440,000 pixel CCD with an effective image area of 795x598 pixels (RGB). The actual captured image area is probably closer to 740x580 pixels (RGB). Thus, if my calculations are correct, you should obtain RGB TIF files which are about 1.29MB in size.
The image processor in the DKC-5000 digital processor system unit expands (interpolates?) the original image to one that is 1520x1144 pixels (RGB). This results in a TIF file that is about 5.21MB in size. However, this resolution is only obtained via the SCSI interface on the back of the system box. This is likely why you are obtaining low resolution results. This is effectively taking a consumer grade RGB video camera and performing an RS-170 frame grab. This will produce up to 800x600 pixels (RGB) maximum. This is OK, but the higher cost of the DKC-5000 was due, I think, to the ability to obtain higher quality images and to do so without any separate/extra image capture hardware. The DKC-5000 was about $14K I think versus about $800 for a really nice RGB video camera with close to 800x600 pixels resolution.
Therefore, if you have any sort of typical Mac, it should have a legacy SCSI-I connector on the rear. This is easily connected to the digital processor box. Then, load the Photoshop plug-in and acquire the higher resolution images using Photoshop and the SCSI bus. You should still be able to use the RGB outputs for framing and focusing. Just don't use the RGB outputs for image capture.
Have you tried this?
gary g.
At 09:57 AM 5/5/00 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Greetings Listers, I am looking for a couple decomissioned electron microscopes to be used as a source for parts. I would greatly appreciate any information or leads. The instruments are the Hitachi 7000 TEM and the Hitachi S-510 (or 515 or 520) SEM. Thank you very much, Alex Greene SCIENTIFIC INSTRUMENTATION SERVICES, INC. PMB-499, 1807 West Slaughter Lane, Number 200 Austin, Texas 78748-6200 Phone: 512/282-5507 Fax: 512/280-0702 QUALITY ELECTRON MICROSCOPE REPAIR
At http://www.couger.com/gcouger/leitz/ are some pictures of a very old Leitz projection microscope. For it's age and rough construction it works extreemly well. It is of cast iorn, steel and brass construction and carries no serial number. It has two objective lenses that are about 6 and 12 x It has a simple condenser in the lamp housing and every thing is adjustable on the stand and the objectives have a heilical ajustment.
I have as several musem curators and infivigules that have a life long assoitions wiht Leitz. No one has seen anything like it. It appears to be all origianl except for the light bulb, its mounting and the cord. They look like they came from the 1920's
} From my decussions with Leitz sholers and deduction I think this was made about the same time as the electric light bulb. It would be possible that it use a gas or lime light but I don't think so. The mechenism for centering a light bulb just looks like it was designed for a bulb.
Any help is welcome. If anyone knows when Leitz started putting serial numbers on their products it would be a great help in dating it.
Thanks Gordon Couger Stillwater, OK 74075 405 624 2855 GMT - 6:00 www.couger.com/gcouger
I am sorry the last post some how escaped my spell check on my email program. It is set to check every email. But it misses some. For Microsoft an occasional miss is pretty good:)
At http://www.couger.com/gcouger/leitz/ are some pictures of a very old Leitz projection microscope. For it's age and rough construction it works extremely well. It is of cast iron, steel and brass construction and carries no serial number. It has two objective lenses that are about 6 and 12 x It has a simple condenser in the lamp housing and every thing is adjustable on the stand and the objectives have a helical adjustment.
I have as several museum curators and individuals that have a life long association with Leitz. No one has seen anything like it. It appears to be all original except for the light bulb, its mounting and the cord. They look like they came from the 1920's
} From my decisions with Leitz scholars and deduction I think this was made about the same time as the electric light bulb. It would be possible that it use a gas or lime light but I don't think so. The mechanism for centering a light bulb just looks like it was designed for a bulb.
Any help is welcome. If anyone knows when Leitz started putting serial numbers on their products it would be a great help in dating it.
Thanks Gordon Couger Stillwater, OK 74075 405 624 2855 GMT - 6:00 www.couger.com/gcouger
Sarah- I can't help you with Cambridge Image Technologies Ltd., but I do know about Nuclide. Nuclide went bankrupt several years ago and was bought up and reborn as Premier American Technologies Corp. which continued making the luminoscopes among other things. I worked for PATCO for a couple of years. I don't know the details, but PATCO has since evolved into Spectru Medix and I imagine they still sell luminoscopes as it was probably their most consistent seller in the past. You can call Spectru Medix at 814-867-8600, they are at 2124 Old Gatesburg Rd., State College, PA. I would suggest you talk to Mike Vollero as I know he still works there and will be able to put you in contact with the proper people. I hope this helps. Matt Ervin
Sarah Lundberg {lundberg-at-nevada.edu} on 05/04/2000 02:10:20 PM
To: MSA listserver {Microscopy-at-sparc5.microscopy.com} cc:
Hello listers- I am hoping that someone might have a current address or phone for "Cambride Image Technologies Ltd." or "Nuclide" A colleague of mine is looking to purchase a cold cathode luminoscope and was given these two names. Or if someone knows of another supplier any information would be appreciated. Thank you, Sarah
Sarah A.W. Lundberg Lab (EPMA) (702) 895-2660 or Electron Microanalysis and (SEM) (702) 895-2462 Imaging Laboratory Office (702) 895-1134 Department of Geoscience, UNLV Fax (702) 895-4064 4505 S. Maryland Parkway Box 454010 Las Vegas, NV 89154-4010 Dept Office (702) 895-3262
We have recently encountered some perplexing problems staining tissue embedded in Spurr's resin with uranyl acetate and calcined lead. The tissue is not taking up the stain although there is some stain precipitate on the sections. Any suggestions?
Sincerely, Harry Grier Stock Enhancement Research Facility Florida Marine Research Institute 14495 Harllee Road Palmetto, FL 34221 harry.grier-at-fwc.state.fl.us
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Apparently I was not clear in my description of our DKC-5000 setup. The RGB signal displayed in a live ATI video window is for focusing and composing. Acqusitition of the image is via the Sony plug-in and SCSI transfer. The resulting image file is about 5MB. It does not meet my standards for a publication quality image. Larry D. Ackerman Lily & Yuh Nung Jan Laboratories Howard Hughes Medical Institute UCSF, Box 0725, Rm U226 533 Parnassus Ave. San Francisco, CA 94143
I have a question about how best to maintain a sputter coater that is used infrequently.
We are a small college with correspondingly small EM facility. Our SEM and prep equipment may go for several weeks without being used, then someone will need it heavily for a class for three or four weeks. Invariably we find that our sputter coater seems to be the weak link in our plans, since it rarely works well when we fire it up after long periods of dormancy. I understand this is common with vacuum equipment: better to use it often rather than shut it off.
Am I correct that we should we have a plan to regularly pump down the sputter coater, and if that is good idea, how often should that be? Every day? Once a week? Once a month?
Or, if we don't need to pump it down regularly, are there other things we should be doing do it the down time instead of letting it just sit there?
and finally, are some designs better able to handle long periods of disuse? Do we just have the wrong sputter coater?
Gary P. Radice gradice-at-richmond.edu Associate Professor of Biology 804 289 8107 (voice) University of Richmond 804 289 8233 (FAX) Richmond VA 23173 http://www.science.richmond.edu/~radice
Does anyone have any old parts for the See-Vac, Inc. Autoconductavac sputter coaters? Especially the power feed-through that connects through the cap to the target. Thanks.
Phil -- }}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{ Philip Oshel Supervisor, AMFSC and BBPIC Dept. of Animal Health and Biomedical Sciences University of Wisconsin 1656 Linden Drive Madison, WI 53706-1581 voice: (608) 263-4162 fax: (608) 262-7420 (dept. fax)
Microscopist: To manage light and electron microscopy facility. Minimal degree requirement BS (MS preferred). Experience with confocal microscopy, computerized image analysis and histological sample preparation essential. Experience in SEM/TEM sample preparation and video microscopy desirable. Must be an interactive person willing to facilitate microscopy experiments for faculty and students with a wide variety of interests in a University setting. Salary commensurate with experience. Full time desired but will consider part time. For further information on the department see http://www.umbc.edu/biosci UMBC is an AA/EOE.
Contact: Dr. Daphne Blumberg, Chair, Microscopist Search Committee, Dept. of Biological Sciences, University of Maryland Baltimore County (UMBC), Baltimore MD 21250.
All vacuum systems are better "stored" under vacuum, with the exception of any systems that cannot vent the rotary pump when power is off. Those systems may suck oil from the pump towards the vacuum chamber.
The more common problem for infrequently used systems is moisture in the pump oil. Particularly in moist climates and when relatively short pumping times are employed a good deal of water is absorbed in the pump. When the pump is not used for a lengthy period this may cause corrosion and certainly lowers the vapour pressure of the contaminated oil and so lower performance results.
I suggest that at the end of your spasmodic activities the pump should be run with the baffle valve partially open for at least 30 minutes. The baffle valve is usually atop the rotary pump. You could also use the sputter coater's needle valve, partially open. Under these conditions the pump will run hotter and throw-out a good deal of oil mist and water vapour. Vent the exhaust into a fume hood since oil mist is not just unpleasant. You will find that the pump performs much better after a baffle run.
SEM and TEM usually do not need this treatment, but they may, for instance if a TEM has been used to "dry" film that was not previously dried in another system. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Tuesday, May 09, 2000 5:11 AM, Gary Radice [SMTP:gradice-at-richmond.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 } -----------------------------------------------------------------------. } } } I have a question about how best to maintain a sputter coater that is used } infrequently. } } We are a small college with correspondingly small EM facility. Our SEM and } prep equipment may go for several weeks without being used, then someone } will need it heavily for a class for three or four weeks. Invariably we } find that our sputter coater seems to be the weak link in our plans, since } it rarely works well when we fire it up after long periods of dormancy. I } understand this is common with vacuum equipment: better to use it often } rather than shut it off. } } Am I correct that we should we have a plan to regularly pump down the } sputter coater, and if that is good idea, how often should that be? Every } day? Once a week? Once a month? } } Or, if we don't need to pump it down regularly, are there other things we } should be doing do it the down time instead of letting it just sit there? } } and finally, are some designs better able to handle long periods of disuse? } Do we just have the wrong sputter coater? } } Gary P. Radice gradice-at-richmond.edu } Associate Professor of Biology 804 289 8107 (voice) } University of Richmond 804 289 8233 (FAX) } Richmond VA 23173 http://www.science.richmond.edu/~radice } }
The new BAL-TEC 'VCT 100' Vacuum-Cryo-Transfer-Equipment is a modular system consisting of:
- Shuttle (Cryo-Vacuum Conditions) - Docking station at any preparation system - Docking station at any analysis system (SEM, ESEM, Cryo-AFM) - Cryo equipment for any analysis system (SEM, ESEM)
The modules can be arranged just to meet your needs.
VCT 100 Cryo equipment has been adapted to a Philips XL30 e.g.
Additional information you will find on www.bal-tec.com --} products--} vct100
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-----Ursprüngliche Nachricht----- Von: Huggins, Bradley J [mailto:HUGGINBJ-at-bp.com] Gesendet: Freitag, 5. Mai 2000 19:23 An: Huggins, Brad; Microscopy listserver Betreff: low temperature microscopy of synthetic fluids
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We are still not set-up well for Cryo-SEM, and although we have made some progress and learned much about these techniques from you all on this Listserver and others, I am still looking for a "cryomicroscopy facility" who has, for example, an E-SEM with Peltier Stage/controlled cooling rate capabilities and with cryostage (capable down to temperatures as low as approximately -75C); who would be interested in working together with one of my clients, in a large Chemical/Oil Company, to investigate/observe the low-temperature structures of various Synthetic Fluids.
Anyone interested, please contact: Brad Huggins at BPAmoco, Naperville, IL hugginbj-at-bp.com 630 420-3668
Gary- I am not sure if you are saying that your sputter coater is pumping down poorly or that it is depositing poor quality films. In either case, it sounds like poor vacuum is the problem. Water vapor adsorbing to the deposition chamber walls over a period of disuse will pump off of the walls very slowly. A "wet" chamber can take 10 times as long to pump down to ultimate vacuum as a "dry" one. If this sounds like it might be causing the symptoms you are seeing, I would suggest you try one of two things:
1. Place a valve between the sputter chamber and the pump. Valve off the chamber and leave it under vacuum between uses. As long as it retains any vacuum this will help when you try to use it again. You don't want to be pumping on the chamber at the pump's ultimate vacuum for any extended period of time because that will allow oil vapor from the mechanical pump to backstream into your sputter chamber and possibly onto your specimen if present! (I am assuming that you have an oil based pump.) It is also nice to spare the pump the wear and tear of pumping continuously over periods of disuse.
OR 2. Several hours or the day before you want to use it, start pumping on the sputter chamber while admitting a small flow of argon. The argon is an important part of this procedure. First of all, the argon will prevent the backstreaming described above. Second, I have been told that the argon helps to desorb the water from the chamber surfaces. I don't know if that is an old wives tale or not, but it does seem reasonable. The argon flow may also help in purging any condensed vapors from the pump's oil. Don't use too much argon though or you may overheat your pump.
I hope that this addresses the problem you are experiencing. Matt Ervin (301)394-0017 U.S. Army Research Laboratory Adelphi MD
Gary Radice {gradice-at-richmond.edu} on 05/08/2000 03:10:46 PM
To: Microscopy-at-sparc5.microscopy.com cc:
I have a question about how best to maintain a sputter coater that is used infrequently.
We are a small college with correspondingly small EM facility. Our SEM and prep equipment may go for several weeks without being used, then someone will need it heavily for a class for three or four weeks. Invariably we find that our sputter coater seems to be the weak link in our plans, since it rarely works well when we fire it up after long periods of dormancy. I understand this is common with vacuum equipment: better to use it often rather than shut it off.
Am I correct that we should we have a plan to regularly pump down the sputter coater, and if that is good idea, how often should that be? Every day? Once a week? Once a month?
Or, if we don't need to pump it down regularly, are there other things we should be doing do it the down time instead of letting it just sit there?
and finally, are some designs better able to handle long periods of disuse? Do we just have the wrong sputter coater?
Gary P. Radice gradice-at-richmond.edu Associate Professor of Biology 804 289 8107 (voice) University of Richmond 804 289 8233 (FAX) Richmond VA 23173 http://www.science.richmond.edu/~radice
I agree. In addition... Given a choice, I would always use the process gas to purge - using the "leak valve" to feed a partial pressure to the system until the vacuum pump heats up. Opening the ballast valve will help (lowers ultimate vacuum when open) purge the pump, but IMHO, it is better to use dry gas than atmosphere.
Woody White McDermott Technology
{SNIP} I suggest that at the end of your spasmodic activities the pump should be run with the baffle valve partially open for at least 30 minutes. The baffle valve is usually atop the rotary pump. You could also use the sputter coater's needle valve, partially open.
Thanks to all who responded to my query about how best to maintain my infrequently used sputter coater. Since some have replied off list and others have asked to see all the responses I thought I would append all of the responses here, (without direct attribution, since strictly speaking I don't have all the authors permission to do this....I hope they understand).
Thanks to all who responded. Based on the comments, I think my problem was accumulation of water in the oil and vacuum surfaces, and dried out gaskets. Greasing the gaskets and running the pump longer got the pump-down time from 20 minutes to 3 minutes. Keeping the chamber under vacuum isn't practical with our coater design, but I can probably solve my problem by arranging to pump down the chamber once a week, paying attention to keeping the seals lightly greased, and changing the pump oil on a regular schedule.
******************
} Hi Gary, } } How are you? Our sputter coater has sat for over a year at a time without } use. It had no problems when it was started. However, that is not an } ideal situation. Vacuum equipment whould be regularly pumped down to } out-gas the chamber, etc. We have a Denton Desk II Sputter Coater. It is } by far the best I have used. It is now used regulary because we have a new } SEM. If I were you I would pump down the chamber on the coater at least } once a month. Change the oil in the pumps once/year. Check the seal } between the chamber and the base and the glass and the seal before the } class use starts. Use fomblin grease or some other non-hydrocarbon vacuum } grease. *************
} There really should not be any heroics needed in order to snap your figers } and have the sputter coater work well. We ship our coaters all over the } world to trade shows, open up the boxes, take them out, put them on the } table, and a few minutes later we are coating samples for prospective } customers. And I think that would probably be the case for most } commercially made coaters today that are used in the SEM market. } } But I will tell you one thing that does happen and that is that the needle } valve can develop a "set" if it is left tightened down real tight over long } periods of time. } } The when you go to use it again, because of the "set", there are problems } controlling its action and therefore the bleed rate of air or inert gas. I } know that conventional wisdom says a vacuum system should be stored "under } vacuum" but the typical coater is sufficiently leaky, that the vacuum is } going to disappear shortly anyhow. } } So you might want to try storing it no under vacuum, that is, with the need } valve open, and see if that does not make your problems disappear.
******************
} Hello Dr. Radice, } Yes, the worst thing you can do to a vacuum system is to not use it. The } least you should do is keep the bell jar under vacuum when it is not being } used. Do you vent with Dry Nitrogen or room air? All high voltage leads } and feed throughs should be kept very clean since they have a tendency to } collect Carbon and crud. It might not be a bad idea to pre-run your high } voltage up rather high (higher than you would normally use it), pror to a } run. This should stabilize things a bit.
***********
} Gary, } After many years of using sputter coaters I have found that it is best } that they are kept under vacuum all the time. Many sputter coaters do } not let you maintain a vacuum when they are off. With this type I have } found that I have to keep the glass cylinder and the metal target very } clean and allow plenty of pump down time when first using the unit after } prolonged shut down. Also any solvent based glues that may be used, } silver dag or colloidal carbon, must be dry before putting the sample in } the coater, at least 2 hours after mounting. **************
} I have a similar use pattern as yours, but I have never had any problems } with my coater. Sometimes, our coater my sit for a period of months before } a pump down is required. Back in 1993 I purchased an Emitech, the same } type that is sold through EMS today, and have had but only one failure. } That failure was due to foil on a circut board that was not heavy enough to } handle the current required by the vacuum pump. The foil had melted, but } was an easy fix for me. Otherwise I have had no failures. } } It certainly can help to pump it down once a week, just like it helps } ink-jet printers to print a test page once a week. Other than keeping } non-contaminated oil in the pump, a light coating of vacuum grease on the 0 } rings to keep them from drying out and getting attacked by ozone, I have } not had other maintenance issues. Perhaps your vacuum problems are related } more to your pump and the need for some maintenance and oil change, perhaps } your seals need replacing, or perhaps your coater is going through a period } where it requires higher than normal maintenance. I'm curious, what type } of problems are you having? } } Good luck with your facility. I always enjoy meeting others who are } running small EM labs. *******************
} All vacuum systems are better "stored" under vacuum, with the exception of } any } systems that cannot vent the rotary pump when power is off. Those systems may } suck oil from the pump towards the vacuum chamber. } } The more common problem for infrequently used systems is moisture in the pump } oil. Particularly in moist climates and when relatively short pumping } times are } employed a good deal of water is absorbed in the pump. When the pump is not } used for a lengthy period this may cause corrosion and certainly lowers the } vapour pressure of the contaminated oil and so lower performance results. } } I suggest that at the end of your spasmodic activities the pump should be run } with the baffle valve partially open for at least 30 minutes. The baffle } valve } is usually atop the rotary pump. You could also use the sputter coater's } needle } valve, partially open. Under these conditions the pump will run hotter and } throw-out a good deal of oil mist and water vapour. Vent the exhaust into a } fume hood since oil mist is not just unpleasant. You will find that the pump } performs much better after a baffle run. } } SEM and TEM usually do not need this treatment, but they may, for instance } if a } TEM has been used to "dry" film that was not previously dried in another } system. *************
} We used to leave the sputter coater sitting for weeks after pumping it down } and never had any problems. It sounds as if you may have a leak at your } sealing surface. We always had to be very careful about cleanliness of the } bell jars surface and the plate it seals on. A small grain of sand or other } material can fracture the glass so that you have leaks and requires } repolishing of the glass bell jar. } } I hope that this helps you. ***************
} Gary- } I am not sure if you are saying that your sputter coater is pumping } down poorly or that it is depositing poor quality films. In either case, } it sounds like poor vacuum is the problem. Water vapor adsorbing to the } deposition chamber walls over a period of disuse will pump off of the walls } very slowly. A "wet" chamber can take 10 times as long to pump down to } ultimate vacuum as a "dry" one. If this sounds like it might be causing } the symptoms you are seeing, I would suggest you try one of two things: } } 1. Place a valve between the sputter chamber and the pump. Valve off the } chamber and leave it under vacuum between uses. As long as it retains any } vacuum this will help when you try to use it again. You don't want to be } pumping on the chamber at the pump's ultimate vacuum for any extended } period of time because that will allow oil vapor from the mechanical pump } to backstream into your sputter chamber and possibly onto your specimen if } present! (I am assuming that you have an oil based pump.) It is also nice } to spare the pump the wear and tear of pumping continuously over periods of } disuse. } } OR } 2. Several hours or the day before you want to use it, start pumping on the } sputter chamber while admitting a small flow of argon. The argon is an } important part of this procedure. First of all, the argon will prevent the } backstreaming described above. Second, I have been told that the argon } helps to desorb the water from the chamber surfaces. I don't know if that } is an old wives tale or not, but it does seem reasonable. The argon flow } may also help in purging any condensed vapors from the pump's oil. Don't } use too much argon though or you may overheat your pump. ******************
} While I have done no experimental protocol to prove this schedule is } optimum, during the off season I try to remember to run the sputter } coaters overnight one night a week. This keeps things in good shape. I } have read that the slow pumpdown of unused vacuum systems is caused mostly } by water vapor adsorbed on interior surfaces, and by traces of moisture in } the pump oil.
Gary P. Radice gradice-at-richmond.edu Associate Professor of Biology 804 289 8107 (voice) University of Richmond 804 289 8233 (FAX) Richmond VA 23173 http://www.science.richmond.edu/~radice
This position is available at the University of Utah. Dr. Erik Jorgensen is the contact person.
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Experience in electron microscopy and a Bachelor's degree in a science or related field required. Degree in a biological science, experience in light microscopy, and familiarity with microcomputers preferred. Operates electron microscopes, prepares specimens for microscopy, produces photographic and digital micrographs, analyzes data and maintains accurate work records. Necessary training will be provided. Applicants must submit a University of Utah Application for Employment.
Erik M. Jorgensen, Ph.D. Assistant Professor Department of Biology University of Utah 257 South 1400 East Salt Lake City, UT 84112-0840 PHN: (801) 585-3517 FAX: (801) 581-4668 jorgensen-at-biology.utah.edu
This position is available at the University of Utah. Dr. Erik Jorgensen is the contact person.
________________________________________
Electron Microscopy Lab Technician
Experience in electron microscopy and a Bachelor's degree in a science or related field required. Degree in a biological science, experience in light microscopy, and familiarity with microcomputers preferred. Operates electron microscopes, prepares specimens for microscopy, produces photographic and digital micrographs, analyzes data and maintains accurate work records. Necessary training will be provided. Applicants must submit a University of Utah Application for Employment.
Erik M. Jorgensen, Ph.D. Assistant Professor Department of Biology University of Utah 257 South 1400 East Salt Lake City, UT 84112-0840 PHN: (801) 585-3517 FAX: (801) 581-4668 jorgensen-at-biology.utah.edu
Yes, we find that sputter coaters do not like being ignored for long periods. Probably, the specimen chamber and vac lines adsorb moisture and other gases from the laboratory and it takes the rotary pump considerably longer to pump down (many hours versus 15-20 minutes).
You should pump the system down at least weekly for at least an hour. After pumping for about 30 minutes, allow the Argon gas to leak through the system. This really purges the residual gases. Then, we find that if you fill the system with Argon, rather than letting it fill with room air, it will take considerably less time to get into a usable range next time.
John B.
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Dear experts,
I know I should be kept out of the lab but I had to do some special resin embedding. The problem is that I didn't read my own instructions and made up Spurr resin by adding all the ingredients together and then mixing. This means I added the DMAE before mixing the other components and have ended up with brittle blocks of inconsistant hardness.
I never thought I would be asking this but is there any way that I can recover these blocks to allow me to examine what I have embedded? Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm
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Hi, all you experts...
In spite of the digital camera on our LEO 912 EFTEM, we have a couple of users who are going through huge quantities of film. I need to set up an evacuated film dessicator (separate from the one on our older TEM), but I find the non-glass, non-clear-plastic vacuum dessicators in the catalogs at hand to be enormously expensive. Does anyone have a favorite vendor and model, or a kludge? I remember one at Berkeley that I think was made out of a pressure cooker hooked to a vacuum pump...
Mahalo! Tina
80 degrees F, sunny blue skies, everything in bloom, and promise of surf.
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
I'm looking for a working used sputter coater for a descent price or donation for a university lab. Please contact me at bcraft-at-uci.edu if you have one available.
While everyone is focussed on the ease with which viruses can be transmitted as and within attachments, maybe it's a good time to ask that postings to the list be only as text messages, not as attachments.
cheers
rtch
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Paul, I am not writing because of your "Dear experts" address. An expert is a squirt under pressure, maybe that suited better the person who mis-mixed the Spurr's. Long time ago I read a note that brittle blocks sectioned better after soaking them overnight in ethanol. I've never tried that, but there is a possibility. Please let us know if that method is any good. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Wednesday, May 10, 2000 4:09 AM, Paul Webster [SMTP:pwebster-at-mailhouse.hei.org] wrote: } } } Dear experts, } } I know I should be kept out of the lab but I had to do some special resin } embedding. } The problem is that I didn't read my own instructions and made up Spurr resin } by adding all the ingredients together and then mixing. This means I added } the DMAE before mixing the other components and have ended up with brittle } blocks of inconsistant hardness. } } I never thought I would be asking this but is there any way that I can recover } these blocks to allow me to examine what I have embedded? } Paul Webster, Ph.D } House Ear Institute } 2100 West Third Street } Los Angeles, CA 90057 } phone:213 273 8026 } fax: 213 413 6739 } e-mail: pwebster-at-hei.org } http://www.hei.org/htm/aemi.htm }
Margaret, Your message struck a chord here, and I would like to extend my heartfelt sympathy. We've been there too. I have reached the point where all the suppliers I contact have a 15-year history of preparing quotations and tenders almost completely in vain. I don't know why the sales reps. (or I) put up with it. That they do is a triumph of hope over realistic expectation, and therefore a credit to all of them. I think there should be a club for stressed out managers of poorly- funded EM facilities. I would be one of the first to join. perhaps some day we should organise a world congress if we can find a venue large enough, thoough whether it would be advisable to have a trade exhibition is open to question. Best wishes Chris
Date sent: Thu, 11 May 2000 14:43:05 -0400 To: Microscopy-at-sparc5.microscopy.com } From: Margaret Dienelt {brannign-at-asrr.arsusda.gov}
Dear Colleagues,
I want to test a new computer program of mine that processes electron diffraction ring-patterns from polycrystalline samples. I did test it with patterns recorded on film. However, I would also like to test it on energy filtered patterns that were recorded with a CCD (or imaging plate).
Could anyone of you send me such patterns from a single phase material with random orientation? If you also characterized the same sample (especially if you proved that the sample is not textured) and you published the results, I could reference this publication of yours.
Thank you in advance.
Dr. Janos L. Labar Research Institute for Technical Physics and Materials Science H-1121 Budapest, Konkoly-Thege u. 29-33 Postal address: H-1525 Budapest-114, Po Box 49 Tel: (36)(1) 392-26-92 Fax: (36)(1) 275-49-96 Fax/phone: (36)(1) 395-92-32 home page: www.mfa.kfki.hu/~labar
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Your concerns and frustrations expressed are a recurrent thread on this site. It would be nice to be able to sit down with other facility managers and discuss common problems and possible solutions. I think our common concerns are such that they effect both materials and biological facilities. I am thinking of things such as: use guidelines, multi-user vs. service functions, formal courses and informal instruction for new users, equipment maintenance costs, justification of new equipment needs to administrators, advisor committee formats, funding sources, etc. Perhaps we could arrange such an informal session at the upcoming M&M meeting. If you would be interested in such a session, take a look at the meeting schedule and suggest a time. Then I will ask the organizing committee to designate a room. Debby
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
--------------------------------------
Margaret, Your message struck a chord here, and I would like to extend my heartfelt sympathy. We've been there too. I have reached the point where all the suppliers I contact have a 15-year history of preparing quotations and tenders almost completely in vain. I don't know why the sales reps. (or I) put up with it. That they do is a triumph of hope over realistic expectation, and therefore a credit to all of them. I think there should be a club for stressed out managers of poorly- funded EM facilities. I would be one of the first to join. perhaps some day we should organise a world congress if we can find a venue large enough, thoough whether it would be advisable to have a trade exhibition is open to question. Best wishes Chris
Date sent: Thu, 11 May 2000 14:43:05 -0400 To: Microscopy-at-sparc5.microscopy.com } From: Margaret Dienelt {brannign-at-asrr.arsusda.gov}
recall reading of ibm's hunt for a silicon wafer cleaving tool capable of handling samples 5mm in diameter. any luck out there?
mark riggs svg lithography wilton, ct 06897 riggsm-at-svg.com
We are contrasting connective tissue filaments by negative stain technique using uranyl formate. Our protocol is to adsorb filaments onto carbon film coated grids, then to wash in two drops of water and two drops of uranyl formate, removing each drop using filter paper but not allowing the grid to dry until after the second drop of UF. We can not charge the grid surface prior to specimen adsorption or our specimens will not adsorb.
My question is to do with making uranyl formate. Our formula is to boil 5ml water, add .0375 g uranyl formate (our solid is very old...), stir for 20 minutes, then add 10 microlitter 5 M NaOH, stir for 20 minutes, then filter through a 0.1 micron before use. We are not getting a consistant staining pattern. Our wish is to see a uniform coating of stain and we seldom see even single grid squares evenly coated. We've tried different concentrations of stain and also different volumes of NaOH. Any suggestions? Does anyone know the purpose of boiling the water?
Thanks in advance,
Doug ---------------------- Douglas R. Keene Associate Investigator Shriners Hospital Research Facilities 3101 S.W. Sam Jackson Park Road Portland, Oregon 97201 phone: 503-221-3434 FAX: 503-412-6894 (9-5 PST) e-mail: DRK-at-shcc.org
As a former user and current vendor of such systems as you are inquiring about I can try to provide a bit of information regarding camera improvements:
There have been a number of improvements, but I am not sure what you are comparing the latest cameras against. Cameras are now usually cooled and provide 12 bits per pixel, the number of pixels has gone up a bit (but not much in general), and cameras read out faster than they used to (up to 20 fps and more). I think all cameras now use a line transfer mechanism, which makes shutters obsolete. On the software side, real-time FFT and real-time shading correction can be done now due to faster computers without special processing boards, and there have been other software developments that make using the cameras and computers easier. Other changes that affect the usability of cameras is the use of pneumatics to insert and retract the phosphors, higher frame rates for live viewing with the camera, etc.
If you have questions, please give me a call, drop me an email, or go to our web site.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== 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: Margaret Dienelt [mailto:brannign-at-asrr.arsusda.gov] Sent: Thursday, May 11, 2000 12:43 PM To: Microscopy-at-sparc5.microscopy.com
Hi,
Year after year I hopefully gather information about digital imaging systems for TEMs (ours is a JEOL 100CX) , only to learn we have no money. This year it looks like it might really happen but I have not kept up with innovations in the field and am wondering the following:
1. Anything new in the last two years -- especially in terms of cameras? I'm most familiar with the Gatan and AMT systems but their web sites don't reflect much in the way of changes over a year ago. 2. With more and more microscopists finally getting their systems -- I'd love to get feedback.
Thanks, Margaret
P.S. Would welcome contacts from vendors.
-- Margaret Dienelt
Plant Pathologist Electron Microscopy Lab
Floral and Nursery Plants Research Unit U.S. National Arboretum/Agricultural Research Service/USDA
B. 010A, Rm. 238, BARC-W 10300 Baltimore Avenue Beltsville MD. 20705 USA
Patrick Echlin from Cambridge UK noted in private message that "strong" agent who may dissolve epoxy is sodium methoxide. Thanks, Patrick.
Sergey.
} Date: Thu, 11 May 2000 16:06:39 -0700 } From: Sergey Ryazantsev {sryazant-at-ucla.edu} } Subject: Re: Spurr resin problem } X-Sender: sryazant-at-pop.ben2.ucla.edu } To: Microscopy-at-sparc5.microscopy.com } X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.6 (32) } } ------------------------------------------------------------------------ } 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
My procedure for uranyl formate is a little bit simpler than yours: 0.05-0.1 g uranyl formate (0.5-1% final) + 10 ml deinozed (cell culture quality or double distilled in the glass) water in the 15 ml plastic tube. It dissolved at the same speed (even better) as acetate salt (UA). Usually I am using rotator to shake slowly the tube with stain. It takes 0.5-1 hour to dissolve salt completely. I do not filter solution yet. The difference between formate and acetate salts of the uranium is that formate is light sensitive (UA - too, but less, less sensitive). You have to avoid direct high intensity light. Usually I wrapped tube in alumina foil and prepared the samples under diffused light moderate intensity (general illumination in the lab, no local lights). Staining procedure is exact the same as for acetate salt. The advantage of formate salt - it generates smaller granularity (and less contrast than UA), spreaded sometime better than acetate salt, and pH is higher. The disadvantage of the formate is that the water solution is not stable: I do prepare fresh solution every time I have to work with it. This is great disadvantage of the uranyl formate. I guess, you may store solution in the dark at +4oC for couple of days, but this is your own risk to experiment with that. As for staining procedure, I would avoid any washes with just water. As a biochemist I am under impression that ionic conditions is important to preserve "native" structure of the sample. Therefore I am using the same buffer as for sample to wash (usually I do not wash at all). Of coarse for any uranium salts you have to avoid any phosphates in the buffer. Any Tris, MES, HEPES buffers may be the good point to start. I don't know exactly how it works, but it seems to me, that buffer in the wash may help spread satin better (don't ask me why, I have no idea). If you have problem to dissolve uranyl formate, you probably have to replace it on the fresh one (it is cheap). Double-carbon technique may also help (you may call off line for details). Good luck and sorry for the long message.
Sergey
} Date: Fri, 12 May 2000 13:00:33 -0700 (Pacific Daylight Time) } From: Douglas Keene {DRK-at-shcc.org} } Subject: Uranyl Formate } Sender: drk-at-shcc.org } To: microscopy-listserver {Microscopy-at-sparc5.microscopy.com} } Reply-to: DRK-at-shcc.org } X-Mailer: Simeon for Win32 Version 4.1.3 Build (39) } Priority: NORMAL } X-Authentication: none } } ------------------------------------------------------------------------ } 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
} From Debby Sherman: } } Your concerns and frustrations expressed are a recurrent thread on this } site. It would be nice to be able to sit down with other facility managers } and discuss common problems and possible solutions. I think our common } concerns are such that they effect both materials and biological } facilities. I am thinking of things such as: use guidelines, multi-user } vs. service functions, formal courses and informal instruction for new } users, equipment maintenance costs, justification of new equipment needs } to administrators, advisor committee formats, funding sources, etc. } Perhaps we could arrange such an informal session at the upcoming M&M } meeting. If you would be interested in such a session, take a look at the } meeting schedule and suggest a time. Then I will ask the organizing } committee to designate a room.
} Debby -
It's too late to add to the program now, but I attended a Long Beach 2001 LAC meeting last week, and there's a committee member there who wants to organize something. It's almost too late to add programming even for that one! The solution that I suggested is to start an annual breakfast or lunch for facility managers, taking care to avoid other large scheduled meetings (which aren't listed in the program summary). It might even be possible to get more time Sunday afternoon, pre-opening reception. Contact the LAC chairs for that: Stacie Kirsch for Philly & Zed Mason for Long Beach.
Caroline Schooley
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/PCI/pci.html
University of Connecticut Institute for Materials Science
Postdoctoral Research Position in Electron Microscopy Studies of Fatigue Crack Initiation Sites in Ti-6-4 Alloys
The Institute for Materials Science (IMS) at UConn is an interdisciplinary center with the threefold mission of fostering education, research and outreach in all areas of the materials sciences. Applications are invited for a Postdoctoral Position to study the microcrystallography of fatigue crack initiation sites in Ti 6-4 alloys. The appointee will be involved in electron microscopy studies of failed test pieces produced at Pratt and Whitney in the previous phase of this program. It is envisaged that this work will involve extensive SEM and TEM studies in the IMS at UConn with some use of the FIB/TEM/STEM and OIM facilities in the High Temperature Materials Laboratory at Oak Ridge National Laboratory. Candidates should hold a PhD in Materials Science, Physics or a related discipline and must have extensive hands-on experience in a broad range of electron microscopy techniques. Experience in the assessment of deformation substructures would also be beneficial. The appointment is for one year in the first instance and is available from June 1st. Screening of the applications will begin immediately and will continue until the post is filled.
Applications from under-represented groups, including minorities, women and people with disabilities are encouraged.
Interested candidates should send a curriculum vitae, including publication list, and the names of at least three referees with postal addresses, telephone numbers and Email addresses to: Dr. M. Aindow, Institute for Materials Science, University of Connecticut, 97 North Eagleville Road, U-3136, Storrs, CT 06269-3136 USA. Email: maindow-at-ims.uconn.edu -- *********************************************************
Mark Aindow, Associate Professor, Department of Metallurgy and Materials Engineering Institute of Materials Science, U-136 University of Connecticut, Storrs, CT 06269-3136, USA
I have a question concerning sample preparation for brightfield and polarized particle size analysis. My customer is trying to devise an experiment to analyze fly ash collected on a filter during smokestack emissions testing. He collects ~1gr. per sample, however, more is easily possible. The sample is clumped and incongruent and he would like a simple protocol for proper sample dispersion on a slide.
Hello there everyone, I am having hard time cutting gold coated polypropylene (pp) wires. I am using a diamond knife, I have embedded the samples into epoxy and hardener, I will be doing TEM. But when I cut, I get good thickness but the slice keep rolling up. I have been trying for the past three days and do not have any luck. I have change the ratio of the epoxy to hardener from10/4 to 10/6 still no luck. Can anyone suggest some changes. Is cryogenic an option here.
Thank you all,
Briget Ngampa 28 Cedar street LOWELL,MA 01852 Tel (978) 970 0433 Fax (978) 937 2297 EMAIL: hdmhos-at-aol.com
Hi, You may want to check SELA. They have an evergrowing line of cleavers for the semiconductor industry. Contact Efrat Raz: efrat-at-sela.com
Caveat: MME has no financial interest in this product
Good hunting Barbara Foster,President Microscopy/Microscopy Education 125 Paridon Street Suite 102 Springfield, MA 01118-2130 PH: 413-746-6931 FX: 413-746-9311 email: MME-at-map.com Website: www.MME-Microscopy.com/education
America's first national consortium of microscopy specialists offering customized on-site training in all areas of microscopy, image analysis, and sample preparation ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^
At 03:01 PM 5/12/00 -0400, Mark Riggs wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } Hildy hello, } } I could not understand, how polymerized (mean crosslinked) epoxy may be } dissolved back in PO? You have to break chemical bonds between polymer's } chains first and than it will become soluble. I believe, there are some } very strong oxidizing agents should be used in order to break chemical } bonds in the epoxies. } } Sergey. } } } } } Date: Thu, 11 May 2000 11:28:43 -0600 (MDT) } } From: HILDEGARD CROWLEY {hcrowley-at-du.edu} } } Subject: Re: Spurr resin problem } } X-Sender: hcrowley-at-odin.cair.du.edu } } To: Paul Webster {pwebster-at-mailhouse.hei.org} } } Cc: MSA listserver submission {Microscopy-at-sparc5.microscopy.com} } } } } ------------------------------------------------------------------------ } } 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 } } -----------------------------------------------------------------------. } } } } } } } } } } On Tue, 9 May 2000, Paul Webster 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 experts, } } } } } } I know I should be kept out of the lab but I had to do some special } resin embedding. } } } The problem is that I didn't read my own instructions and made up Spurr } resin by adding all the ingredients together and then mixing. This means I } added the DMAE before mixing the other components and have ended up with } brittle blocks of inconsistant hardness. } } } } } } I never thought I would be asking this but is there any way that I can } recover these blocks to allow me to examine what I have embedded? } } } Paul Webster, Ph.D } } } House Ear Institute } } } 2100 West Third Street } } } Los Angeles, CA 90057 } } } phone:213 273 8026 } } } fax: 213 413 6739 } } } e-mail: pwebster-at-hei.org } } } http://www.hei.org/htm/aemi.htm } } } } } } } } } } } Hi, } } } } First try this: Get a beaker of water, heat it to the highest temperature } } at which your blocks were polymerized, then subtract 5 deg. After temp } } has been reached, put in one block. Leave it for 15 min. Take it out, } } trim it, and section it immediately. Too brittle? Repeat the water soak } } for 15 min. Repeat again. Not much can be accomplished after one hour of } } soaking, but this may be different in your case. Sometimes in desperation } } when I wanted 4 micrometer thick sections from difficult material I have } } soaked blocks overnight with good success. } } } } One time I was given immensely valuable blocks which were so bad (kind } } expression) that they curled my hair. They could not be cut. I had to } } get the epoxy out and reembed. (The micrographs later ended up in a } } publication in the Comp. Neurol. Journal) } } } } What I did was to rotate the blocks in vials continuously in the REVERSE } } order in which they were embedded, with much elongated times. That is, } } first they went into PO and epoxy (same formulation as the bad embedment) } } 3:1, for hours, the 2:1 for the rest of the day, then 1:1 for overnight, } } then pure PO for a whole day. All the above were done with numerous } } changes. Once I was in PO, pure for a day, most of the epoxy had left the } } tissue. I then remembedded the same way I deembedded. What a pain. The } } blocks were never wonderful, but they sectioned OK and went for } } publication. It should work for Spurr's. Don't give up. When } } deembedding, I left out the accelerator, of course. It helps to have some } } very good chocolate on hand for this maneuver. } } } } Good luck, } } Hildy } } } } Hildegard H. Crowley } } University of Denver } } } } } } } _____________________________________ } } Sergey Ryazantsev Ph. D. } Electron Microscopy } UCLA School of Medicine } Department of Biological Chemistry } Box 951737 } Los Angeles, CA 90095-1737 } } Phone: (310) 825-1144 } Pager: (310) 845-0248 } FAX (departmental): (310) 206-5272 } mailto:sryazant-at-ucla.edu } http://www.bol.ucla.edu/~sryazant } } } } Hi,
Yes, it is a mystery to me that one can get old epoxy out of tissue with PO. However, I have done it at least 3 times with enough success to collect data. The only thing I can think of is that at least 10% of monomers never bind due to the low embed temps we use for our TEM work. Those will surely dissolve out leaving holes. (I have seen this). PO is the simplest of epoxies and a very strong solvent. That is all I know. I never do the "REVERSE" embed unless forced to do it, because it is so difficult dealing with the final cutting and the staining. And then the "new" sections are unstable and uneven. I would rather clean a bathroom! Bye, Hildy
} ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } Patrick Echlin from Cambridge UK noted in private message that "strong" } agent who may dissolve epoxy is sodium methoxide. Thanks, Patrick. } } Sergey. } } } } Date: Thu, 11 May 2000 16:06:39 -0700 } } From: Sergey Ryazantsev {sryazant-at-ucla.edu} } } Subject: Re: Spurr resin problem } } X-Sender: sryazant-at-pop.ben2.ucla.edu } } To: Microscopy-at-sparc5.microscopy.com } } X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.6 (32) } } } } ------------------------------------------------------------------------ } } 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 } } -----------------------------------------------------------------------. } } } } } } Hildy hello, } } } } I could not understand, how polymerized (mean crosslinked) epoxy may be } } dissolved back in PO? You have to break chemical bonds between polymer's } } chains first and than it will become soluble. I believe, there are some } } very strong oxidizing agents should be used in order to break chemical } } bonds in the epoxies. } } } } Sergey. } } } } } } } } } Date: Thu, 11 May 2000 11:28:43 -0600 (MDT) } } } From: HILDEGARD CROWLEY {hcrowley-at-du.edu} } } } Subject: Re: Spurr resin problem } } } X-Sender: hcrowley-at-odin.cair.du.edu } } } To: Paul Webster {pwebster-at-mailhouse.hei.org} } } } Cc: MSA listserver submission {Microscopy-at-sparc5.microscopy.com} } } } } } } ------------------------------------------------------------------------ } } } 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 } } } -----------------------------------------------------------------------. } } } } } } } } } } } } } } } On Tue, 9 May 2000, Paul Webster 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 experts, } } } } } } } } I know I should be kept out of the lab but I had to do some special } } resin embedding. } } } } The problem is that I didn't read my own instructions and made up Spurr } } resin by adding all the ingredients together and then mixing. This means I } } added the DMAE before mixing the other components and have ended up with } } brittle blocks of inconsistant hardness. } } } } } } } } I never thought I would be asking this but is there any way that I can } } recover these blocks to allow me to examine what I have embedded? } } } } Paul Webster, Ph.D } } } } House Ear Institute } } } } 2100 West Third Street } } } } Los Angeles, CA 90057 } } } } phone:213 273 8026 } } } } fax: 213 413 6739 } } } } e-mail: pwebster-at-hei.org } } } } http://www.hei.org/htm/aemi.htm } } } } } } } } } } } } } } } Hi, } } } } } } First try this: Get a beaker of water, heat it to the highest temperature } } } at which your blocks were polymerized, then subtract 5 deg. After temp } } } has been reached, put in one block. Leave it for 15 min. Take it out, } } } trim it, and section it immediately. Too brittle? Repeat the water soak } } } for 15 min. Repeat again. Not much can be accomplished after one hour of } } } soaking, but this may be different in your case. Sometimes in desperation } } } when I wanted 4 micrometer thick sections from difficult material I have } } } soaked blocks overnight with good success. } } } } } } One time I was given immensely valuable blocks which were so bad (kind } } } expression) that they curled my hair. They could not be cut. I had to } } } get the epoxy out and reembed. (The micrographs later ended up in a } } } publication in the Comp. Neurol. Journal) } } } } } } What I did was to rotate the blocks in vials continuously in the REVERSE } } } order in which they were embedded, with much elongated times. That is, } } } first they went into PO and epoxy (same formulation as the bad embedment) } } } 3:1, for hours, the 2:1 for the rest of the day, then 1:1 for overnight, } } } then pure PO for a whole day. All the above were done with numerous } } } changes. Once I was in PO, pure for a day, most of the epoxy had left the } } } tissue. I then remembedded the same way I deembedded. What a pain. The } } } blocks were never wonderful, but they sectioned OK and went for } } } publication. It should work for Spurr's. Don't give up. When } } } deembedding, I left out the accelerator, of course. It helps to have some } } } very good chocolate on hand for this maneuver. } } } } } } Good luck, } } } Hildy } } } } } } Hildegard H. Crowley } } } University of Denver } } } } } } } } } } } _____________________________________ } } } } Sergey Ryazantsev Ph. D. } } Electron Microscopy } } UCLA School of Medicine } } Department of Biological Chemistry } } Box 951737 } } Los Angeles, CA 90095-1737 } } } } Phone: (310) 825-1144 } } Pager: (310) 845-0248 } } FAX (departmental): (310) 206-5272 } } mailto:sryazant-at-ucla.edu } } http://www.bol.ucla.edu/~sryazant } } } } } } } } } _____________________________________ } } Sergey Ryazantsev Ph. D. } Electron Microscopy } UCLA School of Medicine } Department of Biological Chemistry } Box 951737 } Los Angeles, CA 90095-1737 } } Phone: (310) 825-1144 } Pager: (310) 845-0248 } FAX (departmental): (310) 206-5272 } mailto:sryazant-at-ucla.edu } http://www.bol.ucla.edu/~sryazant } } } } Hi,
Absolutely sodium methoxide will take the epoxy out of the tissue. However, in our laboratory it caused so much tissue damage (since epoxies actually bind with proteins in the tissue and not simply throw a net through and around the tissue like the acrylics) that after reembedding it was not useful for collecting data. Somebody else might have better results than myself with that method, so we should not discard the idea.
Briget: Polypropylene has a Tg of about -19C. Whenever I microtome I always cut at least 15-20 degrees below the Tg. If you are trying to cut these samples at room temperature my guess is this is your problem. If you need to embed you can still do this when cutting at cryo temperatures by trimming as much of the epoxy away as possible. If you leave a very thin layer of epoxy around your sample you should still get good sections even though you will get some chatter in the epoxy region. Steve
Hello there everyone, I am having hard time cutting gold coated polypropylene (pp) wires. I am using a diamond knife, I have embedded the samples into epoxy and hardener, I will be doing TEM. But when I cut, I get good thickness but the slice keep rolling up. I have been trying for the past three days and do not have any luck. I have change the ratio of the epoxy to hardener from10/4 to 10/6 still no luck. Can anyone suggest some changes. Is cryogenic an option here.
Thank you all,
Briget Ngampa 28 Cedar street LOWELL,MA 01852 Tel (978) 970 0433 Fax (978) 937 2297 EMAIL: hdmhos-at-aol.com
Stephen McCartney Research Associate 2108 Hahn Hall Materials Institute Virginia Tech Blacksburg, VA 24061-0344 USA
A newly appointed researcher here has asked my advice on several pieces of TEM spec. prep equipment. I turn to you for helpful suggestions.
The research involves serial sectioning biological tissues and many grids. EM is a minor, but essential component of the project. The lab runs through dozens of grids and hundreds of pictures, but only 4 - 6 times a year. Big bursts of activity followed by long periods of analysis and investigations using other techniques. Of the hundreds of pictures taken, they may only use a few for data.
The researcher is looking for ideas on what choices are available, how useful, and approximate costs of the following:
Digital imaging to add to our existing JEOL 1200EX TEM - I have seen other threads on this topic and would welcome any new ideas. Digital imaging will save a lot of time and money since they discard so many pictuers, but the question of image quality is one I am to investigate.
Ultra microtome - We have an older A/O Ultracut (the model before it became the Reichert Ultracut E) which is OK for the sectioning we do in the general lab. But she wants a new one for her exclusive use. The question is whether a new microtome will allow folks in her lab to do serial sectioning any faster or easier, or by less skilled users, than our current system.
Staining machine - Anyone have info on staining machines or systems for lots of TEM grids. I have never had to do so many grids that this was an issue, so I have never kept up on the offerings. If you know of something and/or have experience let me know. Again, this is something she would keep in her lab.
Tissue processing machine - Same as above for me, never did so much at one time that I ever thought I needed one of these. The samples to be processed are C. elegans, anything available that could do these unattended? How about upkeep and volumes of chemicals needed. Also an item to be kept in her lab.
I will filter and pass on your comments. Anything you might offer will, as always, be received with appreciation and thanks.
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
Jon, I can speak to the digital camera at least. We have been using a Gatan BioScan on our microscope and it serves our needs for 95% of the images that we produce. It is a very good system, when it is working. We have had more problems with it than I would have expected. We bought the system early in the production and perhaps they have worked out the bugs by now. -------------------------------------.
} Dear List: } } A newly appointed researcher here has asked my advice on several pieces of } TEM spec. prep equipment. I turn to you for helpful suggestions. } } The research involves serial sectioning biological tissues and many grids. } EM is a minor, but essential component of the project. The lab runs through } dozens of grids and hundreds of pictures, but only 4 - 6 times a year. Big } bursts of activity followed by long periods of analysis and investigations } using other techniques. Of the hundreds of pictures taken, they may only } use a few for data. } } The researcher is looking for ideas on what choices are available, how } useful, and approximate costs of the following: } } Digital imaging to add to our existing JEOL 1200EX TEM - I have seen other } threads on this topic and would welcome any new ideas. Digital imaging will } save a lot of time and money since they discard so many pictuers, but the } question of image quality is one I am to investigate. } } Ultra microtome - We have an older A/O Ultracut (the model before it became } the Reichert Ultracut E) which is OK for the sectioning we do in the } general lab. But she wants a new one for her exclusive use. The question is } whether a new microtome will allow folks in her lab to do serial } sectioning any faster or easier, or by less skilled users, than our current } system. } } Staining machine - Anyone have info on staining machines or systems for } lots of TEM grids. I have never had to do so many grids that this was an } issue, so I have never kept up on the offerings. If you know of something } and/or have experience let me know. Again, this is something she would keep } in her lab. } } Tissue processing machine - Same as above for me, never did so much at one } time that I ever thought I needed one of these. The samples to be processed } are C. elegans, anything available that could do these unattended? How } about upkeep and volumes of chemicals needed. Also an item to be kept in } her lab. } } I will filter and pass on your comments. Anything you might offer will, as } always, be received with appreciation and thanks. } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } }
Gregory W. Erdos, Ph.D. Assistant Director, Biotechnology Program PO Box 110580 University of Florida Gainesville, FL 32611
Hi all - a first attempt at unloading a problem onto the listserver!
I am trying to get thin sections of blue-green algae that are loaded with starch granules. The problem is that every time we do the preps, we end up with cells that lose some granules, leaving a gaping hole, or of starch granules that have holes in the centre. I'm using a standard sort of method - fix in phosphate + 2.5% glut 2 hours, osmium 1% 2 hours, dehydrate in ethanol series, finishing with prop. ox, 3:1 prop. ox/resin around 2 h, 1:1 2 or 3h then another change overnight, a couple of 100% resin changes for 1/2 to one day, then polymerization 48h/60C. The fixing and EtOH steps are done on ice, the rest at room temp, the resin infiltration is done on a rotating mixer. We've tweaked the method around but nothing seems to work. I've tried using LR White as well, but with no luck. Can anybody out there help!?
Mark
******************************************** Mark West, Unidad de Microscopia Electronica, (Electron Microscopy Unit) Instituto de Fisiologia Celular, Universidad Nacional Autonoma de Mexico, 04510 Mexico D.F.
"I am Mark Armogida, VP of Engineering at Ted Pella, Inc. in Redding, California, USA. We have an old (1964) TEM, Hitachi model Hu 11a, that is a decommissioned instrument which we are trying to give away for removal/shipping costs. It was operational a year ago and kept under vacuum since then. We have the instruction manual. This unit consists of the main body with diffusion pumps, a large electronics console, a large oil filled resistor and mechanical pumps. The oil was analyzed and does not contain PCB's. I estimate that this system weighs several thousand pounds. I will have it removed for salvage in a week, so please contact me if you are interested in this system."
Mark Armogida VP, Engineering & Production Ted Pella, Inc.
I am sending this for a colleague. It was a wonderful little scope while I was there. } LifeCell Corporation (The Woodlands, TX and Branchburg, NJ) is closing their facility in The Woodlands. } LifeCell has available for donation a Philips EM300 TEM. The scope was in } working order when shut down in December 1999. } If you are interested please contact: } Sy Griffey, Ph.D. } 908-947-1143 or sgriffey-at-lifecell.com }
At 01:38 PM 5/10/00, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
It seems to me that no digital camera system would work on a SEM in place of a Polaroid or other film-based output device. Since the recording CRT in a SEM is based on a sequential line scan, one would need a camera that would capture each line as it is produced. Most digital backs are single or triple pass units of a single linear set of sensors. There are other cameras that do snapshot capture but even these would not work since the whole image is not present on the record CRT at the time of taking a picture with the digital camera. The final image is generated sequentially, line by line, on the record CRT.
If the SEM image is stored in a frame buffer, the buffer can be converted to RS-170 TV video and frame grabbed. But the best that this would typically do is 640 lines.
Its an interesting problem and dilemma about being in a situation where digital camera products simply won't work in place of film. But since the goal is to obtain a digital file, why not start with a digital interface? For example, a passive digital capture system would transfer the record CRT information to computer and directly result in a nice digital file. Alternatively, for some systems, an active system can be applied to directly control the SEM's beam. In doing so, the range of final digital image resolution is limited only by the attached hardware system.
gary g.
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Modern surfers use PC boards. You can too at http://photoweb.net ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
I am Mark Armogida, VP of Engineering at Ted Pella, Inc. in Redding, California, USA. We have an old (1964) TEM, Hitachi model Hu 11a, that is a decommissioned instrument which we are trying to give away for removal/shipping costs. It was operational a year ago and kept under vacuum since then. We have the instruction manual. This unit consists of the main body with diffusion pumps, a large electronics console, a large oil filled resistor and mechanical pumps. The oil was analyzed and does not contain PCB's. I estimate that this system weighs several thousand pounds. I will have it removed for salvage in a week, so please contact me if you are interested in this system. I can be reached by e-mail or by phone at 530-241-2200 ext 212 between the hours of 8:00am and 5:00pm pacific time zone.
Electron Microscopist / Materials Scientist Materials Science Division, Argonne National Laboratory
The Materials Science Division at Argonne National Laboratory invites applications for a Staff Scientist position in Electron Microscopy. The candidate should have a strong background in the various techniques of electron microscopy and a very strong interest in application of these techniques in materials science. The candidate should have a state-of-the-art knowledge of either analytical transmission electron microscopy including high spatial resolution x-ray and electron spectroscopy or the application of electron holography and Lorentz imaging, with a particular emphasis on field emission (S)TEM. Candidates with exceptional experience in other areas of microscopy will also be considered.
Areas of particular research interest include defects and interfaces in materials, in situ studies of critical phenomena, irradiation and ion implantation effects, and applications of electron holography and Lorentz imaging. A familiarity with research in one or more of the following areas is highly desirable: magnetic and superconducting materials, irradiation effects, ferroelectrics, nanoscale materials, diamond films, and non-crystalline materials. The successful candidate will work closely with Electron Microscopy Center personnel and other research groups within the Materials Science Division to develop strong research efforts in one or more of these areas.
Interested candidates should send a curriculum vitae, a brief statement of research interests and plans, and the names and contact information of three references to: Susan Walker, Employment and Placement Box MSD-210121 9700 S. Cass Avenue Argonne, IL 60439 TDD: 630-252-7722
Questions can be addressed to Dr. Dean J. Miller, Materials Science Division, Argonne National Laboratory, 9700 S. Cass Ave., Argonne, IL 60439, tel. 630-252-4108 (with voice mail), fax 630-252-7529, or miller-at-anl.gov.
Argonne National Laboratory is a multidisciplinary center of energy research and related scientific studies and is operated by the University of Chicago for the U.S. Department of Energy. Argonne National Laboratory is a federal contractor and complies with all federal contractor rules and regulations regarding the maintenance and implementation of our Affirmative Action Program.
--------------------------- Dean J. Miller Materials Science Division Argonne National Laboratory Argonne, IL 60439
Dr. Gary Gaugler 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 } -----------------------------------------------------------------------. } } At 01:38 PM 5/10/00, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } } } I'm curious ... has anyone tried simply installing a digital } } 4x5 back in place of a Polaroid 4x5 back??? For example, see: } } } } http://www.phaseone.com/brochures/powerfx.html } } } } cheerios, shAf } } It seems to me that no digital camera system would work on a SEM } in place of a Polaroid or other film-based output device. Since the } recording CRT in a SEM is based on a sequential line scan, one } would need a camera that would capture each line as it is produced. } Most digital backs are single or triple pass units of a single linear } set of sensors. There are other cameras that do snapshot capture } but even these would not work since the whole image is not present } on the record CRT at the time of taking a picture with the digital } camera. The final image is generated sequentially, line by line, } on the record CRT. } } If the SEM image is stored in a frame buffer, the buffer can be } converted to RS-170 TV video and frame grabbed. But the } best that this would typically do is 640 lines. } } Its an interesting problem and dilemma about being in a situation } where digital camera products simply won't work in place of } film. But since the goal is to obtain a digital file, why not start } with a digital interface? For example, a passive digital capture } system would transfer the record CRT information to computer } and directly result in a nice digital file. Alternatively, for some } systems, an active system can be applied to directly control the } SEM's beam. In doing so, the range of final digital image } resolution is limited only by the attached hardware system. } } gary g. } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } Modern surfers use PC boards. You can too at } http://photoweb.net } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Shaf, Aside from the problems already mentioned, the back alone (without computer) is almost as expensive as the smallest SEMs. It also has about 4 times the resolution of the best recording systems out there (ETEC) so what you'd get is a lot of empty (information-wise) pixels. It might be adaptable to TEM, though.
For an SEM an active digital control could be set up to gather 10K x 10K images, but the stability of the column drivers becomes very important. Some could handle it fairly well while some would again be useless because their drivers have no noise immunity due to the fact that their push-pull mag drivers were designed backwards and all powere supply noise is passed directly to the scan coils.
Ken Converse Quality Images third party SEM service Delta, PA
Embedding and sectioning material containing a lot of starch is always a problem. If you do not need to look at the structure of the mature starch grains, you could try embedding the algae first thing in the morning before they have had time to produce the starch. Most plants have a low starch content after being kept in the dark for approx 12 hours. For instance, active, growing leaves are difficult to section when collected midday, but early morning collecting makes for easy sectioning.
Jan Coetzee
Mark West wrote:
} I am trying to get thin sections of blue-green algae that are loaded with } starch granules. The problem is that every time we do the preps, we end up } with cells that lose some granules, leaving a gaping hole, or of starch } granules that have holes in the centre. I'm using a standard sort of } method - fix in phosphate + 2.5% glut 2 hours, osmium 1% 2 hours, } dehydrate in ethanol series, finishing with prop. ox, 3:1 prop. ox/resin } around 2 h, 1:1 2 or 3h then another change overnight, a couple of 100% } resin changes for 1/2 to one day, then polymerization 48h/60C. The fixing } and EtOH steps are done on ice, the rest at room temp, the resin } infiltration is done on a rotating mixer. We've tweaked the method around } but nothing seems to work. I've tried using LR White as well, but with no } luck. Can anybody out there help!? }
-- Prof Jan Coetzee Lab for Microscopy and Microanalysis University of Pretoria, South Africa. Tel: 012-420-2075, Fax 012-362-5150 www.up.ac.za/science/electron/emunit1.htm
Question: Dear sir In order to test the quality of difraction limited microscope I am looking for reticles that will enable me to produce difraction patterns of about 0.2 micron source can you advice where can I find such reticles . Who can produce them for me if they are not available comercially . Best regards Moshe Marcovitch
I've seen references to using potassium ferrocyanide reduced osmium tetroxide to help preserve glycogen. Karnovsky(1971) Use of ferrocyanide-reduced OsO4 in EM. In Proc.14th Annu.Meet. Am. Soc. Cell Biol., p. 146. Abstract 284.
Tracy Gales Fox Chase Cancer Center Electron Microscopy Facility 7701 Burholme Ave. Philadelphia PA 19111 Tel. 215-728-2484 Fax 215-728-2412
At 04:02 PM 5/15/00 -0700, Dr. Gary Gaugler wrote: } It seems to me that no digital camera system would work on a SEM } in place of a Polaroid or other film-based output device. Since the } recording CRT in a SEM is based on a sequential line scan, one } would need a camera that would capture each line as it is produced.
What you're saying is there must be a system out there that digitizes that single stream of line scan intensities, then processes all that data inside the computer as an image as opposed to trying to digitize the frame buffer.
Try Klarmann Rulings, Inc., they manufacture reticles. If not a stock item they will make one to your specification.
There URL is http://www.reticles.com
Best Regards
Joseph Passero
mailto:jp-at-spacelab.net
On Tuesday the 16th of May, 2000 at 07:39:23 -0500, moshe_marc-at-gohip.com wrote and posted:
} Email: moshe_marc-at-gohip.com } Name: moshe marcovitch } } Question: Dear sir } In order to test the quality of difraction limited microscope I am looking } for reticles that will enable me to produce difraction patterns of about } 0.2 micron source can you advice where can I find such reticles . Who can } produce them for me if they are not available comercially . } Best regards } Moshe Marcovitch } } ---------------------------------------------------------------------------
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Dear colleagues there are new websites names available. It will look like: www.diptera.ws or www.????.ws more information you can find on www.coleoptera.org in section {software house} Keep care and be of good cheer.
Regards
(name) Vratislav Richard Eugene Maria John Baptist (surname) of Bejsak (Bayshark)-Collorado-Mansfeld
website: http://www.coleoptera.org listserver: coleoptera on www.egroup.com/group/coleoptera/info.html Coleoptera - Australia, Tenebrionidae of World (incl. Lagriinae, Alleculinae)
University of Sydney The Wentworth Bldg., Box 62 NSW 2006 AUSTRALIA phone : +61 414 540 465 email: vratislav-at-bigfoot.com ricardo-at-ans.com.au (before Ricardo-at-compuserve.com and ricardo-at-login.cz )
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.
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In response to the current thread on the problems of facility managers, I say count me in as being highly interested! If a discussion group does get together at M&M it would be great to see a report posted on this listserver for those of us who unfortunately can't attend the meeting. If somebody could take notes and post them, I for one would be very appreciative.
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Mark, I have been working with cyanobacteria for over 15 years. We found out in the late 80's that the only way to really hold the starch granules together is by using plunge freezing and freeze substitution techniques. I would substitute in acetone + osmium and embed in Spurr's for normal ultrastructure and use ETOH and embed in HM-20 for immuno. Check out the following references for details of method and contact me for further explanations:
Schneegart, M.A., D. M. Sherman, S. Nayar, and L. A. Sherman (1994). Oscillating Behavior of Carbohydrate Granule Formation and Dinitrogen Fixation in the Cyanobacterium Cyanothese sp. strain ATCC 51142. J. Bacteriology. 176:1586-1597.
Sherman, D. M., T. Troyan, and L. A. Sherman (1994). Localization of Membrane Proteins in the Cyanobacterium Synechococcus sp. PCC7942. Plant Physiol. 106:251-262
Plunge freezing apparatus can be made in a university shop at relatively low cost and works great for many unicellular organisms. Debby
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
--------------------------------------
Hi all - a first attempt at unloading a problem onto the listserver!
I am trying to get thin sections of blue-green algae that are loaded with starch granules. The problem is that every time we do the preps, we end up with cells that lose some granules, leaving a gaping hole, or of starch granules that have holes in the centre. I'm using a standard sort of method - fix in phosphate + 2.5% glut 2 hours, osmium 1% 2 hours, dehydrate in ethanol series, finishing with prop. ox, 3:1 prop. ox/resin around 2 h, 1:1 2 or 3h then another change overnight, a couple of 100% resin changes for 1/2 to one day, then polymerization 48h/60C. The fixing and EtOH steps are done on ice, the rest at room temp, the resin infiltration is done on a rotating mixer. We've tweaked the method around but nothing seems to work. I've tried using LR White as well, but with no luck. Can anybody out there help!?
Mark
******************************************** Mark West, Unidad de Microscopia Electronica, (Electron Microscopy Unit) Instituto de Fisiologia Celular, Universidad Nacional Autonoma de Mexico, 04510 Mexico D.F.
Responding to the message of {Pine.WNT.4.10.10005151651240.-3843205-100000-at-mwest.ifisiol.unam.mx} from Mark West {mwest-at-ifcsun1.ifisiol.unam.mx} : } Mark,
If you don't really care about seeing the starch, perhaps you could "light starve" the algae, put them in the dark for some period of time to deplete or minimize granule size of the starch, without killing them of course. Then process as usual. We do this with plant samples to avoid the embedding problems and the resultant holes in the setions that you describe.
Good luck!
Gib
} Hi all - a first attempt at unloading a problem onto the listserver! } } I am trying to get thin sections of blue-green algae that are loaded with } starch granules. The problem is that every time we do the preps, we end up } with cells that lose some granules, leaving a gaping hole, or of starch } granules that have holes in the centre. I'm using a standard sort of } method - fix in phosphate + 2.5% glut 2 hours, osmium 1% 2 hours, } dehydrate in ethanol series, finishing with prop. ox, 3:1 prop. ox/resin } around 2 h, 1:1 2 or 3h then another change overnight, a couple of 100% } resin changes for 1/2 to one day, then polymerization 48h/60C. The fixing } and EtOH steps are done on ice, the rest at room temp, the resin } infiltration is done on a rotating mixer. We've tweaked the method around } but nothing seems to work. I've tried using LR White as well, but with no } luck. Can anybody out there help!? } } Mark ******************************************** } Mark West, } Unidad de Microscopia Electronica, } (Electron Microscopy Unit) } Instituto de Fisiologia Celular, } Universidad Nacional Autonoma de Mexico, } 04510 Mexico D.F. } } tel (unidad/lab) *(525) 622 5610* } (casa/home) (525) 619 3020 } Fax (525) 616 2282 } ********************************************
Gib Ahlstrand Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN. USA. 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.cdl.umn.edu http://biosci.umn.edu/MIC/consortium.html
Me too, though I would like to attend the meeting in Philadelphia. Elaine } } } Collegues, } } In response to the current thread on the problems of facility managers, I } say count me in as being highly interested! If a discussion group does get } together at M&M it would be great to see a report posted on this listserver } for those of us who unfortunately can't attend the meeting. If somebody } could take notes and post them, I for one would be very appreciative. } } Thanks! } Dee
Dr. Elaine Humphrey Biosciences Electron Microscopy Facility 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
In my experience, two weeks of downtime a year is not bad at all, for any EM. Especially when you are dealing with many users with varying levels of expertise and a heavy usage schedule. EM's are maintenance-intensive instruments (especially TEM's), and even performing preventive maintenance routines can easily eat up a week a year.
In my opinion, you're doing great.
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: "HARRISm-at-esm-semi.co.uk"-at-sparc5.microscopy.com [mailto:"HARRISm-at-esm-semi.co.uk"-at-sparc5.microscopy.com] Sent: Tuesday, May 23, 2000 5:02 AM To: Microscopy-at-sparc5.microscopy.com
A how long is a piece of string ? type question
I would be pleased if anyone could broaden my views on equipment reliability . Basically we have a 3yr old FESEM which I consider to be fairly reliable in that it is on call 24hrs/day , has numerous ( non dedicated users ) and apart from downtime for filament change and the odd wear and tear type problems answers our needs . As we do not have a back up instrument and when we do experience problems it's always at the worse time certain personnel have the impression that it is unreliable .
What do other sem users expect in terms of reliability , apart from my ' subjective ' comments is it quantifiable , would approx 2wks /year downtime including planned maintenance be considered excessive ?
Two weeks a year (avg) seems like a lot of downtime for a FE-SEM to me. Our Hitachi S-800 SEM, for example, is 13 years old, just recently had only it's second emitter replacement, and, not counting building air, water and power problems, has certainly had only about 4 weeks of instrument-related downtime in that 13 year period, including annual PM services.
I guess it's tough to break an anvil... ;-).
Larry PS it's a heavily used multi-user instrument...
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Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
Gib, Have you thought about microwave fixation in the pressence of aldehyes (para/GA) followed by microwave fixation in osmium. each takes seconds and fixations are as good or better (for rapid fixation) than standard fixation. The key is to keep heat off the sample (use water baths and/or ice bath). The theory is that the microwave pulsations increase the penetration speed of the fixatives. Call Ted Pella's tech divison for more info or protocols.
Dear Woody, I had a problem once that really strained the ultimate Z number sensitivity of my BSE detector. This was a case of a small amount of a marker chemical, I think Strontium, being fed to growing fish and trying to detect it in a scale of the fish. I was definintely able to detect the faint, brighter band on my GW BSE detector (solid state), but not on my Robinson (scinntilator) on the other SEM. I had to use more beam current for the GW, but it saw the contrast when the Robinson didn't. More sensitivity does not mean better Z resolution. At 01:36 PM 5/22/00 -0500, you wrote: } } Hello Gary, } } I have never used a sintillator type BSE detector, but the major differences } are } two fold. } } Typically (though diodes are getting better all the time) the Robinson has a } better low energy BSE detection effiency. It follows that for the same } noise } level electronics, it would exhibit better sensitivity. The dynamic range } problem will still exist for very different Zs in the field of view. In the } middle and upper portions of the sensitivity range (low and lower), I would } not } expect much difference between them. ...Any Comments from users of both???? } } I like the 4 quad diodes since I can go differential mode for macro } topography } (like fracture surfaces) and show the gross features while suppressing the } fine } detail. } } The Pt coating can have a profound negative effect on sensitivity if not } extremely thin. If given a choice, I would always use carbon for the best } BSE } sensitivity. ...If not hazardous, I would use Be, BUT!!!! For elements in } the } range of Al & Si, I prefer to use carbon and lower beam voltages to minimize } penetration, especially if they are films or very small features. } } I once presented some data illustrating the BSE signal attenuation as a } function } of sputtered Au thickness. But that data would be hard to retrieve now. } } Woody White } McDermott Technology
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
Dear Listers, I now know that the size of the gold used in the label was 0.8 nm and the DNA in another exp. will be ssDNA (7249 bases, circular). We're prepared to try 1.4 nm and 3.0 nm and darkfield before shadowing. I have copied the repsonses below and will post the results of our efforts. Once again, I am most grateful for your help. Rosemary
I don't think the PT shadowing would obscuring the gold labelling. I did some rotary shadowing of myosin molecules with antibody attached. You couldn't see the actual Y structure but you could see arrowheads. If you can see isolated IgG molecules you should be able to pick up the gold particles. Patty Jansma
Do you mean you want to see a gold particle in a preparation that is shadowed with Pt after the gold labelling step has take place? If so, the answer is probably "yes." The Pt gives such a high-contrast shadow that it may be difficult to pick out the small gold probe against the contrasty background. Carol Heckman
NO, it should enhance the whole image so that you can more readily see exactly where the gold is labelling. Cheers,Marilyn Henderson
I would suggest to use dark field imaging of your labeled DNA-protein complexes in TEM. By using this technique, you do not need to use Pt- shadowing of your samples. Best regards from Prague. O. Benada
Rosemary, The typical "grain" size using Pt/C is on the order of 0.8 nm. So the answer to your question would be dependent on the size of the gold probe you are using. I would think that you would be OK with a gold probe larger than 3 nm, but this is speculation on my part, I have not actually done that exact thing with my own two hands. Chuck (Charles Garber)
Rosemary Walsh, Manager The Electron Microscope Facility for the Life Sciences, A Shared Technology Facility, The Life Sciences Consortium 1 South FrearLab Penn State University University Park, PA 16802 (814) 865-0212 rw9-at-psu.edu http://www.lsc.psu.edu/stf/em/home.html
Sure it is possible to melt samples by sputter coating but it is usually when you are using the old fashioned "brute force" systems that use a variac to adjust the operating voltage.
A big problem with sputter coating is that many of the solutions to one problem are the reasons for others!
1. To help prevent melting increase the working distance (} 5cms) and cut down the current (~10mA) - problem this cuts down the coating thickness so I need to coat for longer!
2. To help prevent melting use a number of short coating periods with a cooling down period between - problem multi coats build structure on the specimen and from tests I have conducted the first 10 seconds of the plasma are the hottest! We actually use the multi coat method to make test specimens ( 5 x 1 minute coats at 20mA 5cms working distance with 1 minute between coats).
3. During the early days of SEM sputter coating I would place the sputter head in a refrigerator for an hour prior to use, part of my method in order to tray and track down the heat problems. There was considerably less heating under these circumstances but we were very very careful with condensation on the then "high voltage" connection. This test led us to talk very seriously about water cooling the sputter head, a route that was taken by Baltzers at one stage.
4. My route for a specimen that melts would be to cut down the current, give a longer coating time and provided you were not working above 5,000X have a few minutes "cooling time" between no more than three coating sessions.
Good luck
Steve Chapman Senior Consultant Protrain For consultancy and professional training in EM world wide Tel 44+ 1280 814774 Fax 814007 www.emcourses.com
I have been involved with clients around the world since FEG systems first became available. I have found this style of equipment to be very reliable with if anything less down time than the conventional W hairpin instruments.
I must say that I have seen a considerable difference in the performance of these instruments with a certain manufacturer's range of FEG microscopes being far less of a problem in the production of very high quality results than any of the others.
This said if you are only averaging two weeks per year down time there should be no one within your organisation who should complain.
As a consultant and ex service engineer I can only suggest that people look at other instruments within your establishment of equal complexity and compare the up time performances.
If we can help a phone call consultation costs nothing?
Steve Chapman Senior Consultant Protrain For consultancy and professional training in EM world wide Tel 44+ 1280 814774 Fax 814007 www.emcourses.com
And another - same as Larry really, we have a four year-old Hitachi 4500 that has had no emitter deterioration and probably 5 days total downtime counting bakeouts. Touch wood. Also in a multiuser facility. But this FESEM series are proverbially reliable and in general two weeks average per year doesnt seem excessive, particularly if the FESEM is only three years old, and some teething problems with any EM column would not be unusual in the first couple of years. Depends on the context what is acceptable I guess, but if absolute reliability is a requirement, 24 hr access by "non-dedicated" users might be the first point to examine? good luck, Sally Stowe
} } } Larry Allard {l2a-at-ornl.gov} 05/24/00 12:21am } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Just another opinion...
Two weeks a year (avg) seems like a lot of downtime for a FE-SEM to me. Our Hitachi S-800 SEM, for example, is 13 years old, just recently had only it's second emitter replacement, and, not counting building air, water and power problems, has certainly had only about 4 weeks of instrument-related downtime in that 13 year period, including annual PM services.
I guess it's tough to break an anvil... ;-).
Larry PS it's a heavily used multi-user instrument...
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Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
I hope that someone can make a suggestion to me. I am looking for a sample to use as a standard for low level nanoindentation.
What would be ideal is a gold film of perhaps 1 micron or more on perhaps polished silicon wafer. This needs to be as smooth as possible, rms roughness of less than a nanometer. I know that this is quite possible, I have samples that are just like this but they are too thin (15nm).
We have a JOEL 5200 SEM with a Link Analytical QX 2000 EDS detector. The problem is that we always get an aluminum peak in every EDS sprectrum regardless of sample & scan size. Does anyone have any suggestions on why this is?
Thanks, Hanson Fong
Univ. of Washington Materials Science & Engineering Department Box 352120 Seattle, WA 98195 USA
This may be off-topic for this listserver but I'll give it a shot for those who are into materials science.
Suppose that you have an etching system based on a small plasma chamber using CF4 and O. Is there some simple/convenient way to measure species during the etching process to indicate that some end point has been reached? i.e., a major etching area has been etched and the nature of the species has dramatically or noticeably changed?
I can think of many x-ray methods, but what I am looking for is basically a sensor of some sort at a port in the etching chamber system. There is no SEM beam--and presumedly, no direct or indirect x-rays.
Are you sure it's Al and not Br? If your accelerating voltage is not high enough to stimulate the emission of Br K radiation, it can be easy to mistake the Br L lines for the K lines of Al which are slightly narrower but at the same peak position. Like the other halogens, Br can initiate corrosion of metals and end up in the corrosion products. And if this metal is part of your detector window.....
John Twilley Art Conservation Scientist
H. Fong 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 have a JOEL 5200 SEM with a Link Analytical QX 2000 EDS detector. The } problem is that we always get an aluminum peak in every EDS sprectrum } regardless of sample & scan size. Does anyone have any suggestions on why } this is? } } Thanks, } Hanson Fong } } Univ. of Washington } Materials Science & Engineering Department } Box 352120 } Seattle, WA 98195 } USA
A change in the absorption spectrum of light could to be used as a cut off. The photon penetration is of the order of the first few monolayers. The difficulty is using sources and filters of the right frequency range that is compatible with the etchants and the material you want to detect (when it becomes exposed).
You would need to find out the optical reflectivity/absorption of the materials involved. Is this feasible? ******************************************************** Dr Jonathan Barnard
Analytical Materials Physics The Angstrom Laboratory, Uppsala University P O Box 534, SE-751 21 Uppsala, Sweden Phone: +46-(0)18-4716838 Fax: +46-(0)18-500131 Phone: Microscope room +46 18 471 6365 http://www.angstrom.uu.se/analytical/home.html ********************************************************
Hi Hanson Fong, A possible explanation is that you have a collimator made of aluminium. Either the collimator is not properly aligned, or, the carbon paint inside the collimator has broken. Another explanation can be high energy x-rays hitting the collimator.
Vennlig Hilsen dr.ing Gunnar Kopstad overingeni¿r Avd f Patologi, Rit
Dr. Gary Gaugler 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 may be off-topic for this listserver but I'll give it } a shot for those who are into materials science. } } Suppose that you have an etching system based } on a small plasma chamber using CF4 and O. } Is there some simple/convenient way to measure } species during the etching process to indicate that some } end point has been reached? i.e., a major etching } area has been etched and the nature of the species } has dramatically or noticeably changed? } } I can think of many x-ray methods, but what I am } looking for is basically a sensor of some sort at a } port in the etching chamber system. There is no } SEM beam--and presumedly, no direct or indirect } x-rays. } } Any ideas? } } gary g.
Gary, There are some small, inexpensive mass-specs available that can operate at pressures up to 25 mT. I believe Ferranti in New Mexico is one that I've seen. I don't know what pressure your plasma system operates at, but if it's too high, you'd only need a roughing pump to operate this system.
When I looked into it a few years ago, it was about $3000 plus a computer to plug it into.
H. Fong 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 have a JOEL 5200 SEM with a Link Analytical QX 2000 EDS detector. The } problem is that we always get an aluminum peak in every EDS sprectrum } regardless of sample & scan size. Does anyone have any suggestions on why } this is? } } Thanks, } Hanson Fong } } Univ. of Washington } Materials Science & Engineering Department } Box 352120 } Seattle, WA 98195 } USA
Hanson, I'm not familiar with the 5200 chamber geometry, but I'm sure there is aluminum in there. The question is: can the detector see the aluminum and can the aluminum be excited by either backscattered electrons or x-rays generated by from the specimen? The detector doesn't care or know WHERE the x-rays come from, only that they are being generated and are within line of sight of the detector. Often, moving the detector closer to the specimen and making sure that the collimator is properly placed on the detector nose will help narrow the field of view.
I would check your electron trap and make sure that it is installed correctly, your detector window is almost certainly coated with Aluminum to keep light out, electrons striking the window can produce the effect you are observing. If this is the case you should also be seeing a large hump in the high end of your continuum. Good luck and let the list know what you discover. Scott
} } We have a JOEL 5200 SEM with a Link Analytical QX 2000 EDS detector. The } problem is that we always get an aluminum peak in every EDS sprectrum } regardless of sample & scan size. Does anyone have any suggestions on why } this is? } } Thanks, } Hanson Fong } } Univ. of Washington } Materials Science & Engineering Department } Box 352120 } Seattle, WA 98195 } USA
------------------- note: new mailing address ------------------------ Scott Wight fax: 301-417-1321 NIST 222/A113 W voice: 301-975-3949 100 Bureau Dr STOP 8371 | email: scott.wight-at-nist.gov Gaithersburg, MD 20899-8371 \|/ disclaimer: Any opinion expressed is my own and does not represent those of my employer.
If you use a tride sputter coater, you should not experience any significant heat build-up during a normal coating process, because the geometry of the sputter target, employing a central magnet element to create a shaped magnetic field, directs the electrons in the plasma away from the sample, thus reducing the "I-square-R" heating. If you are using an older diode sputter coater, it is a simple matter to use a pulsed coating process to reduce heating effects. We used this process many years ago before the triode sputter coaters were introduced. It turns out that a cycle of 1 sec ON, 2 sec OFF (or maybe it was 2 ON, 1 OFF) ended up generating an increase in temperature on an insulating sample only to about 40¡C, or about body temperature (directly measured using thin-wire thermocouples). We found we could coat a piece of styrofoam cup with 200 of gold using this process in a diode sputterer, and see no evidence of melting of the structure.
This is basically the suggestion Steve Chapman makes, to use several brief coating times. The more regulated, very short heating times with some cooling time in between seemed to do the trick for us. Try it, you'll like it... :-).
Larry PS I think we published this somewhere...if I find it, I'll post.
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Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
How big is the Al peak compared to other peaks? Does it vary according to the sample composition? Do you get it even if there is no sample in place? Are you sure you have the sample height correct (the "nominal" height is often not right - you need to check the X-ray count rate vs. sample height)? Are you certain it is Al (the position is accurate, the peak width right)? Do you always use the same voltage, or is the peak there at different beam voltages? Do you have a thin window or Be detector? Is the detector working normally in all other respects? Has this spurious response always been present (i.e. since installation in the '80's) or have you only recently observed it?
Sorry to ask these questions, but they are relevant.
If there really is an Al peak, it means that within the detector's field of view is something made primarily of Al which is being irradiated either with x-rays or electrons. Assuming the working distence (sample height) is correct, this implies some problem with the collimator, because it's purpose is to eliminate exactly this type of spurious x-ray signal. You do have your original collimator and electron trap, do you? Has it been damaged or moved in some accident with the sample stage?
If the Al signal varies strongly with sample composition (for example, much weaker with a carbon sample than with a tungsten sample) then it could well be related to backscattered electrons or secondary x-ray flourescence.
If, on the other hand, the signal varies strongly with operating voltage (especially if the peak moves) then it probably isn't Al at all, but a spurious response related either to electrons getting through the window or to electron noise.
No solutions here (and some of my thoughts are included for completeness as this is a positing going out for everyone to read), but I hope my musings help.
Tony Garratt-Reed.
At 05:48 PM 05/23/2000 -0700, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
** Anthony J. Garratt-Reed ** MIT Room # 13-1027 ** 77 Massachusetts Avenue ** Cambridge, MA 02139-4307 ** USA ** ** Phone: (+) 1-617-253-4622 ** Fax: (+) 1-617-258-6479 **
1) You should be able to get some spectral data from the plasma itself. Hook up a spectrometer and you should be able to see the components in the spectrum. You may have to excite the plasma with some light.
2) Hook up a mass spectrometer (quadrupole). That should be able to give you masses of the components.
Don't people do that on a regular basis? You may check the web or literature for "residual gas analysis" or similar.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== 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: Dr. Gary Gaugler [mailto:gary-at-gaugler.com] Sent: Tuesday, May 23, 2000 9:27 PM To: MSA listserver Cc: just_in_case_I_bounce
This may be off-topic for this listserver but I'll give it a shot for those who are into materials science.
Suppose that you have an etching system based on a small plasma chamber using CF4 and O. Is there some simple/convenient way to measure species during the etching process to indicate that some end point has been reached? i.e., a major etching area has been etched and the nature of the species has dramatically or noticeably changed?
I can think of many x-ray methods, but what I am looking for is basically a sensor of some sort at a port in the etching chamber system. There is no SEM beam--and presumedly, no direct or indirect x-rays.
This is the same thing as I am trying to do. In my case, the passivation will be either sinox or PSG. At present, I remove them using two methods. One is to do a short soak in BOE, rinse & dry. Then plasma etch with CF4 at about 80 mTorr. Power and gas flow have dramatic effects on etch rate. Same for dilution of BOE. The challenge is to nicely get through the passivation and stop at the uppermost SiO2 dry ox layer.
The physical size of my specimens is small. A 3" diameter cylindrical chamber would be fine. I use a sputter coater now in etch mode (quartz chamber of course). It seems to me that there would not be much of a change from a detection/measuring system after the process finishes off the passivation and reaches the SiO2. Maybe not true.
There have been several good suggestions on the list so far. One I will check out right away is the residual gas analyzer. I also may need some different type of etching unit rather than the coater operating in etch mode. Since I am interested in FA too, the specimens are too small to justify the cost of impressive huge etching units.
gary
At 06:27 AM 5/24/00, you wrote: } Hello Gary, } I am also interested in this. We make PLD and I work in the FA group. Being } this as it may I am new to this field and would wish to find a technique to } get through the passivation and intrametal layers. } Thank you for any help. } } Sincerely, } Robb Westby } Associate Reliability Engineer } Lattice Semiconductor } } "Dr. Gary Gaugler" wrote: } } } This may be off-topic for this listserver but I'll give it } } a shot for those who are into materials science. } } } } Suppose that you have an etching system based } } on a small plasma chamber using CF4 and O. } } Is there some simple/convenient way to measure } } species during the etching process to indicate that some } } end point has been reached? i.e., a major etching } } area has been etched and the nature of the species } } has dramatically or noticeably changed? } } } } I can think of many x-ray methods, but what I am } } looking for is basically a sensor of some sort at a } } port in the etching chamber system. There is no } } SEM beam--and presumedly, no direct or indirect } } x-rays. } } } } Any ideas? } } } } gary g.
The weekend workshop on Chemical Microscopy, sponsered by the New York Microscopical Society, originally sheduled for the weekend of May 20 has been rescheduled for the weekend of June 17 & 18.
This is an opportunty to learn some of the fundamentals of Chemical Microscopy from Skip Panenik of Trace Analysis.
The course will be held in West Paterson, NJ.
For further information contact Don O'Leary Phone (201) 797-8849 Fax (425) 988-1415 E-mail donoleary-at-worldnet.att.net
I am looking at this same problem for my own Plasma cleaning process. I just purchased a small, fiber optic, emission spectrometer (USB 2000) from OCEAN OPTICS to examine the light coming from my cleaning plasma. I just setup the software yesterday and will be trying it today. If I get some end point results I will repost to this thread in a few days. There is plasma etch literature that suggests that end points can be observed in the plasma emission lines.
Ronald Vane XEI Scientific (650) 369-0133
-----Original Message----- } From: Dr. Gary Gaugler {gary-at-gaugler.com} To: MSA listserver {Microscopy-at-sparc5.microscopy.com} Cc: just_in_case_I_bounce {zaluzec-at-sparc5.microscopy.com}
Al is present everywhere in the SEM. X-rays are produced by emission (electrons hitting the material) and Fluorescence (X-rays hitting the material). Collimator fluorescence is a major design problem for EDS system designers. High Z materials for stopping X-rays fluoresce strongly, and Low Z materials have no stopping power. Collimators often have high Z material lined with Aluminum to stop X-rays and then filter out the Fluorescence. If some of the AL is displaced it can be excited by either stray electrons or x-rays and cause the Al peak you see.
Ronald Vane XEI Scientific
-----Original Message----- } From: Gunnar Kopstad {gunnar.kopstad-at-medisin.ntnu.no} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
On many scopes I've found this was an artifact of an aluminum sample holder. Painting the surface of the sample holder with colloidal graphite or using a carbon planchet usually eliminates the problem.
Phil Swab Engineering Development Deposition Sciences Inc. Santa Rosa, CA 707-566-3718 phil.swab-at-depsci.com
-----Original Message----- } From: H. Fong [SMTP:hfong-at-u.washington.edu] Sent: Tuesday, May 23, 2000 5:49 PM To: microscopy-at-sparc5.microscopy.com
We have a JOEL 5200 SEM with a Link Analytical QX 2000 EDS detector. The problem is that we always get an aluminum peak in every EDS sprectrum regardless of sample & scan size. Does anyone have any suggestions on why this is?
Thanks, Hanson Fong
Univ. of Washington Materials Science & Engineering Department Box 352120 Seattle, WA 98195 USA
Dear Hanson, I solved that problem by cutting a circle of thin lead (Pb) foil to line the inside of my Al collimator. Poke a hole in the foil for the hole in the collimator. At 05:48 PM 5/23/00 -0700, you wrote:
} } } } We have a JOEL 5200 SEM with a Link Analytical QX 2000 EDS detector. The } problem is that we always get an aluminum peak in every EDS sprectrum } regardless of sample & scan size. Does anyone have any suggestions on why } this is? } } Thanks, } Hanson Fong } 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
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Hello Friends, Does anyone know if one needs to make any adjustments/compromises to Koehler illumination when using a digital camera? We are trying out a few demo's and get circular patterns or edge unevenness at optimum Koehler. If the condenser is defocused things are better...but...isn't the entire purpose of Koehler to get the most out of Abbe's equation that we can?? How much of a loss of resolution can be expected without optimum condenser settings. My sense is that one should always go with great scope alignment. I am just checking to see if any of us old microscopists can be taught a thing or two regarding digital camera set up. Thanks, Linda Fox lfox1-at-wpo.it.lumc.edu
Hi We have a client on an older Leo S200 with the standard solid state BSD fitted who has to look at slag off their stainless steel plant. In this slag there are slight compositional differences which they can only define should they run their filament on first peak. Strange but true! At first peak the slightest difference can be seen very easily, at saturation not a chance. Now I remember that Steve Chapman did give us an explanation for this the first time we mentioned it but, alas age catches up on me too and I have forgotten what it was.
Point is, try this on your system. It works for them. Better resolution on the BSD at first peak than at saturation.
Cheers Luc Harmsen Anaspec, South Africa Technical support on microscopy. www.anaspec.co.za
Remember, ICEM 15 will be in 2002, Durban, South Africa. www.icem15.com
-----Original Message----- } From: Mary Mager [mailto:mager-at-interchange.ubc.ca] Sent: Tuesday, May 23, 2000 6:17 PM To: White, Woody N Cc: Microscopy-at-sparc5.microscopy.com
Dear Woody, I had a problem once that really strained the ultimate Z number sensitivity of my BSE detector. This was a case of a small amount of a marker chemical, I think Strontium, being fed to growing fish and trying to detect it in a scale of the fish. I was definintely able to detect the faint, brighter band on my GW BSE detector (solid state), but not on my Robinson (scinntilator) on the other SEM. I had to use more beam current for the GW, but it saw the contrast when the Robinson didn't. More sensitivity does not mean better Z resolution. At 01:36 PM 5/22/00 -0500, you wrote: } } Hello Gary, } } I have never used a sintillator type BSE detector, but the major differences } are } two fold. } } Typically (though diodes are getting better all the time) the Robinson has a } better low energy BSE detection effiency. It follows that for the same } noise } level electronics, it would exhibit better sensitivity. The dynamic range } problem will still exist for very different Zs in the field of view. In the } middle and upper portions of the sensitivity range (low and lower), I would } not } expect much difference between them. ...Any Comments from users of both???? } } I like the 4 quad diodes since I can go differential mode for macro } topography } (like fracture surfaces) and show the gross features while suppressing the } fine } detail. } } The Pt coating can have a profound negative effect on sensitivity if not } extremely thin. If given a choice, I would always use carbon for the best } BSE } sensitivity. ...If not hazardous, I would use Be, BUT!!!! For elements in } the } range of Al & Si, I prefer to use carbon and lower beam voltages to minimize } penetration, especially if they are films or very small features. } } I once presented some data illustrating the BSE signal attenuation as a } function } of sputtered Au thickness. But that data would be hard to retrieve now. } } Woody White } McDermott Technology
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
Optical Coating Laboratory, Inc. (OCLI) a JDS-Uniphase company is looking to fill a position in the microscopy area in its Analytical Laboratory . The candiddate is to have an MS or BS degree in materials science, physics or related areas with experience in AFM , Light microscopy, FTIR, interferometry...experience in SEM and other electron microscopy, surface chemistry analysis is a plus. The assumption of the position is immediate. OCLI is located in Santa Rosa, California, a leader in thin film products for the telecommunication industry, counter-feiting applications, photonics and a variety of other products (please visit OCLI's web site: http://www.ocli.com/career_opps/index.htm, for more information about the open position and OCLI and its products.) If you or any microscopist you know is interested, please send your resume to Human Resources, Att: Earl Jensen or to me directly by responding to this email or to my address: Said A. Mansour 2789 Northpoint Parkway MS 274-3 Santa Rosa, CA 95407
Our SEM room is a little too noise for high res SEM work, so the service engineering recommended to dampen the noise. I know there is panels I can put up on the wall to do this. What is the cheapest solution for doing this?
I have worked on Noran Systems for 10 years. I have seen this problem several times and always on a JEOL SEM. Most times it was not the detector. It usually was a problem with alignment of the beam. The obvious point here is that detectors do not produce x-rays, they detect them.
} Hello Friends, } Does anyone know if one needs to make any adjustments/compromises to Koehler illumination when using a digital camera? We are trying out a few demo's and get circular patterns or edge unevenness at optimum Koehler. If the condenser is defocused things are better...but...isn't the entire purpose of Koehler to get the most out of Abbe's equation that we can?? How much of a loss of resolution can be expected without optimum condenser settings. } My sense is that one should always go with great scope alignment. I am just checking to see if any of us old microscopists can be taught a thing or two regarding digital camera set up. .} } } } } } } } } } } } } } } } } } .
You are giving up a lot of resolution with a digital camera due to the pixel spacing so the loss of resolution due to non optimal illumination have little effect on the image quality.
You can also solve the problem by increasing the distance from the CCD to the eyepeice so the ragged edge doesn't fall on the the CCD. This would also reduce the loss of resolution due to the spacing of the pixels. Of course is aslo decreases the coverage of the image.
Gordon
Gordon Couger gcouger-at-couger.com
Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
My Amray 1910 FESEM is only used by me. It is up as long as I say so (don't shut it down, put it in failsafe mode, etc.). PM takes about one day and this is done per contract twice a year. The only real bummer is when all 3 apertures have gone bad, one-by-one over time. Vent, pull the holder, change out the apertures and evacuate. This takes me about 45 minutes to accomplish. Each aperture typically lasts about 2 months. Since Amray gold flashes them, they cannot be flamed. Guess that is why they call them "consumables."
The only major down time I experienced was with my 1830 load lock system. The Balzers 240 turbo was going out (high frequency oscillation). That took about 3 days to fix for a total pump exchange. Other than this, both systems are very easy to keep running.
If they were in a mixed user environment, I'd opt for the FESEM over the LaB6. The FESEM is rather tough to screw up....unless of course, someone really worked at it.
gary g.
At 04:37 PM 5/23/00, you wrote: } [snip] } } ------------------------------------------------------------------------ } } 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 } } -----------------------------------------------------------------------. } } } } } } } } A how long is a piece of string ? type question } } } } } } I would be pleased if anyone could broaden my views on equipment } } reliability . } } Basically we have a 3yr old FESEM which I consider to be fairly } } reliable in that it is on call 24hrs/day , has numerous ( non } } dedicated users ) and apart from downtime for filament change and } the } } odd wear and tear type problems answers our needs . } } As we do not have a back up instrument and when we do experience } } problems it's always at the worse time certain personnel have the } } impression that it is unreliable . } } } } What do other sem users expect in terms of reliability , apart from } my } } ' subjective ' comments is it quantifiable , would approx 2wks } /year } } downtime including planned maintenance be considered excessive ? } } } } Regards } } Martyn Harris } } harrism-at-esm-semi.co.uk } } Dr. Lawrence F. Allard } Senior Research Staff Member } High Temperature Materials Laboratory } Oak Ridge National Laboratory } 1 Bethel Valley Road } Bldg. 4515, MS 6064 } PO Box 2008 } Oak Ridge, TN 37831-6064 } } 865-574-4981 } 865-576-5413 Fax } allardlfjr-at-ornl.gov
Assuming that the noise is being generated from equipment in the room then curtains will reduce the noise level quite effectively.
Ron
On Wed, 24 May 2000 19:22:19 -0700 (PDT) Ben Craft {bcraft-at-uci.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 } -----------------------------------------------------------------------. } } } Our SEM room is a little too noise for high res SEM work, so the service } engineering recommended to dampen the noise. I know there is panels I can } put up on the wall to do this. What is the cheapest solution for doing } this? } } } } ####### } #####\_O -Ben Craft- } ####/\/} } #### /" } ### \ } } } }
---------------------- 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
} ------------------------------------------------------------------------ } 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 Sergey and others, } } I want to add my 2 cents as I like the topic. } Most of this is a result of my experience with our TEM 2Kx2K CCD. } We are working in bright field - so I cannot comment on dark field } performance } Several points: } } 1. The CCD is much more convenient - you get your pictures instantly. } } 2. CCD is linear and has larger dynamic range than the film. } } 3. The bad thing about the CCD is resolution - about 4 times lower than that } of the film (in our camera the pixel size is 30 microns).
If You would choose a CCD with smaller pixel size You would get a better resolution.
} So if you want to } work in minimum dose you will do better with film.
Never, a good CCD is much more sensitive than a film.
} As I know the CCD is not } performing well in terms of signal to noise at low doses (and if you have to } work at 4 times higher magnification because of the resolution the things } become much worse).
see above , a smaller pixel gives a better sensitivity and a better resolution.
} } } 4. The CCD has smaller observation area - again loss of information.
use the so called Image mounting, than You get very large images with much more image information due to the larger dynamic range and better sensitivity.
} } } 5. I don't know about the detection efficiency compared to the film - it } depends on the thickness of the phosphorous and the accelerating voltage. If } a photon reaches the CCD chip it will be detected ... the problems are in } the conversion electron-photon.
The currently leading CCD systems reach single electron sensitivity at thin phosphor screens and good resolution.
} There are two sides - if you make the } phosphorous thicker you will get higher detection efficiency but the point } spread also increases so always a compromise is made between detection } efficiency and resolution. When I say detection efficiency this is not only } related to the detection of single electrons (as it detects single } electrons) but more to the actual signal detected on the background of the } noise. Apart from the shot noise additional noise is added due to the } scintillator driven detection and thermal noise in the CCD chip.
In good, highly sensitive CCD systems the Poisson noise of the incoming signal is dominating, not the CCD noise.
} } } The CCDs are now very popular in diffraction work because of the dynamic } range and linearity. } } Here is one reference where a nice comparison between 2Kx2K CCD and film has } been made: } } Downing, K., Hendrickson, F., Ultramicroscopy, 75, (1999), 215-233
Thanks for that.
} } } Best regards, } } Rado } } --------------------------------------------------------------------- } Radostin Danev } Laboratory of Ultrastructure Research } National Institute for Physiological Sciences } Myodaiji-cho, Okazaki 444-8585, JAPAN } e-mail: rado-at-nips.ac.jp } ---------------------------------------------------------------------
-- Best regards / Mit freundlichen Gruessen Dr. Frank Jenichen Proscan elektronische Systeme GmbH Tel.: +49 8195 999 -511 Fax: -512 mailto:Jenichen-at-proscan.de ------------------------------------------------------------ More information concerning our products and services can be found on our website http://www.proscan.de
I have a question about the study of for instance the translocation of a fluorescent labeled cellular component from the cytoplasm to the nucleus. At first it seems that labeling the component you want to study and do a counterstain for the nucleus would be sufficient to give an idea of the migration of a componenent from the cytoplasm to the nucleus or not.
By doing the experiment this way however you do not have a clue about the total cell content, because the cytoplasm is not counterstained with a background stain to show the cell outlines to give an idea of the actual cell extent. Without a cytoplasm stain, you have no idea of the actual size of the cell in which the label for the cellular componenent resides. For an "absolute" idea of the migration I think a background cellular counterstain is necessary ?
Also the nuceus is thicker than the cytoplasm, so for a given focuslevel inside the nucleus there is more light falling in the lens form above and below than in the cytoplasm, which probably will give a non-linear response curve for the quantification of the translocation ?
A clear case for doing the experiment with a fluorescent probe with a confocal or multiphoton microscope! Either way, out-of-focus contributions to intensity are not important. Your confocal image is also a sample from a well-defined volume of cell or tissue. Therefore, provided you can regard each compartment as homogeneously labelled the total compartment (e.g. cytoplasm, nucleus) volume does not need to be determined.
} From: "Van Osta, Peter [JanBe]" {PVOSTA-at-janbe.jnj.com} To: Microscopy-at-sparc5.microscopy.com
McMaster-Carr has some of the best selection for sound deadening material and generally a better price than specialty dealers or other distributors. Their Web site is very functional http://www.mcmaster.com Delivery has always been more than prompt and they have more indespensible items for any microscope lab. No lab should go without one of their catalogs.
Sound control products are in my catalog on page 2777-2779 products ranging from flat foam to sculptured foam to "sono-tech" foam to acoustical quilts to acoustical cylinders. Looking through this stuff isn't cheap but of the products I have seen offered other places the prices here are competitive.
Of course you could always head to a carpet store and dig through their dumpsters for throw-away remnants and hang them on the walls in the scope room. Two or three layers might work well enough - not sure about the smell though. . .
Good luck Geoff
Ben Craft wrote:
} Our SEM room is a little too noise for high res SEM work, so the service } engineering recommended to dampen the noise. I know there is panels I can } put up on the wall to do this. What is the cheapest solution for doing } this? } } ####### } #####\_O -Ben Craft- } ####/\/} } #### /" } ### \
-- Geoff Williams,
Electron Microscope Facility Supervisor Biology Department Brooks Hall Central Michigan University Mt. Pleasant, MI 48859
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Linda, Look for a digital camera that allows you to capture an image of the illumination pattern with your specimen slide removed from the beam path. This "background" image should then be automatically subtracted from your final image. This will eliminate not only uneven illumination but also small light distortions due to dirt on lenses (that you cannot get off by cleaning external surfaces), etc. Using this feature permits good Koehler illumination and very even illumination on your final image file. As an example, the SPOT RT software has a feature called "Flatscreen". I capture images from each objective after checking for proper Koehler illumination. These are stored and easily called up as needed. However, the microscope alignment should still be rechecked prior to capturing images. This is a very important feature if you do Nomarski/DIC imaging. You can easily smooth out the very directional illumination pattern by capturing the lighting pattern without the sample and then subtracting it automatically when capturing the sample image.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
--------------------------------------
Hello Friends, Does anyone know if one needs to make any adjustments/compromises to Koehler illumination when using a digital camera? We are trying out a few demo's and get circular patterns or edge unevenness at optimum Koehler. If the condenser is defocused things are better...but...isn't the entire purpose of Koehler to get the most out of Abbe's equation that we can?? How much of a loss of resolution can be expected without optimum condenser settings. My sense is that one should always go with great scope alignment. I am just checking to see if any of us old microscopists can be taught a thing or two regarding digital camera set up. Thanks, Linda Fox lfox1-at-wpo.it.lumc.edu
In a message dated 5/25/00 10:26:43 AM, sherman-at-btny.purdue.edu writes:
} Look for a digital camera that allows you to capture an image of the } illumination pattern with your specimen slide removed from the beam path. } This "background" image should then be automatically subtracted from your } final image. This will eliminate not only uneven illumination but also } small light distortions due to dirt on lenses (that you cannot get off } by cleaning external surfaces), etc. Using this feature permits good Koehler } illumination and very even illumination on your final image file.
One additional note. Using a background image captured with a log-response camera (e.g., a Vidicon) does call for subtraction. For a linear response camera (most CCDs unless you are using some built-in gamma circuitry) you want to divide by the background (ratio of signal to background). Also, the problem with this method is that it uses some of your dynamic range, so you effectively cannot handle as great a range from bright to dark.
Ben:Ê we had the same problem in in our SEM/FIB rooms.Ê We got large sheets of egg-crate foam and glued them to the walls.Ê It gives the place sort of a "rubber room" appearance, but it works really well.
Ben Craft wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } ToÊ Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Our SEM room is a little too noise for high res SEM work, so the service } engineering recommended to dampen the noise.Ê I know there is panels I can } put up on the wall to do this. What is the cheapest solution for doing } this? } } ####### } #####\_OÊÊÊÊÊÊÊÊÊÊÊ -Ben Craft- } ####/\/} } #### /" } ###Ê \
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky HoldfordÊ (r-holdford-at-ti.com) 972-598-1291 (pager) KFAB Physical Analysis Labs--SEM/FIB/FA Kilby Center West Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Ê
I received an AO fluorescence microscope from a kind colleague but need to find an instruction manual for it. The microscope is an AO model 10 or 20 and it has a vertical fluorescence illuminator (model 2071) attached. Also, I'm looking for a trinocular head for the microscope to do photography. Any assistance in locating these would be greatly appreciated.
David A. Doe -- Dr. David A. Doe Biology Department Westfield State College Westfield, MA 01086 413/572-5291 fax: 413-562-3613
The amount of backscatter generated from a specimen depends on the kV, the probe size and the composition of the material under investigation. If the level of backscatter is insufficient under "normal" saturation conditions i.e. the gun is correctly saturated, then by de-saturating the larger source will result in a larger probe dimension on the specimen; increasing the probe diameter increases the volume of material involved in the production of BSE.
Steve Chapman Senior Consultant Protrain For consultancy and professional training in EM world wide Tel 44+ 1280 814774 Fax 814007 www.emcourses.com
I have been following off and on this discussion on CCD cameras for EM. Much of this discussion has been concerned with "sensitivity". I am being naive here, but when we talk about "sensitivity" of CCDs, isn't this the same thing as the QE of camera/chip? I don't think I have ever seen a QE for em CCD's for various accelerating voltages/wavelenghts. Does the electron beam directly hit the silicon photodyodes or it there an interface that converts the incoming electrons to different (longer?) wavelengts? I know em films are sensitive to specific acc voltages. Are CCD cameras for em the same. For long exposures it may not matter, but for short exposures or low level intensity does the QE of the camera come into play?
} ===== Original Message From Jenichen {Jenichen-at-proscan.de} ===== } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hank Adams Manager Integrated Microscopy Core Molecular and Cellular Biology Baylor College of Medicine Houston, Tx 77025
Microscopy Experts, We have recently inherited a Varian 936-40 Porta-Test leak detector. It came with out any documentation. Big surprise, I know. I called Varian and they offered to sell me an operators manual for $185.00. This seems just a bit excessive. If anyone has one, I would be happy to pay a Xeroxing and shipping fee. TIA Kim DeRuyter Electron Microscopy Technician 308 Natural Science Facility P.O. Box 755780 University of Alaska Fairbanks, AK 99775-5780 907-474-5452 907-474-5163 fax
} } I have worked on Noran Systems for 10 years. I have seen this problem several } times and always on a JEOL SEM. Most times it was not the detector. It } usually was a problem with alignment of the beam. The obvious point here is } that detectors do not produce x-rays, they detect them. } } Regards, } } Craig Theberge }
Did you ever figure out what sort of alignment problem it was, and from exactly what the Al X-rays were being produced?
cheers
rtch
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Looking for spare parts for an ISI-40 SEM. Particularly, column pieces, apertures, filament assembly and wehnelt cylinders. Any suggestions would be welcome.
Kim DeRuyter Electron Microscopy Technician Room 308 Natural Sciences Facility P.O. Box 755780 University of Alaska Fairbanks, AK 99775-5780 907-474-5452 907-474-5163 fax
Hi friends I agree Steve, sometimes (I seem it depends on electron gun design and distance between wenelt and anode, wenelt and filament) the first peak on saturation curve is more than saturation level even in secondary emission signal. Victor Sidorenko, ANTRON Co.Ltd., Moscow, Russia
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Has anyone knowledge of a method to rapidly fix adherent cell cultures so as to prevent the loss of small hydrophobic molecules? The problem involves subsequent diffusion of the antibody marker into the cytoplasm. Material is examined using fluorescence/confocal microscopy The organelles of interest in this case are mitochondia but general recommendations would be most appreciated too.
Regards
Andrew McNaughton
______________________________________________________________________________ Andrew McNaughton South Campus Electron Microscope Unit C/-Department of Anatomy and Structural Biology School of Medical Sciences University of Otago PO Box 913, Dunedin NEW ZEALAND
At 05:07 PM 25-05-2000 +0100, you wrote: Steve Chapman wrote,
} Hi } } Yes Luc is right I did give him an explanation. } } The amount of backscatter generated from a specimen depends on the kV, the } probe size and the composition of the material under investigation. If the } level of backscatter is insufficient under "normal" saturation conditions } i.e. the gun is correctly saturated, then by de-saturating the larger source } will result in a larger probe dimension on the specimen; increasing the } probe diameter increases the volume of material involved in the production } of BSE. } But how does this lead to an increase in the BSE differentiation (implying more signal and hence less noise). Surely just defocussing would do the same thing. Defocussing as such should not effect the BSE coeffecient, unless you had sub surface charging or some other artefact.
Very interesting! Ken.
Moran Scientific Pty Ltd P.O. Box 651 Goulburn NSW 2580 Australia Tel 02 4844 4234 (International, 61 2 48444234) Fax 02 4844 4291 (International, 61 2 48444291) Email {kmoran-at-goulburn.net.au} Web Page http://www.goulburn.net.au/~kmoran/ "Patience accomplishes its object, while hurry speeds to its ruin. Gulistan 1258"
} ------------------------------------------------------------------------ } 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 following off and on this discussion on CCD cameras for EM. } Much of this discussion has been concerned with "sensitivity". I am being } naive here, but when we talk about "sensitivity" of CCDs, isn't this the same } thing as the QE of camera/chip?
No, sensitivity means how many electrons You need for a digital response from the CCD.
} I don't think I have ever seen a QE for em } CCD's for various accelerating voltages/wavelenghts. Does the electron beam } directly hit the silicon photodyodes or it there an interface that converts } the incoming electrons to different (longer?) wavelengts?
For TEM investigations the high energy electrons (80 - 400 keV) hit a scintillator (YAG- or Phosphor-screen). These screen emits visible photons (energy in the region 2eV) which are detected by the CCD. If we would use the high energy electrons directly onto the CCD the CCD would be damaged.
} I know em films are } sensitive to specific acc voltages. Are CCD cameras for em the same.
The response for a phosphor scintillator rises linear with the energy, but reaches saturation for high energies (higher than 200keV). This response depends also on the material You use and on the thickness of the screen.
} For long } exposures it may not matter, but for short exposures or low level intensity } does the QE of the camera come into play?
If You want to get a good statistics of Your signal a response of one digital count for one electron would be very good. If the application gives You only a small amount of electrons to detect (Filter applications, low contrast applications, biological application and other) the sensitivity (reponse) of the camera should be higher to avoid long exposure times to overcome problems with drift an sample damage. So the best cameras optimized for high sensitivity (the screen is directly coupled with a fibreoptic to a cooled slow-scan CCD with up to 16bit digitization) reach more than one digital count per incident electron to have a good compromise between sensitivity and good statistics. Standard system with optical coupling do not reach this sensitivity and can be used only for applications with high beam density.
} } } } ===== Original Message From Jenichen {Jenichen-at-proscan.de} ===== } } ------------------------------------------------------------------------ } } 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 } } -----------------------------------------------------------------------. } } } } } } } } } } Radostin Danev schrieb: } } } } } ------------------------------------------------------------------------ } } } 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 Sergey and others, } } } } } } I want to add my 2 cents as I like the topic. } } } Most of this is a result of my experience with our TEM 2Kx2K CCD. } } } We are working in bright field - so I cannot comment on dark field } } } performance } } } Several points: } } } } } } 1. The CCD is much more convenient - you get your pictures instantly. } } } } } } 2. CCD is linear and has larger dynamic range than the film. } } } } } } 3. The bad thing about the CCD is resolution - about 4 times lower than } that } } } of the film (in our camera the pixel size is 30 microns). } } } } If You would choose a CCD with smaller pixel size You would get a better } } resolution. } } } } } So if you want to } } } work in minimum dose you will do better with film. } } } } Never, a good CCD is much more sensitive than a film. } } } } } As I know the CCD is not } } } performing well in terms of signal to noise at low doses (and if you have } to } } } work at 4 times higher magnification because of the resolution the things } } } become much worse). } } } } see above , a smaller pixel gives a better sensitivity and a better } resolution. } } } } } } } } } } } 4. The CCD has smaller observation area - again loss of information. } } } } use the so called Image mounting, than You get very large images with much } more } } image information due to the larger dynamic range and better sensitivity. } } } } } } } } } } } 5. I don't know about the detection efficiency compared to the film - it } } } depends on the thickness of the phosphorous and the accelerating voltage. } If } } } a photon reaches the CCD chip it will be detected ... the problems are in } } } the conversion electron-photon. } } } } The currently leading CCD systems reach single electron sensitivity at thin } } phosphor screens and good resolution. } } } } } There are two sides - if you make the } } } phosphorous thicker you will get higher detection efficiency but the point } } } spread also increases so always a compromise is made between detection } } } efficiency and resolution. When I say detection efficiency this is not only } } } related to the detection of single electrons (as it detects single } } } electrons) but more to the actual signal detected on the background of the } } } noise. Apart from the shot noise additional noise is added due to the } } } scintillator driven detection and thermal noise in the CCD chip. } } } } In good, highly sensitive CCD systems the Poisson noise of the incoming } signal } } is dominating, not the CCD noise. } } } } } } } } } } } The CCDs are now very popular in diffraction work because of the dynamic } } } range and linearity. } } } } } } Here is one reference where a nice comparison between 2Kx2K CCD and film } has } } } been made: } } } } } } Downing, K., Hendrickson, F., Ultramicroscopy, 75, (1999), 215-233 } } } } Thanks for that. } } } } } } } } } } } Best regards, } } } } } } Rado } } } } } } --------------------------------------------------------------------- } } } Radostin Danev } } } Laboratory of Ultrastructure Research } } } National Institute for Physiological Sciences } } } Myodaiji-cho, Okazaki 444-8585, JAPAN } } } e-mail: rado-at-nips.ac.jp } } } --------------------------------------------------------------------- } } } } -- } } Best regards / Mit freundlichen Gruessen } } Dr. Frank Jenichen } } Proscan elektronische Systeme GmbH } } Tel.: +49 8195 999 -511 Fax: -512 } } mailto:Jenichen-at-proscan.de } } ------------------------------------------------------------ } } More information concerning our products } } and services can be found on our website } } http://www.proscan.de } } Hank Adams } Manager } Integrated Microscopy Core } Molecular and Cellular Biology } Baylor College of Medicine } Houston, Tx 77025
-- Best regards / Mit freundlichen Gruessen Dr. Frank Jenichen Proscan elektronische Systeme GmbH Tel.: +49 8195 999 -511 Fax: -512 mailto:Jenichen-at-proscan.de ------------------------------------------------------------ More information concerning our products and services can be found on our website http://www.proscan.de
I suppose in principle a line-scanner type of CCD could be made to do this, but the practical difficulties in setting it up would be enormous - the scan system would have to be synchronously stepping with the vertical progression of the SEM scan, and the alignment and line geometry of the system would also have to be exactly right. Not at all easy to achieve. Also, it is fairly well understood that the resolution of the record screen underrepresents the resolution of the raw signal fed to it (partly to ensure that lines are not visible in the image). Therefore this just seems to be the wrong approach. There are plenty of low-cost (~10% of the cost of this camera) image grabbers for SEM that digitise the stream of analogue data fed to the record tube.
It would be more interesting to consider whether this type of camera can contribute to high quality TEM imaging. I am not clear what is limiting the resolution of current TEM digital cameras - could a high-resolution CCD camera approach the resolution of TEM film more closely than the current generation of these, or is the phosphor/YAG not good enough to make it worthwhile.
Chris
Date sent: Tue, 16 May 2000 09:06:16 -0500 To: {Microscopy-at-sparc5.microscopy.com} } From: John Foust {jfoust-at-threedee.com}
Dear List Members,
Does anyone know of a source for glass coverslips which have been treated with something to optimize cell growth? A colleague of mine is looking for some, particularly round ones.
Thanks.
Matthew J. Schibler Ph.D. UCLA Brain Research Institute 1524A Gonda (Goldschmied) Center for Neuroscience and Genetics Los Angeles, CA 90095-1761
On Mon, 15 May 2000 11:19:12 -0700, Jon Krupp 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: } } A newly appointed researcher here has asked my advice on several pieces of } TEM spec. prep equipment. I turn to you for helpful suggestions. } } The research involves serial sectioning biological tissues and many grids. } EM is a minor, but essential component of the project. The lab runs through } dozens of grids and hundreds of pictures, but only 4 - 6 times a year. Big } bursts of activity followed by long periods of analysis and investigations } using other techniques. Of the hundreds of pictures taken, they may only } use a few for data. } } The researcher is looking for ideas on what choices are available, how } useful, and approximate costs of the following: } } Digital imaging to add to our existing JEOL 1200EX TEM - I have seen other } threads on this topic and would welcome any new ideas. Digital imaging will } save a lot of time and money since they discard so many pictuers, but the } question of image quality is one I am to investigate. } } Ultra microtome - We have an older A/O Ultracut (the model before it became } the Reichert Ultracut E) which is OK for the sectioning we do in the } general lab. But she wants a new one for her exclusive use. The question is } whether a new microtome will allow folks in her lab to do serial } sectioning any faster or easier, or by less skilled users, than our current } system. } } Staining machine - Anyone have info on staining machines or systems for } lots of TEM grids. I have never had to do so many grids that this was an } issue, so I have never kept up on the offerings. If you know of something } and/or have experience let me know. Again, this is something she would keep } in her lab. } } Tissue processing machine - Same as above for me, never did so much at one } time that I ever thought I needed one of these. The samples to be processed } are C. elegans, anything available that could do these unattended? How } about upkeep and volumes of chemicals needed. Also an item to be kept in } her lab. } } I will filter and pass on your comments. Anything you might offer will, as } always, be received with appreciation and thanks. } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } } } Jon:
A few comments about the equipment. I have done literally thousands of serial sections, so I think I can offer some first hand comments.
First: My microtome of choice for serial sectioning has been the old Reichert OMU-3--the predecessor of the Ultracut E. I have to admit that part of the reason was not wanting to take the time for a learning curve. The other reason was that I found the illumination system on the old OMU-3 to be outstanding (I did repetitive serial thins and thicks over several hundred microns to about a millimeter).
Second:Staining machine--I use/have used the LKB/Leice Ultrastainer. The original LKB unit was a marvel--we did over 300 grids at one point, losing only one (stuck the forceps through the formvar!!!), and no precipitates. Can't say the same for the Leice unit, although it should be pretty much the same. The stains are the biggest variable, as is a really rigorous cleaning regime. Check it out.
Third: Processor--I use/love/hate the Lynx unit (currently available through EM Sciences). I had an early unit (from Australia)--no problems over 3 or 4 years. Have a Leica unit here--it took 4 or 5 years to get it to work consistently, but now seems ok, and it gets substantial use in bursts. Check out the RMC/Ventana unit--it is the progeny of the old LKB unit, and is probably a worthy competitor. (I just can't get one for demo--they've been promising for nearly 4 years, but I think they gave up after last year.) Maintenance on both is fairly routine, and consumables are not prohibitive. Both use very small volumes, and can be used osmium through pure epoxy (if your protocol is limited to 20 or so steps). Personally wouldn't be without one.
Hope this helps.
Roger C Moretz, Ph.D. Dept of Toxicology BI Pharmaceuticals
Disclaimer: I have no financial interest in any of the products or suppliers. Just a long time user.
_______________________________________________________ Get 100% FREE Internet Access powered by Excite Visit http://freelane.excite.com/freeisp
Do the cells need some sort of support matrix (e.g., collagen) or are they just not fond of glass? We do our own treating....collagen, poly-lysine, there is a mussel protein (not a spelling error - I think it is the byssal thread stuff from Mytilus sp.) that is sticky (CellTak?)...you can also dissolve PS culture dishes in solvent and coat glass coverslips with the resulting goo.
Tamara Howard CSHL
On Tue, 16 May 2000, Schibler, Matthew 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, } } Does anyone know of a source for glass coverslips which have been treated } with something to optimize cell growth? A colleague of mine is looking for } some, particularly round ones. } } Thanks. } } } Matthew J. Schibler Ph.D. } UCLA Brain Research Institute } 1524A Gonda (Goldschmied) Center } for Neuroscience and Genetics } Los Angeles, CA 90095-1761 } } (310) 825-9783 } FAX (310) 206-5855 } E-mail: mschibler-at-mednet.ucla.edu } } }
I just got off the phone with Stacie Kirsch, and discovered that I was the 3rd caller today asking about 'something in Philaldelphia'! Debby Sherman was first and is to talk to the Meeting Arrangements person, or some such, but I (and probably some accompanying colleagues) would be happy to participate if something informal were organized for the Sunday afternoon. Our Group at this Canadian federal lab:
- has 8 different beam instruments in 7 distinct 'labs' (and formally accesses another 2 at an on-site private sector service provider), - is manned by 18 scientists and technologists, the majority of which are permanent, - does a ~50-100 project/yr mix of contract work and core research projects for and with internal programs and external clients (academics, industrial, other federal), - has a significant number (too many!) of internal and external operators, and - has a reasonable operating budget (but currently no capital), so - we have likely encountered some variation of almost every conceivable problem (and solved only a fraction).
If something comes to pass, now or next year, I would gladly share experiences with others, especially as this is an aspect of delivering science that is commonly overlooked. A couple of thoughts:
- this could easily turn into a 'gripe-athon'. Someone should be prepared to lead it and direct it towards problem-solving if this occurs. - Ron Anderson and I have made sporadic attempts over the years to lead something called " Factors Influencing the Establishment of a New TEM Facility" at numerous workshops, often with great success. This also touched upon many of the real-world issues of an EM lab, though from the slightly more positive viewpoint of actually some hard cash in hand. -
---------- From: Elaine Humphrey [SMTP:ech-at-unixg.ubc.ca] Sent: May 16, 2000 11:39 AM To: Microscopy-at-sparc5.microscopy.com Subject: Re: SEM facility managers
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Me too, though I would like to attend the meeting in Philadelphia. Elaine } } } Collegues, } } In response to the current thread on the problems of facility managers, I } say count me in as being highly interested! If a discussion group does get } together at M&M it would be great to see a report posted on this listserver } for those of us who unfortunately can't attend the meeting. If somebody } could take notes and post them, I for one would be very appreciative. } } Thanks! } Dee
Dr. Elaine Humphrey Biosciences Electron Microscopy Facility 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
Guess it depends on what you mean by optimize. The intestinal cell lines i work with prefer bare glass (i wash in acetone, then ethanol, then lots of dH2O, then boil in dH2O, then place each one on a piece of filter paper so they aren't overlapping and then autoclave - a pain in the neck but I think it really matters). i seed the cells in serum containing medium without any other pre-treatement
I have coated with collagen, fibronectin, etc by placing coverslips in 24 well trays and adding the matrix material. this makes no difference or reduces differentiation of my cells.
many labs report cell lines that differentiate better on permeable filters so nutrients have access to the basolateral membrane. } } Dear List Members, } } Does anyone know of a source for glass coverslips which have been treated } with something to optimize cell growth? A colleague of mine is looking for } some, particularly round ones. } } Thanks. } } } Matthew J. Schibler Ph.D. } UCLA Brain Research Institute } 1524A Gonda (Goldschmied) Center } for Neuroscience and Genetics } Los Angeles, CA 90095-1761 } } (310) 825-9783 } FAX (310) 206-5855 } E-mail: mschibler-at-mednet.ucla.edu
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)
I very much agree with others in this issue. Unfortunately, I won't be able to attend this year's meeting, but I will love to find out what others had to say about managing a multi-user EM facility. Please post the highlights of that meeting if it takes place in Philadelphia.
Thanks in advance,
Soumitra
***************************************************************** Soumitra Ghoshroy Ph.D. Electron Microscopy Lab Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3600 Fax: 505-646-5665 e-mail: ghoshroy-at-nmsu.edu http://confocal.nmsu.edu/eml
Hi all, I believe to be on track with mounting clay rich rocks (that disagregate readily in perturbed water (i.e. shaken or sonicated), but not in ethylene glycol or ethanol (at rest). My goal is to mount the samples (whole, not seperates) for analysis in TEM. I have been soaking them in LR-White at 60C, where fixation occurs without accelerator in less than 24 hours. I am planning to (but have not yet) microtome the samples, or potentially ion-mill. Thoughts for the future are to use a set of glassware with a Millipore filter and vacuum to pull ethanol(cleanser and dilator), ethylene glycol(for swelling clay component), and an LR White "chaser" to view "saturated" textures, as opposed to compacted. My question is this: Can anyone point out pitfalls with this approach that I am not seeing, again I haven't tried the whole thing yet, but am in a position to start prepping the samples. In particular, is microtoming preferred to ion-milling for weak, soft samples that rely on epoxy for reinforcement? Does LR-white readily pull through a sample given a weak vacuum and a porous plate (its viscous, but not as viscous as water, for example)? Does swelling the clays with ethylene glycol and the like, and then directly mounting, introduce volatiles into the column under 120-200 kV? Any recommendations are welcome, the literature helps a bit, but is usually sketchy about these fine details of preparation procedure. thanks in advance, N
_____________________________ Nicholas W. Hayman \ Dept. of Geological Sci., U.W.\ http://www.geology.washington.edu/~hayman Box 351310, Seattle WA 98195 \_________________________________________
Dear John, That is exactly what passive image capture systems like Quartz PCI do. At 09:06 AM 5/16/00 -0500, you wrote:
} At 04:02 PM 5/15/00 -0700, Dr. Gary Gaugler wrote: } } It seems to me that no digital camera system would work on a SEM } } in place of a Polaroid or other film-based output device. Since the } } recording CRT in a SEM is based on a sequential line scan, one } } would need a camera that would capture each line as it is produced. } } What you're saying is there must be a system out there that } digitizes that single stream of line scan intensities, then } processes all that data inside the computer as an image as } opposed to trying to digitize the frame buffer. } } - John 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
some of my users think it is tome to retire our ancient but still functional cryostat. Could anyone who has bought one in the recent past give me some info about who is making them these days as well as any pros and cons of these new-fangled models.
} From: "Chris Jeffree" {cjeffree-at-srv0.bio.ed.ac.uk} } } } I suppose in principle a line-scanner type of CCD could be made to } do this, but the practical difficulties in setting it up would be } enormous - the scan system would have to be synchronously } stepping with the vertical progression of the SEM scan, and the } alignment and line geometry of the system would also have to be } exactly right. Not at all easy to achieve. Also, it is fairly well } understood that the resolution of the record screen underrepresents } the resolution of the raw signal fed to it (partly to ensure that lines } are not visible in the image). } Therefore this just seems to be the wrong approach. There are } plenty of low-cost (~10% of the cost of this camera) image } grabbers for SEM that digitise the stream of analogue data fed to } the record tube. } } It would be more interesting to consider whether this type of } camera can contribute to high quality TEM imaging. I am not clear } what is limiting the resolution of current TEM digital cameras - } could a high-resolution CCD camera approach the resolution of } TEM film more closely than the current generation of these, or is } the phosphor/YAG not good enough to make it worthwhile. }
If you control the scan you only need a single light sensitive element. You step the beam digitize the light, step the beam, wait for the last spot to go out and repeat. You don't need a camera. You do need a very fast responding system for changing electron beams to light.
This system has the resolution of the scan beam. It would not be particualy fast but you could increase the number of bits resolution to as large a number as you wanted.
Gordon
Gordon Couger gcouger-at-couger.com
Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
At 10:08 AM 5/16/00, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
This is true except that the record CRT has a fixed resolution. These are typically 2000 horizontal lines. Different scan rates change the pixel dwell time. But the final image is still not much more than 2000 lines. This is fine for a Polaroid print. But if using real film, it is less than ideal or optimum. I find that film has much more resolution than a Polaroid print. A good alternative is the Polaroid PN (positive/negative). But one would still have to scan the negative to get a digital file. This is another topic, all together.
} It would be more interesting to consider whether this type of } camera can contribute to high quality TEM imaging. I am not clear } what is limiting the resolution of current TEM digital cameras - } could a high-resolution CCD camera approach the resolution of } TEM film more closely than the current generation of these, or is } the phosphor/YAG not good enough to make it worthwhile.
Not having TEM experience, I cannot comment on this type of application. But I am experienced with SEM usage. It would seem to me at first blush that TEM images are continuous whereas the SEM images are discrete. This would mean that CCD imaging devices would work for TEM applications but not for SEM. The common denominator remains the Polaroid and silver emulsion film.
gg
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Modern surfers use PC boards. You can too at http://photoweb.net ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
It's called a "passive" or "listening" digital image acquisition system, such as our ADDA II or similar devices from other manufacturers. Benefits: fairly easy to set up and use. Disadvantages (as opposed to an "active" or "talking" system): You're still limited to what the microscope can provide in terms of resolution, dwell time, etc.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: John Foust [mailto:jfoust-at-threedee.com] Sent: Tuesday, May 16, 2000 8:06 AM To: Microscopy-at-sparc5.microscopy.com
At 04:02 PM 5/15/00 -0700, Dr. Gary Gaugler wrote: } It seems to me that no digital camera system would work on a SEM } in place of a Polaroid or other film-based output device. Since the } recording CRT in a SEM is based on a sequential line scan, one } would need a camera that would capture each line as it is produced.
What you're saying is there must be a system out there that digitizes that single stream of line scan intensities, then processes all that data inside the computer as an image as opposed to trying to digitize the frame buffer.
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Thanks to all who indicated an interest in meeting to discuss some common problems associated with managing microscopy facilities at the M&M meeting. I am in the process of arranging this and will send details once we are a bit further along. Debby
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
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Like Gary and Ken have pointed out, there are several reasons why 4x5 backs won't work. Even if they did, there is another issue. A 4x5 back would incorporate the photo CRT with its phosphor into the loop. That means the electronic signal from the PMT or BSE detector has to be converted back into an analog brightness via the photo CRT, then the digital 4x5 back would be used to digitize the signal, and that in a most unwieldly manner.
As a rule, if you don't have to convert signals back and forth or pass them through extra stages of processing, don't do it. I understand photo CRTs are quite good, but there are extra focus, noise, and calibration factors involved passing the signal through the CRT. It is far better to take the signal straight over to digital using a good, single channel A/D converter for the video (plus one for X and Y position rather than using the millions of A/D converters in a CCD. Makes for a lot cheaper system, too.
Warren S.
At 04:02 PM 5/15/2000 -0700, Gary Gaugler wrote: } At 01:38 PM 5/10/00, you wrote: } } } } I'm curious ... has anyone tried simply installing a digital } } 4x5 back in place of a Polaroid 4x5 back??? For example, see: } } } } http://www.phaseone.com/brochures/powerfx.html } } } } cheerios, shAf } } It seems to me that no digital camera system would work on a SEM } in place of a Polaroid or other film-based output device. Since the } recording CRT in a SEM is based on a sequential line scan, one } would need a camera that would capture each line as it is produced. } Most digital backs are single or triple pass units of a single linear } set of sensors. There are other cameras that do snapshot capture } but even these would not work since the whole image is not present } on the record CRT at the time of taking a picture with the digital } camera. The final image is generated sequentially, line by line, } on the record CRT. } } If the SEM image is stored in a frame buffer, the buffer can be } converted to RS-170 TV video and frame grabbed. But the } best that this would typically do is 640 lines. } } Its an interesting problem and dilemma about being in a situation } where digital camera products simply won't work in place of } film. But since the goal is to obtain a digital file, why not start } with a digital interface? For example, a passive digital capture } system would transfer the record CRT information to computer } and directly result in a nice digital file. Alternatively, for some } systems, an active system can be applied to directly control the } SEM's beam. In doing so, the range of final digital image } resolution is limited only by the attached hardware system. } } gary g.
} Date: Tue, 16 May 2000 14:27:27 -0700 } To: John Foust {jfoust-at-threedee.com} } From: "Dr. Gary Gaugler" {gary-at-gaugler.com} } Subject: Re: SEM: digital 4x5 backs } } No. What I was saying was that there is not a digital } camera system out there that will capture the line-by-line } recording CRT output--as far as I see it at present. } } To get a digital file from the SEM, the method needs to } be digital but either passively attached to the record CRT } or actively connected as a replacement for the SEM's } internal scan generator. } } gary } } } } At 07:06 AM 5/16/00, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
ELECTRON MICROSCOPE TECHNICIAN Naval Research Laboratory
Announcement Number 74-0448-00 http://amp.nrl.navy.mil/code1800/74-0448.htm Job Title: Physical Scientist, NP-1301-II, $22,563* to $49,794* (*Includes locality pay)
Description: The Marine Geosciences Division, Naval Research Laboratory (NRL), Stennis Space Center, MS, seeks an electron microscope technician to perform technical duties in support of the Marine Geosciences Electron Microscopy Center.
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Required qualifications include a degree in physical science, engineering, or mathematics that included 24 semester hours in physical science and/or related engineering science such as mechanics, dynamics, properties of materials, and electronics. The candidate must also have one year of specialized experience equivalent to the Career Level I (GS-Equivalent 1-4).
All candidates will be rated on the following factors: 1) Knowledge in the preparation of transmission electron microscope (TEM) thin specimens (preferably from fine-grained sediment/soil specimens and /or other geological samples). 2) Knowledge in the operation of TEMâs and scanning electron microscopes (SEMâs) and their analytical detectors. 3) Knowledge in the basic interpretation of TEM data. 4) Ability to communicate technical concepts orally and in writing.
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or visit the web page http://www.nrl.navy.mil/hro.htm
-- _____________________________________________________ Tyrone L. Daulton Director: Marine Geosciences - Electron Microscopy Center Physicist and Senior Electron Microscopist
Naval Research Laboratory Marine Geosciences Division (Code 7400) Stennis Space Center, MS 39529-5004
} If you control the scan you only need a single light sensitive } element. You step the beam digitize the light, step the beam, } wait for the last spot to go out and repeat. You don't need } a camera. You do need a very fast responding system for } changing electron beams to light. } } This system has the resolution of the scan beam. It would not } be particualy fast but you could increase the number of bits } resolution to as large a number as you wanted. } } Gordon } } Gordon Couger gcouger-at-couger.com } } Stillwater, OK www.couger.com/gcouger } 405 624-2855 GMT -6:00
The light sensitive element is already in-place. It is the Everhart-Thornley scintillator detector, or any other SE or BSE detector on a SEM. Just passively tap into the record CRT signals, digitize the detected video (SE/BSE), and save the resulting image as a digital file. Changing scan rates will change pixel dwell time. But the final image ought to still be a digital representation of what was intended to go to the film/Polaroid camera.
Again, the other option is active control of the SEM's beam. This just swaps the internal scan generator with an external one. The video system and digitization is the same as for a passive system. A robust active system will be capable of producing higher resolution digital images than a passive system. This is because a good active system can go up to 4096 horizontal pixels (12-bit D/A converter) and the pixel dwell time is usually adjustable. The only down side is the total frame time based on pixel dimensions and dwell time. At extremes, one could be waiting a very long time for a single image. If the beam is not stable, and as pointed out in an earlier posting, the drive circuits are not stable, there would be some upper limit on pixel dimensions and dwell time. Beyond this limit, the image would appear to shift as the beam shifted during the capture.
Several of the current generation SEMs have incorporated digital control and digital image capture. But these SEMs are of course at today's prices. Its rather easy to breathe new life into an older SEM by adding active or passive third party digital control/capture systems. One gets essentially a "new" modern SEM at a fraction of the cost of buying a new one.
We are exploring the possibility of purchasing a freeze fracture system to do FFTEM of emulsions/microemulsions. Does anyone have any recommendations? Does anyone have a used system available?
Hello Nicholas, I've looked at several soils similar to your easily disaggregated clay rich rocks in the TEM and find that the best procedure is to imbed aggregates in 2% agar to hold the aggregates together. The aggregates are then cut out as agar-soil cubes and chemical processing (fixation, Os-fix, buffer rinse, dehydration, resin-solvent washes, resin infiltration) done directly onto the agar cubes. The cubes are placed into molds and after curing, microtomed to 40-60nm sections. The sections come out nicely, but if you have a high primary mineral content (quartz, feldspars } 15%) then you'll have a lot of torn sections. Use of a diamond knife rather than glass gives you significantly better results.
My methods used in my thesis are on the web at: http://wilfred.berkeley.edu/~gordon/PHD Look particularly at chapter four which concentrates on TEM methods and results. If it would be of use, I'll be glad to send a cdrom copy of my thesis to you.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ 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 Tue, 16 May 2000, N. Hayman 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 all, } I believe to be on track with mounting clay rich rocks (that } disagregate readily in perturbed water (i.e. shaken or sonicated), but not } in ethylene glycol or ethanol (at rest). My goal is to mount the samples } (whole, not seperates) for analysis in TEM. I have been soaking them in } LR-White at 60C, where fixation occurs without accelerator in less than 24 } hours. I am planning to (but have not yet) microtome the samples, or } potentially ion-mill. } Thoughts for the future are to use a set of glassware with a } Millipore filter and vacuum to pull ethanol(cleanser and dilator), } ethylene glycol(for swelling clay component), and an LR White "chaser" to } view "saturated" textures, as opposed to compacted. } My question is this: Can anyone point out pitfalls with this } approach that I am not seeing, again I haven't tried the whole thing yet, } but am in a position to start prepping the samples. In particular, is } microtoming preferred to ion-milling for weak, soft samples that rely on } epoxy for reinforcement? Does LR-white readily pull through a sample given } a weak vacuum and a porous plate (its viscous, but not as viscous as } water, for example)? Does swelling the clays with ethylene glycol and the } like, and then directly mounting, introduce volatiles into the column } under 120-200 kV? Any recommendations are welcome, the literature helps a } bit, but is usually sketchy about these fine details of preparation } procedure. } thanks in advance, } N } } _____________________________ } Nicholas W. Hayman \ } Dept. of Geological Sci., U.W.\ http://www.geology.washington.edu/~hayman } Box 351310, Seattle WA 98195 \_________________________________________ } } } }
Dear All, Does anyone know of an EDS mineral spectrum database or mineral identification program? Is there such a thing available? Possibly a program that would allow the input of an image file of the unknown mineral spectrum thereby generating a list of possible 'best-fit' candidates. Or maybe it would work by predicting what the spectrum of a certain mineral should look like for a particular beam voltage etc. Regards Martin Roe
Martin J. Roe Macaulay Land Use Research Institute Craigiebuckler Aberdeen Scotland UK Phone 01224 318611 e-mail m.roe-at-mluri.sari.ac.uk
I will wait for info about sensitivity. I think, for EM in particular, sensitivity is very important parameter of the system. I am surprised that manufacturers don't have data on this matter. If sensitivity, say 10 times higher than SO-163 film - it may be a great reason to switch from film to the CCD. The major disadvantage of the CCD cameras for TEM is their price in my point of view. Thanks for your respond.
Sergey
} Date: Wed, 17 May 2000 11:34:05 -0600 } From: Michael Bode {mb-at-Soft-Imaging.com} } Subject: RE: new developments in imaging systems? } To: 'Sergey Ryazantsev' {sryazant-at-ucla.edu} } X-Mailer: Internet Mail Service (5.0.1457.3) } } Sergey, } } I will try to find out what we have regarding the relative } sensitivities. One thing that I can say now is, that we acquire images } with a 50 or 100 msec exposure of the digital camera when the film } requires an exposure of about 2 seconds. This would mean a 20 to 40 } times better sensitivity of the CCD camera. But to answer your question } in more detail would require to compare also the resolution of film and } camera and that is where it becomes very difficult, as it is not easy to } determine the resolution of film in terms of spatial resolution and } dynamic range, as both are interwoven. I will try to find some answers } for you. } } Regarding the linearity: As the CCDs simply count Photons, they have } almost perfect linearity over their complete dynamic range. Even more so } for TEMs, where all Photons have the same energy and the quantum } efficiency does not change from photon to photon. The Phosphor is a part } of a system that could theoretically introduce some non-linearities. } However, I did measure the linearity of some TEM camera systems a few } years back and did not find any significant deviations from linearity. } } Michael } } Michael Bode, Ph.D. } Soft Imaging System Corp. } 1675 Carr St., #105N } Lakewood, CO 80215 } =================================== } phone: (888) FIND SIS } (303) 234-9270 } fax: (303) 234-9271 } email: mailto:info-at-soft-imaging.com } web: http://www.soft-imaging.com } =================================== } } } } -----Original Message----- } From: Sergey Ryazantsev [mailto:sryazant-at-ucla.edu] } Sent: Wednesday, May 17, 2000 1:29 AM } To: Michael Bode } Subject: RE: new developments in imaging systems? } } } Mickhael hello } } I have question for you. I am thinking about adding CCD camera to my } JEM1200EX. The information I gathered from Internet is not so } optimistic. } The standard CCD resolution is about 1-1.5 million pixels, 2 million is } a } max as I understand. The price for cheaper camera is about 20-30 K$ - } much } more that I expect to spend for "film" process. There are two things } may } attract me to the modern CCD camera: dynamic range and sensitivity. I } am } pretty sure that dynamic range for CCD itself is a few orders better } than } any film available. But what about phosphorus screen? Does it reduce } dynamic range for the EM images? How dramatic? This is my first } question. } The next question is: could you tell me something about sensitivity of } the } modern CCD cameras used in EM? I am using dark field imaging at x80K } magnification and the exposure time for SO-163 (non deluded D-19, 7 min, } 20oC) is about 2 sec. I called GATAN, but they did not say anything } useful. } Could you provide some comparison of your side-mount camera with } sensitivity of the SO-163 film at condition I mentioned? I will greatly } appreciate any information in this matter. Thanks. Sergey. } } } } Date: Fri, 12 May 2000 13:46:01 -0600 } } From: Michael Bode {mb-at-Soft-Imaging.com} } } Subject: RE: new developments in imaging systems? } } To: "'Microscopy-at-MSA.Microscopy.Com'" } {Microscopy-at-sparc5.microscopy.com} } } Cc: 'Margaret Dienelt' {brannign-at-asrr.arsusda.gov} } } X-Mailer: Internet Mail Service (5.0.1457.3) } } } } ----------------------------------------------------------------------- } - } } 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 no digital imaging guru, but I have been following this thread with interest. This whole discussion has been of interest to me because of the idea of the passive 4x5 (camera back) "detector" that could provide an extremely simple user venue for converting those countless analog SEMs into a very usable digital output format.
We have a 10 year old JEOL 840 microanalysis system that we would like to see in operation for another 10 years, and I think it may be possible. However, digital imaging, although "doable", is not "convenient" for us, at this point for this instrument. Too much time and effort is required in digital acquisition, maintenance of instrument condition info with the image, labeling the image, and archiving. Then on top of it all, the digital image is often not quite as good as the analog Polaroid shot that also has the Mag, Bar Scale, KeV, WD, and other info permanently incorporated (We use Type 53 for much of our work.) Don't get me wrong, we do some digital imaging with it, but "it ain't the same" as using a newer digital scope with integrated digital control and digital interface. If someone put such a passive digital detector into a 4x5 camera back-type mount, and it was capable of passively detecting at least 2500 horizontal lines and an approximately equivalent vertical resolution, I believe it would be a huge success with the analog SEMs. The 840 and many other instruments like it are great conventional SEMs, and such a simple interface (if affordable) would greatly extend their value. Many of these scanners have exceptional imaging capability, but often the easiest/best way to record that impressive image is by Polaroid film.
Is it doable? Brad Huggins
} ---------- } From: Michael Bode[SMTP:mb-at-Soft-Imaging.com] } Sent: Wednesday, May 17, 2000 1:36 AM } To: 'Microscopy-at-MSA.Microscopy.Com' } Cc: 'John Foust' } Subject: RE: SEM: digital 4x5 backs } } ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } Yes, there is. } } It's called a "passive" or "listening" digital image acquisition system, } such as our ADDA II or similar devices from other manufacturers. } Benefits: fairly easy to set up and use. Disadvantages (as opposed to an } "active" or "talking" system): You're still limited to what the } microscope can provide in terms of resolution, dwell time, etc. } } Michael } } Michael Bode, Ph.D. } Soft Imaging System Corp. } 1675 Carr St., #105N } Lakewood, CO 80215 } =================================== } phone: (888) FIND SIS } (303) 234-9270 } fax: (303) 234-9271 } email: mailto:info-at-soft-imaging.com } web: http://www.soft-imaging.com } =================================== } } } } -----Original Message----- } } From: John Foust [mailto:jfoust-at-threedee.com] } Sent: Tuesday, May 16, 2000 8:06 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: SEM: digital 4x5 backs } } } ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } At 04:02 PM 5/15/00 -0700, Dr. Gary Gaugler wrote: } } It seems to me that no digital camera system would work on a SEM } } in place of a Polaroid or other film-based output device. Since the } } recording CRT in a SEM is based on a sequential line scan, one } } would need a camera that would capture each line as it is produced. } } What you're saying is there must be a system out there that } digitizes that single stream of line scan intensities, then } processes all that data inside the computer as an image as } opposed to trying to digitize the frame buffer. } } - John } }
} Dear All, } Does anyone know of an EDS mineral spectrum database or } mineral identification program? Is there such a thing available? } Possibly a program that would allow the input of an image file of } the unknown mineral spectrum thereby generating a list of possible } 'best-fit' candidates. Or maybe it would work by predicting what the } spectrum of a certain mineral should look like for a particular beam } voltage etc. } Regards
} Martin Roe
Martin,
I know of no publicly available database of mineral EDX spectra or identification software. However, I believe several commercial EDX systems have a facility for generating your own database by collecting spectra from standard mineral samples. The software can then compare a spectrum from an unknown with those in the database.
One must be very careful of comparing like with like. All instrument parameters such as kV, tilt, etc, etc must be equivalent for a reasonable chance of a correct match.
I use the Desk Top Spectrum Analyser (DTSA) program from NIST to simulate EDX spectra from minerals, ceramics etc from the known composition. It requires a fair bit of effort to master the program but I feel it is worth the effort. Of course it does a lot more than just simulate spectra, including analysis of real measured spectra with a huge amount of flexibility in choosing analysis parameters.
DTSA is available free from the NIST web site:
http://www.cstl.nist.gov/div837/837.02/dtsa.html
Hope this helps,
Mark Blackford TEM Group Materials Division, ANSTO PMB 1, Menai, N.S.W. Australia 2234
Phone 61 2 9717 3027 Fax 61 2 9543 7179
Disclaimer: The views expressed in this E-mail message do not necessarily represent the official views of ANSTO from which this message was conveyed.
For what it's worth - if anything, there is a company out here in California called Silicon Film Technologies that has developed a technology that converts a traditional 35mm SLR camera into a digital camera. You essentially just replace the film with something they call (e)film. I don't know if they have any interest in developing something for this application, but it may be worth a look. It's a pretty nifty concept for people who want to do both traditional film photography and also digital photography with the same equipment.
I have copied a blurb from their website to give you an overview of what they do:
"Our EFS product suite is a digital photo system comprised of an electronic film cartridge and a carrier/adapter. The user simply inserts the cartridge, called (e)film, into the film cavity in the back of a conventional 35mm SLR camera. After recording up to 24 images, the cartridge is placed into a carrier/adapter, called (e)port, which may then be connected directly to a personal computer, enabling images to be downloaded quickly into the computer and then printed, sent via email, or modified into a photo end product using photo management software such as Adobe PhotoShop LE, which comes bundled with the EFS system. We will also market a digital photo storage module, called (e)box, which can store and transport hundreds of digital images in the field when the photographer does not have access to a computer.
The EFS system transforms conventional camera equipment into a digital image capture system. It is the only system currently available that provides the convenience and flexibility of choosing between conventional and electronic film formats with the same camera body. The product is aimed toward the large, installed base of 35mm camera owners who would like to enjoy the benefits of digital imaging while not giving up the cameras, lenses, and photo accessories with which they are familiar."
You can check out their website for more information at www.siliconfilm.com.
DISCLAIMER: I do own a small amount of stock in their parent company, Irvine Sensors Corporation. Of course, even if all of you bought a dozen of their neat little cameras, I'd still have to keep my day job, but I thought I should disclose it.
David Henriks Vice President 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
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by "Huggins, Bradley J" } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I'm no digital imaging guru, but I have been following this thread with interest. This whole discussion has been of interest to me because of the idea of the passive 4x5 (camera back) "detector" that could provide an extremely simple user venue for converting those countless analog SEMs into a very usable digital output format.
We have a 10 year old JEOL 840 microanalysis system that we would like to see in operation for another 10 years, and I think it may be possible. However, digital imaging, although "doable", is not "convenient" for us, at this point for this instrument. Too much time and effort is required in digital acquisition, maintenance of instrument condition info with the image, labeling the image, and archiving. Then on top of it all, the digital image is often not quite as good as the analog Polaroid shot that also has the Mag, Bar Scale, KeV, WD, and other info permanently incorporated (We use Type 53 for much of our work.) Don't get me wrong, we do some digital imaging with it, but "it ain't the same" as using a newer digital scope with integrated digital control and digital interface. If someone put such a passive digital detector into a 4x5 camera back-type mount, and it was capable of passively detecting at least 2500 horizontal lines and an approximately equivalent vertical resolution, I believe it would be a huge success with the analog SEMs. The 840 and many other instruments like it are great conventional SEMs, and such a simple interface (if affordable) would greatly extend their value. Many of these scanners have exceptional imaging capability, but often the easiest/best way to record that impressive image is by Polaroid film.
Dear Martin: An interesting, but difficult idea. Besides the problem of instrument parameters, and settings, there is the problem of mineral compositions being in many cases very similar, but having different structures. For example, there are several iron oxide/hydroxide phases that would be difficult to tell apart with an EDS analysis. More complex are the silicates which have a number of different major structural families, many having the same or similar chemical compositions. There are sites on the web that would allow you to input an element list and it will output all the minerals with those elements and their formulas. It won't identify the minerals, but it would give you a start. To identify them you need to do optical or x-ray diffraction or some other more appropriate technique. Michael L. Boucher Sr. mboucher-at-isd.net http://www.isd.net/mboucher -----Original Message----- } From: Martin J. Roe {m.roe-at-mluri.sari.ac.uk} To: microscopy-at-sparc5.microscopy.com {microscopy-at-sparc5.microscopy.com}
If sensitivity was ten times higher, one would in some cases save some beam damage and in most cases suffer excessive electron noise. As a rough guide, electron noise becomes apparent at 30x enlargement, rarely a problem. Magnification is a linear function and electron density relates to an area, but clearly very short exposure images would be much noisier and could not be enlarged nearly as much. When not enough electrons form an image it appears grainy, which makes it unsuitable for further enlarging. Photo enlarging utilises higher depths-of-field and without this, very high power TEM is much, much harder. By nature, slower emulsions have finer grain and higher resolution and contrast. It is fortuitous that these desirable characteristics run in tandem with relatively long exposures, so the image is formed by more electrons. TEM films rate about 6 ISO, so X-ray film at about ISI 3000 is about a 10x faster than TEM films. Unfortunately its rather grainy, but if the exposure was well adjusted, chances are that electron noise would be more bothersome than the film's grain. Digital is inevitable and already fairly common, but some aces remain with conventional film. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Thursday, May 18, 2000 5:06 AM, Sergey Ryazantsev [SMTP:sryazant-at-ucla.edu] wrote: } } } Thank you Mickhael. } } I will wait for info about sensitivity. I think, for EM in particular, } sensitivity is very important parameter of the system. I am surprised that } manufacturers don't have data on this matter. If sensitivity, say 10 times } higher than SO-163 film - it may be a great reason to switch from film to } the CCD. The major disadvantage of the CCD cameras for TEM is their price } in my point of view. } Thanks for your respond. } } Sergey } } } } Date: Wed, 17 May 2000 11:34:05 -0600 } } From: Michael Bode {mb-at-Soft-Imaging.com} } } Subject: RE: new developments in imaging systems? } } To: 'Sergey Ryazantsev' {sryazant-at-ucla.edu} } } X-Mailer: Internet Mail Service (5.0.1457.3) } } } } Sergey, } } } } I will try to find out what we have regarding the relative } } sensitivities. One thing that I can say now is, that we acquire images } } with a 50 or 100 msec exposure of the digital camera when the film } } requires an exposure of about 2 seconds. This would mean a 20 to 40 } } times better sensitivity of the CCD camera. But to answer your question } } in more detail would require to compare also the resolution of film and } } camera and that is where it becomes very difficult, as it is not easy to } } determine the resolution of film in terms of spatial resolution and } } dynamic range, as both are interwoven. I will try to find some answers } } for you. } } } } Regarding the linearity: As the CCDs simply count Photons, they have } } almost perfect linearity over their complete dynamic range. Even more so } } for TEMs, where all Photons have the same energy and the quantum } } efficiency does not change from photon to photon. The Phosphor is a part } } of a system that could theoretically introduce some non-linearities. } } However, I did measure the linearity of some TEM camera systems a few } } years back and did not find any significant deviations from linearity. } } } } Michael } } } } Michael Bode, Ph.D. } } Soft Imaging System Corp. } } 1675 Carr St., #105N } } Lakewood, CO 80215 } } =================================== } } phone: (888) FIND SIS } } (303) 234-9270 } } fax: (303) 234-9271 } } email: mailto:info-at-soft-imaging.com } } web: http://www.soft-imaging.com } } =================================== } } } } } } } } -----Original Message----- } } From: Sergey Ryazantsev [mailto:sryazant-at-ucla.edu] } } Sent: Wednesday, May 17, 2000 1:29 AM } } To: Michael Bode } } Subject: RE: new developments in imaging systems? } } } } } } Mickhael hello } } } } I have question for you. I am thinking about adding CCD camera to my } } JEM1200EX. The information I gathered from Internet is not so } } optimistic. } } The standard CCD resolution is about 1-1.5 million pixels, 2 million is } } a } } max as I understand. The price for cheaper camera is about 20-30 K$ - } } much } } more that I expect to spend for "film" process. There are two things } } may } } attract me to the modern CCD camera: dynamic range and sensitivity. I } } am } } pretty sure that dynamic range for CCD itself is a few orders better } } than } } any film available. But what about phosphorus screen? Does it reduce } } dynamic range for the EM images? How dramatic? This is my first } } question. } } The next question is: could you tell me something about sensitivity of } } the } } modern CCD cameras used in EM? I am using dark field imaging at x80K } } magnification and the exposure time for SO-163 (non deluded D-19, 7 min, } } 20oC) is about 2 sec. I called GATAN, but they did not say anything } } useful. } } Could you provide some comparison of your side-mount camera with } } sensitivity of the SO-163 film at condition I mentioned? I will greatly } } appreciate any information in this matter. Thanks. Sergey. } } } } } } } Date: Fri, 12 May 2000 13:46:01 -0600 } } } From: Michael Bode {mb-at-Soft-Imaging.com} } } } Subject: RE: new developments in imaging systems? } } } To: "'Microscopy-at-MSA.Microscopy.Com'" } } {Microscopy-at-sparc5.microscopy.com} } } } Cc: 'Margaret Dienelt' {brannign-at-asrr.arsusda.gov} } } } X-Mailer: Internet Mail Service (5.0.1457.3) } } } } } } ----------------------------------------------------------------------- } } - } } } 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 } } } ----------------------------------------------------------------------- } } . } } } } } } } } } Margaret, } } } } } } As a former user and current vendor of such systems as you are } } inquiring } } } about I can try to provide a bit of information regarding camera } } } improvements: } } } } } } There have been a number of improvements, but I am not sure what you } } are } } } comparing the latest cameras against. Cameras are now usually cooled } } and } } } provide 12 bits per pixel, the number of pixels has gone up a bit (but } } } not much in general), and cameras read out faster than they used to (up } } } to 20 fps and more). I think all cameras now use a line transfer } } } mechanism, which makes shutters obsolete. } } } On the software side, real-time FFT and real-time shading correction } } can } } } be done now due to faster computers without special processing boards, } } } and there have been other software developments that make using the } } } cameras and computers easier. } } } Other changes that affect the usability of cameras is the use of } } } pneumatics to insert and retract the phosphors, higher frame rates for } } } live viewing with the camera, etc. } } } } } } If you have questions, please give me a call, drop me an email, or go } } to } } } our web site. } } } } } } Michael } } } } } } } } } Michael Bode, Ph.D. } } } Soft Imaging System Corp. } } } 1675 Carr St., #105N } } } Lakewood, CO 80215 } } } =================================== } } } 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: Margaret Dienelt [mailto:brannign-at-asrr.arsusda.gov] } } } Sent: Thursday, May 11, 2000 12:43 PM } } } To: Microscopy-at-sparc5.microscopy.com } } } Subject: TEM: new developments in imaging systems? } } } } } } } } } ----------------------------------------------------------------------- } } - } } } 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, } } } } } } Year after year I hopefully gather information about digital imaging } } } systems for TEMs (ours is a JEOL 100CX) , only to learn we have no } } } money. This year it looks like it might really happen but I have not } } } kept up with innovations in the field and am wondering the following: } } } } } } 1. Anything new in the last two years -- especially in terms of } } } cameras? I'm most familiar with the Gatan and AMT systems but their } } } web sites don't reflect much in the way of changes over a year ago. } } } 2. With more and more microscopists finally getting their systems -- } } } I'd love to get feedback. } } } } } } Thanks, } } } Margaret } } } } } } P.S. Would welcome contacts from vendors. } } } } } } -- } } } Margaret Dienelt } } } } } } Plant Pathologist } } } Electron Microscopy Lab } } } } } } Floral and Nursery Plants Research Unit } } } U.S. National Arboretum/Agricultural Research Service/USDA } } } } } } B. 010A, Rm. 238, BARC-W } } } 10300 Baltimore Avenue } } } Beltsville MD. 20705 USA } } } } } } (301) 504-6097 } } } Fax: (301) 504-5096 } } } } } } } } _____________________________________ } } } } Sergey Ryazantsev Ph. D. } } Electron Microscopy } } UCLA School of Medicine } } Department of Biological Chemistry } } Box 951737 } } Los Angeles, CA 90095-1737 } } } } Phone: (310) 825-1144 } } Pager: (310) 845-0248 } } FAX (departmental): (310) 206-5272 } } mailto:sryazant-at-ucla.edu } } http://www.bol.ucla.edu/~sryazant } } } } } _____________________________________ } } Sergey Ryazantsev Ph. D. } Electron Microscopy } UCLA School of Medicine } Department of Biological Chemistry } Box 951737 } Los Angeles, CA 90095-1737 } } Phone: (310) 825-1144 } Pager: (310) 845-0248 } FAX (departmental): (310) 206-5272 } mailto:sryazant-at-ucla.edu } http://www.bol.ucla.edu/~sryazant } }
I can identify with your situation. While I do not know the intricacies of the JEOL instruments, some SEMs are made to accept external drive for x-ray analysis. This same input scheme works perfectly for active control of the SEM and direct digital capture of images. At the worst, you can replicate the resolution of your record CRT using a passive mode. How easy either of these modes are depends greatly on how the SEM system was designed.
gary g.
At 01:44 PM 5/17/00, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} Dear Martin: } An interesting, but difficult idea. Besides the problem of instrument } parameters, and settings, there is the problem of mineral compositions being } in many cases very similar, but having different structures.
And then there is also the converse: the problem of mineral compositions being in many cases very different, but having the same structures. One of a huge number of examples would be the feldspar series where Na and Ca can substitute for each other continuously from Na to Ca endmembers.
Various other bits of crystallographic information, some intangibles such as crystal shape and growth habit, associations with other minerals and so on, would have to be taken into account in such software for it to really zero in on an unambiguous ID for you.
Arthur Day, Electron Microscope Unit Phone: 61-2-9717-3457 Ansto Materials Division Fax: 61-2-9543-7179 PMB 1, Menai (Sydney), NSW, 2234 Email: ard-at-ansto.gov.au Australia www: http://www.ansto.gov.au/
Hello all ..... we have an old ETEC Omniscan SEM, and we need to contact with any people that can supply manuals ....( fundamentally electric and electronic schematic )
any help is welcome
best regards
=================================================== Fernando D. Balducci Laboratorio de Microscopia Electrnica Facultad de Ingeniera - Bioingenieria Universidad Nacional de Entre Rios Argentina e-mail: microsc-at-fi.uner.edu.ar tel: 54 43 975100 fax: 54 43 975077
At 07:02 PM 5/17/00, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I think that their aim is a bit off the target. Its nice to not have to give up the hardware but their offering makes the user give up the benefits of 35mm film--frame size and selection of speed. And, a field of view factor of 2.8 is absurd.
I beta tested what must have been an initial offering of this product early last year. It was by a different company. Same idea though. Same deficiencies. The sensor is 1.3M pixels and has an odd aspect ratio of 1.25 (35mm frame is 1.5). Also, and most importantly, the product does not image the entire 35mm frame, only a small central portion. 1.3M pixel point and shoot cameras are a lot cheaper than this silicon film thing. A really dedicated user would have to buy more than one silicon film insert. At $600 each, that would buy a lot of P&S cameras. But there is no need to do that since one just pops out a SmartMedia or Compact Flash module ($150 or so each).
I still say wait and see how the Fuji Finepix S1 Pro performs. If it lives up to spec, I'd say that it will be a raging success and a major turning point in digital cameras which are based on the 35mm format.
gary g.
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Modern surfers use PC boards. You can too at http://photoweb.net ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Dear Friends, Can anyone give me advice regarding the dismantling and the packing for shipment of a Philips 201 TEM? I need to pay particular attention to not disturbing the alignment. I plan to move the TEM from Univ. of Delaware to Naples, Florida in an enclosed U-Haul Trailer. Any help or suggestions will be most welcome. If anyone has hands-on experience in doing this sort of thing and is willing to give me a hand at U-Del, please contact me and name your price. I'll gladly pay for the assistance. Best regards, Mike Urbanik www.crystalguru.com
I will reiterate my point and that of others. An SEM technically has video only coming from one point in the image at a time. Sure, you could pay lots of $$ for a big CCD with the resolution you desire, or you could simply digitize the one (or more) video signals as the beam scans the screen. You have the choice of either asserting the x-y positions through active digital control or you can read them off passively.
From a hardware perspective, it is a much easier (and cheaper) task to build an active or passive system like those on the market than to build a 4x5 camera back detector. For both systems you would still need the software and computer
At 10:58 PM 5/17/2000 -0700, Gary Gaugler wrote:
} I can identify with your situation. While I do not know the intricacies } of the JEOL instruments, some SEMs are made to accept external } drive for x-ray analysis. This same input scheme works perfectly } for active control of the SEM and direct digital capture of images. } At the worst, you can replicate the resolution of your record CRT } using a passive mode. How easy either of these modes are depends } greatly on how the SEM system was designed. } } gary g. } } At 01:44 PM 5/17/00, you wrote: } } } } I'm no digital imaging guru, but I have been following this thread with } } interest. This whole discussion has been of interest to me because of the } } idea of the passive 4x5 (camera back) "detector" that could provide an } } extremely simple user venue for converting those countless analog SEMs into } } a very usable digital output format. } } } } We have a 10 year old JEOL 840 microanalysis system that we would like to } } see in operation for another 10 years, and I think it may be possible. } } However, digital imaging, although "doable", is not "convenient" for us, at } } this point for this instrument. Too much time and effort is required in } } digital acquisition, maintenance of instrument condition info with the } } image, labeling the image, and archiving. Then on top of it all, the } } digital image is often not quite as good as the analog Polaroid shot that } } also has the Mag, Bar Scale, KeV, WD, and other info permanently } } incorporated (We use Type 53 for much of our work.) Don't get me wrong, we } } do some digital imaging with it, but "it ain't the same" as using a newer } } digital scope with integrated digital control and digital interface. If } } someone put such a passive digital detector into a 4x5 camera back-type } } mount, and it was capable of passively detecting at least 2500 horizontal } } lines and an approximately equivalent vertical resolution, I believe it } } would be a huge success with the analog SEMs. The 840 and many other } } instruments like it are great conventional SEMs, and such a simple interface } } (if affordable) would greatly extend their value. Many of these scanners } } have exceptional imaging capability, but often the easiest/best way to } } record that impressive image is by Polaroid film. } } } } Is it doable? } } Brad Huggins
I fuss with mineral identification via EDS spectra every day. I keep the program "Mineral" running at all times. It is a Windows Filemaker Pro database which includes the chemical formula, physical properties, type locale and 7 to 10 x-ray diffraction lines for 4500 accredited mineral species and many un-named ones. Searches can be constructed from any single or combination of fields. The software is available from Aleph Enterprises in Livermore, CA (510 443 7319) The price was around $550 a few years ago. Less expensive DOS based, but similar software is available from the Fersmann Institute.
More complete chemistry (but no spectra) is offered by Alexander Holzel's MDAT program. It includes a database of chemical analyses of many minerals and can perform a search based on weight per cents of elements present as well the unknown's physical properties and x-ray diffraction data. This software is $1000 and up depending upon options. Dr. Holzel's e-mail address was Compuserve100333,2771 as of last August. He is in Ober-Olm, Germany.
None of the software programs work directly from spectra and only MDAT contains sample analyses so you will likely be estimating peak heights from chemical formula. For more definitive results you will need to start building your own spectrum library from your own reference materials.
Even with a high elemental analysis correlation, one will almost always need supplemental methods for a definitive ID. This is usually x-ray diffraction or optical microscopy.
Some day affordable EBSP retrofitted into SEMS will allow the analyst to distinguish, in situ in polished section between orthorhombic FeS2 (marcasite) and cubic FeS2 (pyrite). Currently available systems are $90,000+, I think.
If you need help obtaining grains of some of the less common minerals, I can help. I supply a catalog which includes hundreds of well identified reference quality mineral grains in addition to synthetic probe standards.
Bart Cannon Cannon Microprobe 1041 NE 100th Street Seattle, WA 98125 206 522 9233 (3947 fax)
In evaluating some electron beam damage data, I am trying to determine the electron dose to the sample using a small screen current reading from a JEOL 2010 TEM.
The screen current density is given in pA/cm^2 on the microscope terminal, and can be converted to dose (electrons/cm^2*s) in the sample. I have spoken to JEOL, and they indicated that the actual current density on the screen is the reading multiplied by a factor of 10.
I am unable to use a borrowed stage with a Farraday cup for calibration on this microscope because it is suspect for internal radioactive contamination. Thus, I would be grateful for insights from other users of 2010's who may have checked the accuracy of the reading (whether or not the factor of 10 is right, etc.) on their instruments.
Many Thanks, Wharton -- ++++++++++++++++++++++++++++++++++++++++++++++++++ Wharton Sinkler Argonne National Laboratory West P. O. Box 2528 Idaho Falls, ID 83403 Tel: (208) 533-7724
Dear Martin, As the other responders have stated, minerals can be difficult to identify with EDS only. There is no substitute for experience (and some diffraction data). My rule of thumb is to group minerals as do most mineral classification schemes; is it a silicate, oxide, carbonate, sulfide, phosphate, salt, organic, or metalic phase? Sometimes it's hard to decide even at this level (i.e., If you don't have a light element detector, you can't differentiate between oxides and carbonates, P and Si substitute for each other). Most times you can. Since I often work with silicates, my second observation is to look at Si/Al. This will help limit the choices greatly, but is not usually diagnostic by itself. There are relatively few minerals with Al} Si. Knowledge of the approximate Si/Al (the most common tetrahedral cations) combined with the ratio of Si and Al with other elements like Mg, Ti, Cr, Mn and Fe (sometimes Ca) will give you an idea of the tetrahedral to octahedral ratio. The alkali elements Na and K, and also Ca in significant abundances are very important for identifying feldspars and sheet silicates, but again, there are no hard and fast rules that I've come up with. The best thing is to know your mineral compositions (and their structures) or hire a mineralogist!;-) I also agree that even the mineralogist can find the DTSA program very useful and worth investing in a MAC. Hope this helps. Ciao for now, Ken
Kenneth JT Livi Department of Earth and Planetary Sciences 34th and Charles Streets Johns Hopkins University Baltimore, Maryland 21218 USA Phone: (410) 516-8342 Fax: (410) 516-7933 e-mail: klivi-at-jhu.edu
Due to the price of today's microscopes I am in need of advice concerning the quality of Russian microscopes. I had one in Spain and it was excellent. Please, advice. Thanks, Jose
Hello Folks, In reading the third edition of Hayat's Principles and Techniques of Electron Microscopy, there was mention of a 'Safety Chart, Chemicals in Electron Microscopy', for free distribution, by EMscope Laboratories Ltd (Kingsnorth Industrial Estate, Ashford, Kent). Does anyone out there have it? Or can anyone tell me how to get it or any wall chart that lists chemicals/hazards specific to EM? Thanks, Winnie
Lou, Very true. However, not one instrument can unambigously identify ALL minerals! In the case of my interests, PLM is not very useful since most of the crystals in shale are less than 2 um. Martin's original question was regarding how to identify EDS patterns, not what is the best way to identify a mineral. Ciao for now, Ken
} Colleagues; } } My question is why have you forgotten that polarized light microscopes were } devised for a need to identify and classify minerals? It almost seems } intentional to ignore it. Perhaps I am beginning to sound like Dr. McCrone } as I get older, but you dont have to throw a million dollar instrument at a } problem to solve it. If people have a problem with a $20k light microscope } doing a job better, that is there problem not mine. Just dont neglect the } fact that PLM is still a powerful technique in the hands of a competent } microscopist. PLM is still the standard for characterizing new crystal } compounds at the be.....visit any pharmaceutical companys R&D department and } you will see that it is so. } } Lou Solebello } } } ----- Original Message ----- } From: Kenneth JT Livi {klivi-at-jhu.edu} } To: {microscopy-at-sparc5.microscopy.com} } Cc: {m.roe-at-mluri.sari.ac.uk} } Sent: Thursday, May 18, 2000 8:15 AM } Subject: Re: EDS mineral identification and database } } } } ------------------------------------------------------------------------ } } 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 Martin, } } As the other responders have stated, minerals can be difficult to identify } } with EDS only. There is no substitute for experience (and some diffraction } } data). My rule of thumb is to group minerals as do most mineral } } classification schemes; is it a silicate, oxide, carbonate, sulfide, } } phosphate, salt, organic, or metalic phase? Sometimes it's hard to decide } } even at this level (i.e., If you don't have a light element detector, you } } can't differentiate between oxides and carbonates, P and Si substitute for } } each other). Most times you can. Since I often work with silicates, my } } second observation is to look at Si/Al. This will help limit the choices } } greatly, but is not usually diagnostic by itself. There are relatively few } } minerals with Al} Si. Knowledge of the approximate Si/Al (the most common } } tetrahedral cations) combined with the ratio of Si and Al with other } } elements like Mg, Ti, Cr, Mn and Fe (sometimes Ca) will give you an idea } of } } the tetrahedral to octahedral ratio. The alkali elements Na and K, and } also } } Ca in significant abundances are very important for identifying feldspars } } and sheet silicates, but again, there are no hard and fast rules that I've } } come up with. The best thing is to know your mineral compositions (and } } their structures) or hire a mineralogist!;-) I also agree that even the } } mineralogist can find the DTSA program very useful and worth investing in } a } } MAC. Hope this helps. } } Ciao for now, } } Ken } } } } Kenneth JT Livi } } Department of Earth and Planetary Sciences } } 34th and Charles Streets } } Johns Hopkins University } } Baltimore, Maryland 21218 USA } } Phone: (410) 516-8342 } } Fax: (410) 516-7933 } } e-mail: klivi-at-jhu.edu } } } } } }
Yes, we are aware of the problems in reaching our web site in the last few days. We have posted our new web site of microscopy supplies at a new URL and it is now up and running. Please change your bookmarks to:
http://www.laddresearch.com
Thank you,
John Arnott --
LADD RESEARCH 131 Dorset Lane Williston, VT 05495 USA
TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net
Warren's point is right on. The JEOL 840 or any other SEM that has a record CRT for 4x5 film or Polaroid is the signal source of passive information. It is not a big technical deal to do this. Depending on how readily available the CRT signals are (blank, frame, etc.), it may not be convenient or really simple. Some systems use BNC connectors to snake the signals throughout the system. T-ing off of these makes digitizing the SEM quite easy. If the signals are hardwired, it is more work to install the capture system. But in either case, it is a one time effort.
A passive system will record all legends just like the Polaroid does. But it will do it a lot better. The reason is that the tonal range and exposure latitude of a Polaroid is rather poor compared to real film. Digital capture systems are typically 10-bits; or 12-bits for ones with really exceptional dynamic range. Either of these are vastly superior to Polaroid prints.
The number of horizontal lines that a passive system will capture is the same as a Polaroid. This is because the record CRT circuitry fixes this dimension. However, the scan rate alters the pixel dwell time. Slower scan rates produce images that have less noise--be it Polaroid or digital capture. If the Polaroid print works OK for you, I would suggest that a digital capture system would be even better.
Compared to the cost of a modern computerized SEM, a digitizing attachment can be the key factor in keeping a good old SEM.
gg
At 06:42 AM 5/18/00, you wrote:
} I will reiterate my point and that of others. An SEM technically has video } only coming from one point in the image at a time. Sure, you could pay } lots of $$ for a big CCD with the resolution you desire, or you could } simply digitize the one (or more) video signals as the beam scans the } screen. You have the choice of either asserting the x-y positions through } active digital control or you can read them off passively. } } From a hardware perspective, it is a much easier (and cheaper) task to } build an active or passive system like those on the market than to build } a 4x5 camera back detector. For both systems you would still need the } software and computer } } At 10:58 PM 5/17/2000 -0700, Gary Gaugler wrote: } } } I can identify with your situation. While I do not know the intricacies } } of the JEOL instruments, some SEMs are made to accept external } } drive for x-ray analysis. This same input scheme works perfectly } } for active control of the SEM and direct digital capture of images. } } At the worst, you can replicate the resolution of your record CRT } } using a passive mode. How easy either of these modes are depends } } greatly on how the SEM system was designed. } } } } gary g. } } } } At 01:44 PM 5/17/00, you wrote: } } } } } } I'm no digital imaging guru, but I have been following this thread with } } } interest. This whole discussion has been of interest to me because of the } } } idea of the passive 4x5 (camera back) "detector" that could provide an } } } extremely simple user venue for converting those countless analog SEMs into } } } a very usable digital output format. } } } } } } We have a 10 year old JEOL 840 microanalysis system that we would like to } } } see in operation for another 10 years, and I think it may be possible. } } } However, digital imaging, although "doable", is not "convenient" for us, at } } } this point for this instrument. Too much time and effort is required in } } } digital acquisition, maintenance of instrument condition info with the } } } image, labeling the image, and archiving. Then on top of it all, the } } } digital image is often not quite as good as the analog Polaroid shot that } } } also has the Mag, Bar Scale, KeV, WD, and other info permanently } } } incorporated (We use Type 53 for much of our work.) Don't get me wrong, we } } } do some digital imaging with it, but "it ain't the same" as using a newer } } } digital scope with integrated digital control and digital interface. If } } } someone put such a passive digital detector into a 4x5 camera back-type } } } mount, and it was capable of passively detecting at least 2500 horizontal } } } lines and an approximately equivalent vertical resolution, I believe it } } } would be a huge success with the analog SEMs. The 840 and many other } } } instruments like it are great conventional SEMs, and such a simple interface } } } (if affordable) would greatly extend their value. Many of these scanners } } } have exceptional imaging capability, but often the easiest/best way to } } } record that impressive image is by Polaroid film. } } } } } } Is it doable? } } } Brad Huggins }
I am in need of a way to modify my current method of preparing TEM cross-sections as to introduce no heating. Specifically, I need to find new adhesive materials. We currently use thermally cured M-bond, Gatan G1 epoxy and crystal bond wax - all of which require heat.
If anyone has information on an adhesive that ion mills at a rate to similar Si, is stable under the electron beam, will cure at room temperature within about 24 hours, and once cured is impervious to solvents such as acetone, please respond. It would be an additional plus if the adhesive has low viscosity, so it can be used to secure TEM grids to the specimens.
Also I am looking for a replacement for the wax that we currently use to affix the samples to the polishing studs. This should cure quickly, bond strongly enough to endure the mechanical stresses of grinding, and be readily soluble in a solvent other than water so the samples can be removed from the studs.
Any help would be greatly appreciated.
Brenda
Brenda I. Prenitzer, Ph.D. Member of Technical Staff Cirent Semiconductor (Lucent Technologies) 9333 S. John Young Parkway 6D-Lab Orlando, FL 32819-8612
Our management has finally agreed to connect my electron microscopes to a recirculating water system.
I need some ideas what type of systems are on the market and the Australian agent for these units. We need to attach 2 SEMs, 1 TEM plus 1 coating unit to the new reciculating line.
Thank you
Hans Brinkies Senior Lecturer Swinburne, University of Technology School of Engineering and Science P.O.Box 218 - Hawthorn - Vic -3122 - Australia Phone: +61 3 9214 8657 Fax: +61 3 9214 8264 Email: Hbrinkies-at-swin.edu.au
1) Sensitivity: The CCD cameras are sensitive enough to see single electrons striking the phosphor. You can't get much more sensitive than that.
2) Price: I had a discussion about this with George McAuliffe in this forum about that theme a while ago. That thread was also printed in Microscopy Today. You may want to check the archives of this list server for "digital archiving/cost". I think George would agree with me, that the calculations of cost can swing in one or the other direction, depending on how you define cost, what you need to include and how many images you take (remember, a negative is on the order of $1 for the material alone, not labor time or anything else).
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Sergey Ryazantsev [mailto:sryazant-at-ucla.edu] Sent: Wednesday, May 17, 2000 1:06 PM To: Microscopy-at-sparc5.microscopy.com
Thank you Mickhael.
I will wait for info about sensitivity. I think, for EM in particular, sensitivity is very important parameter of the system. I am surprised that manufacturers don't have data on this matter. If sensitivity, say 10 times higher than SO-163 film - it may be a great reason to switch from film to the CCD. The major disadvantage of the CCD cameras for TEM is their price in my point of view. Thanks for your respond.
Sergey
} Date: Wed, 17 May 2000 11:34:05 -0600 } From: Michael Bode {mb-at-Soft-Imaging.com} } Subject: RE: new developments in imaging systems? } To: 'Sergey Ryazantsev' {sryazant-at-ucla.edu} } X-Mailer: Internet Mail Service (5.0.1457.3) } } Sergey, } } I will try to find out what we have regarding the relative } sensitivities. One thing that I can say now is, that we acquire images } with a 50 or 100 msec exposure of the digital camera when the film } requires an exposure of about 2 seconds. This would mean a 20 to 40 } times better sensitivity of the CCD camera. But to answer your question } in more detail would require to compare also the resolution of film and } camera and that is where it becomes very difficult, as it is not easy to } determine the resolution of film in terms of spatial resolution and } dynamic range, as both are interwoven. I will try to find some answers } for you. } } Regarding the linearity: As the CCDs simply count Photons, they have } almost perfect linearity over their complete dynamic range. Even more so } for TEMs, where all Photons have the same energy and the quantum } efficiency does not change from photon to photon. The Phosphor is a part } of a system that could theoretically introduce some non-linearities. } However, I did measure the linearity of some TEM camera systems a few } years back and did not find any significant deviations from linearity. } } Michael } } Michael Bode, Ph.D. } Soft Imaging System Corp. } 1675 Carr St., #105N } Lakewood, CO 80215 } =================================== } phone: (888) FIND SIS } (303) 234-9270 } fax: (303) 234-9271 } email: mailto:info-at-soft-imaging.com } web: http://www.soft-imaging.com } =================================== } } } } -----Original Message----- } From: Sergey Ryazantsev [mailto:sryazant-at-ucla.edu] } Sent: Wednesday, May 17, 2000 1:29 AM } To: Michael Bode } Subject: RE: new developments in imaging systems? } } } Mickhael hello } } I have question for you. I am thinking about adding CCD camera to my } JEM1200EX. The information I gathered from Internet is not so } optimistic. } The standard CCD resolution is about 1-1.5 million pixels, 2 million is } a } max as I understand. The price for cheaper camera is about 20-30 K$ - } much } more that I expect to spend for "film" process. There are two things } may } attract me to the modern CCD camera: dynamic range and sensitivity. I } am } pretty sure that dynamic range for CCD itself is a few orders better } than } any film available. But what about phosphorus screen? Does it reduce } dynamic range for the EM images? How dramatic? This is my first } question. } The next question is: could you tell me something about sensitivity of } the } modern CCD cameras used in EM? I am using dark field imaging at x80K } magnification and the exposure time for SO-163 (non deluded D-19, 7 min, } 20oC) is about 2 sec. I called GATAN, but they did not say anything } useful. } Could you provide some comparison of your side-mount camera with } sensitivity of the SO-163 film at condition I mentioned? I will greatly } appreciate any information in this matter. Thanks. Sergey. } } } } Date: Fri, 12 May 2000 13:46:01 -0600 } } From: Michael Bode {mb-at-Soft-Imaging.com} } } Subject: RE: new developments in imaging systems? } } To: "'Microscopy-at-MSA.Microscopy.Com'" } {Microscopy-at-sparc5.microscopy.com} } } Cc: 'Margaret Dienelt' {brannign-at-asrr.arsusda.gov} } } X-Mailer: Internet Mail Service (5.0.1457.3) } } } } ---------------------------------------------------------------------- - } - } } 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
A few months ago, I asked the same question and received the answers I copied below. By the way, I would like to thank again to all who respond my question. I do appreciate it. I still use these procedures and am planning to prepare a study comparing these techniques. All of them work very well.
Hope these helps. Good luck.
Ranan Gulhan AKTAS, M.D. Trakya University, Faculty of Medicine Pathology Department Edirne 22030 TURKEY
Dear Dr. Ranan Gulhan Aktas Protocol as follows: De-wax in 100% xylene - small pieces of tissue - 1mm3 (3X) Re-hydrate to water xylene/ethanol 100% each 100% ethanol 96% ethanol 70% ethanol H2O GA in buffer} Normal EM processing from here on Buffer }
Contact me if you have any further queries
Kind regards John
Mr John F. Putterill Electron Microscopy Unit Tel: (Int) 27-12-529-9174 Pathology Section Fax: (Int) 27-12-529-9165 Onderstepoort Veterinary Institute E-mail: john-at-moon.ovi.ac.za Private Bag X05 http://www.ovi.ac.za Onderstepoort 0110 South Africa
The basic procedure for re-embedding paraffin embedded is relatively simple. We had a Lynx Tissue Processor set up to do it automatically. Cut the paraffin tissue into 1-2 mm cubes with a warm scalpel. Trim as much excess paraffin away as possible. De-paraffinize with 6 alternately warm/room temp xylene washes with agitation for 30 minutes each. Reverse the dehydration process from 100% EtOH to your normal EM buffer. Refix and postfix with appropriate EM fixatives, dehydrate, and embed. The smaller the paraffin pieces, typically the better the deparaffinization. Your tissue will never look great though. The initial fixation for LM is a poor TEM fixative. good Luck.
Chuck Butterick Engineered Carbons, Inc.
Ranan,
Kai Chein did some nice work in this in the 1970's. He dissolved osmium in xylene(1 percent)and then deparaffinized in that solution. This took care of osmicating and depariffinization in one step (assuming you want to use osmium). After several changes in this solution you can go right into resin.. I've done this many times and it works very well and saves a lot of time.
Tim Morken, B.S., EMT(MSA), HTL(ASCP) Infectious Disease Pathology Centers for Disease Control MS-G32 1600 Clifton Rd. Atlanta, GA 30333 USA
email: tim9-at-cdc.gov timcdc-at-hotmail.com
FAX: (404)639-3043
MICROSCOPY AND IMAGING SERVICE CENTER CELL BIOLOGY AND NEUROSCIENCES
PROCEDURE FOR PRELIMINARY PREPARATION OF PARAFFIN EMBEDDED TISSUE FOR ELECTRON MICROSCOPY:
1. PLACE 0.5-1.0 mm CUBES OF TISSUE FROM PARAFFIN BLOCK IN XYLENE FOR 3 CHANGES OF 30 MINUTES EACH.
2. PUT IN 100% ETHANOL FOR 2 CHANGES OF 15 MINUTES WITH AGITATION.
3. PUT IN 95% ETHANOL FOR 2 CHANGES FOR 15 MINUTES WITH AGITATION.
4. PUT IN 70% ETHANOL FOR 2 CHANGES FOR 15 MINUTES WITH AGITATION.
5. PUT IN 50% ETHANOL FOR 2 CHANGES FOR 15 MINUTES WITH AGITATION.
6. RINSE IN PBS FOR 2 CHANGES OF 5 MINUTES EACH WITH AGITATION.
7. FOLLOW ROUTINE PREPARATION OF TISSUE FOR ELECTRON MICROSCOPY.
REFERENCE:
PIERCE, A., 1972. "A MANUAL FOR HISTOLOGICAL TECHNICIANS", LITTLE AND BROWN, BOSTON.
Your morphology will not be great since the tissue was probably fixed in formalin and not glut. I've done this many times and have even had publications with this method. Make sure you get all of the paraffin out.
George Lawton Chief Electron Microscopist Microscopy and Imaging Service Center UT Southwestern Medical Center at Dallas Phone: 214-648-7291 eMail: George.Lawton-at-email.swmed.edu
Hi: This is Bob Mixon ( I work with Bob Kayton on the PNEMS society and am on the OHSU campus) . I have re-embedded tissues from paraffin many times and many ways. I would recommend cutting out the piece of tissue from the paraffin block and putting through at least two changes of xylene for about 30 minutes each. Then you could put the tissue through either absolute alcohol or acetone to remove the rest of the paraffin. Probably 30 minutes and two changes. Some folks continue to run the tissue through a series of alcohol and try to refix in EM fix and post-fix in osmium. THIS IS FUTILE! I have never found any enhancement by doing this. You can simply dissolve osmium in the acetone and try fixing in this (two percent). and then running through by your routine into plastic (through alcohol, acetone, or propylene oxide etc). Thanks Bob Mixon
Hello Ranan.
Here is the method used by the pathology people around here.
- Remove tissue of interest from parafin embedded specimen, cut in 1-2 mm cubes. - Leave the specimens in fresh 1% OsO4 in XYLENE for 1,5 - 2 hrs. - Wash in xylene 2 times 10 min - One part resin(we use Epon-Araldite) + two parts xylene for 1 hr. - 1 part resin + 1 part xylene for 1 hr. - Resin for 30 min to 1 hour without a lid on the glass
All this steps on a carousel.
Embedd as usual, but keep the specimens in the resin over night before polymerization.
Good luck!
Best regards Randi Olsen
Randi Olsen Department of Electron Microscopy Faculty of Medicine University of Tromso MH-Breivika N-9037 TROMSO NORWAY
The following procedure may sound a little obvious, but I imagine you may be facing a rather limited access to all those books and journals, and I would feel really happy if this could help. I do wish you all the best in your EM lab-raising mission!
First, you will have to select smaller, "EM-size", portions of your paraffin-embedded specimens, cut them out and thoroughly deparaffinate. I would recommend at least 2 hours in xylene with frequent changes of xylene and some agitation; you should be able to see when it's all gone. Then several changes of 100% (anhydrous) ethanol, at least 1-1.5 h total time, and then gradually down the ethanol concentrations, like 95-85-70-50, 30 min or more at each step. Rinse 3x5 min in your choice of phosphate or cacodylate buffer, and then osmicate for 1-2 h, 1% OsO4 in the same buffer. Then dehydrate and embed for EM as you normally would.
Needless to say, even with the best original fixation the material will look pityful, but most of those diagnostically significant cell-to-cell junctions, filaments, etc. must still be there.
Best of luck! Sincerely, Vlad.
Vladislav V. Speransky Postdoctoral Research Associate School of Marine Sciences University of Maine 5722 Deering Hall Orono ME 04469-5722 Ph: 207 581 2998 FAX: 207 581 2969 Email: vladis-at-maine.maine.edu
Hi Keith,
At least for the specimens I happened to deal with so far (vertebrate tissues, cell culture, bacteria, microalgae), the material normally will not really blacken until you wash and start dehydrating it AFTER osmication in traditional aqueous OsO4 solutions. While in aqueous OsO4, it will remain brownish. (Unless, of course, you are using that simultaneous, single solution glutaraldehyde/OsO4 fixation, or a reducing buffer like PIPES, and have passed the time when it all turns black...)
The explanation used to be that, at the stage of osmication, while some of the OsO4, indeed, oxidizes nonsaturated lipids, etc., a lot of it simply dissolves in the lipids of the specimen as nonreduced OsO4. Then, say, the ethanol reduces that specifically accumulated OsO4, to form the so called "osmium black".
As for making FRESH solutions of OsO4 in xylene (or CCl4, etc.), I would just never have a spare OsO4 ampule (and enough paraffine blocks to EM-reevaluate) to try that! :-)
Sincerely, Vlad.
Vladislav V. Speransky Postdoctoral Research Associate School of Marine Sciences University of Maine 5722 Deering Hall Orono, ME 04469-5722 Ph: 207 581 2998 FAX: 207 581 2969 Email: vladis-at-maine.maine.edu
} I was interested in the use of osmium in xylene that you described. Does the tissue blacken as } in a water-solution or does it simply become yellowish/brown? I am curious because the latter is } what happens in the absence of water during freeze-substitution using e.g. 1% in acetone at -80 } *C, although the colour change probably happens when warming up from low temperature.
Hello Keith.
According to the people here that most often works with this (Irene Lund) the blocks are not as dark as with standard methods, but light brown. We haven't done freeze-substitution with osmium, so it's not easy to compare directly. For this purpose we buy ampoulas with 0,1 gram OsO4.
Best regards Randi Olsen Randi Olsen Department of Electron Microscopy Faculty of Medicine University of Tromso MH-Breivika N-9037 TROMSO NORWAY
At least for the specimens I happened to deal with so far (vertebrate tissues, cell culture, bacteria, microalgae), the material normally will not really blacken until you wash and start dehydrating it AFTER osmication in traditional aqueous OsO4 solutions. While in aqueous OsO4, it will remain brownish. (Unless, of course, you are using that simultaneous, single solution glutaraldehyde/OsO4 fixation, or a reducing buffer like PIPES, and have passed the time when it all turns black...)
The explanation used to be that, at the stage of osmication, while some of the OsO4, indeed, oxidizes nonsaturated lipids, etc., a lot of it simply dissolves in the lipids of the specimen as nonreduced OsO4. Then, say, the ethanol reduces that specifically accumulated OsO4, to form the so called "osmium black".
As for making FRESH solutions of OsO4 in xylene (or CCl4, etc.), I would just never have a spare OsO4 ampule (and enough paraffine blocks to EM-reevaluate) to try that! :-)
Sincerely, Vlad.
Vladislav V. Speransky Postdoctoral Research Associate School of Marine Sciences University of Maine 5722 Deering Hall Orono, ME 04469-5722 Ph: 207 581 2998 FAX: 207 581 2969 Email: vladis-at-maine.maine.edu
Dear HistoNetters,
Dr. Aktas asked about the re-embedding of paraffin embedded tissues in resin for observation by electron microscopy. There are two papers at our web site which describe this "Pop-Off" technique. Go to the URL provided in my signature file and follow the links to the JB-4 microtomy system, then to "Technical Information"
If anyone needs reprints of these articles and has no WWW access, send a note to Sonja White in my office (sales-at-ebsciences.com) {mailto:sales-at-ebsciences.com)} , and she'll s-mail you the dead tree version.
Best regards, Steven E. Slap, Vice-President
******************************************* Energy Beam Sciences, Inc. The Laboratory Microwave Company Adding Brilliance to Your Vision http://www.ebsciences.com {http://www.ebsciences.com} Ranan,
Kai Chein did some nice work in this in the 1970's. He dissolved osmium in xylene(1 percent)and then deparaffinized in that solution. This took care of osmicating and depariffinization in one step (assuming you want to use osmium). After several changes in this solution you can go right into resin.. I've done this many times and it works very well and saves a lot of time.
Tim Morken, B.S., EMT(MSA), HTL(ASCP) Infectious Disease Pathology Centers for Disease Control MS-G32 1600 Clifton Rd. Atlanta, GA 30333 USA
email: tim9-at-cdc.gov timcdc-at-hotmail.com
FAX: (404)639-3043 The basic procedure for re-embedding paraffin embedded is relatively simple. We had a Lynx Tissue Processor set up to do it automatically. Cut the paraffin tissue into 1-2 mm cubes with a warm scalpel. Trim as much excess paraffin away as possible. De-paraffinize with 6 alternately warm/room temp xylene washes with agitation for 30 minutes each. Reverse the dehydration process from 100% EtOH to your normal EM buffer. Refix and postfix with appropriate EM fixatives, dehydrate, and embed. The smaller the paraffin pieces, typically the better the deparaffinization. Your tissue will never look great though. The initial fixation for LM is a poor TEM fixative. good Luck.
Chuck Butterick Engineered Carbons,
Inc."Michelle.Taurino-at-aventis.com"-at-sparc5.microscopy.com wrote: ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Any suggestions on re-embedding paraffin sections in epon? Has anyone worked with the "pop out" method using BEEM capsules?
Any suggestions would be helpful...
Michelle Taurino Aventis Pharmaceuticals Bioimaging and Molecular Histology Michelle.Taurino-at-aventis.com 908-231-3357 "Michelle.Taurino-at-aventis.com"-at-sparc5.microscopy.com wrote: ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Any suggestions on re-embedding paraffin sections in epon? Has anyone worked with the "pop out" method using BEEM capsules?
Any suggestions would be helpful...
Michelle Taurino Aventis Pharmaceuticals Bioimaging and Molecular Histology Michelle.Taurino-at-aventis.com 908-231-3357
____________________________________________________________________ Get free email and a permanent address at http://www.netaddress.com/?N=1
I was talking about very special EM case: dark-field TEM. At this point we are talking about a few electrons per square angstrom, even less. In high resolution EM crystallography people have deal with 0.6 e/A2. At such conditions, high sensitivity and linearity of the modern digital cameras may be a plus. Talking about long exposures --what about drift? I never had good pictures at x100K with exposure longer that a few seconds. I don't understand your point about noise. In case of digital camera the noise is a function of camera. Current cameras has a very low level of noise (they uses cooling, etc) and we have to pay for that astronomical price. This is a life. I am not friendly with TEM cameras, but I do know that in the light microscopy people count individual photons using CCD cameras. In TEM camera we have phosphorous screen as a source of image and my questions to Mickhael were addressed mostly to the problem how effectively (and correctly) information is traveled thought that funny screen. The Mickhael's answer is that screen does not affect dynamic range and sensitivity is much higher than on convention films. Am I correct, Mickhaels?
Sergey
} Date: Thu, 18 May 2000 14:15:02 +1000 } From: jim {jim-at-proscitech.com.au} } Subject: RE: new developments in imaging systems? } To: 'Sergey Ryazantsev' {sryazant-at-ucla.edu} , } "Microscopy-at-sparc5.microscopy.com" {Microscopy-at-sparc5.microscopy.com} } Reply-to: "jim-at-proscitech.com.au" {jim-at-proscitech.com.au} } Organization: ProSciTech } X-Mailer: Microsoft Internet E-mail/MAPI - 8.0.0.4211 } } ------------------------------------------------------------------------ } 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
Hi We have a JEOL 733 with a two-sector PN type BS detector. Anyone out there have any idea on the resolution of these things in terms of mean atomic number. What I want to know basically is the sensitivity of these things to compositional variation. Cheers, Malc.
-- Dr MP Roberts Phone: [+27](0)46 603 8313 Dept of Geology Fax: [+27](0)46 622 9715 Rhodes University Cell: 083 4060 262 (usually off) 6140 Grahamstown e-mail: malc-at-rock.ru.ac.za SOUTH AFRICA
Thanks for all the responses to this posting both on and off the list. I routinely analyse minerals found in soils and reservoir sandstones and can easily recognise most of the common minerals by their EDS spectra. The problem for me recently has been looking at samples containing some of the more 'exotic' minerals, or put simply minerals I'm not used to looking at, e.g. sphene and clinpyroxenes. This has been more of a problem because we had no supporting XRD data for these particular analyses. What I was asking for, perhaps naively, was a program that would help identify an unknown mineral from its spectrum and give a list of possible alternatives. This is evidently not available. What is available: Thereis a free Mac program DTSA that can simulate spectra of a mineral for a particular detector and kV etc. Scott Wight/Mark Blackford). This can be downloaded from http://www.cstl.nist.gov/div837/837.02/dtsa.html Thereis a EDS spectrum database with search capabilities (based on archived spectra) written for the US FBI which may be commercially available soon. I will certainly follow up this up soon (thanks Dennis Ward and Nicholas Ritchie.) Thereseems to be various databases including MDAT that allow a search by chemical elements but not actual analyses. (thanks Bart Cannon) About a year ago, I started building a spectrum library collected on our system recording the various beam operating parameters, preparation (rough or polished, C or Au coated) etc. I agree with Bart Cannon that this is probably the best way forward. Just one thing - my database needs many more spectra added to it, especially different variations of many of the solid solution minerals. I suppose the idea of the database is that it should be used as a reference guide only. Although some minerals could never be identified by their EDS spectrum alone this is where an extensive reference library of my own would come in so useful and help answer the question: is this spectrum a Ca feldspar or Ca-rich zeolite? Although the best answer to this specific question would probably be the unknown spectrum indicates it may be a Zeolite because it is more similar to the zeolite reference we collected under similar operating parameters. Of course there would be no substitute if there were XRD data telling you in the same sample of zeolite and no Ca-feldspar. Thanks again to all of you who responded. Best regards Martin Roe
Martin J. Roe Macaulay Land Use Research Institute Craigiebuckler Aberdeen Scotland UK Phone 01224 318611 e-mail m.roe-at-mluri.sari.ac.uk
---------- } From: Hans Brinkies {HBrinkies-at-groupwise.swin.edu.au} } To: microscopy-at-sparc5.microscopy.com } Subject: Our management has finally agreed to connect my electronmicroscopes to a recirculating water system } Date: May 18, 2000 9:49 PM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Our management has finally agreed to connect my electron microscopes to } a recirculating water system. } } I need some ideas what type of systems are on the market and the } Australian agent for these units. We need to attach 2 SEMs, 1 TEM plus 1 } coating unit to the new reciculating line. } } Thank you } } Hans Brinkies } Senior Lecturer } Swinburne, University of Technology } School of Engineering and Science } P.O.Box 218 - Hawthorn - Vic -3122 - Australia } Phone: +61 3 9214 8657 } Fax: +61 3 9214 8264 } Email: Hbrinkies-at-swin.edu.au } } I don't really know what companies distribute such systems in Australia, but one thing occurs to me....if you set up all your instruments on one circulator, it's nice and cost-effective, but when the pump crashes or motor burns out, all your toys are down until the one circulation system is repaired. Separate systems for each 'scope would be better, (that'll probably give your managers a heart attack) or at least try and retain the capability of switching back to the old constant-loss system during emergencies. Our own ESEM was recently down for nearly three weeks while I was trying to replace the circulator pump motor. Turned out to be kind of a hard one to find...
Frank Thomas MicroAnalysis Facility Geological Survey of Canada Atlantic Bedford Institute of Oceanography Dartmouth, Nova Scotia
For the past 20 years I have used the Philips TEM heating holder furnace and heater elements to build hotstages for a range of microscopes. Unfortunately, Philips have told me that they can no longer supply the Pt furnace bodies and I cannot get information on possible replacements.
If anyone has a spare furnace (or more?) - Philips part number 5322 265 70028 - that they wish to part with I am willing to pay a `Philips' price. I am also willing to consider lightly used furnaces or complete holders that are now surplus to requirements and will pay a price depending on condition and history.
If anyone has any information on replacements to the previous style or on present day heaters I would be most grateful.
Thanks, Ron
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
Atomic number contrast sensitivity using BSE is dependent on several variables in addition to the equipment. I may not cover everything, but what quickly comes to mind....
Specimen surface condition - If a specimen is well polished, compo imaging is best. Topography of rough specimens (like a fracture) will generate feature contrast, "diluting" the compo contrast.
Beam current and potential affect sensitivity.
Conductive films will reduce sensitivity. Use carbon rather than sputtered Au, Pd etc. to minimize the effect if the specimen must be coated.
Depending on the detector geometry, working distance can affect the solid angle of collection thus changing BSE collection effiency.
The absloute atomic numbers present in the specimen will affect sensitivity in two ways.
Sensitivity will be different for a composition of lighter elements compared to a specimen composed of higher atomic numbers.
Also, if it is desired to NOT saturate the video, the range of atomic numbers can limit sensitivity. This is a typically a result of limited dynamic range in the image capture device/photo. For example, if C and W are present as pure inclusions in a brass, it will be difficult (to say the least) to contrast the difference between Cu and Zn phases whild not loosing C & W contrast variability.
Considering the above, any single statement of sensitivity is difficult, but if I had to generalize... My 4 diode GW Electronics BSE system will resolve 0.1 atomic number differencees under "good" conditions.
Woody White McDermott Technology
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Hi We have a JEOL 733 with a two-sector PN type BS detector. Anyone out there have any idea on the resolution of these things in terms of mean atomic number. What I want to know basically is the sensitivity of these things to compositional variation. Cheers, Malc.
-- Dr MP Roberts Phone: [+27](0)46 603 8313 Dept of Geology Fax: [+27](0)46 622 9715 Rhodes University Cell: 083 4060 262 (usually off) 6140 Grahamstown e-mail: malc-at-rock.ru.ac.za SOUTH AFRICA
I want to add my 2 cents as I like the topic. Most of this is a result of my experience with our TEM 2Kx2K CCD. We are working in bright field - so I cannot comment on dark field performance Several points:
1. The CCD is much more convenient - you get your pictures instantly.
2. CCD is linear and has larger dynamic range than the film.
3. The bad thing about the CCD is resolution - about 4 times lower than that of the film (in our camera the pixel size is 30 microns). So if you want to work in minimum dose you will do better with film. As I know the CCD is not performing well in terms of signal to noise at low doses (and if you have to work at 4 times higher magnification because of the resolution the things become much worse).
4. The CCD has smaller observation area - again loss of information.
5. I don't know about the detection efficiency compared to the film - it depends on the thickness of the phosphorous and the accelerating voltage. If a photon reaches the CCD chip it will be detected ... the problems are in the conversion electron-photon. There are two sides - if you make the phosphorous thicker you will get higher detection efficiency but the point spread also increases so always a compromise is made between detection efficiency and resolution. When I say detection efficiency this is not only related to the detection of single electrons (as it detects single electrons) but more to the actual signal detected on the background of the noise. Apart from the shot noise additional noise is added due to the scintillator driven detection and thermal noise in the CCD chip.
The CCDs are now very popular in diffraction work because of the dynamic range and linearity.
Here is one reference where a nice comparison between 2Kx2K CCD and film has been made:
Downing, K., Hendrickson, F., Ultramicroscopy, 75, (1999), 215-233
Best regards,
Rado
--------------------------------------------------------------------- Radostin Danev Laboratory of Ultrastructure Research National Institute for Physiological Sciences Myodaiji-cho, Okazaki 444-8585, JAPAN e-mail: rado-at-nips.ac.jp --------------------------------------------------------------------- ----- Original Message ----- } From: Sergey Ryazantsev {sryazant-at-ucla.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Friday, May 19, 2000 2:03 PM
If your samples are not site specific, then I would recommend a modified version of the small angle cleavage technique. You can get a detailed description of this technique in the MRS TEM Sample Prep IV book, vol 480.
Instead of the low temperature melting wax to hold the samples down, you use superglue. It takes longer to soak the samples off in acetone, but there is no heat. Then instead of the silver epoxy that is normally used, you need to use a slow curing viscous epoxy to mount the samples on the copper grid. There may be a temperature spike in the curing, but you would have to experiment and find out yourself. The amount of epoxy that is needed is very small and I doubt that it would raise the temperature an appreciable amount. The down side is that it will take a day to fully cure the epoxy. The net result is that the only heating the sample really sees is the heat generated during grinding the back side of the samples down.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
} -----Original Message----- } From: prenitzer,brenda s [mailto:prenitzer-at-lucent.com] } Sent: Thursday, May 18, 2000 7:06 PM } To: 'List Server' } Subject: Room Temperature TEM Prep } } } -------------------------------------------------------------- } ---------- } 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/MicroscopyListse} rver/FAQ.html } } } -------------------------------------------------------------- } ---------. } } } Hello all, } } I am in need of a way to modify my current method of preparing TEM } cross-sections as to introduce no heating. Specifically, I } need to find new } adhesive materials. We currently use thermally cured M-bond, } Gatan G1 epoxy } and crystal bond wax - all of which require heat. } } If anyone has information on an adhesive that ion mills at a } rate to similar } Si, is stable under the electron beam, will cure at room } temperature within } about 24 hours, and once cured is impervious to solvents such } as acetone, } please respond. It would be an additional plus if the } adhesive has low } viscosity, so it can be used to secure TEM grids to the specimens. } } } Also I am looking for a replacement for the wax that we } currently use to } affix the samples to the polishing studs. This should cure } quickly, bond } strongly enough to endure the mechanical stresses of grinding, and be } readily soluble in a solvent other than water so the samples } can be removed } from the studs. } } Any help would be greatly appreciated. } } Brenda } } Brenda I. Prenitzer, Ph.D. } Member of Technical Staff } Cirent Semiconductor (Lucent Technologies) } 9333 S. John Young Parkway } 6D-Lab } Orlando, FL 32819-8612 } } Phone: 407 371 7108 } Fax: 407 371 6999 } } prenitzer-at-lucent.com } bsp101-at-worldnet.att.net } }
as I said in my response, a direct comparison between the sensitivities of film and a CCD camera are very complicated. The film has a non-linear response curve, it reacts to electrons directly, there is grain size to take into account, etc. All I wanted to say is the following: For a film camera setting of 2 seconds, we acquire an image in approx. 50 to 100 msec with similar contrasts to that of film.
Regarding the linearity of the phosphor-CCD system: I have measured the linearity by measuring the beam current and the image intensity and I found it to be linear over the entire range that I measured. The deviations were insignificant and probably due to noise (I would give you the numbers, but this was in a different life and I don't have them). There may be nonlinearities that show up if you reach extreme illumination levels of the phosphor, but I did not measure them and it will probably depend on the phosphor itself.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Sergey Ryazantsev [mailto:sryazant-at-ucla.edu] Sent: Thursday, May 18, 2000 11:04 PM To: Microscopy-at-sparc5.microscopy.com
Jim hello
I was talking about very special EM case: dark-field TEM. At this point we are talking about a few electrons per square angstrom, even less. In high resolution EM crystallography people have deal with 0.6 e/A2. At such conditions, high sensitivity and linearity of the modern digital cameras may be a plus. Talking about long exposures --what about drift? I never had good pictures at x100K with exposure longer that a few seconds. I don't understand your point about noise. In case of digital camera the noise is a function of camera. Current cameras has a very low level of noise (they uses cooling, etc) and we have to pay for that astronomical price. This is a life. I am not friendly with TEM cameras, but I do know that in the light microscopy people count individual photons using CCD cameras. In TEM camera we have phosphorous screen as a source of image and my questions to Mickhael were addressed mostly to the problem how effectively (and correctly) information is traveled thought that funny screen. The Mickhael's answer is that screen does not affect dynamic range and sensitivity is much higher than on convention films. Am I correct, Mickhaels?
Sergey
} Date: Thu, 18 May 2000 14:15:02 +1000 } From: jim {jim-at-proscitech.com.au} } Subject: RE: new developments in imaging systems? } To: 'Sergey Ryazantsev' {sryazant-at-ucla.edu} , } "Microscopy-at-sparc5.microscopy.com" {Microscopy-at-sparc5.microscopy.com} } Reply-to: "jim-at-proscitech.com.au" {jim-at-proscitech.com.au} } Organization: ProSciTech } X-Mailer: Microsoft Internet E-mail/MAPI - 8.0.0.4211 } } ----------------------------------------------------------------------- - } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am new to this list. I would like to know if anyone has information I could use to write an article on the analytical chemistry techniques used to authenticate art objects using light microscopes or any other type of microscope. Also, if anyone has information on any of the other techniques used to authenticate art objects such as x-ray diffraction, x-ray fluorescence, accelerator mass spectrometry, that would be helpful too.
Sincerely,
Margaret M. Mitchell Assistant Editor AOAC INTERNATIONAL 301-924-7077 (tel) 301-924-7089 (fax) mmitchell-at-aoac.org
I'm looking for information regarding final polishing solutions for preparing metallic cross-section samples for TEM analysis.
I've looked into colloidal silica suspensions such as Ludox and Syton, and am concerned that their alkilinity may chemically etch my samples. I'm analyzing different compositions of NiFe on copper substrates, so the specimens are prone to preferential etching of different phases.
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With that agreement you also have a few new problems to consider. I suggest that: In Australia you could not buy a large system like that as a supplied item. Several smaller plants would be much too expensive and cumbersome. Any imported system will result in problems with fittings and parts
I expect that it would be best (been there, done that a couple of times) to design your own large system with a bit of spare capacity.
Beer chillers as used in hotels are most suitable. You want at least two large chillers, so if one fails you still could run more than half of the equipment). Calculate total heater element wattage with some spare capacity.
Diffusion pumps are designed to work best at 16 degrees. At that temperature the TEM column will condense water. Depending on ambient temperature, it might be sufficient to use the dif pump return line to cool the column and so avoid that problem. The chillers should be installed under an open shelter as they generate a lot of heat. Chiller temperature cut-in and out requires a thermostat each.
I suggest that a centrifugal pump of 1hp capacity will give enough pressure at the relative small flow rates. Chose a very common pump make so you can install a spare quickly without replacing fittings. Other pump types give more pressure, but they are more expensive and generally less reliable.
12mm reinforced garden hoses with hose clamps to and from the instruments make installation easy, pretty secure and cause minimal pressure drop.
Required is a SS tank (insulated) or a fiberglass lined tank to act as a heat sink. 100 liters would be a reasonable minimum capacity.
A good inline filter is required and a bypass, so the system can run during filter change.
If a mains water line is maintained, it is very important to have the returning water going to a drain whenever mains water is in use. I have lived through some horrid floods, which resulted from people opening mains but not the bypass to the drain.
If you have the components any plumber can install the system in under a day. The plumber would supply gate valves, temperature and pressure gauges etc.
Hans, you may fax your rough design to me, I'll be happy to make any suggestions. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Friday, May 19, 2000 10:50 AM, Hans Brinkies [SMTP:HBrinkies-at-groupwise.swin.edu.au] wrote: } } Our management has finally agreed to connect my electron microscopes to } a recirculating water system. } } I need some ideas what type of systems are on the market and the } Australian agent for these units. We need to attach 2 SEMs, 1 TEM plus 1 } coating unit to the new reciculating line. } } Thank you } } Hans Brinkies } Senior Lecturer } Swinburne, University of Technology } School of Engineering and Science } P.O.Box 218 - Hawthorn - Vic -3122 - Australia } Phone: +61 3 9214 8657 } Fax: +61 3 9214 8264 } Email: Hbrinkies-at-swin.edu.au }
John, The solutions that the colloidal silica are in have a ph of 8.5 to 9.8 depending on the formulation. I suggest you try our 0.05 micron water based polycrystalline diamond suspension. Allied High Tech http://www.alliedhightech.com sells this product. I would also suggest our Final A cloth for this polishing step. If you would like samples of either of these products please let me know and I will be happy to send it to you.
If you need further technical assistance please contact me off-line and I will be happy to help or you may contact our main office at (800)675-1118 located in CA.
I hope this helps. Ed Please note, I have a financial interest in providing you with these products and other sample preparation equipment and consumable items.
************************************************* Edward A. Hirsch Product Application Specialist Allied High Tech Products 2376 East Pacifica Place Rancho Dominguez, CA 90220 ph: (919) 846-9628 vm:(800)675-1118 x245 fx: (310)762-6808 http://www.alliedhightech.com
Equipment and Consumables for Metallurgical Sample Preparation *************************************************
-----Original Message----- } From: john david whitaker [mailto:jwhitake-at-u.washington.edu] Sent: Friday, May 19, 2000 4:01 PM To: Microscopy Lister Server
I'm looking for information regarding final polishing solutions for preparing metallic cross-section samples for TEM analysis.
I've looked into colloidal silica suspensions such as Ludox and Syton, and am concerned that their alkilinity may chemically etch my samples. I'm analyzing different compositions of NiFe on copper substrates, so the specimens are prone to preferential etching of different phases.
Dear Friends, Can anyone give me advice regarding the dismantling and the packing for shipment of a Philips 201 TEM? I need to pay particular attention to not disturbing the alignment. I plan to move the TEM from Univ. of Delaware to Naples, Florida in an enclosed U-Haul Trailer. Any help or suggestions will be most welcome. If anyone has hands-on experience in doing this sort of thing and is willing to give me a hand at U-Del, please contact me and name your price. I'll gladly pay for the assistance. Best regards, Mike Urbanik www.crystalguru.com
I find what you are trying to do interesting, but there are things that may never work and this may be one.
We had it confirmed by Michael Bode that good digital system "see "single photons - so that is not likely to improve much. Film too registers single electrons. We were told that in digital 1 electron can expose one pixel. On film one electron initiates the exposure of a silver halide and subsequent electrons in a near identical location increase the size of the grain.
Low noise (cooled cameras) hopefully do not add their own noise, but they cannot make up for lack of information in the image.
My case (below) was that in high resolution TEM at least, less exposed images will be inferior because there are simply not enough electrons to produce a good image. A more sensitive digital system uses fewer electrons and so makes matters worse.
In dark field EM we are using the beam indirectly and such images too suffer particularly from electron noise. Here too the use of a digital system would only make this worse. Furthermore, the slower 4489 film would be better than SO-163.
If you don't like long exposures (4 to 8 seconds I found a practical limit), you cannot ignore the reality of electron noise and hope to correct that with yet fewer electrons, albeit processed in a "noise-free" digital system. I think that the answer is more electrons, i.e. a field emission source or a slower digital camera. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Friday, May 19, 2000 3:04 PM, Sergey Ryazantsev [SMTP:sryazant-at-ucla.edu] wrote: } } } Jim hello } } I was talking about very special EM case: dark-field TEM. At this point we } are talking about a few electrons per square angstrom, even less. In high } resolution EM crystallography people have deal with 0.6 e/A2. At such } conditions, high sensitivity and linearity of the modern digital cameras } may be a plus. Talking about long exposures --what about drift? I never } had good pictures at x100K with exposure longer that a few seconds. I } don't understand your point about noise. In case of digital camera the } noise is a function of camera. Current cameras has a very low level of } noise (they uses cooling, etc) and we have to pay for that astronomical } price. This is a life. I am not friendly with TEM cameras, but I do know } that in the light microscopy people count individual photons using CCD } cameras. In TEM camera we have phosphorous screen as a source of image and } my questions to Mickhael were addressed mostly to the problem how } effectively (and correctly) information is traveled thought that funny } screen. The Mickhael's answer is that screen does not affect dynamic range } and sensitivity is much higher than on convention films. Am I correct, } Mickhaels? } } Sergey } } } Date: Thu, 18 May 2000 14:15:02 +1000 } } From: jim {jim-at-proscitech.com.au} } } Subject: RE: new developments in imaging systems? } } To: 'Sergey Ryazantsev' {sryazant-at-ucla.edu} , } } "Microscopy-at-sparc5.microscopy.com" {Microscopy-at-sparc5.microscopy.com} } } Reply-to: "jim-at-proscitech.com.au" {jim-at-proscitech.com.au} } } Organization: ProSciTech } } X-Mailer: Microsoft Internet E-mail/MAPI - 8.0.0.4211 } } } } ------------------------------------------------------------------------ } } 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 } } -----------------------------------------------------------------------. } } } } } } If sensitivity was ten times higher, one would in some cases save some beam } } damage and in most cases suffer excessive electron noise. } } As a rough guide, electron noise becomes apparent at 30x enlargement, } rarely a } } problem. Magnification is a linear function and electron density relates } to an } } area, but clearly very short exposure images would be much noisier and } could } } not be enlarged nearly as much. When not enough electrons form an image it } } appears grainy, which makes it unsuitable for further enlarging. Photo } } enlarging utilises higher depths-of-field and without this, very high } power TEM } } is much, much harder. } } By nature, slower emulsions have finer grain and higher resolution and } } contrast. It is fortuitous that these desirable characteristics run in } tandem } } with relatively long exposures, so the image is formed by more electrons. } } TEM films rate about 6 ISO, so X-ray film at about ISI 3000 is about a 10x } } faster than TEM films. Unfortunately its rather grainy, but if the } exposure was } } well adjusted, chances are that electron noise would be more bothersome than } } } } the film's grain. } } Digital is inevitable and already fairly common, but some aces remain with } } conventional film. } } Cheers } } Jim Darley } } ProSciTech Microscopy PLUS } } PO Box 111, Thuringowa QLD 4817 Australia } } Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com } } Great microscopy catalogue, 500 Links, MSDS, User Notes } } www.proscitech.com } } } } On Thursday, May 18, 2000 5:06 AM, Sergey Ryazantsev } [SMTP:sryazant-at-ucla.edu] } } wrote: } } } } } } } } } Thank you Mickhael. } } } } } } I will wait for info about sensitivity. I think, for EM in particular, } } } sensitivity is very important parameter of the system. I am surprised } } } that } } } manufacturers don't have data on this matter. If sensitivity, say 10 } } } times } } } higher than SO-163 film - it may be a great reason to switch from film to } } } the CCD. The major disadvantage of the CCD cameras for TEM is their price } } } in my point of view. } } } Thanks for your respond. } } } } } } Sergey } } } } } } } } } } Date: Wed, 17 May 2000 11:34:05 -0600 } } } } From: Michael Bode {mb-at-Soft-Imaging.com} } } } } Subject: RE: new developments in imaging systems? } } } } To: 'Sergey Ryazantsev' {sryazant-at-ucla.edu} } } } } X-Mailer: Internet Mail Service (5.0.1457.3) } } } } } } } } Sergey, } } } } } } } } I will try to find out what we have regarding the relative } } } } sensitivities. One thing that I can say now is, that we acquire images } } } } with a 50 or 100 msec exposure of the digital camera when the film } } } } requires an exposure of about 2 seconds. This would mean a 20 to 40 } } } } times better sensitivity of the CCD camera. But to answer your question } } } } in more detail would require to compare also the resolution of film and } } } } camera and that is where it becomes very difficult, as it is not easy to } } } } determine the resolution of film in terms of spatial resolution and } } } } dynamic range, as both are interwoven. I will try to find some answers } } } } for you. } } } } } } } } Regarding the linearity: As the CCDs simply count Photons, they have } } } } almost perfect linearity over their complete dynamic range. Even more so } } } } for TEMs, where all Photons have the same energy and the quantum } } } } efficiency does not change from photon to photon. The Phosphor is a part } } } } of a system that could theoretically introduce some non-linearities. } } } } However, I did measure the linearity of some TEM camera systems a few } } } } years back and did not find any significant deviations from linearity. } } } } } } } } Michael } } } } } } } } Michael Bode, Ph.D. } } } } Soft Imaging System Corp. } } } } 1675 Carr St., #105N } } } } Lakewood, CO 80215 } } } } =================================== } } } } phone: (888) FIND SIS } } } } (303) 234-9270 } } } } fax: (303) 234-9271 } } } } email: mailto:info-at-soft-imaging.com } } } } web: http://www.soft-imaging.com } } } } =================================== } } } } } } } } } } } } } } } } -----Original Message----- } } } } From: Sergey Ryazantsev [mailto:sryazant-at-ucla.edu] } } } } Sent: Wednesday, May 17, 2000 1:29 AM } } } } To: Michael Bode } } } } Subject: RE: new developments in imaging systems? } } } } } } } } } } } } Mickhael hello } } } } } } } } I have question for you. I am thinking about adding CCD camera to my } } } } JEM1200EX. The information I gathered from Internet is not so } } } } optimistic. } } } } The standard CCD resolution is about 1-1.5 million pixels, 2 million is } } } } a } } } } max as I understand. The price for cheaper camera is about 20-30 K$ - } } } } much } } } } more that I expect to spend for "film" process. There are two things } } } } may } } } } attract me to the modern CCD camera: dynamic range and sensitivity. I } } } } am } } } } pretty sure that dynamic range for CCD itself is a few orders better } } } } than } } } } any film available. But what about phosphorus screen? Does it reduce } } } } dynamic range for the EM images? How dramatic? This is my first } } } } question. } } } } The next question is: could you tell me something about sensitivity of } } } } the } } } } modern CCD cameras used in EM? I am using dark field imaging at x80K } } } } magnification and the exposure time for SO-163 (non deluded D-19, 7 min, } } } } 20oC) is about 2 sec. I called GATAN, but they did not say anything } } } } useful. } } } } Could you provide some comparison of your side-mount camera with } } } } sensitivity of the SO-163 film at condition I mentioned? I will greatly } } } } appreciate any information in this matter. Thanks. Sergey. } } } } } } } } } } } } } Date: Fri, 12 May 2000 13:46:01 -0600 } } } } } From: Michael Bode {mb-at-Soft-Imaging.com} } } } } } Subject: RE: new developments in imaging systems? } } } } } To: "'Microscopy-at-MSA.Microscopy.Com'" } } } } {Microscopy-at-sparc5.microscopy.com} } } } } } Cc: 'Margaret Dienelt' {brannign-at-asrr.arsusda.gov} } } } } } X-Mailer: Internet Mail Service (5.0.1457.3) } } } } } } } } } } ----------------------------------------------------------------------- } } } } - } } } } } 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 } } } } } ----------------------------------------------------------------------- } } } } . } } } } } } } } } } } } } } } Margaret, } } } } } } } } } } As a former user and current vendor of such systems as you are } } } } inquiring } } } } } about I can try to provide a bit of information regarding camera } } } } } improvements: } } } } } } } } } } There have been a number of improvements, but I am not sure what you } } } } are } } } } } comparing the latest cameras against. Cameras are now usually cooled } } } } and } } } } } provide 12 bits per pixel, the number of pixels has gone up a bit (but } } } } } not much in general), and cameras read out faster than they used to (up } } } } } to 20 fps and more). I think all cameras now use a line transfer } } } } } mechanism, which makes shutters obsolete. } } } } } On the software side, real-time FFT and real-time shading correction } } } } can } } } } } be done now due to faster computers without special processing boards, } } } } } and there have been other software developments that make using the } } } } } cameras and computers easier. } } } } } Other changes that affect the usability of cameras is the use of } } } } } pneumatics to insert and retract the phosphors, higher frame rates for } } } } } live viewing with the camera, etc. } } } } } } } } } } If you have questions, please give me a call, drop me an email, or go } } } } to } } } } } our web site. } } } } } } } } } } Michael } } } } } } } } } } } } } } } Michael Bode, Ph.D. } } } } } Soft Imaging System Corp. } } } } } 1675 Carr St., #105N } } } } } Lakewood, CO 80215 } } } } } =================================== } } } } } 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: Margaret Dienelt [mailto:brannign-at-asrr.arsusda.gov] } } } } } Sent: Thursday, May 11, 2000 12:43 PM } } } } } To: Microscopy-at-sparc5.microscopy.com } } } } } Subject: TEM: new developments in imaging systems? } } } } } } } } } } } } } } } ----------------------------------------------------------------------- } } } } - } } } } } 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, } } } } } } } } } } Year after year I hopefully gather information about digital imaging } } } } } systems for TEMs (ours is a JEOL 100CX) , only to learn we have no } } } } } money. This year it looks like it might really happen but I have not } } } } } kept up with innovations in the field and am wondering the following: } } } } } } } } } } 1. Anything new in the last two years -- especially in terms of } } } } } cameras? I'm most familiar with the Gatan and AMT systems but their } } } } } web sites don't reflect much in the way of changes over a year ago. } } } } } 2. With more and more microscopists finally getting their systems -- } } } } } I'd love to get feedback. } } } } } } } } } } Thanks, } } } } } Margaret } } } } } } } } } } P.S. Would welcome contacts from vendors. } } } } } } } } } } -- } } } } } Margaret Dienelt } } } } } } } } } } Plant Pathologist } } } } } Electron Microscopy Lab } } } } } } } } } } Floral and Nursery Plants Research Unit } } } } } U.S. National Arboretum/Agricultural Research Service/USDA } } } } } } } } } } B. 010A, Rm. 238, BARC-W } } } } } 10300 Baltimore Avenue } } } } } Beltsville MD. 20705 USA } } } } } } } } } } (301) 504-6097 } } } } } Fax: (301) 504-5096 } } } } } } } } } } } } } } _____________________________________ } } } } } } } } Sergey Ryazantsev Ph. D. } } } } Electron Microscopy } } } } UCLA School of Medicine } } } } Department of Biological Chemistry } } } } Box 951737 } } } } Los Angeles, CA 90095-1737 } } } } } } } } Phone: (310) 825-1144 } } } } Pager: (310) 845-0248 } } } } FAX (departmental): (310) 206-5272 } } } } mailto:sryazant-at-ucla.edu } } } } http://www.bol.ucla.edu/~sryazant } } } } } } } } } } } _____________________________________ } } } } } } Sergey Ryazantsev Ph. D. } } } Electron Microscopy } } } UCLA School of Medicine } } } Department of Biological Chemistry } } } Box 951737 } } } Los Angeles, CA 90095-1737 } } } } } } Phone: (310) 825-1144 } } } Pager: (310) 845-0248 } } } FAX (departmental): (310) 206-5272 } } } mailto:sryazant-at-ucla.edu } } } http://www.bol.ucla.edu/~sryazant } } } } } } } } } } } } } _____________________________________ } } Sergey Ryazantsev Ph. D. } Electron Microscopy } UCLA School of Medicine } Department of Biological Chemistry } Box 951737 } Los Angeles, CA 90095-1737 } } Phone: (310) 825-1144 } Pager: (310) 845-0248 } FAX (departmental): (310) 206-5272 } mailto:sryazant-at-ucla.edu } http://www.bol.ucla.edu/~sryazant } }
Dear Malc! Some years ago I got the resolution 0.1 in the middle of periodic table (around Z=28) on standard 733 BSE. But for this purpose it is necessary to balance additionally the preamplifier to remove completely the rest of topo signal, which becomes comparable by value with a very low signal of differential compo. There are no means in the preamplifier for this additional balancing, therefore I have soldered the additional variable resistor in one of shoulders of a differential amplifier in the preamplifier. Frankly speaking I have forgotten the details already, but I can restore them, if you wait about two weeks. But basically it is very simple. Regards. Victor Sidorenko, ANTRON Co. Ltd., scientific service, Moscow, Russia.
-----é ----- ë: Dr Malcolm Roberts {malc-at-rock.ru.ac.za} æ: Microprobe discussion group {Microscopy-at-sparc5.microscopy.com} : 19 Ø 2000 . 16:13 ñ: BSE resolution
} --------------------------------------------------------------------- --- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Polishing suspensions that does not interact chemically with your samples include diamond suspensions (available down to 0.1µm at least) and alumina suspensions (down to 0.05µm). Many vendors have information available on the World-Wide-Web.
Cheers, Paul =================== Paul Baggethun Alcoa Technical Center Alcoa Center, PA 15069 USA =================== + 724 - 337-1760 (tel) + 724 - 337-2044 (fax) ===================
} ---------- } From: john david whitaker[SMTP:jwhitake-at-u.washington.edu] } Sent: Friday, May 19, 2000 5:01 PM } To: Microscopy Lister Server } Subject: final polishing for metals (Cu, Ni, Fe, NiFe) } } ------------------------------------------------------------------------ } 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'm looking for information regarding final polishing } solutions for preparing metallic cross-section samples } for TEM analysis. } } I've looked into colloidal silica } suspensions such as Ludox and Syton, and am concerned } that their alkilinity may chemically etch my samples. } I'm analyzing different compositions of NiFe } on copper substrates, so the specimens are prone to } preferential etching of different phases. } } Does anyone have any experience with this? } } Any input would be appreciated. } } John } }
On Thu, 18 May 2000 ComCryLab1-at-aol.com-at-sparc5.microscopy.com wrote:
} Dear Friends, } Can anyone give me advice regarding the dismantling and } the packing for shipment of a Philips 201 TEM? I need to } pay particular attention to not disturbing the alignment. } I plan to move the TEM from Univ. of Delaware to Naples, Florida } in an enclosed U-Haul Trailer. Any help or suggestions } will be most welcome.
I will make sure Ron Veil, a very experienced independent EM service tech, sees this and has a chance to reply. It was with his advice that we (my hubby and I and its new owner) packed up and shipped a Philips 201. Like you, we wanted to ship it intact, column on and all. We built a heavy duty skid and added a strong base to which we bolted the instrument. Then we wrapped it with whatever that plastic packing tape that is like Saran Wrap is called, making sure to secure any part of the column we didn't want to move, but avoiding putting any pressure on things, such as the aperture drives. Wrap the bottom, wrap the column, wrap the column to the bottom and back again, etc. Build a crate up around the instrument. I think I remember putting a wood insert with a crescent cut out near the top of, but not touching the column, but hubby thinks not. The idea is NOT to let any shock to the crate get transferred to the column, so perhaps we didn't. Add some packing material, such as old egg crate foam from your bed (it needed replacing anyway) and whatever. Then, and this was the fun part, buy that stuff that when you mix it with a catalyst, produces huge volumes of foam that hardens in a few minutes. I think you can also buy spray cans of similar stuff, but since hubby had this on hand for other things, we had the bulk stuff. Fill the voids in the crate with it. It will easily chip off later. Add a top, and away you go. The scope made it in great shape.
I think we packed the rotary pump and a couple of other things separately. Get it into the truck and TIE IT DOWN. I say this because an SEM we once packed for shipping with the base and saran, but no closed crate, made it from Hawaii to Ohio, then slid in the shippers truck in the last mile and slammed into the driver's seat. The driver was OK, the ion getter pump was bent, and the column needed a bit of work, but it could have been worse. Duh.
I've also packed up a couple of Denton vcuum evaporators and a couple of ultramicrotomes. The saran stuff and blow foam are a good way to stabilize the instruments.
Good luck!
Aloha, Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
} From: "Tina Carvalho" {tina-at-pbrc.hawaii.edu} } On Thu, 18 May 2000 ComCryLab1-at-aol.com-at-sparc5.microscopy.com wrote: } } } Dear Friends, } } Can anyone give me advice regarding the dismantling and } } the packing for shipment of a Philips 201 TEM? I need to } } pay particular attention to not disturbing the alignment. } } I plan to move the TEM from Univ. of Delaware to Naples, Florida } } in an enclosed U-Haul Trailer. Any help or suggestions } } will be most welcome. } } I will make sure Ron Veil, a very experienced independent EM service tech, } sees this and has a chance to reply. It was with his advice that we (my } hubby and I and its new owner) packed up and shipped a Philips 201. Like } you, we wanted to ship it intact, column on and all. We built a heavy } duty skid and added a strong base to which we bolted the instrument. Then } we wrapped it with whatever that plastic packing tape that is like Saran } Wrap is called, making sure to secure any part of the column we didn't } want to move, but avoiding putting any pressure on things, such as the } aperture drives. Wrap the bottom, wrap the column, wrap the column to the } bottom and back again, etc. Build a crate up around the instrument. } I think I remember putting a wood insert with a crescent cut out near the } top of, but not touching the column, but hubby thinks not. The idea is } NOT to let any shock to the crate get transferred to the column, so } perhaps we didn't. Add some packing material, such as old egg crate foam } from your bed (it needed replacing anyway) and whatever. Then, and this } was the fun part, buy that stuff that when you mix it with a catalyst, } produces huge volumes of foam that hardens in a few minutes. I think you } can also buy spray cans of similar stuff, but since hubby had this on hand } for other things, we had the bulk stuff. Fill the voids in the crate with } it. It will easily chip off later. Add a top, and away you go. The } scope made it in great shape. } } I think we packed the rotary pump and a couple of other things } separately. Get it into the truck and TIE IT DOWN. I say this because an } SEM we once packed for shipping with the base and saran, but no closed } crate, made it from Hawaii to Ohio, then slid in the shippers truck in the } last mile and slammed into the driver's seat. The driver was OK, the ion } getter pump was bent, and the column needed a bit of work, but it could } have been worse. Duh. } } I've also packed up a couple of Denton vcuum evaporators and a couple of } ultramicrotomes. The saran stuff and blow foam are a good way to } stabilize the instruments. } Since you are using a U haul you don't have to worry about sides and a top on the crate. Make sure it is tied down real well. Dry wall screws through the crate and into the floor work well for locking it down but make sure you have it braced with timbers to the front and sides of the trailer in case you make a panic stop.
Also make sure that the weight is centers in front of the trailer axle if you use a trailer. Negitive weight on the trailer tounge at the very least makes driving very interesting. You have no Idea how fast a trailer can pass you while it is still tied on to the truck. I have had the privilege of experiencing this and I can promise you that you won't enjoy it:).
The foam in place stuff is great. Just cover the instrument in plastic and foam away. If you can get foam between the instrument and the Support the foam will dissipate most of those little shocks that get things out of line.
Last of all make sure you understand what you insurance covers and what it doesn't on moving equipment. You should be fine on liability but the value of the contents is probably not covered. Rental truck companies have contents insurance as an extra.
Good luck Gordon
Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
I would like to announce an exciting job opportunity to the Microscopy community.
JOB ANNOUNCEMENT
POSITION TITLE: SR. SCIENTIST/ENGINEER LIGHT MICROSCOPY NIKON INC. MELVILLE, NY Posted: May 18, 2000
As seen in the 19 May issue of Science: http://recruit.sciencemag.org/cgi/show/5469/5469x03207
Sr. Applications Scientist/Engineer Bring Your Light Microscopy Experience To The BioScience Division Of Nikon Inc.
Our world renowned company has an excellent opportunity for a professional with extensive experience in light microscopy, specifically in advanced BioScience technologies in our USA headquarters located in Melville, Long Island , NY. Today, scientists at world-renowned biological institutions are making tremendous advances in Cancer, AIDS, Alzheimer's, in-vitro fertilization and other leading-edge research. Nikon is proud to be playing a role in this enormously important work. Our ongoing commitment to optical excellence and technological advancement is allowing researchers to view objects never before seen by the human eye. We would like to invite you to investigate the new opportunities and technologies available at Nikon Inc. BioSciences.
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Selected candidate will act as liaison for technical information and feedback between the end user/sales network and Nikon factory engineers on all matters pertaining to our BioScience technical product line. Applicants must have a Ph.D. in BioSciences, BioEngineering or Physics with a specialty in optics, or equivalent experience. Knowledge of advanced microscopy techniques and optical principals including fluorescence applications, imaging and confocal applications required. Excellent communications skills in English a must, along with good writing and public speaking abilities. Excellent benefits and compensation provided.
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I've had to coat some PLGA polymer samples and we think the coater may be melting them. The samples have a rolled or beaded edge and large cracks. I have jpeg images if anyone wants me to send them for a look. Are we correct in our assessment or should we look elsewhere for the source of cracks, shrinkage, beading around edges. These artifacts, BTW do not appear when the uncoated samples are checked under a light microscope.
Dear friends, I have just finished writing the programs for TEMAlert system(http://www6.ewebcity.com/temalert), which serves as a pre-print announcement system. Currently it serves only TEM-related region. Do you think this is very useful for all TEM users? The system was designed to be totally self-maintaining and everyone can post messages to announce his/her newly published (accepted) paper there.
I hope that TEMAlert will become a widespread blackboard in TEM region and tighten the relationship between electron microscopists allover the world.
I am sorry that the server is somewhat slow - because it is a free internet host. I will be very grateful if anyone of you can supply me a space (should support ASP).
With best regards, Shu-You Li ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Please visit my new homepage - a personal website on transmission electron microscopy (TEM) - at http://syli.homepage.com at your convenience! It contains my current research work, my resume TEM-related journals, instruction for authors link to on-line EELS database, periodic table, physical constants JOB list and RESUMEs (only for TEM region) TEMAlert - a self-maintaining preprint announcement system ************************************************** Shu-You Li, Dr. Institut fuer Physikalische Chemie Johannes Guttenberg Universitaet Jakob-Welder-Weg 11 D-55099 Mainz, Germany
} In Australia you could not buy a large system like that as a supplied item.
Don't know about that...We've had excellent results from a locally made system we've been using here now on the SEMs for several years. Service and parts are readily available and it has heaps of spare capacity. Contact Mark Blackford (mgb-at-postoffice.ansto.gov.au) for details on who to contact about this system because they probably have a branch in Melbourne.
} } Beer chillers as used in hotels are most suitable.
Certainly a mass produced item in Australia.
Arthur Day, Electron Microscope Unit Phone: 61-2-9717-3457 Ansto Materials Division Fax: 61-2-9543-7179 PMB 1, Menai (Sydney), NSW, 2234 Email: ard-at-ansto.gov.au Australia www: http://www.ansto.gov.au/
I have a batch of images (~300) that I would like to present as a QuickTime movie. Does anybody have experience how to do this? I am using a Macintosh. Comments welcome. Thanks,
Edgar
________________ Dr. Edgar Voelkl ORNL Bldg 4515 1 Bethel Valley Road P.O. Box 2008 Oak Ridge, TN 37831-6064
As a metallographic equipment and consumables manufacturer, BUEHLERš offers a number of products which would work for your application. However, I would suggest our MASTERPREP* (Part No. 40-6377-032) product. This is a 0.05 micron alumina suspension. What makes it unique is the fact that it is produced through the seeded gel process instead of by calcining. In the seeded gel process, the alumina is precipitated from a liquid phase. This results in a better controlled particulate size distribution, higher particulate density, and more consistent particle geometry. We've found this product to be superior to all of the other 0.05 micron alumina products that we sell for preparation of ductile metals.
I hope this helps. If you need further information, you can email me privately, call me at the numbers listed below, or contact our sales department for pricing.
Best regards, Scott D. Holt BUEHLERš , LTD. PO Box 1 41 Waukegan Rd. Lake Bluff, IL 60044 (847)295-4546 or (800) 323-9330 www.buehler.com
I am also trying to figure out how to make a QuickTime video from a series of images. I am using a PC, and the images are from a Zeiss 510 Confocal, in LSM format, which have been converted to Tiff. The number I am dealing with is 64 images. Any help would be greatly appreciated Linda Barthel Research Associate II Department of Cell and Developmental Biology University of Michigan barthel-at-umich.edu
We have a client, here in South Africa, who has recently purchased a Philips XL30 ESEM LaB6 with a cryo system, hot stage and CL detectors. We would like to now run a workshop, here on this system in South Africa, on the applications of ESEM. This system is a national facility and would therefore like to introduce the local EM users to the exciting world of ESEM.
We realise that there are a few friends of ours in Australia who would be ideal, but then we have various contacts in England and the USA too. In this way we feel we should get a chance at the best choice of getting a really exciting workshop set up or possibly a series of workshops. We expect to keep the delegates riveted for at least 4 days of workshop. The idea of the workshop is to show as many applications for ESEM as possible. Then to specialise in some of the biological areas, as this would be some of the more regular users for this system. Those who could assist should please contact us and we will pass your information on to the client. Please indicate your availability, field of interest and the, always important, costs involved.
Thanks for your time.
Luc Harmsen Anaspec, South Africa Technical support on microscopy. Tel + 27 (0) 11 476 3455 Fax + 27 (0) 11 476 7290 anaspec-at-icon.co.za www.anaspec.co.za
Remember, ICEM 15 will be in 2002, Durban, South Africa. www.icem15.com
We have a sample of 6 micrometers Ni-Zn-P electroless plated onto 1 mm thick Al and we would like to prepare cross sections for TEM examination. It looks very difficult to use the tripod polishing technique for metals.So,we are thinking at an electropolishing procedure with our double-jet Tenupol apparatus. This would be done on slices cutted from a sample initially thicked up to 3mm by electroplating.But how to avoid differential material removal problems? Which material for plating?Which electrolyte for electropolishing?Other method? Does anyone have suggestions for us?
Thanks in advance for your help.
Jean Dille
Materials Science and Electrochemistry Free University of Brussels CP 194/3 Avenue F.Roosevelt 50 1050 BRUSSELS BELGIUM tel:32-2-6502723 fax:32-2-6502786 e-mail: jdille-at-ulb.ac.be
The results are determined by the largest source of noise, be it the initial electron statistics or subsequent noise introduced by conversions or recording techniques. In this case (dark field imaging), the largest source for noise is probably the statistical nature of the electron beam. The lower you go in exposure (i.e., the fewer electrons you record per pixel), the higher the relative noise. And there is no way to get around this either by film or digital camera. The only way to improve the noise is to go to more electrons, i.e., longer exposure or brighter beam.
If the sample is so sensitive that even a dark field exposure damages the sample, there is probably not much you can do.
If the problem is drift, a digital system can help you: Instead of 1 exposure at, for example, 30 seconds, acquire 3 exposures at 10 seconds. Each one of them will be very noisy, but one can add them to get the same noise figure as a 30 sec exposure. And the 10 sec exposure of each one cuts down on drift. All you need to do is to align the 3 images.
I used to take dark field images with a digital camera and (of course) I loved it. It allowed me to acquire the images and immediately see them. I did not have to wait hours to develop the negatives and then find out the exposure was not long enough (or too long).
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== 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: jim [mailto:jim-at-proscitech.com.au] Sent: Sunday, May 21, 2000 2:38 AM To: 'Sergey Ryazantsev'; Microscopy-at-sparc5.microscopy.com
I find what you are trying to do interesting, but there are things that may never work and this may be one.
We had it confirmed by Michael Bode that good digital system "see "single photons - so that is not likely to improve much. Film too registers single electrons. We were told that in digital 1 electron can expose one pixel. On film one electron initiates the exposure of a silver halide and subsequent electrons in a near identical location increase the size of the grain.
Low noise (cooled cameras) hopefully do not add their own noise, but they cannot make up for lack of information in the image.
My case (below) was that in high resolution TEM at least, less exposed images will be inferior because there are simply not enough electrons to produce a good image. A more sensitive digital system uses fewer electrons and so makes matters worse.
In dark field EM we are using the beam indirectly and such images too suffer particularly from electron noise. Here too the use of a digital system would only make this worse. Furthermore, the slower 4489 film would be better than SO-163.
If you don't like long exposures (4 to 8 seconds I found a practical limit), you cannot ignore the reality of electron noise and hope to correct that with yet fewer electrons, albeit processed in a "noise-free" digital system. I think that the answer is more electrons, i.e. a field emission source or a slower digital camera. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Friday, May 19, 2000 3:04 PM, Sergey Ryazantsev [SMTP:sryazant-at-ucla.edu] wrote: } } } Jim hello } } I was talking about very special EM case: dark-field TEM. At this point we } are talking about a few electrons per square angstrom, even less. In high } resolution EM crystallography people have deal with 0.6 e/A2. At such } conditions, high sensitivity and linearity of the modern digital cameras } may be a plus. Talking about long exposures --what about drift? I never } had good pictures at x100K with exposure longer that a few seconds. I } don't understand your point about noise. In case of digital camera the } noise is a function of camera. Current cameras has a very low level of } noise (they uses cooling, etc) and we have to pay for that astronomical } price. This is a life. I am not friendly with TEM cameras, but I do know } that in the light microscopy people count individual photons using CCD } cameras. In TEM camera we have phosphorous screen as a source of image and } my questions to Mickhael were addressed mostly to the problem how } effectively (and correctly) information is traveled thought that funny } screen. The Mickhael's answer is that screen does not affect dynamic range } and sensitivity is much higher than on convention films. Am I correct, } Mickhaels? } } Sergey } } } Date: Thu, 18 May 2000 14:15:02 +1000 } } From: jim {jim-at-proscitech.com.au} } } Subject: RE: new developments in imaging systems? } } To: 'Sergey Ryazantsev' {sryazant-at-ucla.edu} , } } "Microscopy-at-sparc5.microscopy.com" {Microscopy-at-sparc5.microscopy.com} } } Reply-to: "jim-at-proscitech.com.au" {jim-at-proscitech.com.au} } } Organization: ProSciTech } } X-Mailer: Microsoft Internet E-mail/MAPI - 8.0.0.4211 } } } } ----------------------------------------------------------------------- - } } 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 } } -----------------------------------------------------------------------
My thanks to all those who responded to my question about treating coverslips to make cells stick to them better. I have used one or two of these in the past, but my colleague wanted to try some different approaches. Thanks again.
Matthew J. Schibler Ph.D. UCLA Brain Research Institute 1524A Gonda (Goldschmied) Center for Neuroscience and Genetics Los Angeles, CA 90095-1761
Linda, if you get quicktime pro (its an upgrade to regular quicktime, costs about $30 from Apple) you can import a numbered series of tiffs directly into quicktime. which then can be made into a movie Simon
-----Original Message----- } From: Linda Barthel [mailto:barthel-at-umich.edu] Sent: Monday, May 22, 2000 10:44 AM To: Microscopy listserver
I am also trying to figure out how to make a QuickTime video from a series of images. I am using a PC, and the images are from a Zeiss 510 Confocal, in LSM format, which have been converted to Tiff. The number I am dealing with is 64 images. Any help would be greatly appreciated Linda Barthel Research Associate II Department of Cell and Developmental Biology University of Michigan barthel-at-umich.edu
I have a functional, but old, Balzers Freeze Fracture machine. It is headed for the scrap pile unless someone out there would like to have it. I will explore the possibilities if anyone shows an interest.
Greg Gregory W. Erdos, Ph.D. Assistant Director, Biotechnology Program PO Box 110580 University of Florida Gainesville, FL 32611
Thanks for the input. I was hesitant to use diamond abrasives on my samples due to previous advice. I was told that due to the hardness of diamond, it tends to embed itself in soft metals such as copper, which results in "smearing" more than polishing. Do you have any input regarding the smearing issue?
John
On Fri, 19 May 2000, Edward Hirsch wrote:
} John, } The solutions that the colloidal silica are in have a ph of 8.5 to 9.8 } depending on the formulation. I suggest you try our 0.05 micron water based } polycrystalline diamond suspension. Allied High Tech } http://www.alliedhightech.com sells this product. I would also suggest our } Final A cloth for this polishing step. If you would like samples of either } of these products please let me know and I will be happy to send it to you. } } If you need further technical assistance please contact me off-line and I } will be happy to help or you may contact our main office at (800)675-1118 } located in CA. } } I hope this helps. } Ed } Please note, I have a financial interest in providing you with these } products and other sample preparation equipment and consumable items. } } ************************************************* } Edward A. Hirsch } Product Application Specialist } Allied High Tech Products } 2376 East Pacifica Place } Rancho Dominguez, CA 90220 } ph: (919) 846-9628 } vm:(800)675-1118 x245 } fx: (310)762-6808 } http://www.alliedhightech.com } } Equipment and Consumables for Metallurgical Sample Preparation } ************************************************* } } -----Original Message----- } From: john david whitaker [mailto:jwhitake-at-u.washington.edu] } Sent: Friday, May 19, 2000 4:01 PM } To: Microscopy Lister Server } Subject: final polishing for metals (Cu, Ni, Fe, NiFe) } } } ------------------------------------------------------------------------ } 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'm looking for information regarding final polishing } solutions for preparing metallic cross-section samples } for TEM analysis. } } I've looked into colloidal silica } suspensions such as Ludox and Syton, and am concerned } that their alkilinity may chemically etch my samples. } I'm analyzing different compositions of NiFe } on copper substrates, so the specimens are prone to } preferential etching of different phases. } } Does anyone have any experience with this? } } Any input would be appreciated. } } John } } } }
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Our software, 4D turnaround will make quicktime movies out of image stacks that are in biorad format, pics, or tiff. Please see our website for more details and to download: http://www.loci.wisc.edu/4d/native/4d.html
Currently we have only a mac version available, however we will be releasing a java version next month that works cross platform.
Best regards, kevin
Kevin W. Eliceiri Project Director Laboratory for Optical and Computational Instrumentation http://www.loci.wisc.edu 159 Animal Sciences 1675 Observatory Dr. Madison, WI 53706 608-263-6288 voice 608-265-4076 fax
A view days ago I found your message in my mailbox. Regarding your interest about digital imaging systems I can provide you some information on our imaging plate system for TEM.
DIBIS Imaging Plate Technology is adapted for TEMs from, FEI/Philips, LEO/Zeiss, JEOL and HITACHI. The Imaging Plate (81 x 100 mm) is inserted into the sheet film cameras of the various TEMs and is directly exposed by electrons. After that the imaging plate reader creates digital images directly from the plates without involving chemical processing thus providing extraordinary image quality with 3600 x 3200 pixel at 25 µm and 16 or 20 bit dynamic range with true linearity. The high pixel count supports printouts in real photographic quality, also on larger formats as you are used to from photographic film. One Instrument in your lab will serve all your TEMs with highest quality digital imaging technology, no matter what type and manufacturer. The high performance, resolution, sensitivity and dynamic range makes Imaging Plate Technology first choice for life science and material sciences imaging, low dose applications and high dynamic diffraction patterns.
So, DIBIS introduces MICRON Digital Imaging Plate Technology as the alternative to overcome the limits of CCD technology for TEM.
For more details visit our homepage.
R. Kuenzler ---------------------------------- DIBIS Digital Biomedical Imaging Systems AG Gewerbestra§e 11; D-75217 Birkenfeld Tel.: +49 (0)7082 940639 Fax : +49 (0)7082 940076 E-Mail: contact-at-dibis.de E-Mail: kuenzler-at-dibis.de Internet: http://www.dibis.de
Margaret Dienelt schrieb:
Hi,
} } Year after year I hopefully gather information about digital imaging } systems for TEMs (ours is a JEOL 100CX) , only to learn we have no } money. This year it looks like it might really happen but I have not } kept up with innovations in the field and am wondering the following: } } 1. Anything new in the last two years -- especially in terms of } cameras? I'm most familiar with the Gatan and AMT systems but their } web sites don't reflect much in the way of changes over a year ago. } 2. With more and more microscopists finally getting their systems -- } I'd love to get feedback. } } Thanks, } Margaret } } P.S. Would welcome contacts from vendors. } } -- } Margaret Dienelt } } Plant Pathologist } Electron Microscopy Lab } } Floral and Nursery Plants Research Unit } U.S. National Arboretum/Agricultural Research Service/USDA } } B. 010A, Rm. 238, BARC-W } 10300 Baltimore Avenue } Beltsville MD. 20705 USA } } (301) 504-6097 } Fax: (301) 504-5096
GraphicConverter will convert a folder of numbered tiffs into QT. Just be aware that after choosing the folder with your tiffs, and clicking on Convert, the next window has an easily overlooked choice at its top to save the files as ONE movie. Forget this selection and you will have converted your 300 tiffs into 300 moov files. GraphicConverter has many options for compression and output size.
NIH Image can open a folder of numbered tiffs with the 'Open All' option. Use the 'Windows to Stack' command then save as QT. Image will place your tiffs into the stack in the order in which they were opened. So if your filenames don't end with an incrementing number you could open them manually in the desired order.
Regards, Glen
Glen MacDonald Research Scientist Hearing Research Laboratories of the Virginia Merrill Bloedel Hearing Research Center Box 35-7923 University of Washington Seattle, WA 98195-7923 (206) 616-4156 glenmac-at-u.washington.edu
********************************************************************* C:} The box said "Requires Windows95 or better". So I bought a Macintosh. *********************************************************************
I am unsure what versions are available for the Mac, but I have made videos using ULEAD video editing software for the PC. TIFF images can be imported and displayed for "N" number of frames. Special effects (like a wipe or fade) can be added between sets fo (still) frame sets.
Do beware this process can tax the resources of most PCs. If the video is uncompressed, the data rate can be 10+ MB/sec. I use a Matrox video card which has MJPEG hardware compression. At full VHS resolution and 15 frames/sec (not 29.9) the data rate is below 2 MB/sec. With MJPEG, I can get about 10 minutes of "fair" quality video on one CD-R (650 MB). Mpeg compression can make a smaller file, but I havent tried a compression program I like - Lousy quality - Jumps, etc. I haven't tried the MPEG program from Xing Tech. which is rated better thatn the sharewares I have tried.
Another issue is the conversion time using software mpeg convertors is that 10 minutes of MJPEG video to mpeg on my PC (350 MHz/256 Meg Ram) takes well over an hour.
E.A. Fischione Instruments, Inc., a rapidly growing company specializing in the development and manufacture of TEM specimen preparation instrumentation, is seeking a highly motivated individual for the position of a Application Scientist. The candidate's responsibilities will include:
* Obtain and prepare TEM specimens of customer's material for sales purposes. * Analyze customers' specimens in Fischione's TEM. * Library research to support design and application work activities. * Answer customers' questions regarding use of Fischione products. * Instrument training (when required). * Provide technical support at major tradeshows. * Give product demonstrations. * Collaborate with potential customers on experiments. * Write applications notes. * Give scientific presentations. * Conduct short courses and workshops on specimen preparation and other Fischione related technology. * Write refereed journal articles for publication. * Generate information for Website. * Revise instrument instruction manuals. * Evaluate emerging microscopy related technologies and market opportunities. * Convey market opportunities to Fischione management. * Obtain input from customers on possible new products and on improvements to existing products. * Work with the product design team on new product developments. * Provide design support from the microscopist's standpoint. * Prototype and test both existing product improvements and new products. * Procure needed instrumentation to get specimen preparation facility running appropriately. * Optimize Fischione's TEM laboratory. * Work with architects to design new EM facility when building expansion occurs. * Travel approximately 10%-20%.
The candidate should have a Ph.D in Material Science, Engineering, or Physics with a specialization in Electron Microscopy.
Salary is commensurate with experience.
E.A. Fischione Instruments, Inc. is an equal opportunity employer. Please send your resume and salary requirements to:
Human Resources Director E.A. Fischione Instruments, Inc. 9003 Corporate Circle Export, PA 15632 Phone (724) 325-5444 FAX (724) 325-5443 E-mail: info-at-fischione.com
I have never used a sintillator type BSE detector, but the major differences are two fold.
Typically (though diodes are getting better all the time) the Robinson has a better low energy BSE detection effiency. It follows that for the same noise level electronics, it would exhibit better sensitivity. The dynamic range problem will still exist for very different Zs in the field of view. In the middle and upper portions of the sensitivity range (low and lower), I would not expect much difference between them. ...Any Comments from users of both????
I like the 4 quad diodes since I can go differential mode for macro topography (like fracture surfaces) and show the gross features while suppressing the fine detail.
The Pt coating can have a profound negative effect on sensitivity if not extremely thin. If given a choice, I would always use carbon for the best BSE sensitivity. ...If not hazardous, I would use Be, BUT!!!! For elements in the range of Al & Si, I prefer to use carbon and lower beam voltages to minimize penetration, especially if they are films or very small features.
I once presented some data illustrating the BSE signal attenuation as a function of sputtered Au thickness. But that data would be hard to retrieve now.
Woody White McDermott Technology
=================================================== I use a Robinson Model 6 which is specified at a Z contrast of 0.003 at } = 2KV. From your work with a 0.1Z detector, what would you say is the qualitative effect that a higher Z contrast detector would offer? I am especially interested in imaging Al/Si alloys and I typically sputter coat them with Pt. I may try carbon one of these days to see what the difference might be.
Yes, heating in the sputter coater is likely the cause.
Confirm this by checking the coated samples under the optical microscope before putting them in the SEM. This will eliminate from consideration any artifacts caused by higher SEM vacuum and electron beam damage.
At 2:56 AM -0400 5/22/00, lherault wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
====================================== Carl Henderson Electron Microbeam Analysis Laboratory University of Michigan 2501 C.C. Little Bldg. 425 E. University Ave. Ann Arbor, MI 48109-1063 USA (734) 936-1550 FAX (734) 763-4690 ======================================
Are you using a magnetron sputter coater? If so, then you should not be getting melting, etc. But ...
For how long did you coat your samples? You might try coating for short periods, with some rest in between -- say 30 sec, turn off (maybe add some argon to help carry away any heat), coat, rest, etc. If your samples are very sensitive, use 10 sec. coat times. Rest for 30 sec more or less (empirically). I've done Teflon fabrics this way, using 90 sec. coat periods without problems, but if your samples are thick, this would increase the problems.
Phil
} I've had to coat some PLGA polymer samples and we think the coater may be } melting them. The samples have a rolled or beaded edge and large cracks. I } have jpeg images if anyone wants me to send them for a look. Are we } correct in our assessment or should we look elsewhere for the source of } cracks, shrinkage, beading around edges. These artifacts, BTW do not appear } when the uncoated samples are checked under a light microscope. } } Thanks in advance. } } Ron L } lherault-at-bu.edu
-- }}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{ Philip Oshel Supervisor, AMFSC and BBPIC Dept. of Animal Health and Biomedical Sciences University of Wisconsin 1656 Linden Drive Madison, WI 53706-1581 voice: (608) 263-4162 fax: (608) 262-7420 (dept. fax)
How long of running time, approximately? Do you want dissolve from image to image or simple image swap? Any audio?
gary g.
At 07:44 AM 5/22/00, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
If your samples are particularly heat sensitive, you should probably look into an Ion Beam Sputtering System. The heating effects in such a system are negligible. Of course, you have the added advantages of thinner, more uniform coatings etc. also. We do make the IBS/e Ion Beam Sputter Deposition and Etching System. If this is an infrequent application for you, perhaps I can put you in touch with one of our local users who could help you out.
David Henriks Vice President 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
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy. Message text written by "lherault" } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I've had to coat some PLGA polymer samples and we think the coater may be melting them. The samples have a rolled or beaded edge and large cracks. I have jpeg images if anyone wants me to send them for a look. Are we correct in our assessment or should we look elsewhere for the source of cracks, shrinkage, beading around edges. These artifacts, BTW do not appear when the uncoated samples are checked under a light microscope.
We are moving to a new building and need to get rid of some old equipment. The following are available in New Jersey, USA. All items were fully functional when last used, most have all accessories and owners manuals. Any reasonable offer considered.
For sale:
-GW Electronics attachments to go on Hitachi HS-510: Dual magnification unit, Graphics generator, and Homomorphic Processor
-LKB 8800 Ultrotome III (thermal advance, includes ALL accessories)
-Denton Critcal Point Dryer (CPD-1) With extra baskets, seals, and coupling for CO2 tank
-Ames Lab-Tek cryostat (slow leak in refrigerant line)
-Zeiss microspectrophotometer (MPM microscope photometer for cytospectrophotometry) for Zeiss Photo-microscope, Ultraphot, Standard Universal microscopes (includes light chopper, power supply, monochromator, photometer head, photomultiplier housing, indicator unit, and coupling for microscope head)
-Printz automatic print dryer model JET 260158 ferrotyper drum and canvas belt in like-new condition)
-Kodak Ektamatic Model 214-K automatic print processor -Accessories for AO/Reichert Microstar compound scope (available with or without microscope): AO Expostar photomicrographic system (lens and shutter, control unit model 1190, polaroid and 35 mm film backs), AO verical illuminator for incident light fluorescence microscopy (includes mercury lamp model 2054A, filter housing, AND microscope), Camera lucida attanchment AO #1030
- 2 ea Omni-mixer homogenizer with stand model # 17105, 16,000 rpm, no impellers -Lightnin Mixer Model F (no impellor)
-B&L Dynazoom microscope with integrated 4x5 (polaroid) and 35 mm camera backs
**Items looking for a good home (shipping cost plus a little extra for us to be able to say that we actually sold them):
-B&L # 42-63-89 projecting compound scope (used to do camera lucida)
-stainless steel frames for holding individual sheets/plates of 3 1/4 x 4 EM film
--Arthur Thomas Co., paraffin for 50-52oc (33 lib pkg of 1/4 lb bars)
Dealers are welcomed to contact us. Perhaps we could give you some items plus a cash allowance to purchase some used items that you may have that we want.
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
You can get NIH Image to do this and a great deal more. For PC's it's called Scion Image, and it's available free from www.scioncorp.com. Hope this helps.
Lesley Weston.
On Mon, 22 May 2000, Kevin W. Eliceiri 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 } -----------------------------------------------------------------------. } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } } } I am also trying to figure out how to make a QuickTime video from a series } } of images. I am using a PC, and the images are from a Zeiss 510 Confocal, } } in LSM format, which have been converted to Tiff. The number I am dealing } } with is 64 images. Any help would be greatly appreciated } } Linda Barthel } } Research Associate II } } Department of Cell and Developmental Biology } } University of Michigan } } barthel-at-umich.edu } } Our software, 4D turnaround will make quicktime movies out of image } stacks that are in biorad format, pics, or tiff. Please see our } website for more details and to download: } http://www.loci.wisc.edu/4d/native/4d.html } } Currently we have only a mac version available, however we will be } releasing a java version next month that works cross platform. } } Best regards, } kevin } } Kevin W. Eliceiri } Project Director } Laboratory for Optical and Computational Instrumentation } http://www.loci.wisc.edu } 159 Animal Sciences } 1675 Observatory Dr. } Madison, WI 53706 } 608-263-6288 voice } 608-265-4076 fax } } }
: Frieda Lim } Barbaric Yawp } 224 west 13th Street #5r } NYC, NY 10011-7775 } 212.206.0683 } barbaricyawp-at-att.net } } Dear MSA Members } } It was suggested to me that this may be the perfect spot to list my query. } I would appreciate your perusal of the below queries and am anxious to hear from you. } } Sincerely, } Frieda Lim } } } } 1st query: } } Dear MSA Members : } } I invite you to take a close look at our query. We are a film production } company seeking existing moving microscopic organism film or video footage. } We will utilize this footage for our single purpose of making a feature } film. The film we are creating is a microscopic fantasia. Zooming in on } your microscopic world, we will spin a tale through imagery and music. } } We would appreciate your help with obtaining some footage directly or any } referring leads. Please contact us at your convenience to discuss further } details. } } Sincerely, } Frieda Lim } } } } } Elaborating 2nd Query: } } Dear MSA Members: } } I thank you for your prompt response to my query for microscopic } microorganism footage. Our intended use of your existing footage is to edit } & generate a full feature film comprised solely of microscopic imagery and } music with which we hope, eventually, will be distributed to the general } public at large--large in fact by mesmerizing all ages, the young & the } young of heart. Our microscopic fantasia will bring to life a single } narrative, the classic myth of Cupid & Psyche. } } In our original query, we had cast a wide net being intentionally } non-specific with regard to the kind of microlife we are seeking. At this } stage, we have no restrictions as to what microworld would be best suited } for our story. However, our intent is to portray our story within a } scientific veritable reality. We don't want micro species colliding that } would never interact in truth. Right now we are in our initial phase of } investigative hunting & gathering-a casting call for microscopic actors. We } actually want to cast organisms as actual characters and to use others as } metaphorical imagery. My suspicion is that you have already created footage } for your scientific research and discovered many a talented organism } awaiting their big break. We further suspect that we will need to pool } microscopy resources and see what footage offered is most varied in look & } movements that will best fill the wide assortment of roles required. Since } we are approaching your world as layman, we would appreciate your expertise } in advising what microorganism microcosm might be most readily available. } } To note, this is a low budget project with big hopes and dreams. We } believe our film can achieve success similar to that of "Microcosmos"-the ' } 96 Cannes Film Festival jury prize winner. That movie was testament that } science has mass marketable enthusiasm in the entertainment world. } Comparatively, we hope to explode your frontier world onto the big screen } and excite a wide audience. } } So, if you know of some microorganisms wanting to make it big, we are } anxious to hear from you. Thank you. } } Sincerely, } Frieda Lim
Firstly a confession, that my taxonomy is badly in need of repairing - the alga we have been working with is not a blue-green algae at all - it's actually Polytomella, a eukaryote, and I think is a relative of Chlamydomonas, but doesn't have a cell wall or photosynthetic apparatus.
Anyway, embedding in Spurr's seems to have done the trick as far as stopping the starch granules dropping out. I've just been looking at grid-fulls of cells full of starch granules, and they're all there! Many thanks for the suggestions. I'll be back with more problems soon!
Mark
******************************************** Mark West, Unidad de Microscopia Electronica, (Electron Microscopy Unit) Instituto de Fisiologia Celular, Universidad Nacional Autonoma de Mexico, 04510 Mexico D.F.
We've applied microscopic techniques (e.g., polarized light microscopy, fluorescence microscopy, FTIR microspectroscopy, and SEM-EDS) and non-microscopic techniques to more than 700 projects involving historic and artistic works, from Egyptian antiquity to contemporary. Material evidence of authenticity -- or more likely, inauthenticity -- has been or become an objective of many of these studies.
Scientific investigation plays an important role in the authentication process, often providing indirect or direct evidence of date, and an objective basis by which to compare materials and techniques to works of unquestioned authenticity. Given sampling limitations and the layered structure of many art objects and decorative finishes, microscopy is ideally and elegantly suited to their study of the former - that is, structure and composition. We use a new infinity-corrected optical/FTIR microscopy system, that we had custom-fitted for polarized light microscopy, fluorescence microscopy, and FTIR microspectroscopy (narrow-band MCTA and wide-band MCTB detectors).
Your article sounds very interesting -- please feel free to call.
Jamie
James Martin Orion Analytical, LLC Post Office Box 550 Williamstown, MA 01267 phone: 413-458-0233 e-mail: martin-at-orionanalytical.com website: www.orionanalytical.com
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Hello,
I am new to this list. I would like to know if anyone has information I could use to write an article on the analytical chemistry techniques used to authenticate art objects using light microscopes or any other type of microscope. Also, if anyone has information on any of the other techniques used to authenticate art objects such as x-ray diffraction, x-ray fluorescence, accelerator mass spectrometry, that would be helpful too.
Sincerely,
Margaret M. Mitchell Assistant Editor AOAC INTERNATIONAL 301-924-7077 (tel) 301-924-7089 (fax) mmitchell-at-aoac.org
Jean Dille wrote: ===================================================== We have a sample of 6 micrometers Ni-Zn-P electroless plated onto 1 mm thick Al and we would like to prepare cross sections for TEM examination. It looks very difficult to use the tripod polishing technique for metals.So, we are thinking at an electropolishing procedure with our double-jet Tenupol apparatus.
This would be done on slices cutted from a sample initially thicked up to 3mm by electroplating.But how to avoid differential material removal problems? Which material for plating?Which electrolyte for electropolishing?Other method?
Does anyone have suggestions for us?
Thanks in advance for your help. ==================================================== If the aluminum substrate can be thinned down a bit more, then a coating of this thickness should be able to be thin sectioned using diamond knife ultramicrotomy. It is hard to predict whether better results will be obtained embedded or unembedded (there is a tendency for the coating to separate from the substrate). We usually find that such separation is less likely to occur if the sample is embedded. However, structure within the coating is less well preserved when embedded. Naturally our experience has been with our own SPI-Pon 812 epoxy embedding resin and SPI diamond knives, but I would expect that at least most of the other "Epon substitutes" (and knives) would work just as well. With regard to the substrate separation, the tendency for this to happen can be reduced by using a knife included angle that is smaller rather than larger (we were successful with 45 deg.). The use of larger included angles, at least in our experience, seemed to produce a level of compression artifacts in the sections that we found unacceptable.
In any case, we have found the diamond knife thin sectioning approach to often times offer certain advantages over the alternatives for sample preparation.
Disclaimer: SPI Supplies offers materials science diamond knives and also our preferred embedding resin, SPI-Pon 812 Embedding System. Our Structure Probe laboratory services division performs this kind of diamond knife thin sectioning as a service for commercial clients.
Chuck
PS: Remember that we are trying to become 100% paperless and the only way that we can manage this kind of correspondence, in a paperless environment, is for our correspondents to always reply by way of "reply" on their software so that the entire string of correspondence on that topic gets returned to us, and the entire correspondence history can be kept in one place.
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.spi.cc ######################## ============================================
Ronald J. L'Herault wrote: ==================================================================== I've had to coat some PLGA polymer samples and we think the coater may be melting them. The samples have a rolled or beaded edge and large cracks. I have jpeg images if anyone wants me to send them for a look. Are we correct in our assessment or should we look elsewhere for the source of cracks, shrinkage, beading around edges. These artifacts, BTW do not appear when the uncoated samples are checked under a light microscope. ==================================================================== If by PLGA you mean polylactic glycolic acid, which in certain forms could be a (dissolvable) surgical suture material, then the polymer could be quite hygroscopic and if it has had time to absorb enough moisture, it could be evolving moisture, and that could be the reason for your problems.
Some sputter coaters have a "test mode" which enables the user to momentarily expose in a gentle way the polymer surface to the glow of the plasma. If moisture is evolving, then there will be an immediate diminution of the quality of the vacuum. When confronted with that kind of situation, we go through the cycle of "test mode", then pump down, test mode, etc, until hitting the test mode button results in no more deflection of the vacuum. Ten or more cycles might be needed but in the end, the surface moisture is removed.
Then you are ready to coat.
If you are not talking about polylactic glycolic acid, I apologize for taking up the extra bandwidth.
I realize that not all sputter coaters have such a test mode, but that is the perfect kind of example where such a "test mode" has its greatest value.
Disclaimer: SPI Supplies manufactures sputter coaters with "test modes".
Chuck
PS: Remember that we are trying to become 100% paperless and the only way that we can manage this kind of correspondence, in a paperless environment, is for our correspondents to always reply by way of "reply" on their software so that the entire string of correspondence on that topic gets returned to us, and the entire correspondence history can be kept in one place.
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.spi.cc ######################## ============================================
I've applied microscopic techniques (e.g., polarized light microscopy, fluorescence microscopy, FTIR microspectroscopy, and SEM-EDS) and non-microscopic techniques to more than 700 projects involving historic and artistic works, from Egyptian antiquity to contemporary. Material evidence of authenticity -- or more likely, inauthenticity -- has been or become an objective of many of these studies.
Scientific investigation plays an important role in the authentication process, often providing indirect or direct evidence of date, and an objective basis by which to compare materials and techniques to works of unquestioned authenticity. Given sampling limitations and the layered structure of many art objects and decorative finishes, microscopy is ideally and elegantly suited to their study of the former - that is, structure and composition. We use a new infinity-corrected optical/FTIR microscopy system, that we had custom-fitted for polarized light microscopy, fluorescence microscopy, and FTIR microspectroscopy (narrow-band MCTA and wide-band MCTB detectors).
Your article sounds very interesting -- please feel free to call.
Jamie
James Martin Orion Analytical, LLC Post Office Box 550 Williamstown, MA 01267 phone: 413-458-0233 e-mail: martin-at-orionanalytical.com website: www.orionanalytical.com
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello,
I am new to this list. I would like to know if anyone has information I could use to write an article on the analytical chemistry techniques used to authenticate art objects using light microscopes or any other type of microscope. Also, if anyone has information on any of the other techniques used to authenticate art objects such as x-ray diffraction, x-ray fluorescence, accelerator mass spectrometry, that would be helpful too.
Sincerely,
Margaret M. Mitchell Assistant Editor AOAC INTERNATIONAL 301-924-7077 (tel) 301-924-7089 (fax) mmitchell-at-aoac.org
When I purchased a copy of Adobe Photoshop 5.5, it came bundled with their "ImageReady 2.0" This package can be used to create animated GIF images from layered images, or the animation exported in QuickTime format.
When I purchased a copy of Adobe Photoshop 5.5, it came bundled with their "ImageReady 2.0" This package can be used to create animated GIF images from layered images, or the animation exported in QuickTime format.
What I ended up doing in your situation was downloading Confocal Assistant, and reconstructing the TIFF series using that. You first need to add a BioRad header... the software and tricks can be found at:
http://www.cs.ubc.ca/spider/ladic/source.html
You can export the animation from CA as an .avi file. Then if you have Image Ready (Photoshop 5.5), you can convert the .avi to .mov. (I think Graphics Converter will also work) A roundabout way to get the job done, but it works.
All the best,
Angela
} I am also trying to figure out how to make a QuickTime video from a series } of images. I am using a PC, and the images are from a Zeiss 510 Confocal, } in LSM format, which have been converted to Tiff. The number I am dealing } with is 64 images. Any help would be greatly appreciated } Linda Barthel } Research Associate II } Department of Cell and Developmental Biology } University of Michigan } barthel-at-umich.edu } } } } } } --------------------------------------------- Angela V. Klaus
Laboratory Manager, Interdepartmental Laboratories American Museum of Natural History Central Park West at 79th Street New York, NY 10024-5192 USA
I would be pleased if anyone could broaden my views on equipment reliability . Basically we have a 3yr old FESEM which I consider to be fairly reliable in that it is on call 24hrs/day , has numerous ( non dedicated users ) and apart from downtime for filament change and the odd wear and tear type problems answers our needs . As we do not have a back up instrument and when we do experience problems it's always at the worse time certain personnel have the impression that it is unreliable .
What do other sem users expect in terms of reliability , apart from my ' subjective ' comments is it quantifiable , would approx 2wks /year downtime including planned maintenance be considered excessive ?
I have done this procedure many times using many different programs. Most often I was using TIFF files in sequence. I can't remember all the names of the programs that I tried, most were freeware or shareware including NIH image and the results were never to my satisfaction, I wanted control over Frames per second, compression, output format, light levels, you know complete control. The program that I wound up using was Adobe After Effects. An incredible program, capabilities way beyond what was needed for creating simple sequence movies from 3D volumes captured in a LSCM and rendered on a SGI. The benefit is unlimited file type importation (well nearly unlimited - hey it supports SGI .rgb files!) and complete control over the creation process. I could make an animated gif with any frame rate I wanted or a full compression-less Quicktime (.mov) or Microsoft Video for Windows format (.avi) with sound and special effects.
Most of you have used Adobe photoshop at one time or another, Adobe After Effects has the same intuitive interface and many of the same filters and image adjusting features. The educational discount for the lab brought the price down to less the $300! An incredible value for anyone doing lots of routine slice parade movies or animations for presentations.
If you have the ability to record sound on your computer you can even record a whole seminar presentation. Imagine showing up at a meeting plugging in your laptop computer (with sound) to the projection system and clicking play. You can sit down and listen to your own presentation, and then answer questions at the end. No more forgetting to mention a point, you can make sure that you are making sense. Of course this all takes away from the art of the presentation. I have never done this mind you all but the idea is intriguing isn't it?
Adobe also has a software program called Premier that is used in the film industry to do even more digital effects and post production (I think the last guess I heard was that about 75% of all rolling credits and film intros are done with this program and many of the same features are in After Effects).
-Geoff
-- Geoff Williams, M.S.
Electron Microscope Facility Supervisor Biology Department Brooks Hall Central Michigan University Mt. Pleasant, MI 48859
Gordon Couger 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: "Tina Carvalho" {tina-at-pbrc.hawaii.edu} } } On Thu, 18 May 2000 ComCryLab1-at-aol.com-at-sparc5.microscopy.com wrote: } } } } } Dear Friends, } } } Can anyone give me advice regarding the dismantling and } } } the packing for shipment of a Philips 201 TEM? I need to } } } pay particular attention to not disturbing the alignment. } } } I plan to move the TEM from Univ. of Delaware to Naples, Florida } } } in an enclosed U-Haul Trailer. Any help or suggestions } } } will be most welcome. } } } } I will make sure Ron Veil, a very experienced independent EM service tech, } } sees this and has a chance to reply. It was with his advice that we (my } } hubby and I and its new owner) packed up and shipped a Philips 201. Like } } you, we wanted to ship it intact, column on and all. We built a heavy } } duty skid and added a strong base to which we bolted the instrument. Then } } we wrapped it with whatever that plastic packing tape that is like Saran } } Wrap is called, making sure to secure any part of the column we didn't } } want to move, but avoiding putting any pressure on things, such as the } } aperture drives. Wrap the bottom, wrap the column, wrap the column to the } } bottom and back again, etc. Build a crate up around the instrument. } } I think I remember putting a wood insert with a crescent cut out near the } } top of, but not touching the column, but hubby thinks not. The idea is } } NOT to let any shock to the crate get transferred to the column, so } } perhaps we didn't. Add some packing material, such as old egg crate foam } } from your bed (it needed replacing anyway) and whatever. Then, and this } } was the fun part, buy that stuff that when you mix it with a catalyst, } } produces huge volumes of foam that hardens in a few minutes. I think you } } can also buy spray cans of similar stuff, but since hubby had this on hand } } for other things, we had the bulk stuff. Fill the voids in the crate with } } it. It will easily chip off later. Add a top, and away you go. The } } scope made it in great shape. } } } } I think we packed the rotary pump and a couple of other things } } separately. Get it into the truck and TIE IT DOWN. I say this because an } } SEM we once packed for shipping with the base and saran, but no closed } } crate, made it from Hawaii to Ohio, then slid in the shippers truck in the } } last mile and slammed into the driver's seat. The driver was OK, the ion } } getter pump was bent, and the column needed a bit of work, but it could } } have been worse. Duh. } } } } I've also packed up a couple of Denton vcuum evaporators and a couple of } } ultramicrotomes. The saran stuff and blow foam are a good way to } } stabilize the instruments. } } } Since you are using a U haul you don't have to worry about sides } and a top on the crate. Make sure it is tied down real well. Dry } wall screws through the crate and into the floor work well for } locking it down but make sure you have it braced with timbers } to the front and sides of the trailer in case you make a panic } stop. } } Also make sure that the weight is centers in front of the trailer } axle if you use a trailer. Negitive weight on the trailer tounge } at the very least makes driving very interesting. You have no } Idea how fast a trailer can pass you while it is still tied on to the } truck. I have had the privilege of experiencing this and I can promise } you that you won't enjoy it:). } } The foam in place stuff is great. Just cover the instrument in plastic } and foam away. If you can get foam between the instrument and the } Support the foam will dissipate most of those little shocks that get } things out of line. } } Last of all make sure you understand what you insurance covers and } what it doesn't on moving equipment. You should be fine on liability but } the value of the contents is probably not covered. Rental truck companies } have contents insurance as an extra. } } Good luck } Gordon } } Gordon Couger gcouger-at-couger.com } Stillwater, OK www.couger.com/gcouger } 405 624-2855 GMT -6:00
Gordon, Now I guess I know who to blame when I get U-Haul trailers that are in p----poor shape to move SEMs. Where do you get off running screws through the floor of a trailer you don't own?
You could probably expose your CCD chip directly to the electron bean -- and buy a new chip every few hundred exposures or so! Electrons have mass and carry momentum, which gets transformed into force as they are stopped in the crystal, especially at higher keV. This can lead to damage. Photons only heat up the target.
As far as I know ALL TEM cameras use a medium to convert electrons into light, and this light is then detected. The medium is either a phosphor screen or a very thin YAG crystal. From there on it is different. Some cameras use a fiber-optic to guide the photons to the chip, others use mirrors and lenses. So, it is not simply a matter of comparing the QE of the chip, the entire system has to taken into account. Also, each electron creates many photons in the phosphor or YAG, and in principle only one photon is enough to create a signal in the CCD. In other words, a CCD could theoretically detect "fractions" of an electron, whereas the film is directly exposed to the electron. Of course the electron can "rattle" around in the film and activate several grains, but as you can see, the question of comparing sensitivities now becomes one of comparing two different physical processes. It can definitely be done, but it's not easy.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== 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: hpadams [mailto:hpadams-at-bcm.tmc.edu] Sent: Thursday, May 25, 2000 12:30 PM To: microscopy-at-sparc5.microscopy.com
I have been following off and on this discussion on CCD cameras for EM. Much of this discussion has been concerned with "sensitivity". I am being naive here, but when we talk about "sensitivity" of CCDs, isn't this the same thing as the QE of camera/chip? I don't think I have ever seen a QE for em CCD's for various accelerating voltages/wavelenghts. Does the electron beam directly hit the silicon photodyodes or it there an interface that converts the incoming electrons to different (longer?) wavelengts? I know em films are sensitive to specific acc voltages. Are CCD cameras for em the same. For long exposures it may not matter, but for short exposures or low level intensity does the QE of the camera come into play?
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Hello all, I have a researcher that is interested in utilizing florescent probes to label material with the desire to visualize with a 50 angstrom resolution. Are there any service facilities out there that have a cathodoluminescence detector on their SEM or STEM that would be able to assist him (via contract)? Thank you!
Tracey Pepper Supervisor Bessey Microscopy Facility Iowa State University ph: 515-294-3872 fax: 515.294.1337
} You could probably expose your CCD chip directly to the electron bean -- } and buy a new chip every few hundred exposures or so! Electrons have
There is an even worse problem with exposing the CCD directly the electron beam. The p-n junctions in the CCD chip have a certain "well capacity" - a number of electron/hole pairs they can hold before they saturate. Fast (keV) electrons are much more efficient at producing electron/hole pairs than photons - so much so that a pixel on the CCD would saturate after about 15 fast electrons hit it. Just the square root N shot noise at that level is about 25% - larger than typical TEM micrograph contrast of 10-20%. That's why a phosphor or scintillator is necessary to transform the fast electrons into photons.
In case anyone is interested, you can learn more than you ever possibly want to know about CCD cameras in:
"Applications of slow-scan CCD cameras in transmission electron microscopy" O. L. Krivanek and P. E. Mooney, Ultramicroscopy V. 49 p. 95-108 (1993). First description of Gatan CCD cameras, including some design issues, and measurements of the linearity, modulation transfer function (MTF), and detector quantum efficiency (DQE).
"Methods to measure the properties of slow-scan CCD cameras for electron detection" W. J. de Ruijter and J. K. Weiss, Rev. Sci. Instrum. V. 63, p. 4314 - 4321 (1992). Another early paper, includes comparisons to photographic plates. Uses a slightly different definition of the MTF from everyone else.
J. M. Zuo "Electron detection characteristics of slow-scan CCD camera", Ultramicroscopy V. 66 p. 21-33, (1996). Describes gain, MTF, and DQE measurements, measurement techniques based on stochastic noise (blank beam) images, and theory. This is a good place to start.
"Quantitative characterization of point spread function and detection quantum efficiency for a YAG scintillator slow scan CCD camera" A. L. Weickenmeier, W. Nuchter, and J. Mayer, Optik, V. 99, p. 147-154, (1995). Line-scan method of measuring the MTF.
Happy reading, Paul
Paul Voyles, voyles-at-research.nj.nec.com voice: (609) 951-2627, fax: (609) 951-2496 NEC Research Institute 4 Independence Way Princeton, NJ 08540 USA {http://www.neci.nj.nec.com/homepages/voyles/fluct.html}
On which systems have you observed higher focused beam current from the false (first) saturation peak rather than "true" saturation? I have not observed this in 18 years with an Etec nor on the new Hitachi.
Woody White
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Hi friends I agree Steve, sometimes (I seem it depends on electron gun design and distance between wenelt and anode, wenelt and filament) the first peak on saturation curve is more than saturation level even in secondary emission signal. Victor Sidorenko, ANTRON Co.Ltd., Moscow, Russia
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I would also be interested in learning how/why. I understand that if the larger spot size delivers more incident beam current, signal to noise is improved, but this does not seem to be the parameter to which Steve is referring.
Woody White McDermott Technology
-----------------------------------
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At 05:07 PM 25-05-2000 +0100, you wrote: Steve Chapman wrote, SNIP , then by de-saturating the larger source } will result in a larger probe dimension on the specimen; increasing the } probe diameter increases the volume of material involved in the production } of BSE. } Ken Wrote: But how does this lead to an increase in the BSE differentiation (implying more signal and hence less noise). Surely just defocussing would do the same thing. Defocussing as such should not effect the BSE coeffecient, unless you had sub surface charging or some other artefact.
Dear Walter, yes we have started one. We continue to gather as time goes on many of them are stored at the moment as the library section that will house them will not be finished until November. we are planning to offer some of them online in pdf format when we have permission from the various companies on the really old ones that they seem not to care about.
We need anything we can get our hands on and also include in this pursuit their older equipment brochures and catalogs. Any assistance of originals or good Xerox copies is great!
thanks Ed Sharpe archivist for SMECC
{ { Subj: Service Manuals Date: 5/25/00 10:54:00 PM US Mountain Standard Time From: Corvos-at-aol.com-at-sparc5.microscopy.com To: microscopy-at-sparc5.microscopy.com
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All,
I would like to know if any has started an extensive collection of Microscopy Service Manuals?
I am an author and platform presenter at the M&M 2000 conference at Philadelphia in August,2000. I am a scientist at the National Geophysical Research Insitute , India. I am looking for living accomodation during my 5 day stay at Philadelphia. I was wondering if any of you will be able to provide me with free accomodation or would be aware of any other resources who may be able to help me in this regard. Due to the limited fiancial resources avaliable to me from my lab, I am looking forward to help from my fellow researchers. Hope you can help me in this regard. I am eagerly looking forward to contributing to the M&M 2000 conference.
With kind regards R. Natarajan Scientist, National Geophysical Research Institute, Hyderabad, India e-mail: rnataraj-at-hd2.dot.net.in Phone: 91-40-7170141 X 2430 (W) Fax: 91-40-7170564
A hearty thank you to everyone who replied to my request for help. I had sources for everything I needed before the evening was out. It is the generosity of the subscribers that makes this list-serve a great resource. Thanks again.
Kim DeRuyter Electron Microscopy Technician 308 Natural Science Facility P.O. Box 755780 University of Alaska Fairbanks, AK 99775-5780 907-474-5452 907-474-5163 fax
I am an author and platform presenter at the M&M 2000 conference at Philadelphia in August,2000. I am a male scientist at the National Geophysical Research Insitute , India. I am looking for living accomodation during my 5 day stay at Philadelphia. I was wondering if any of you will be able to provide me with free accomodation or would be aware of any other resources who may be able to help me in this regard. Due to the limited fiancial resources avaliable to me from my lab, I am looking forward to help from my fellow researchers. Hope you can help me in this regard. I am eagerly looking forward to contributing to the M&M 2000 conference.
With kind regards Mr. R. Natarajan Scientist, National Geophysical Research Institute, Hyderabad, India e-mail: rnataraj-at-hd2.dot.net.in Phone: 91-40-7170141 X 2430 (W) Fax: 91-40-7170564
Dear Fellow Microscopists - Someone in the lab in which I work wants to do SEM of yeast colonies. I was able to locate some references and a general protocol, which seems rather straight forward. However, the authors two of the papers refer to an "Edwards Pearce Tissue Drier, EPD3". I contacted Edwards to find out what this tissue drier is, however they said they could not give me any info because it is no longer manufactured. I am thinking that this tissue drier is simply a freeze drier, but I am not sure. Does anyone out there have any insight into what this thing is or what it does? Thanks for your help!!
Tony
Tony Kowal Research Assistant for Dr. Susan Lindquist
Howard Hughes Medical Institute The University of Chicago 5841 S. Maryland MC 1028, Room N339 Chicago, IL 60637
A student has asked me if I can find a way to tell whether small plant seeds have been charred or not. She is examining seeds from an anthropological dig on a site probably inhabited both in historic times (170 years ago) and prehistoric (1000+ years ago). It seems that for seeds to date from the earlier inhabitation, they would need to be charred, which apparently preserves them.
I looked at some (uncoated) in the SEM, along with some 'fresh' seeds of same species as controls. The seeds of unknown age showed a lot more charging than the fresh ones. Could this be evidence that they are uncharred (i.e., would charring make them more conductive/better secondary electron producers?) Does anyone know of a method of differentiating between charred & uncharred seeds?
Thank you thank you thank you :0)
Paul Grover Chief Microscopist and Bottle Washer Microvista Laboratory Lafayette, IN
The Edwards Pearce Tissue drier was a freeze drier with a small peltier cooled specimen stage. The peltier device was a water- cooled 3-stage stack, achieving about -60oC on a stage about the size of a 35mm negative. There was provision for a small tray of phosphorus pentoxide, and the specimen chamber was a simple Pyrex glass bell, pumped by a rotary pump.
Date sent: Sat, 27 May 2000 12:38:36 -0500 To: Microscopy-at-sparc5.microscopy.com } From: Tony Kowal {askowal-at-midway.uchicago.edu}
How about using the low angle cleaving method of Rafferty? Polish with the SiC close to say the {110} planes (e.g. ten degrees) and then cleave along both the scratches and the {110} plane to get a thin acute wedge. It will take some practice, but should work generally with most Perovskites.
references: 1/Thin Solid Films Vol308-309 (1997) pp399-405 S.D.Walck & J.P.McCaffrey: The small angle cleavage technique applied to coatings and thin films
2/Thin Solid Films Vol304 (1997) pp157-159 Suli Suder, C.A.Faunce & S.E.Donelly: Thin solid film preparation by a small-angle cleavage for transmission electron microscopy
-I hope thses can provide some help. By the way this is not a definitive list, but I am sure Scott Walck can give you a far greater insight to this method.
Regards, Jon
******************************************************** Dr Jonathan Barnard
Analytical Materials Physics The Angstrom Laboratory, Uppsala University P O Box 534, SE-751 21 Uppsala, Sweden Phone: +46-(0)18-4716838 Fax: +46-(0)18-500131 Phone: Microscope room +46 18 471 6365 http://www.angstrom.uu.se/analytical/home.html ********************************************************
Chuck CPD certainly did become the most popular method by a country mile for SEM specimen drying, but freeze-drying still has advantages over it in some situations. These include, for example, specimens where lipid content or lipid structures must be retained (e.g. plant and insect epicuticles), where the specimen is an aggregate of objects loosely bound by a fluid or a mucilaginous matrix (e.g. it could be an advantage in Tony Kowal's yeast and bacterial colonies, soils and clays), or where the specimen is mechanically fragile and the components would be dispersed on submersion in baths of liquid during fixation and solvent drying and CPD (soils & clays, fungal sporangia, yeast and bacterial colonies).
The down side of freeze-drying in most of these contexts is that some shrinkage and distortion almost always results. Consequently, for almost all the situations listed above, and a host of others as well, Low-temperature SEM became the method of choice. In LTSEM the specimen can be viewed fully frozen- hydrated, but most commercially-available LTSEM systems have specimen temperature control, and full or partial freeze-drying can be undertaken either on the SEM specimen stage or in the cryo- preparation unit if required.
Anyone seeking a freeze-drier unit for EM specimens should contact Emitech who make a peltier-cooled unit (K750) which operates around -60oC (and is not unlike the Edwards-Pearce tissue drier) and a turbo molecular pumped Liquid nitrogen cooled low-temperature freeze drier (K775) which operates {-80oC.
http://www.Emitech.co.uk/
I have no financial interest in this company
Chris
} Hi Chris, } } Am I not remembering correctly, or was this earlier approach sort of a } precursor to the critical point drying technique? I mean, did not people use } this earlier technique because that was all they knew and then when the } option of CPD came along, everyone switched over to it? } } I myself was reluctant to say that because that was a very long time ago and } the memory does dull at the edges. } } Chuck } -------- REPLY, Original message follows -------- } } } Date: Sunday, 28-May-00 08:57 PM } } } } From: Chris Jeffree \ Internet: (cjeffree-at-srv0.bio.ed.ac.uk) } } To: Tony Kowal \ Internet: (askowal-at-midway.uchicago.edu } ) } } cc: MICROSCOPY BB \ Internet: } } (microscopy-at-sparc5.microscopy.com) } } } } Subject: Re: Tissue Drier } } } } ------------------------------------------------------------------------ } } 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 } } -----------------------------------------------------------------------. } } } } } } The Edwards Pearce Tissue drier was a freeze drier with a small peltier } cooled } } specimen stage. The peltier device was a water- cooled 3-stage stack, } achieving } } about -60oC on a stage about the size of a 35mm negative. There was } provision } } for a small tray of phosphorus pentoxide, and the specimen chamber was a } } simple Pyrex glass bell, pumped by a rotary pump. } } } } Date sent: Sat, 27 May 2000 12:38:36 -0500 } } To: Microscopy-at-sparc5.microscopy.com } } } From: Tony Kowal {askowal-at-midway.uchicago.edu} } } Subject: Tissue Drier } } } } } ------------------------------------------------------------------------ } } } 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 Fellow Microscopists - } } } Someone in the lab in which I work wants to do SEM of yeast colonies. I } } } was able to locate some references and a general protocol, which seems } } } rather straight forward. However, the authors two of the papers refer } to } } } an "Edwards Pearce Tissue Drier, EPD3". I contacted Edwards to find out } } } what this tissue drier is, however they said they could not give me any } } } info because it is no longer manufactured. } } } I am thinking that this tissue drier is simply a freeze drier, but I am } not } } } sure. Does anyone out there have any insight into what this thing is or } } } what it does? Thanks for your help!! } } } } } } Tony } } } } } } } } } Tony Kowal } } } Research Assistant } } } for } } } Dr. Susan Lindquist } } } } } } Howard Hughes Medical Institute } } } The University of Chicago } } } 5841 S. Maryland } } } MC 1028, Room N339 } } } Chicago, IL 60637 } } } } } } Phone: (773) 702-8795 } } } Fax: (773)702-7254 } } } e-mail: askowal-at-midway.uchicago.edu } } } Pager: on campus - 188 - 9668 (YNOT) } } } off campus - (773)753-1880 - 9668 } } } } } } } } } } } } } } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } } Dr Chris Jeffree } } University of Edinburgh } } Biological Sciences EM Facility } } Daniel Rutherford Building } } King's Buildings EDINBURGH EH9 3JH } } Tel: +44 (0) 131 650 5345 } } FAX: +44 (0) 131 650 6563 } } } } Inveresk Cottage, 26 Carberry Road, } } Inveresk, Musselburgh, Midlothian EH21 8PR, UK } } Tel. +44 (0) 131 665 6062 / Mobile 0410 585 401 } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } } } } -------- REPLY, End of original message -------- } }
===================================================================== DR CHRIS JEFFREE BIOSEM - BIOLOGICAL SCIENCES EM FACILITY UNIVERSITY OF EDINBURGH Daniel Rutherford Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JH, Scotland, UK Tel. #44 131 650 5345 FAX. #44 131 650 6563 Mobile 0410 585 401 email c.jeffree-at-ed.ac.uk SEM / TEM bookings sem-at-ed.ac.uk =====================================================================
When using published methodology, some things matter, others do not. How to know which and what? I don't know, other then a good understanding of the field. Anyway, I have been amused over time with researchers insisting on outdated or cumbersome methods, because "it was published". In this particular case I would like to note that the tissue dryer is a freeze dryer designed for microscopy samples. Several other such instruments would do equally well and I expect that rather more researchers have prepared yeast by the critical point method. Freeze drying or critical point drying both work well for numerous samples. There will be a few specimens that are better prepared by one means or the other, but I wonder how frequently the claim "this gives better preservation" should have the disclaimer "in my hands" added.
Point is that you want to dry the yeast for SEM and you may be wasting your time chasing a particular instrument, when another, perhaps already in the department would do equally well. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Sunday, May 28, 2000 3:39 AM, Tony Kowal [SMTP:askowal-at-midway.uchicago.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 } -----------------------------------------------------------------------. } } } Dear Fellow Microscopists - } Someone in the lab in which I work wants to do SEM of yeast colonies. I } was able to locate some references and a general protocol, which seems } rather straight forward. However, the authors two of the papers refer to } an "Edwards Pearce Tissue Drier, EPD3". I contacted Edwards to find out } what this tissue drier is, however they said they could not give me any } info because it is no longer manufactured. } I am thinking that this tissue drier is simply a freeze drier, but I am not } sure. Does anyone out there have any insight into what this thing is or } what it does? Thanks for your help!! } } Tony } } } Tony Kowal } Research Assistant } for } Dr. Susan Lindquist } } Howard Hughes Medical Institute } The University of Chicago } 5841 S. Maryland } MC 1028, Room N339 } Chicago, IL 60637 } } Phone: (773) 702-8795 } Fax: (773)702-7254 } e-mail: askowal-at-midway.uchicago.edu } Pager: on campus - 188 - 9668 (YNOT) } off campus - (773)753-1880 - 9668 } }
Has she looked at them with conventional light microscopy, either stereo or compound? Following Oxam's Razor, this approach seems to be the simplest and should be tried first.
Best regards Barbara Foster,President Microscopy/Microscopy Education 125 Paridon Street Suite 102 Springfield, MA 01118-2130 PH: 413-746-6931 FX: 413-746-9311 email: MME-at-map.com Website: www.MME-Microscopy.com/education
America's first national consortium of microscopy specialists offering customized on-site training in all areas of microscopy, image analysis, and sample preparation ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^
At 04:19 PM 5/27/00 EDT, Pbgrover-at-aol.com"-at-sparc5.microscopy.com wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
It's been awhile since I was involved in this kind of research (i.e., graduate work in palaeoethnobotany, but I do not recall that seeds need to be "charred", per se, to be preserved over large periods of time. I believe that seeds and other plant remains can also become carbonized without fire, as a result of oxidation, etc., in the depositional environment.
To detect charring in the SEM might be a real problem, especially if trying to differentiate it from carbonization by means other than fire. I suppose that "charred" seeds might show physical damage more than other seeds, but even that might not always be true. The charging effects you observed, on the other hand, might possibly be the effects of mineralization of the seeds as the original seed components become replaced with non-conductive soil constituents over time. If so, this might indicate that these seeds are indeed quite old.
I guess if I was faced with this problem as an archaeologist I would try to identify other indications of fire in the context in which the seeds were found, such as wood charcoal, hearths, etc. On the EM side, if I had access to WDS, I might try comparing amounts of carbon in the old seeds, versus fresh ones.
Finally, the simplest thing to try might just be to toast some fresh seeds and compare them to non-toasted ones of the same type. Try a couple different charring methods, like throwing some in a campfire and collecting them later, and toasting them on a frying pan. A reference you might look at is "Palaeoethnobotany", a book by Dr. Deborah Pearsall published by Academic Press. I think it just came out in a new edition.
Good luck.
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: "Pbgrover-at-aol.com"-at-sparc5.microscopy.com [mailto:"Pbgrover-at-aol.com"-at-sparc5.microscopy.com] Sent: Saturday, May 27, 2000 3:19 PM To: Microscopy-at-sparc5.microscopy.com
Esteemed colleagues,
A student has asked me if I can find a way to tell whether small plant seeds
have been charred or not. She is examining seeds from an anthropological dig on a site probably inhabited both in historic times (170 years ago) and prehistoric (1000+ years ago). It seems that for seeds to date from the earlier inhabitation, they would need to be charred, which apparently preserves them.
I looked at some (uncoated) in the SEM, along with some 'fresh' seeds of same species as controls. The seeds of unknown age showed a lot more charging than the fresh ones. Could this be evidence that they are uncharred (i.e., would charring make them more conductive/better secondary electron producers?) Does anyone know of a method of differentiating between charred
& uncharred seeds?
Thank you thank you thank you :0)
Paul Grover Chief Microscopist and Bottle Washer Microvista Laboratory Lafayette, IN
The continuing discussion on whether film is better than digital, and whether we can directly compare their resolving ability is very interesting and throwing up some really useful technical stuff. But aren't we missing the point a bit? Surely the image quality is user defined - if you or your customer, are satisfied with the end product then the equipment has done its job. How often does one need to examine a micrograph to assess its limits? If are getting that close to a picture then you may be taking things out of context a bit and losing the whole concept.
The future is with digital image capture, let's hope the price of the equipment comes down to something more attainable!
Pete
Peter Bond Plymouth Electron Microscopy Unit University of Plymouth Drake Circus Plymouth Devon UK PL4 8AA Tel/Fax: 01752 233092 email: pbond-at-plymouth.ac.uk
A discussion is developing from my comment that a de-saturated W hairpin source provides a better BSE image. This "problem" first came to light when a South African client commented that they have far better atomic number contrast images if they run on "first peak" rather than gun saturation.
I based my explanation on some work by C-R Peters, he relates BSE and SE signals to probe size. The bigger the probe the bigger the reaction volume that is the source of the BSE contribution to an image.
In the de-saturated state a normal W hairpin system has a very much bigger source which reflects in a very much larger probe being placed on the specimen than would be obtained under a "normal saturation" situation and therefore provides the operator with more BSE.
Some instruments do give a higher signal at their false peak than at what we would know as saturation. I have seen this on Cambridge (Leica, Leo) instruments and no matter how hard you try to "correct" what you feel is a mis alignment you just do not win. On some occasions I have also seen this on a Philips but it is much more rare.
I put this down to gun design as you do not see this in a Japanese designed instrument. In their case a higher false peak indicates a definite mis alignment!
I base my comments on running courses with a different SEM in a different laboratory almost every week, not on what happens with one laboratory on one instrument.
Steve Chapman Senior Consultant Protrain For consultancy and professional training in EM world wide Tel 44+ 1280 814774 Fax 814007 www.emcourses.com
Laboratories who have to do pathology work and research also, have particular problems in that they are faced with a wide variety of specimens not all of which are suitable for a single epoxy formulation mixture during embedment.
Many pathology laboratories in the US use the medium hard formulation by Luft. This is indeed a good, nearly all purpose formulation, however, in laboratories that have to use glass knives for thick sectioning the NMA contained in this formulation deterioates the edge of the glass knife very quickly. If glass knives have to be used for thin sectioning, then this formulation cannot be used at all. Otherwise with diamond knives it cuts very well, assuming of course, that the embedding has been done well. The original formulation by Luft required two mixtures to be made up seperately. Years ago I combined this into a single mixture which can be easily frozen (without accelerator!) and kept for months at -80 or -20.
Eponate 12 148 ml DDSA 100 ml NMA 76 ml
This makes 324 ml. Accelerate it with 1.5% DMP-30 before use. If you are interested in this formulation, please print it out now, because this is the last time I will address it.
Sometimes laboratories are faced with processing floating cells. These ar e best enrobed in agarose, hopefully after osmication. In order to avoid enormous cell loss during spinning down repeatedly into embedding medium, it is fortuitous to draw off all buffer after osmium, and spin the cells into a mixture of 3% Ultra-low temperature agarose, Type IX (Sigma). This agarose type gells at 15 deg C, and it has the property of not being so dense and fibrous that embedding medium cannot penetrate it. After the gel is cooled (in the refrigerator, or on ice), the blocks are cut of desired size. It is important to note that one is no longer dealing with cells, but actual blocks and the protocol for dehydration and embedding must fit the requirements of the block size. The resulting blocks also are very well embedded in the above formulation.
The above formulation is good for skin and muscle, but the protocol must be adjusted to allow adequate penetration. We do 2 hours for dehydration, use propylene oxide for an intermediate, and start infiltrating with mixtures of PO and Epoxy, 2:1, 1:1, 1:3, pure, pure, pure, etc. When the tissue goes into the newly accelerated pure mixtures, I put an ordinary 60W bulb in the vicinity so that the mixture can heat to about 37 deg. At this point the formulation becomes very liquid and infiltration is enhanced. After 1.5 hours the lamp is turned off as minor polymerization begins about 40 deg C, which is undesirable during infiltration.
A problem arises when laboratories have to produce a large number of thick sections with glass knives, or they have to also use glass knives for thin sectioning. Then the above formulation cannot be used. A new embedding medium containing no NMA is needed. Many years ago, Mollenhauer invented a "Mixed Embedding" system which contained Araldite 502, Epon, DDSA, and dibutyl pthalate. This is an extremely useful system - a joy to section with glass knives. The downside is that it is more viscous than media without Araldite, and it takes longer to infiltrate. Here is the formulation, again reconfigured by me years ago into a single mixture.
Araldite 502 30ml Eponate 12 50 ml DDSA 55 ml Dibutyl (EMS) 0.75% (mix in with the 3 items above DMP-30 1.5% (add at time of use)
Freeze mixture without the accelerator. Will keep many months at -80. Can also keep at -20. For thick sectioning I polymerize just 24 hours. If blocks are to be thin sectioned, then I put them back into the oven for another 24 or 48 hours. This formulation has the capability of being soft and stiff, as you wish. If only thick sections are to be done, use 1% dibutyl. If blocks are too soft for your liking, just put them into a 60 deg C oven for several days, or use 95 deg C for an hour or two. One time I forgot some blocks in the 95 deg oven for a week. They still sectioned well.
Again, anyone interested please print this out now. I will not be answering these questions again, nor write these formulations again.
This is fun! Keep accurate records of what you do, particularly the infiltration and dehydration times. Finally you will have every block a joy to handle. No frustrations!
Bye, Hildy Crowley Sr. Electron Microscopy Specialist University of Denver Denver, CO
P.S. It is assumed that all processing is done with vials in constant motion! Once infiltration starts, vials must be on a rotator, not on a rocker or a shaker, as the monomers of the formulations will seperate and poor embedding will result.
} The continuing discussion on whether film is better than digital, and } whether we can directly compare their resolving ability is very interesting } and throwing up some really useful technical stuff. But aren't we missing } the point a bit? Surely the image quality is user defined - if you or your } customer, are satisfied with the end product then the equipment has done } its job. How often does one need to examine a micrograph to assess its } limits? If are getting that close to a picture then you may be taking } things out of context a bit and losing the whole concept. } } The future is with digital image capture, let's hope the price of the } equipment comes down to something more attainable! }
Dear Pete, If one is doing quantitative image processing, the better resolution available from film can be relevant. For example, if one wants to do corellation averaging, one needs as many objects in the picture as possible, and also as good resolution as needed--e.g., for 1 nm resolution of the reconstruction, a pixel size of 0.25 nm times the magnification is necessary, so the recording medium must have resolution equal to or better than that. The pixel size of the scanner must also be this small--obviously irrelevant for digital recording. A 5 micrometer scanning-pixel size at 20kx mag gives a pixel resolution of 0.25 nm, and for 6.5 by 9 cm film, one will have about 12,000 by 16,000 (taking the header into account) pixels of useful information. This will give a broader area than that available at equal resolution from any CCD chip now out there. I can assure you that I have often had to determine the information limits in a particular micrograph. Yours, Bill Tivol
Has anyone ever cured Epon-Araldite with UV? I was given a cobbled-together protocol by a non-microscopist post-doc; the protocol calls for a dry-ice, UV cure of Epon-Araldite. It says it can take from 3-7 days. I'm wondering if this is either a resin mix-up *or* the resin just set up all by itself (over that time frame I've seen it happen), just because it was a thin layer and left alone - nothing to do with the UV exposure. She has no ref. for this technique and isn't sure now where she got it. Sigh. I've only been able to find heat-cure protocols...anyone have any leads or thoughts on this UV thing?
(Sorry about the cross-post for those of you on both of these servers)
Thanks!
Tamara (Planning to use the oven.....) Howard CSHL
} ... } } I based my explanation on some work by C-R Peters, he } relates BSE and SE signals to probe size. } The bigger the probe the bigger the reaction volume } that is the source of the BSE contribution to an image. } } In the de-saturated state a normal W hairpin system has a } very much bigger source which reflects in a very much } larger probe being placed on the specimen than would be } obtained under a "normal saturation" situation and } therefore provides the operator with more BSE.
I have seen the 1st peak provide more beam current, but I fail to see how a larger probe diameter at similar beam currents can provide better z contrast. Do you know what the beam current is ... "1st peak" vs "saturated"??
I might also suggest the 1st peak may provide emission from several areas of the filament instead of just from the tip. This would manifest as "ghost" images or double images ... leastwise, I hope this type of phenomenon couldn't be confused with "better z contrast" :o)
Does anyone know of anybody who sells motors with suitable mechanical interfaces to drive X and Y of a JEOL 840A stage?
Replies from suppliers very welcome.
thanks
rtch
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
This is my last posting, I promise, looking for a Probe Current Detector (PCD) for a JEOL 840. The pneumatically-powered Faraday Cup which shoots into the beam just below the objective aperture, samples the beam, and retracts.
Does anyone have one which is redundant or otherwise spare?
Alternatively, does anyone know of a third-party manufacturer of this sort of thing?
cheers
rtch
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
At 12:17 PM 5/30/00, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
The best CCD imagers are typically 6u in size. Complicating this is that they may be square or rectangular. Fuji's new hex-shaped pixels may be a major improvement. We'll see.
Our civil engineering department is trying to look at the bonding between bitumen and aggregate with the aid of a SEM. However, we do not have ESEM facilities (as I suggested). Now I was ask to see if anyone in the EM fraternity has any experience in using standard SEMs for examining bitumen containing specimens (I have not!!; and I don't want to ruin my microscopes).The supplied samples have been cut into slices of approximately 4mm thickness.
Any ideas out there? Thanks
Hans Brinkies Senior Lecturer Swinburne, University of Technology School of Engineering and Science P.O.Box 218 - Hawthorn - Vic -3122 - Australia Phone: +61 3 9214 8657 Fax: +61 3 9214 8264 Email: Hbrinkies-at-swin.edu.au
Peter Bond is right in saying "The future is with digital image capture" if we were at some future date to count heads of digital versus film TEM users. That change will be for reasons of convenience and to save labour. These are powerful and valid reasons.
Bill Tivol has given one set of applications where digital currently does not always measure up to film. Here are another couple of such applications. 1 When great enlargements are required film is superior. This is because of film's greater resolution, but more importantly, because its much, much easier to take images at moderate powers and highly enlarge. That way we take advantage of the TEM's greater depths of focus at low powers and of film's higher resolution/ image detail. 2 Whenever a TEM image is taken at low brightness (to avoid beam damage or at very high powers or in dark-field) relatively few electrons form the image and make that image grainy. Film is very slow and requires then a longer exposure, thus boosting the quantity of electrons used and improving the image. Digital is much more sensitive and so the exposure must be shortened. As a result the best digital camera will record, quiet faithfully the grainy, unsharp image.
Many labs rarely or never use such applications and for them the reasons to change to digital now may be overwhelming. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Wednesday, May 31, 2000 5:18 AM, William Tivol [SMTP:tivol-at-wadsworth.org] 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 } -----------------------------------------------------------------------. } } } Peter Bond wrote: } } } The continuing discussion on whether film is better than digital, and } } whether we can directly compare their resolving ability is very interesting } } and throwing up some really useful technical stuff. But aren't we missing } } the point a bit? Surely the image quality is user defined - if you or your } } customer, are satisfied with the end product then the equipment has done } } its job. How often does one need to examine a micrograph to assess its } } limits? If are getting that close to a picture then you may be taking } } things out of context a bit and losing the whole concept. } } } } The future is with digital image capture, let's hope the price of the } } equipment comes down to something more attainable! } } } } Dear Pete, } If one is doing quantitative image processing, the better } resolution } available from film can be relevant. For example, if one wants to do } corellation averaging, one needs as many objects in the picture as possible, } and also as good resolution as needed--e.g., for 1 nm resolution of the } reconstruction, a pixel size of 0.25 nm times the magnification is necessary, } so the recording medium must have resolution equal to or better than that. } The pixel size of the scanner must also be this small--obviously irrelevant } for digital recording. A 5 micrometer scanning-pixel size at 20kx mag } gives a pixel resolution of 0.25 nm, and for 6.5 by 9 cm film, one will have } about 12,000 by 16,000 (taking the header into account) pixels of useful } information. This will give a broader area than that available at equal } resolution from any CCD chip now out there. I can assure you that I have } often had to determine the information limits in a particular micrograph. } Yours, } Bill Tivol } }
I have a question about the structure of 2H-SiC (x-ray card No.29-1130), which belong to No. 186 space group (P63mc). (a=0.3076 nm, c=0.5048 nm, alfa=beta=120 degrees, gama=90 degrees)
My question is as below:
(A) Based on the symmetry of space group No. 186, the coordinates of the atoms in a unit cell should be
Si 2a : (0,0,0), and (0,0,0.5)
C 2b : (2/3, 1/3, 0.125) and (1/3,2/3,0.625)
(B) However, acoording to the real stacking sequence, the coordinates of the Si and C atoms in a unit cell are
Si sites: (0, 0, 0) and (2/3, 1/3, 0.5) C sites: (2/3, 1/3, 0.125) and (0, 0, 0.625)
It seems that there are some conflict between the two sets of coordinates, could someone help me to solve such a problem.
} Does anyone know of anybody who sells motors with suitable mechanical } interfaces to drive X and Y of a JEOL 840A stage? } } Replies from suppliers very welcome.
Ritchie,
Deben UK Ltd, 11-15 High St, Stowmarket, Suffolk UK IP14 6QL make very nice motors and controllers for X, Y and Z control of SEM stages. You can also contact them at : info-at-deben.co.uk or via their webpage : http://www.deben.co.uk
Cheers,
David Vowles Electron Microscopy Unit Dept of Materials Science and Metallurgy University of Cambridge Pembroke St Cambridge UK CB2 3QZ Tel: +44 1223 334325 Fax: +44 1223 334567 Email: djv23-at-msm.cam.ac.uk
Dear Gao, we have used simulation software for CBED for both ZnO and GaN which are isomorphic with 2H-SiC (same space group:P6_3mc). For our software we use the following atom locations (I will use Si & C instead):
Si positions: (1/3, 2/3, 0) and (2/3, 1/3, 1/2)
C positions: (1/3, 2/3, u) and (2/3, 1/3, u + 1/2)
The Si-C bond length is parameterized by the quantity u. For the ideal (perfect stacking) arrangement u=3/8. However I am reliably informed that for SiC the u parameter is a great mystery at the present. However using u=3/8 should prove to be a good starting point.
Please note that to place Si at (0,0,0) you will have to remove (1/3,2/3,0) from each coordinate above which gives the following locations:
Si: (0,0,0) and (1/3, 2/3, 1/2)
C: (0,0,u) and (1/3, 2/3, u + 1/2)
Does this make sense?
Regards, Jonathan ******************************************************** Dr Jonathan Barnard
Analytical Materials Physics The Angstrom Laboratory, Uppsala University P O Box 534, SE-751 21 Uppsala, Sweden Phone: +46-(0)18-4716838 Fax: +46-(0)18-500131 Phone: Microscope room +46 18 471 6365 http://www.angstrom.uu.se/analytical/home.html ********************************************************
Dear List, I'm looking for methods for measuring the roughness of a sheet of polyethylene. The film consists of grains (80 to 150 microns in diameter) which are loosely packed. Large area analysis is prefered since the surface is very irregular locally.
Hello everybody: I will like to do cryomicrotomy of PP wires. My question is : can anyone suggest what kind of epoxy is appropriate for cryomicrotomy? I used Epoxy Mount and it was chattered during cryo process. The manufacturer confirm to me that epoxy mount is not appropriate for cryomicrotomy. Can anynone help me?
HI Hildy, You are always a great source of information....thanks.
Lee
Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Confocal Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
2 Whenever a TEM image is taken at low brightness (to avoid beam damage or at very high powers or in dark-field) relatively few electrons form the image and make that image grainy. Film is very slow and requires then a longer exposure, thus boosting the quantity of electrons used and improving the image. Digital is much more sensitive and so the exposure must be shortened. As a result the best digital camera will record, quiet faithfully the grainy, unsharp image.
Dear Jim,
Your points are good; however, exposure times are not, by
themselves, relevant. An exception to this is if the damage depends
on the dose rate; i.e., if there are mechanisms to dissipate the absorbed
beam energy which limit damage at low illumination levels. The
image quality depends on how much information is carried by each elec-
tron (basically, the number of photons produced in the scintillator times
the quantum efficiency of the system for these photons for digital, and the
probability of a darkened film grain for film or image plates) times the
number of electrons. The damage--except for dose-rate-dependence--
is linear with the number of electrons, so it is the efficiency of the pro-
cess of obtaining the information from each electron which determines
the ultimate resolution limit. Also, the use of image averaging can make
one clear image from many grainy ones in the case that one has a large
Randy, Don't forget about other analytical techniques, such as XPS. Some well engineered surface analysis may reveal significant chemical differences between "charred", new, and aged or mineralized specimens. In addition to Randy's good advice on the microscopy, I would also utilize the power of both TEM and EELS in comparing morphology and carbon and oxygen chemistry. Electron energy loss spectroscopy combined with EDS is one of my favorites for carbon micro-analyses. Looking at both the low loss and the core loss structure of EELS spectra, there is tremendous detail and differentiation to be obtained. If TEM and "PEELS" are available, and you don't mind sacrificing a bit of the artifact, a series of control specimens and analysis by "AEM" (TEM, EDS, and PEELS) would be worth a shot. Sounds like fun, Good Luck, Brad Huggins BP Amoco, Analytical Naperville IL
} ---------- } From: Tindall, Randy D.[SMTP:TindallR-at-missouri.edu] } Sent: Tuesday, May 30, 2000 9:13 AM } To: '"Pbgrover-at-aol.com"-at-sparc5.microscopy.com' } Cc: 'microscopy-at-sparc5.microscopy.com' } Subject: RE: charred plant seeds } } ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } Paul, } } It's been awhile since I was involved in this kind of research (i.e., } graduate work in palaeoethnobotany, but I do not recall that seeds need to } be "charred", per se, to be preserved over large periods of time. I } believe } that seeds and other plant remains can also become carbonized without } fire, } as a result of oxidation, etc., in the depositional environment. } } To detect charring in the SEM might be a real problem, especially if } trying } to differentiate it from carbonization by means other than fire. I } suppose } that "charred" seeds might show physical damage more than other seeds, but } even that might not always be true. The charging effects you observed, on } the other hand, might possibly be the effects of mineralization of the } seeds } as the original seed components become replaced with non-conductive soil } constituents over time. If so, this might indicate that these seeds are } indeed quite old. } } I guess if I was faced with this problem as an archaeologist I would try } to } identify other indications of fire in the context in which the seeds were } found, such as wood charcoal, hearths, etc. On the EM side, if I had } access } to WDS, I might try comparing amounts of carbon in the old seeds, versus } fresh ones. } } Finally, the simplest thing to try might just be to toast some fresh seeds } and compare them to non-toasted ones of the same type. Try a couple } different charring methods, like throwing some in a campfire and } collecting } them later, and toasting them on a frying pan. A reference you might look } at is "Palaeoethnobotany", a book by Dr. Deborah Pearsall published by } Academic Press. I think it just came out in a new edition. } } Good luck. } } 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: "Pbgrover-at-aol.com"-at-sparc5.microscopy.com } [mailto:"Pbgrover-at-aol.com"-at-sparc5.microscopy.com] } Sent: Saturday, May 27, 2000 3:19 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: charred plant seeds } } } ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } Esteemed colleagues, } } A student has asked me if I can find a way to tell whether small plant } seeds } } have been charred or not. She is examining seeds from an anthropological } dig } on a site probably inhabited both in historic times (170 years ago) and } prehistoric (1000+ years ago). It seems that for seeds to date from the } earlier inhabitation, they would need to be charred, which apparently } preserves them. } } I looked at some (uncoated) in the SEM, along with some 'fresh' seeds of } same } species as controls. The seeds of unknown age showed a lot more charging } } than the fresh ones. Could this be evidence that they are uncharred } (i.e., } would charring make them more conductive/better secondary electron } producers?) Does anyone know of a method of differentiating between } charred } } & uncharred seeds? } } Thank you thank you thank you :0) } } Paul Grover } Chief Microscopist and Bottle Washer } Microvista Laboratory } Lafayette, IN }
Could it be that moisture escaping from the fresh seeds somehow helps to dissipate the charge so that new seeds do not charge like the old ones?
I would also be interested in the x-ray spectra of the two seeds. Perhaps there is partial mineralization as Randy Tindall suggested. A light element detector should also reveal a difference in O/C ratio due to differences in moisture content, or perhaps due to charring.
Warren S.
At 09:13 AM 5/30/2000 -0500, you wrote: } Esteemed colleagues, } } A student has asked me if I can find a way to tell whether small plant seeds } } have been charred or not. She is examining seeds from an anthropological } dig } on a site probably inhabited both in historic times (170 years ago) and } prehistoric (1000+ years ago). It seems that for seeds to date from the } earlier inhabitation, they would need to be charred, which apparently } preserves them. } } I looked at some (uncoated) in the SEM, along with some 'fresh' seeds of } same } species as controls. The seeds of unknown age showed a lot more charging } than the fresh ones. Could this be evidence that they are uncharred (i.e., } would charring make them more conductive/better secondary electron } producers?) Does anyone know of a method of differentiating between charred } } & uncharred seeds? } } Thank you thank you thank you :0) } } Paul Grover } Chief Microscopist and Bottle Washer } Microvista Laboratory } Lafayette, IN
Of course there are situations where one or the other Technique may be better because of the limitations of film or digital imaging. And especially for a task like the one Bill talks about below, where field of view AND resolution are important, images taken on film may be superior to images from a digital camera. On the other hand, digital imaging may have something to offer in those cases as well:
Instead of taking one image and do the corellation averaging, why not have the computer do it? I could probably set up a system that acquires the image at a high enough resolution, extract all the necessary data, then move the stage to an adjacent area and continue the measurements there. This technique would even have an advantage over film: I can set the lower limits for the accuracy before taking the images and the system continues to measure until these limits are satisfied. This could be done without user intervention. So, instead of taking one image, scanning it in with a scanner, and then being "limited" by the field of view to a certain measurement accuracy, one could start the measurement with a predetermined accuracy, go drink a coffee, work on that paper, do the travel expense report, then go back to the microscope and collect the spreadsheet with the measurement data. Of course this requires the setup of a fairly complex system, but those are technical problems, not fundamental ones.
Just a thought ....
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: William Tivol [mailto:tivol-at-wadsworth.org] Sent: Tuesday, May 30, 2000 1:18 PM To: microscopy-at-sparc5.microscopy.com
Peter Bond wrote:
} The continuing discussion on whether film is better than digital, and } whether we can directly compare their resolving ability is very interesting } and throwing up some really useful technical stuff. But aren't we missing } the point a bit? Surely the image quality is user defined - if you or your } customer, are satisfied with the end product then the equipment has done } its job. How often does one need to examine a micrograph to assess its } limits? If are getting that close to a picture then you may be taking } things out of context a bit and losing the whole concept. } } The future is with digital image capture, let's hope the price of the } equipment comes down to something more attainable! }
Dear Pete, If one is doing quantitative image processing, the better resolution available from film can be relevant. For example, if one wants to do corellation averaging, one needs as many objects in the picture as possible, and also as good resolution as needed--e.g., for 1 nm resolution of the reconstruction, a pixel size of 0.25 nm times the magnification is necessary, so the recording medium must have resolution equal to or better than that. The pixel size of the scanner must also be this small--obviously irrelevant for digital recording. A 5 micrometer scanning-pixel size at 20kx mag gives a pixel resolution of 0.25 nm, and for 6.5 by 9 cm film, one will have about 12,000 by 16,000 (taking the header into account) pixels of useful information. This will give a broader area than that available at equal resolution from any CCD chip now out there. I can assure you that I have often had to determine the information limits in a particular micrograph. Yours, Bill Tivol
your remark 1) What you can do of course with digital imaging (provided you have the necessary hardware), is to automatically collect larger areas by taking several images that overlap, reducing the advantage that film has in this area and perhaps offer other possibilities as I just explained in another posting as a response to Bill Tivol's posting.
your remark 2) I am not sure you are not comparing apples and oranges. What you are saying is, that because of the "slowness" of film you need longer exposures, thereby averaging out the statistical noise of the electron, which is not the case for CCD cameras at short exposures, hence they appear more noisy. In essence what you are doing is to compare a short exposure image to a long exposure image. What you can do with a CCD camera is the following: You can get (perhaps) real-time dark field images and position your sample and/or decide if you want to actually take the image. Then you take a SERIES of images, let's say 10, each at an exposure time of 1/10 of the film exposure. This can be done automatically, of course. Finally, you add or average all of these images using a pattern recognition to align them first. The result: A dark field image that should be as noisy as the film image, with much less problems due to drift during long exposures, a higher dynamic range and visible immediately on the viewing screen.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== 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: jim [mailto:jim-at-proscitech.com.au] Sent: Tuesday, May 30, 2000 7:13 PM To: 'William Tivol' Cc: microscopy-at-sparc5.microscopy.com
Peter Bond is right in saying "The future is with digital image capture" if we were at some future date to count heads of digital versus film TEM users. That change will be for reasons of convenience and to save labour. These are powerful and valid reasons.
Bill Tivol has given one set of applications where digital currently does not always measure up to film. Here are another couple of such applications. 1 When great enlargements are required film is superior. This is because of film's greater resolution, but more importantly, because its much, much easier to take images at moderate powers and highly enlarge. That way we take advantage of the TEM's greater depths of focus at low powers and of film's higher resolution/ image detail. 2 Whenever a TEM image is taken at low brightness (to avoid beam damage or at very high powers or in dark-field) relatively few electrons form the image and make that image grainy. Film is very slow and requires then a longer exposure, thus boosting the quantity of electrons used and improving the image. Digital is much more sensitive and so the exposure must be shortened. As a result the best digital camera will record, quiet faithfully the grainy, unsharp image.
Many labs rarely or never use such applications and for them the reasons to change to digital now may be overwhelming. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Wednesday, May 31, 2000 5:18 AM, William Tivol [SMTP:tivol-at-wadsworth.org] 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 } -----------------------------------------------------------------------. } } } Peter Bond wrote: } } } The continuing discussion on whether film is better than digital, and } } whether we can directly compare their resolving ability is very interesting } } and throwing up some really useful technical stuff. But aren't we missing } } the point a bit? Surely the image quality is user defined - if you or your } } customer, are satisfied with the end product then the equipment has done } } its job. How often does one need to examine a micrograph to assess its } } limits? If are getting that close to a picture then you may be taking } } things out of context a bit and losing the whole concept. } } } } The future is with digital image capture, let's hope the price of the } } equipment comes down to something more attainable! } } } } Dear Pete, } If one is doing quantitative image processing, the better } resolution } available from film can be relevant. For example, if one wants to do } corellation averaging, one needs as many objects in the picture as possible, } and also as good resolution as needed--e.g., for 1 nm resolution of the } reconstruction, a pixel size of 0.25 nm times the magnification is necessary, } so the recording medium must have resolution equal to or better than that. } The pixel size of the scanner must also be this small--obviously irrelevant } for digital recording. A 5 micrometer scanning-pixel size at 20kx mag } gives a pixel resolution of 0.25 nm, and for 6.5 by 9 cm film, one will have } about 12,000 by 16,000 (taking the header into account) pixels of useful } information. This will give a broader area than that available at equal } resolution from any CCD chip now out there. I can assure you that I have } often had to determine the information limits in a particular micrograph. } Yours, } Bill Tivol } }
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} Instead of taking one image and do the corellation averaging, why not } have the computer do it? I could probably set up a system that acquires } the image at a high enough resolution, extract all the necessary data, } then move the stage to an adjacent area and continue the measurements } there. This technique would even have an advantage over film: I can set } the lower limits for the accuracy before taking the images and the } system continues to measure until these limits are satisfied. This could } be done without user intervention. So, instead of taking one image, } scanning it in with a scanner, and then being "limited" by the field of } view to a certain measurement accuracy, one could start the measurement } with a predetermined accuracy, go drink a coffee, work on that paper, do } the travel expense report, then go back to the microscope and collect } the spreadsheet with the measurement data. } Of course this requires the setup of a fairly complex system, but those } are technical problems, not fundamental ones. }
Dear Mike, That is an excellent suggestion. Of course, the beam should be
restricted to the area of the detector to avoid damaging the specimen except where inevitable. You are correct that setting the accuracy limits prior to taking the image avoids mixing the required data with those from excess exposure, when the specimen has undergone some radiation damage--this could perhaps be done with film, but it is simpler with electronic data collection. There one can even make sure that those pixels which are most important are the ones optimized, and, with a small enough beam, like that used in spot-scan imaging, one could expose different parts of the image for different times, so that as much as possible of the image is optimally collected. This also avoids damaging the area of the specimen not yet under investigation.
Anyone aware of software capable of generating depth profiles from digital SEM stereo pair images? I am aware of the Oxford ISIS system/program, but wondered if other companies offer similar software that would run 'stand-alone' in a PC and work with any digital files.
Any help or suggestions along these lines would be most appreciated.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Greetings! } } Has anyone ever cured Epon-Araldite with UV? I was given a } cobbled-together protocol by a non-microscopist post-doc; the protocol } calls for a dry-ice, UV cure of Epon-Araldite. It says it can take from } 3-7 days. I'm wondering if this is either a resin mix-up *or* the resin } just set up all by itself (over that time frame I've seen it happen), just } because it was a thin layer and left alone - nothing to do with the UV } exposure. She has no ref. for this technique and isn't sure now where she } got it. Sigh. I've only been able to find heat-cure protocols...anyone } have any leads or thoughts on this UV thing? } } (Sorry about the cross-post for those of you on both of these servers) } } Thanks! } } Tamara (Planning to use the oven.....) Howard } CSHL } } } Hi,
Try it, but not on anything valuable! Resins set up by themselves at rt. I don't remember ever reading anything about UV in the Handbook of Epoxy Resins. Why do you want to do it? That is the question. What would be the advantage of that major fiddle?
When I worked for Zeiss, we did a lot of coal analysis using microspectrophotometry. I've also done some polarized light work on a sample or two for customers. Is the bonding you are looking for on the level where light microscopy might work? Just for your information, I also recently took a look at the "tie" layers between polymer films in ketsup bottles, perhaps a distant but related application. Polarized light and DIC did a great job in that instance.
.. just a thought, but hopefully helpful.
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At 12:13 PM 5/31/00 +1000, Hans Brinkies wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Have you considered structured lighting? A flat beam of light is illuminates the subject from a 45 degree angle. A camera is views it at 90 degrees. The height and size of the roughness can be reconstructed from the shadows the light makes.
Gordon
Gordon Couger gcouger-at-couger.com
Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00 ----- Original Message ----- } From: "DAVID I SAXON" {DISAXON-at-prodigy.net} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, May 31, 2000 7:19 AM
In a message dated 5/31/00 2:52:24 PM, ferreira1-at-llnl.gov writes:
} Anyone aware of software capable of generating depth profiles from digital } SEM stereo pair images? I am aware of the Oxford ISIS system/program, } but wondered if other companies offer similar software that would run 'stand-alone' } in a PC and work with any digital files.
One of the (many) functions in Fovea Pro (http://members.aol.com/FoveaPro), a set of Photoshop-compatible plug-ins intended for the analysis of images including those from microscopes such as SEMs, is a routine that fuses stereo pair images to obtain the elevation of points on the surface. This can then be used to measure elevation profiles, or to reconstruct 3D surface images. Examples of both are included in the tutorial and are illustrated on the web site. It isn't stand-alone (you need Photoshop or a compatible program), but it will do what you ask.
The tissue drier you referred to had a "cold finger" which acted as a vapor trap to remove the water vapor produced from the sublimation of the specimen ice. This is a very important feature for achieving "distortion-free drying" which common type of freeze driers lack I believe. Check out Pearse's book Histochemistry Theoretical and Applied, vol.1. It contains an excellent chapter on this technique and others. However, I would try CPD, also.
Hank Adams Manager Integrated Microscopy Core Molecular and Cellular Biology Baylor College of Medicine Houston, Tx 77030
Hello, there, Does anyone out here know that can we put alkylamine to the turbo pump when carbon supporting film is being glow discharge? Thanks.
Peiyi
Krebs Institute for Biomolecular Research University of Sheffield Firth Court Western Bank Sheffield Yorkshire S10 2UH United Kingdom Tel: +44 (0)114 222 2000 Direct: +44 (0)114 222 2739 FAX: +44 (0)114 272 8697 E-mail: p.wang-at-sheffield.ac.uk
Briget wrote: =========================================================== I will like to do cryomicrotomy of PP wires. My question is : can anyone suggest what kind of epoxy is appropriate for cryomicrotomy? I used Epoxy Mount and it was chattered during cryo process. The manufacturer confirm to me that epoxy mount is not appropriate for cryomicrotomy. Can anynone help me? =========================================================== I am assuming you mean polypropylene coated wires and not polypropylene monofilament. We have generally found that for coated wires samples, we like to Pt coat it first (by sputtering), then embed in SPI-Pon 812 resin. I would expect that any of the other "Epon® 812 substitutes", available from the other major EM supplies firms would work just as well.
You will definitely want to use a diamond knife on this and you can vary the hardness of the resin in way that gives you the best sections. Use a knife angle that is not larger than 45°, the lower the temperature (usually) the better.
Disclaimer: SPI Supplies offers for sale the resin and diamond knives mentioned and performs this kind of cryoultramicrotomy for clients.
Chuck
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Hamamatsu recently introduced a new camera called the electron bombardment ccd where accelerated electrons directly bombard a back thinned, peltier cooled ccd. The electrons are emitted from a photocathode for applications particularly in low light microscopy. With a full well capacity of 300,000 electrons, I wonder if this approach could be used in direct exposure of the ccd to the electron beam. Shane
K. Shane Collins Scientific Instrument Company 805.444.4953 cell 310.568.9188 office 310.568.9189 fax
-----Original Message----- } From: Paul Voyles [mailto:voyles-at-research.nj.nec.com] Sent: Friday, May 26, 2000 9:11 AM To: Microscopy-at-sparc5.microscopy.com
} You could probably expose your CCD chip directly to the electron bean -- } and buy a new chip every few hundred exposures or so! Electrons have
There is an even worse problem with exposing the CCD directly the electron beam. The p-n junctions in the CCD chip have a certain "well capacity" - a number of electron/hole pairs they can hold before they saturate. Fast (keV) electrons are much more efficient at producing electron/hole pairs than photons - so much so that a pixel on the CCD would saturate after about 15 fast electrons hit it. Just the square root N shot noise at that level is about 25% - larger than typical TEM micrograph contrast of 10-20%. That's why a phosphor or scintillator is necessary to transform the fast electrons into photons.
Yes, it could be done "in theory". Somebody would need to figure out the software and perhaps modify the hardware. Then we would find that the total exposure of the specimen to the electron beam maybe a muliple of the film's exposure. Afterall, an 8 sec film exposure would not amount in digital to 10x0.8, but we would require considerable time in between exposures. Since the problems in the discussed circumstances are specimen movement and beam damage, it seems that taking multiple exposures is a poor option.
Digital cameras are for some situation too sensitive to electron exposure. Cutting back on electrons is no option since its the electrons that form the image in the first instance. Much easier in light microscopy . . . insert a neutral density filter. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
} Hello Jim, } } just a couple of remarks to your email. } } your remark 1) What you can do of course with digital imaging (provided } you have the necessary hardware), is to automatically collect larger } areas by taking several images that overlap, reducing the advantage that } film has in this area and perhaps offer other possibilities as I just } explained in another posting as a response to Bill Tivol's posting. } } your remark 2) I am not sure you are not comparing apples and oranges. } What you are saying is, that because of the "slowness" of film you need } longer exposures, thereby averaging out the statistical noise of the } electron, which is not the case for CCD cameras at short exposures, } hence they appear more noisy. In essence what you are doing is to } compare a short exposure image to a long exposure image. What you can do } with a CCD camera is the following: You can get (perhaps) real-time dark } field images and position your sample and/or decide if you want to } actually take the image. Then you take a SERIES of images, let's say 10, } each at an exposure time of 1/10 of the film exposure. This can be done } automatically, of course. Finally, you add or average all of these } images using a pattern recognition to align them first. The result: A } dark field image that should be as noisy as the film image, with much } less problems due to drift during long exposures, a higher dynamic range } and visible immediately on the viewing screen. } } Michael } } Michael Bode, Ph.D. } Soft Imaging System Corp. } 1675 Carr St., #105N } Lakewood, CO 80215 } =================================== } 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: jim [mailto:jim-at-proscitech.com.au] } Sent: Tuesday, May 30, 2000 7:13 PM } To: 'William Tivol' } Cc: microscopy-at-sparc5.microscopy.com } Subject: RE: Film vs Digital } } } ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } Peter Bond is right in saying "The future is with digital image capture" } if we } were at some future date to count heads of digital versus film TEM } users. } That change will be for reasons of convenience and to save labour. These } are } powerful and valid reasons. } } Bill Tivol has given one set of applications where digital currently } does not } always measure up to film. } Here are another couple of such applications. } 1 When great enlargements are required film is superior. This is } because of } film's greater resolution, but more importantly, because its much, much } easier } to take images at moderate powers and highly enlarge. That way we take } advantage of the TEM's greater depths of focus at low powers and of } film's } higher resolution/ image detail. } 2 Whenever a TEM image is taken at low brightness (to avoid beam } damage or at } very high powers or in dark-field) relatively few electrons form the } image and } make that image grainy. Film is very slow and requires then a longer } exposure, } thus boosting the quantity of electrons used and improving the image. } Digital } is much more sensitive and so the exposure must be shortened. As a } result the } best digital camera will record, quiet faithfully the grainy, unsharp } image. } } Many labs rarely or never use such applications and for them the reasons } to } change to digital now may be overwhelming. } Cheers } Jim Darley } ProSciTech Microscopy PLUS } PO Box 111, Thuringowa QLD 4817 Australia } Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com } Great microscopy catalogue, 500 Links, MSDS, User Notes } www.proscitech.com } } On Wednesday, May 31, 2000 5:18 AM, William Tivol } [SMTP:tivol-at-wadsworth.org] } 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 } } } -----------------------------------------------------------------------. } } } } } } Peter Bond wrote: } } } } } The continuing discussion on whether film is better than digital, } and } } } whether we can directly compare their resolving ability is very } interesting } } } and throwing up some really useful technical stuff. But aren't we } missing } } } the point a bit? Surely the image quality is user defined - if you } or your } } } customer, are satisfied with the end product then the equipment has } done } } } its job. How often does one need to examine a micrograph to assess } its } } } limits? If are getting that close to a picture then you may be } taking } } } things out of context a bit and losing the whole concept. } } } } } } The future is with digital image capture, let's hope the price of } the } } } equipment comes down to something more attainable! } } } } } } } Dear Pete, } } If one is doing quantitative image processing, the better } } resolution } } available from film can be relevant. For example, if one wants to do } } corellation averaging, one needs as many objects in the picture as } possible, } } and also as good resolution as needed--e.g., for 1 nm resolution of } the } } reconstruction, a pixel size of 0.25 nm times the magnification is } necessary, } } so the recording medium must have resolution equal to or better than } that. } } The pixel size of the scanner must also be this small--obviously } irrelevant } } for digital recording. A 5 micrometer scanning-pixel size at 20kx mag } } gives a pixel resolution of 0.25 nm, and for 6.5 by 9 cm film, one } will have } } about 12,000 by 16,000 (taking the header into account) pixels of } useful } } information. This will give a broader area than that available at } equal } } resolution from any CCD chip now out there. I can assure you that I } have } } often had to determine the information limits in a particular } micrograph. } } Yours, } } Bill Tivol } } } } }
The electron energies used in this "intensifier" will be in the order of one or several kV. In TEM however the energies are much higher thus one incident electron will generate for example hundred electron-hole pairs in the CCD thus saturating it very fast. Another factors are damage to the CCD chip and X-Rays.
I was thinking about another approach. A chip consisting of matrix of thermo-sensitive elements. Above each element there will be a metal block with height equal or larger than the stop path for the electron energy used. The whole thing will be cooled in a similar way as the CCDs. When this assembly is exposed to the beam each block will increase its temperature depending on the number of electrons stopped. After the exposure the matrix is scanned and the temperature increase at each element is measured (ofcourse before each exposure a reference image has to be taken).
The benefits: - Very high efficiency. Almost every incident electron will contribute to the image. - Huge dynamic range. - Linearity (after the temperature-signal characteristic of each element has been calibrated) - Narrow point spread function (maybe).
Problems: - Difficult to manufacture (the metal blocks should be insulated from each other) - Maybe low sensitivity. I haven't calculated how much the temperature increase will be (for example 5x5x30um Cu block hit by one 300 kV electron) but I suspect it will be very small. Also It will depend on the sensitivity of the thermo-measuring elements. - Saturation. After each exposure one has to wait some time for the thing to cool down again.
There are maybe other difficulties which do not come in mind now.
Hmmm now I start thinking about X-Rays. Actually the main part of the incident energy will go into X-Rays thus making this device useless.
Rado
--------------------------------------------------------------------- Radostin Danev Laboratory of Ultrastructure Research National Institute for Physiological Sciences Myodaiji-cho, Okazaki 444-8585, JAPAN e-mail: rado-at-nips.ac.jp --------------------------------------------------------------------- ----- Original Message ----- } From: Shane Collins {kshanec-at-gte.net} To: Paul Voyles {voyles-at-research.nj.nec.com} ; {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, June 01, 2000 9:15 AM
AnalySIS has a module which does this see Soft Imaging's web site at http://www.soft-imaging.de Chris
To: Microscopy-at-sparc5.microscopy.com } From: Jim Ferreira {ferreira1-at-llnl.gov}
===================================================================== DR CHRIS JEFFREE BIOSEM - BIOLOGICAL SCIENCES EM FACILITY UNIVERSITY OF EDINBURGH Daniel Rutherford Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JH, Scotland, UK Tel. #44 131 650 5345 FAX. #44 131 650 6563 Mobile 0410 585 401 email c.jeffree-at-ed.ac.uk SEM / TEM bookings sem-at-ed.ac.uk =====================================================================
In a message dated 5/31/00 2:52:24 PM, ferreira1-at-llnl.gov writes:
Anyone aware of software capable of generating depth profiles from digital SEM stereo pair images? I am aware of the Oxford ISIS system/program, but wondered if other companies offer similar software that would run 'stand-alone' in a PC and work with any digital files.
Jim I have been using uMex software for the last 6 months which does exactly what you want. Given a stereo pair it will produce a 3D surface reconstruction. It has a function that enables images to the two images to be manually aligned prior to the calculation. This function when used correctly seems to result in faster calculations.
Another other nice feature is that images can be output in VRML file format. So you can view the images with shareware 3D packages. We use a SGI for viewing as its rendering speed is significantly faster than a PC.
I suggest you check out the following web site. http://www.alicona.com/en/products.htm
Regards Colin MacRae
************************************************************************ Manager of Electron Microscopy Group (Clayton)
I've been following the thread on film vs digital. We all know that the resolving power of digital CCD faceplates is approaching that of conventional film emulsion, but isn't equivalent yet. Can someone remind me of the number of mega-pixels that a CCD will need to equate to ISO 100 print film, ISO 64 or ISO100 slide film and the highest resolving B&W film of all, Technical Pan at ISO 25 and ISO 100? If anyone out there can lead me through the logic and the maths, I'm sure others will also find it helpful. I'll repost a summary of the replies that I get.
Regards, Jeremy Sanderson
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} Hamamatsu recently introduced a new camera called the electron bombardment } ccd where accelerated electrons directly bombard a back thinned, peltier } cooled ccd. The electrons are emitted from a photocathode for applications } particularly in low light microscopy. With a full well capacity of 300,000 } electrons, I wonder if this approach could be used in direct exposure of the } ccd to the electron beam. } Shane
This certainly could be a significant improvement in CCD imaging. Most of the width of the point spread function of current CCD camera is due to photon spread in the scintillator, so presumably removing the scintillator would allow digital images at resolutions very close to the pixel size of the CCD chip.
I'm not familiar with the Hamamatsu camera, but I know that the Gatan camera I currently use has a CCD with a full well capacity of ~500,000 electrons. In order for the Hamamatsu chip to work they would have to find some way to reduce the electron/hole yield of the incident fast electrons - maybe with a very thin CCD chip?
Paul Voyles
} K. Shane Collins } Scientific Instrument Company } 805.444.4953 cell } 310.568.9188 office } 310.568.9189 fax } } } -----Original Message----- } From: Paul Voyles [mailto:voyles-at-research.nj.nec.com] } Sent: Friday, May 26, 2000 9:11 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: RE: new developments in imaging systems? } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} Dear All, } } I've been following the thread on film vs digital. } We all know that the resolving power of digital CCD } faceplates is approaching that of conventional film } emulsion, but isn't equivalent yet. } Can someone remind me of the number of mega-pixels } that a CCD will need to equate to ISO 100 print film, } ISO 64 or ISO100 slide film and the highest resolving } B&W film of all, Technical Pan at ISO 25 and ISO 100? } If anyone out there can lead me through the logic and } the maths, I'm sure others will also find it helpful. } I'll repost a summary of the replies that I get. } } Regards, Jeremy Sanderson }
About twenty-five million pixels in a 35mm Kodachrome slide is the number I have heard from several sources, none of which I can remember now.
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 **********************************************
No, it's not really a problem. It's been done with low density microscopy all the time. Granted, there are some technical aspects to be overcome, but (and I can only speak for ourselves) we have done that on a number of microscopes. You are of course correct, that 10 images at 0.8 seconds take longer than 8 seconds as the image has to be transferred, etc. BUT: that's what beam blankers are for. It is pretty straightforward to take an image at 0.8 seconds, then blank the beam very quickly before taking the next image. That way you get pretty close to the 8 sec total exposure. If there is no beam blanker on the microscope, in most cases it can be added.
I am not sure what you mean by "too sensitive". The cameras are usually constructed so that 1 electron from the beam creates between a few tenth to a few counts (these are all statistical data, of course). The well width divided by this sensitivity then determines, how many primary electrons are needed to fully expose one pixel. For example, if the well width is 50,000 electrons and the sensitivity is 1 count/electron, one needs 50,000 primary electrons to fill the well. This translates into roughly a 0.4% statistical error.
} From a practical standpoint: You can take images with most cameras when the exposure meter on the microscope reads a couple of seconds without overexposing the camera. On the other hand, you can reduce the intensity of the beam until you see single electron events.
The one area where CCD cameras may be too sensitive is diffraction. The normally huge intensity in the transmitted beam often leads to saturation. In CCDs this can lead to blooming (the intensity spills over into neighboring pixels). This can be taken care of with special chips that have anti-blooming features, but this usually has some other drawbacks. Again, this can also be overcome somewhat with multiple exposures. Film behaves more civilized here, as it simply stops responding to the electrons, but this makes film more or less useless for quantitative measurements of diffraction patterns. I have done diffraction with CCDs many times and though it does require some tweaking, one can get very good results from them.
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== 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: jim [mailto:jim-at-proscitech.com.au] Sent: Wednesday, May 31, 2000 9:37 PM To: 'Mike Bode'; 'Microscopy-at-MSA.Microscopy.Com'
Yes, it could be done "in theory". Somebody would need to figure out the
software and perhaps modify the hardware. Then we would find that the total exposure of the specimen to the electron beam maybe a muliple of the film's exposure. Afterall, an 8 sec film exposure would not amount in digital to 10x0.8, but we would require considerable time in between exposures. Since the problems in the discussed circumstances are specimen movement and beam damage, it seems that taking multiple exposures is a poor option.
Digital cameras are for some situation too sensitive to electron exposure. Cutting back on electrons is no option since its the electrons that form the image in the first instance. Much easier in light microscopy . . . insert a neutral density filter. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
Jim- I saw an ad a while back for such software. I have since moved on to other things before I had a chance to obtain the demo software. Below is the contact information I have for the company selling the stereo image analysis software. Since this is an edited version of the message I received several months ago, the terms and conditions may have changed. If you do try this software, I, and I am sure the rest of the listserve would be very interested in hearing how well it works, as I do have an occational need for this capability.
Here is the information (edited) that I received from the company offering the software: now the evaluation version of MeX is available. The test period of MeX is limited to 6 weeks.
The evaluation version is delivered with a small database and the complete analysis tools. You can process your own images without restrictions.
We also offer to preinclude your SEM-images into the database of the evaluation version. You just have to send us an email and we give you detailed information on how you should capture your images.
If you have any questions, please do not hesitate to contact us.
Jim, you may want to check out our web site for the Stereo software. It has some images and examples of stereo evaluations from SEM images. The 'Stereo' part is not a stand-alone software, but can be combines with our analySIS Docu software for a stand-alone application. Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
Hello folks Anyone aware of software capable of generating depth profiles from digital SEM stereo pair images? I am aware of the Oxford ISIS system/program, but wondered if other companies offer similar software that would run 'stand-alone' in a PC and work with any digital files. Any help or suggestions along these lines would be most appreciated. Thanks, Jim
Do not give up if you do not have an ESEM or LVSEM there may be hope yet? Do you have a SEM with a rear manifold that connects directly to the DP and do you have a BSE detector? Sorry but those SEM that have the pump directly attached to the specimen chamber are no good for this procedure. AND I must give all the credit to Viv Robinson who spawned this idea back in the 80's.
If you do have a manifold system you are in luck. Find a rubber bung that will fit into the manifold at the rear of the specimen chamber. Drill (use LN2) a 1/4" hole in the bung and then place it in the rear manifold. Switch off or better still unplug your Everhart-Thornley detector (high voltage plus poor vacuum = arcing!). Place you "wet" specimen in the microscope and pump down. The bung will spoil the vacuum in the chamber for about 20 minutes or so and imaging with the BSE detector you will have your own LV SEM.
To retain the moisture longer you may quench the specimen in LN2 before putting it into the microscope. The frost will sublime away and you will be able to watch what happens to the moisture etc.
We use this technique on all sorts of samples. A rubber bung is cheaper than buying an LVSEM and the results, if you work quickly, are pretty good.
Try it?
Steve Chapman Senior Consultant Protrain For consultancy and professional training in EM world wide Tel 44+ 1280 814774 Fax 814007 www.emcourses.com
Steve Chapman Senior Consultant Protrain For consultancy and professional training in EM world wide Tel 44+ 1280 814774 Fax 814007 www.emcourses.com
Is it just my problem, or isn't this all getting a little bit unfocussed! As far as TEM is concerned, the CCD is not directly exposed to the beam at all, so its effective resolution depends on the nature of the system that presents the image to the CCD. The two predominating technologies for transfer of the image to the CCD in TEM cameras are a fibre-optic linkagage between a phosphor or YAG scintillator and the CCD, or an optical coupling via a lens (for example the excellent f1.2 50mm Zuiko macro lens by Olympus). In both instances, the ultimate resolution of the system is probably set by the electron sensor, which is the phosphor or YAG scintillator. I suspect that fibre optic couplings probably degrade that resolution, but I say that without reference to the facts, so please correct me if I am mistaken. Optical coupling could in principle project the spatial data recorded by the phosphor or YAG to any desired magnification. It can thus be recorded by a CCD using many pixels or few depending on the optical configuration. So what do we mean by resolution in this context, when the smallest object which can be imaged by a TEM can be projected onto any desired quantity of CCD pixels?
To begin to answer Jeremy's question directly, we need to know how much detail a Technical Pan negative can record. The figures depend on processing technique and the test object luminance and contrast, but the modulation transfer function figures published by Kodak indicate that a spatial frequency in excess of 200 cycles per mm is easily recordable. For a test object with contrast 100:1 they quote 320 line pairs per mm. The CCD pixel spacing required to achieve this feat would be 640 pixels per mm. That equates to a requirement for 15360 x 23040 pixels to match the resolving power of a 24x36mm Technical Pan exposure. That's 0.35 Giga pixels in round numbers.
So what was that I heard about the death of silver imaging? I don't think so. Not for a while yet.
Best wishes Chris
} Dear All, } } I've been following the thread on film vs digital. } We all know that the resolving power of digital CCD } faceplates is approaching that of conventional film } emulsion, but isn't equivalent yet. } Can someone remind me of the number of mega-pixels } that a CCD will need to equate to ISO 100 print film, } ISO 64 or ISO100 slide film and the highest resolving } B&W film of all, Technical Pan at ISO 25 and ISO 100? } If anyone out there can lead me through the logic and } the maths, I'm sure others will also find it helpful. } I'll repost a summary of the replies that I get. } } Regards, Jeremy Sanderson } } __________________________________________________ } Do You Yahoo!? } Send instant messages & get email alerts with Yahoo! Messenger. } http://im.yahoo.com/ }
===================================================================== DR CHRIS JEFFREE BIOSEM - BIOLOGICAL SCIENCES EM FACILITY UNIVERSITY OF EDINBURGH Daniel Rutherford Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JH, Scotland, UK Tel. #44 131 650 5345 FAX. #44 131 650 6563 Mobile 0410 585 401 email c.jeffree-at-ed.ac.uk SEM / TEM bookings sem-at-ed.ac.uk =====================================================================
We at Ted Pella Inc. have used our Pelco UVC2 UV Cryo Chamber to cure epoxies but most of our UV curing has been with Acrylic resins. The literature we have on Eponate Araldite does not show it to be UV curable but our chemist wouldn't be surprised if it didn't accelerate the cure. The epoxy mixture should cure overnight at 60 degrees C. We have a technical note on the use of Epon-Araldite for embedding specimens and literature on our Cryo Chamber that may help. I would be happy to fax them to you and/or have you talk with our chemist.
Mark J Armogida VP Engineering and Production Ted Pella Inc.
Tamara Howard 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 } -----------------------------------------------------------------------. } } Greetings! } } Has anyone ever cured Epon-Araldite with UV? I was given a } cobbled-together protocol by a non-microscopist post-doc; the protocol } calls for a dry-ice, UV cure of Epon-Araldite. It says it can take from } 3-7 days. I'm wondering if this is either a resin mix-up *or* the resin } just set up all by itself (over that time frame I've seen it happen), just } because it was a thin layer and left alone - nothing to do with the UV } exposure. She has no ref. for this technique and isn't sure now where she } got it. Sigh. I've only been able to find heat-cure protocols...anyone } have any leads or thoughts on this UV thing? } } (Sorry about the cross-post for those of you on both of these servers) } } Thanks! } } Tamara (Planning to use the oven.....) Howard } CSHL
I am seeking to speak to individuals who know about designing and developing applications for indentification of rare cells in microscopic biological preparations. Someone who knows how to optimize microscopy autofocusing procedures. I am more interested in speaking to an electrical engineer who is more into digital image processing. Can anyone suggest what direction to take?
We are trying to help a client find and identify a bacteriophage. His bacteria (Pasturella sp.) are plated out on agar and show well-defined clear areas with sharp edges where the phage are supposed to be. So far, we have taken carbon-coated grids and placed them gently onto the surface of the clear areas, then lifted them off and negative-stained with PTA or uranyl acetate. We have also run buffer across the clear areas, then pipetted it onto the grids and stained it. A microbiologist who works with phage in another lab has taken samples from the clear areas and concentrated them down and we have stained those also.
So far we have found exactly two phage-like organisms in a total of about 10 grids. Not a stellar performance.
We figure the possibilities are: 1) the bacteria are being killed by something other than phage; 2) we're looking for a particular type of phage that may not be there, and we're just not seeing what's actually causing the clear areas, or 3) for some reason we're just not getting the things adhering to the grids, although we've used these methods successfully many times before.
Does anyone else have any ideas that might help us out, especially on technique? Our client is almost certain that phage are present. We just can't find them.
Thanks in advance.
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/
One of our SEM users would like to label biofilms with lectins. Three of them. At once. I have used gold conjugated to goat anti-biotin to label biotinylated lectins for TEM in the past, so I'm hoping to follow the same kind of procedure. I have not done any immunolabeling for SEM, although our FESEM has been used for such. I would be grateful for any advice! If he wants to triple label, what sizes of gold would be useful? If he wants to quantify the three, what kinds of controls for labeling efficiency should we run? All hints ahd tips gratefully accepted!
Mahalo, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
I have not generally followed any discussion on gold enhancement for light microscopy (photons? I don't do photons), but now I need to ask for someone what people suggest for immunogold enhancement for confocal microscopy. We have 10nm gold left over from TEM, and it would be useful to use it for the confocal experiment. Yes, we're being cheap!
Mahalo, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
} From: "Chris Jeffree" {cjeffree-at-srv0.bio.ed.ac.uk} To: "Jeremy Sanderson" {jb_sanderson-at-yahoo.com} {snip} } To begin to answer Jeremy's question directly, we need to know } how much detail a Technical Pan negative can record. The figures } depend on processing technique and the test object luminance and } contrast, but the modulation transfer function figures published by } Kodak indicate that a spatial frequency in excess of 200 cycles per } mm is easily recordable. For a test object with contrast 100:1 they } quote 320 line pairs per mm. The CCD pixel spacing required to } achieve this feat would be 640 pixels per mm. That equates to a } requirement for 15360 x 23040 pixels to match the resolving power } of a 24x36mm Technical Pan exposure. That's 0.35 Giga pixels in } round numbers. } At 16 bits this is per image this is 5,662,310,400 or 5.5 gig. I don't think film is in any danger for a long time. We need at least 1 order of magnitude for storage and 2 or 3 for processing. I remember taking 4 days to process an image. And then work on the program and trigger it again.
Gordon
Gordon Couger gcouger-at-couger.com
Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
Hello all, This is the last call for a second post-doc vacancy at the University of Barcelona (Spain) to work in the frame of a TMR programme concerning UV coatings. (More details below).
Since we are expecting the Mid-Term evaluation of the project, the starting date has been delayed to next September, so we have extended the deadline for applications.
Any one interested please reply directly to paqui-at-el.ub.es and/or send applications and a CV by mail before 30 June 2000.
Kind regards
F. Peir
************************************************************************** Laboratory: Electronic Materials and Engineering, Department of Electronics, University of Barcelona.
Duration: 12-months, starting September 2000.
Hi Jim, some comments to Your remarks within Your text:
jim schrieb:
} ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } Yes, it could be done "in theory". Somebody would need to figure out the } software and perhaps modify the hardware. Then we would find that the total } exposure of the specimen to the electron beam maybe a muliple of the film's } exposure. Afterall, an 8 sec film exposure would not amount in digital to } 10x0.8, but we would require considerable time in between exposures.
Simply choose a CCD with higher readout performance (faster), but same quality.
} Since the } problems in the discussed circumstances are specimen movement and beam damage, } it seems that taking multiple exposures is a poor option. } } Digital cameras are for some situation too sensitive to electron exposure.
Not correct at all. The only thing is to choose the correct camera for the application You work on. It is not complicated to make a digital system which collects one count per incident electron to achieve the same signal to noise as in the electron beam. This system will be less sensitive than normally sold systems but the main advantage of digital systems that You see what You get remains and You get instant results of Your work. The only problem You have to solve for these systems is to use a not very sensitive scintillator but a very high performance slow-scan CCD. So You get less visible photons from Your electron which have to be converted in one digital count. But these digital counts must be more than presently available to achieve a good statistic. Thats the reason for our new 16bit CCD (dynamic up to 65536 digits) for high performance TEM applications.
} } Cutting back on electrons is no option since its the electrons that form the } image in the first instance. } Much easier in light microscopy . . . insert a neutral density filter. } Cheers } Jim Darley } ProSciTech Microscopy PLUS } PO Box 111, Thuringowa QLD 4817 Australia } Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com } Great microscopy catalogue, 500 Links, MSDS, User Notes } www.proscitech.com
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} About twenty-five million pixels in a 35mm Kodachrome slide is the } number I have heard from several sources, none of which I can remember now. } } Geoff Geoff Analysing this a little further: A 25 Mbyte image = 8.3 M pixels to cover 24x36mm or 864mm^2 this represents 98.2 pixels per mm, or 49 line pairs per mm. Pixel size 10.2 um. Resolution about 50 line pairs per mm at best.
A 25M pixels image used to capture a 24x36mm Kodachrome slide represents 28,935 pixels per mm^2 or 170 pixels per mm. This is equivalent to a maximum resolution of 85 line pairs per mm, which may be on the conservative side for Kodachrome. pixel size = 5.88um. The image will be approx. 6120x4082 pixels, generating a file size of approx 75Mb for a 24-bit (8+8+8bit) RGB image.
To record 120 line pairs per mm, which many top 35mm camera lenses can achieve, a minimum of 240 pixels per mm are required, each 4.2 um wide. This equates to 8640x5760 pixels for a 24x36mm frame = 50M-pixels or 50Mb in 8-bit greyscale, or 150Mb in 24-bit RGB.
At 320 line pairs per mm (Technical Pan) the minimum required 640 pixels per mm is a pixel size of 1.56 um
Presumably for a light image the diffraction limited resolution is approx 1/2 lambda which at 540nm is 0.27um. So looking to the future of ultimate-performance CCDs, direct recording of a diffraction limited light image projected onto the sensor requires at the very least 3703 pixels per image mm or 13,717,421 pixels per mm^2 (greyscale 8-bit)
However, if we are doing light microscopy with an NA 1.4 x100 objective, how much resolving power do we need on CCD or film?
Data is at 0.27um resolution (lambda = 540nm). Let's round this to 0.3um. Magnification at 24x36mm film image is x100, so pixels must be an absolute maximum of 30um wide to record the significant data = 33.3 pixels per mm, equivalent to 800x1200 pixels to record the whole 35mm negative area. However, most CCDs are much smaller than 35 mm frames, typically 1/3 inch. So an 800x1200 pixel CCD at 8x12 mm, 1/3 of the linear dimensions of a 35mm frame would use 9 pixels to record the smallest image details. This is about right from the point of view of resolution, but to record the whole 35mm frame we need about 2400x3600 pixels on our CCD.
Note also that Technical Pan has (depending on the criterion used to assess its performance) up to 10 times the resolving power required to record all there is to see in a diffraction-limited LM image made with a 100x NA 1.4 lens. So you can comfortably afford to use a 60x NA 1.4 lens, thereby getting the same resolution with a bigger field of view.
Many years ago, I took a photograph of a street scene using a Canon 35mm SLR loaded with Kodak Recordak (I think this was a single layer microfilm emulsion). Examined in a light microscope, the image clearly, legibly recorded the brand-name of a child's push chair. I tried to print this brand name using a DeVere point- source enlarger with an image size of 20x30 inches produced with a Schneider Componon lens, but was completely unable to produce a legible image. The point I am making here is that the combination of some high performance films, with high quality lenses of the standard produced by the leading camera manufacturers can record more detail on the film than you can easily get back out by conventional printing. I suspect the same is true of EM exposures.
So there is no contest - film beats CCDs for resolution hands down. And you can process the image on the cheap. No money goes to Intel or Microscoft, Adobe or Epson. But resolution is not primarily what we buy CCDs for. We buy them primarily for instant image capture in a format suitable for digital storage, digital transmission quantitative data recording and image processing.
Chris
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Gordon's and Chris' contribution look very bleak for digital, but the comparison is not really fair, as we should look at the practical aspects too. The unaided eye resolves lines 0.1mm apart, so to see the full (200 black+200 white) 400+ lines/mm that may be recorded on TEM film we would need to enlarge over 40x. This requires an enlarger with a very wide angle lens to print small portions of the negative and it is technically difficult to so enlarge a whole negative since a 4" negative becomes 160" or over 4m in size. That degree of enlargement is not fully useful since in a well exposed TEM negative at just under 30x electron noise becomes the problem, meaning not enough electrons contributed to the image. So enlargements beyond 30x are empty- and provide no further information. Truly not a serious problem. The practical part is that few people ever find it useful to enlarge more than 15x and most TEM images reproduced are barely the size of the original negative, however, they are enlarged from a smaller portion thereof. For these most common applications, digitals with 10+ megabytes provide excellent image details and tonal gradation. However, that size image can only cover the equivalent of a small 35mm negative equivalent and does not allow high enlargement or choosing of an adjacent field. It appears that the best of both worlds is the use of conventional TEM negatives for archiving and scanning those as required for printing.
It should be noted that this discussion concerns TEM. Digitals in SEM are less daunting and it is not problematical to produce excellent digitals, comparable with film. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Friday, June 02, 2000 11:41 AM, Gordon Couger [SMTP:gcouger-at-couger.com] wrote: } } } } From: "Chris Jeffree" {cjeffree-at-srv0.bio.ed.ac.uk} } To: "Jeremy Sanderson" {jb_sanderson-at-yahoo.com} } {snip} } } To begin to answer Jeremy's question directly, we need to know } } how much detail a Technical Pan negative can record. The figures } } depend on processing technique and the test object luminance and } } contrast, but the modulation transfer function figures published by } } Kodak indicate that a spatial frequency in excess of 200 cycles per } } mm is easily recordable. For a test object with contrast 100:1 they } } quote 320 line pairs per mm. The CCD pixel spacing required to } } achieve this feat would be 640 pixels per mm. That equates to a } } requirement for 15360 x 23040 pixels to match the resolving power } } of a 24x36mm Technical Pan exposure. That's 0.35 Giga pixels in } } round numbers. } } } At 16 bits this is per image this is 5,662,310,400 or 5.5 gig. I don't } think film is in any danger for a long time. We need at least 1 order } of magnitude for storage and 2 or 3 for processing. I remember } taking 4 days to process an image. And then work on the program and } trigger it again. } } Gordon } } Gordon Couger gcouger-at-couger.com } } Stillwater, OK www.couger.com/gcouger } 405 624-2855 GMT -6:00
The world is full of possible solutions, but are they practical.? To produce high-resolution, dark-field or any others TEM images that require more electrons to form a clear image, Mike Bode would use multiple digital exposures. The exposures could be layered and combined into one superior image. This image would be made up of more pixel and is formed by more electrons and so would be noise-free and hence could be further enlarged then otherwise possible. Perhaps. Beam blanking would largely save the specimen from beam damage and drift could be compensated for by matching up the digitals. Great. How much time is required between exposures to transfer a minimum 10mb image per exposure? What would be the total time from focusing to the last exposure? What about Z-drift in the interim requiring objective changes and what about the total cost of this additional get-up. The mind boggles at a through focus series. When pushing the limits a piece of film seems more effective, cheaper and fa ster. Again, I don't doubt that there is now a large place for digital in TEM, but its no panacea. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Friday, June 02, 2000 1:24 AM, Mike Bode [SMTP:mb-at-soft-imaging.com] wrote: } } No, it's not really a problem. It's been done with low density } microscopy all the time. Granted, there are some technical aspects to be } overcome, but (and I can only speak for ourselves) we have done that on } a number of microscopes. You are of course correct, that 10 images at } 0.8 seconds take longer than 8 seconds as the image has to be } transferred, etc. BUT: that's what beam blankers are for. It is pretty } straightforward to take an image at 0.8 seconds, then blank the beam } very quickly before taking the next image. That way you get pretty close } to the 8 sec total exposure. If there is no beam blanker on the } microscope, in most cases it can be added. } } I am not sure what you mean by "too sensitive". The cameras are usually } constructed so that 1 electron from the beam creates between a few tenth } to a few counts (these are all statistical data, of course). The well } width divided by this sensitivity then determines, how many primary } electrons are needed to fully expose one pixel. For example, if the well } width is 50,000 electrons and the sensitivity is 1 count/electron, one } needs 50,000 primary electrons to fill the well. This translates into } roughly a 0.4% statistical error. } } } From a practical standpoint: You can take images with most cameras when } the exposure meter on the microscope reads a couple of seconds without } overexposing the camera. On the other hand, you can reduce the intensity } of the beam until you see single electron events. } } The one area where CCD cameras may be too sensitive is diffraction. The } normally huge intensity in the transmitted beam often leads to } saturation. In CCDs this can lead to blooming (the intensity spills over } into neighboring pixels). This can be taken care of with special chips } that have anti-blooming features, but this usually has some other } drawbacks. Again, this can also be overcome somewhat with multiple } exposures. Film behaves more civilized here, as it simply stops } responding to the electrons, but this makes film more or less useless } for quantitative measurements of diffraction patterns. I have done } diffraction with CCDs many times and though it does require some } tweaking, one can get very good results from them. } } Michael Bode, Ph.D. } Soft Imaging System Corp. } 1675 Carr St., #105N } Lakewood, CO 80215 } =================================== } 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: jim [mailto:jim-at-proscitech.com.au] } Sent: Wednesday, May 31, 2000 9:37 PM } To: 'Mike Bode'; 'Microscopy-at-MSA.Microscopy.Com' } Subject: RE: Film vs Digital } } } ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } Yes, it could be done "in theory". Somebody would need to figure out the } } software and perhaps modify the hardware. Then we would find that the } total } exposure of the specimen to the electron beam maybe a muliple of the } film's } exposure. Afterall, an 8 sec film exposure would not amount in digital } to } 10x0.8, but we would require considerable time in between exposures. } Since the } problems in the discussed circumstances are specimen movement and beam } damage, } it seems that taking multiple exposures is a poor option. } } Digital cameras are for some situation too sensitive to electron } exposure. } Cutting back on electrons is no option since its the electrons that form } the } image in the first instance. } Much easier in light microscopy . . . insert a neutral density filter. } Cheers } Jim Darley } ProSciTech Microscopy PLUS } PO Box 111, Thuringowa QLD 4817 Australia } Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com } Great microscopy catalogue, 500 Links, MSDS, User Notes } www.proscitech.com }
if possible, select plates obtained from serial dilutions which show confluent lysis (i.e., where plaques touch each other). They are a good source for obtaining high-titer bacteriophage lysates (i.g., 10E10 / 10E11 plaque-forming units per ml). Harvest the phage by scraping the top agar and transfer it into a test tube or equivalent. Rinse the plates with some ml of phage diluent (i.g., 5 ml). Shake this supension containing phage, host cells and agar for some while before spinning down the cells and the agar. It is also a good idea to pick up a single plaque. Resuspend it in a small volume of broth, and prepare a fresh lysate in some ml of broth with fresh host cells. For rapid screening, we sometimes pipette a drop of } buffer onto a plaque and float a piece of carbon film into the drop directly from a mica support. After some minutes, we pick up the film with a grid and do the routine negative staining. But you are right, the number of phage particles is low in this case. Best regards Horst Neve
} At 15:14 01.06.00 -0500, you wrote: } Hi, } } We are trying to help a client find and identify a bacteriophage. His } bacteria (Pasturella sp.) are plated out on agar and show well-defined clear } areas with sharp edges where the phage are supposed to be. So far, we have } taken carbon-coated grids and placed them gently onto the surface of the } clear areas, then lifted them off and negative-stained with PTA or uranyl } acetate. We have also run buffer across the clear areas, then pipetted it } onto the grids and stained it. A microbiologist who works with phage in } another lab has taken samples from the clear areas and concentrated them } down and we have stained those also. } } So far we have found exactly two phage-like organisms in a total of about 10 } grids. Not a stellar performance. } } Thanks in advance. } } Randy } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility } W122 Veterinary Medicine } University of Missouri } Columbia, MO 65211
**************************************************************************** **** Dr. Horst Neve Institut fuer Mikrobiologie / Institute for Microbiology Bundesanstalt fuer Milchforschung / Federal Dairy Research Centre Postfach / P.O. Box 6069, D-24121 Kiel Hermann-Weigmann-Str. 1, D-24103 Kiel **************************************************************************** **** Tel. / Phone: +49 (0) 431 609 2343 {} Fax: +49 (0) 431 609 2306 E-mail: neve-at-bafm.de {} Internet: http://www.bafm.de **************************************************************************** ****
if possible, select plates obtained from serial dilutions which show confluent lysis (i.e., where plaques touch each other). They are a good source for obtaining high-titer bacteriophage lysates (i.g., 10E10 / 10E11 plaque-forming units per ml). Harvest the phage by scraping the top agar and transfer it into a test tube or equivalent. Rinse the plates with some ml of phage diluent (i.g., 5 ml). Shake this supension containing phage, host cells and agar for some while before spinning down the cells and the agar. It is also a good idea to pick up a single plaque. Resuspend it in a small volume of broth, and prepare a fresh lysate in some ml of broth with fresh host cells. For rapid screening, we sometimes pipette a drop of } buffer onto a plaque and float a piece of carbon film into the drop directly from a mica support. After some minutes, we pick up the film with a grid and do the routine negative staining. But you are right, the number of phage particles is low in this case. Best regards Horst Neve
} At 15:14 01.06.00 -0500, you wrote: } Hi, } } We are trying to help a client find and identify a bacteriophage. His } bacteria (Pasturella sp.) are plated out on agar and show well-defined clear } areas with sharp edges where the phage are supposed to be. So far, we have } taken carbon-coated grids and placed them gently onto the surface of the } clear areas, then lifted them off and negative-stained with PTA or uranyl } acetate. We have also run buffer across the clear areas, then pipetted it } onto the grids and stained it. A microbiologist who works with phage in } another lab has taken samples from the clear areas and concentrated them } down and we have stained those also. } } So far we have found exactly two phage-like organisms in a total of about 10 } grids. Not a stellar performance. } } Thanks in advance. } } Randy } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility } W122 Veterinary Medicine } University of Missouri } Columbia, MO 65211
**************************************************************************** **** Dr. Horst Neve Institut fuer Mikrobiologie / Institute for Microbiology Bundesanstalt fuer Milchforschung / Federal Dairy Research Centre Postfach / P.O. Box 6069, D-24121 Kiel Hermann-Weigmann-Str. 1, D-24103 Kiel **************************************************************************** **** Tel. / Phone: +49 (0) 431 609 2343 {} Fax: +49 (0) 431 609 2306 E-mail: neve-at-bafm.de {} Internet: http://www.bafm.de **************************************************************************** ****
I'm looking for a (preferably) bench top incubator. Non-water jacketed, not using B.O.D. bottles, unit needs to be using CFC free refrigeration system. Does anyone know of such a unit or where one can be purchased? Need to order one ASAP. We are a small research lab and the one we have is a monster (48"Hx46"Wx28"deep and weighs almost 500lbs.). I'd appreciate any info.
The Microscopy & Microanalysis 2000 Local Arrangements Committee is pleased to announce an additional social event at M&M2000---
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Date sent: Fri, 02 Jun 2000 11:03:03 -0400 To: c.jeffree-at-ed.ac.uk } From: joe fu {jofu-at-nist.gov}
I have a researcher here at USU who would like to have some freeze fracture preps made of lysosome. Is there anyone out there who is currently doing FF preps and would be willing to assist us? I can image the preps here in Logan.
William McManus Supervisor Electron Microscopy Facility Department of Biology Utah State University Logan UT 84322-5305
Having worked with a variety of phages (Candida, Streptococcus, E. coli) I can tell you that finding phages from an agar plaque is very difficult--as you have determined. The best way is to do a liquid culture and then high speed followed by ultracentrifugation to concentrate the particles. FYI, as I recall, on a 200 mesh grid each virus particle is roughly equivalent to 3.4 x 10E6 vp/ml. So, you need a lot of particles to even find one of them using this approach.
JB #################################################################### John J. Bozzola, Ph.D., Director Micro-Imaging and Analysis Center 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 ####################################################################
My EM lab is going to be moved and I have just seen the plans. Apparently two large equipment room are going to be built, one right across the hall and the other two doors down. I have been told that the equipment room will house -80 freezers, centrifuges, etc. Does anyone out there have experience with this type of equipment near their TEM? I will not be able to test the room before the move because nothing has been built yet and we all scheduled to move in at the same time. I am hoping to have some influence on the architects now. They have been told (and I shall keep reminding them) that no electrical circuits are to be passed around the EM room.
Thanks.
Ruth
********************************************** Ruth Yamawaki Department of Comparative Medicine Building 330, Quad 7, RAF-1 Stanford CA 94305 (650) 723-3457 **********************************************
Mike Bode would use multiple digital exposures. The exposures could be layered and combined into one superior image. This image would be made up of more pixel and is formed by more electrons and so would be noise-free and hence could be further enlarged then otherwise possible. Perhaps.
No, I did not talk about further enlargements. All I wanted to say is, that a more noise-free image can be achieved by adding multiple images, and that this also to some extent helps with drift of the sample during acquisition.
Jim wrote:
How much time is required between exposures to transfer a minimum 10mb image per exposure?
How did you arrive at 10 MB? A 1280x1024 image with 16 bit pixel information is about 2.5 MB (uncompressed). We acquire about 10 of those per second and transfer them across the PC bus to the display. Putting them on them into Memory might add a few tenth of a second. Writing to HD can be done after all images are acquired.
Jim wrote:
What would be the total time from focusing to the last exposure? What about Z-drift in the interim requiring objective changes
Why would we have to worry about that, if we don't have to worry about that when taking the image on film? In fact, we could take care of this by looking at the image between exposures and correct for z-drift. However, as you said, that would add to the overall time and exposure. I was comparing a normal dark field image taken on film at 8 seconds with acquiring the same image on a "too sensitive" CCD camera by adding up 10 consecutive .8 second images. Why would the sample drift (in x, y or z) substantially more in 8+delta seconds than in 8?
Jim wrote:
what about the total cost of this additional get-up
That of course depends on the microscope and there is no general answer. For example on a LEO 912 I believe the blanker is standard. The additional cost to use an acquisition scheme like this with our software is $0 plus perhaps a bit of time to write a small macro. On other microscopes one might have to add a beam blanker and perhaps a control mechanism for the beam blanker. But I would guess, that this cost is not very high. All modern microscopes are computer controller anyway, so it is most likely just a control command that needs to be sent to the microscope over a serial port if the beam blanker is installed. Piece of cake.
Jim wrote:
The mind boggles at a through focus series.
You're right here. But I don't think we were talking about through-focus series. Incidentally, we do through-focus series on light microscopes and reconstruction routinely. Takes a few images at different focus (or for a light microscope: stage) settings. The rest is done off-line. Takes maybe a couple of minutes for about 20 images of about 1kx1k. I agree that TEM is different here and much more complicated due to the complicated Contrast Transfer Function. However, this could in principle be sorted out.
Jim wrote:
Again, I don't doubt that there is now a large place for digital in TEM, but its no panacea.
I also agree with you on that one. But using the additional computer possibilities of digital imaging might take you further than expected.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== 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: jim [mailto:jim-at-proscitech.com.au] Sent: Friday, June 02, 2000 5:15 AM To: 'Mike Bode'; 'Microscopy-at-MSA.Microscopy.Com' Cc: 'jim-at-proscitech.com.au'
This can be hard to predict. Once we were having trouble with rather huge (100 eV) energy fluctuations in our GIF 200 energy filter. We traced the source to an adjoining room filled with constant-temperature ovens, fans, and other high-current equipment. The unlikely source finally turned out to be a $50 hot plate-stirrer!
Keeping the AC circuitry from running near the lab is a very good start- I know this has devastated other labs. Ask for your own independent electrical ground for the 'scope, and that all electrical circuits to your lab remain independent of other rooms. I doubt that the equipment you describe will be a big problem as long as you don't share AC circuits, ground, or a common wall. Good luck.
"The chief source of problems is solutions." -Eric Sevareid
........................................................................... ...................................................... Jeffrey A. Fortner Argonne National Laboratory CMT/205 9700 S. Cass Avenue Argonne, IL 60439-4837
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} ---------- } From: Ruth Yamawaki } Sent: Friday, June 2, 2000 11:48 AM } To: 'Microscopy-at-sparc5.microscopy.com' } Subject: Outside electrical near the EM lab } } ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } My EM lab is going to be moved and I have just seen the plans. Apparently } two large equipment room are going to be built, one right across the hall } and the other two doors down. I have been told that the equipment room } will } house -80 freezers, centrifuges, etc. Does anyone out there have } experience } with this type of equipment near their TEM? I will not be able to test } the } room before the move because nothing has been built yet and we all } scheduled } to move in at the same time. I am hoping to have some influence on the } architects now. They have been told (and I shall keep reminding them) } that } no electrical circuits are to be passed around the EM room. } } Thanks. } } Ruth } } ********************************************** } Ruth Yamawaki } Department of Comparative Medicine } Building 330, Quad 7, RAF-1 } Stanford CA 94305 } (650) 723-3457 } ********************************************** } }
There is a standard technique for phage isolation and purification in Maniatis. I have found it to work better than any kit and can be modified accordingly.
Horst Neve 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 } -----------------------------------------------------------------------. } } Randy, } } if possible, select plates obtained from serial dilutions which show } confluent lysis (i.e., where plaques touch each other). They are a good } source for obtaining high-titer bacteriophage lysates (i.g., 10E10 / 10E11 } plaque-forming units per ml). Harvest the phage by scraping the top agar } and transfer it into a test tube or equivalent. Rinse the plates with some } ml of phage diluent (i.g., 5 ml). Shake this supension containing phage, } host cells and agar for some while before spinning down the cells and the } agar. It is also a good idea to pick up a single plaque. Resuspend it in a } small volume of broth, and prepare a fresh lysate in some ml of broth with } fresh host cells. For rapid screening, we sometimes pipette a drop of } } buffer onto a plaque and float a piece of carbon film into the drop } directly from a mica support. After some minutes, we pick up the film with } a grid and do the routine negative staining. But you are right, the number } of phage particles is low in this case. } Best regards } Horst Neve } } } At 15:14 01.06.00 -0500, you wrote: } } Hi, } } } } We are trying to help a client find and identify a bacteriophage. His } } bacteria (Pasturella sp.) are plated out on agar and show well-defined clear } } areas with sharp edges where the phage are supposed to be. So far, we have } } taken carbon-coated grids and placed them gently onto the surface of the } } clear areas, then lifted them off and negative-stained with PTA or uranyl } } acetate. We have also run buffer across the clear areas, then pipetted it } } onto the grids and stained it. A microbiologist who works with phage in } } another lab has taken samples from the clear areas and concentrated them } } down and we have stained those also. } } } } So far we have found exactly two phage-like organisms in a total of about 10 } } grids. Not a stellar performance. } } } } Thanks in advance. } } } } Randy } } } } Randy Tindall } } EM Specialist } } Electron Microscopy Core Facility } } W122 Veterinary Medicine } } University of Missouri } } Columbia, MO 65211 } } **************************************************************************** } **** } Dr. Horst Neve } Institut fuer Mikrobiologie / Institute for Microbiology } Bundesanstalt fuer Milchforschung / Federal Dairy Research Centre } Postfach / P.O. Box 6069, D-24121 Kiel } Hermann-Weigmann-Str. 1, D-24103 Kiel } **************************************************************************** } **** } Tel. / Phone: +49 (0) 431 609 2343 {} Fax: +49 (0) 431 609 2306 } E-mail: neve-at-bafm.de {} Internet: http://www.bafm.de } **************************************************************************** } ****
I spent about two years trying to make good pictures of my NanoGold labeled protein-DNA complexes. Doing this job I find two main problems: the sample is unstable under the beam as any biological sample; NanoGold is much more bright than protein core in dark-field. I find that it is impossible to record equally perfect signals from NanoGold and protein core because of short dynamic range for SO-163 film, I believe. I was trying to make two pictures with different exposure, but it is tricky: in dark-field mode the automatic exposure meter usually does not work and we have to set exposure manually, in this case it is difficult to get "right" exposure time in the right moment, you know. Again, because of sample's short life under the beam, it is impossible to make a couple pictures at the different conditions sometime. Keep in mind, please, that to change the film in the microscope it takes about 10 seconds. Your idea about increasing signal-noise ratio by collecting more electrons is bright but not practical. For biological samples (I am talking about non-fixed, non-stained samples of proteins, DNA or RNA-protein complexes etc) the electron damage is a huge problem. People are trying to solve it in different ways. Some using cryo temperature (to stabilize the biological structure). I was using freeze-drying (I find that freeze-dried samples are more stable under the beam). But in any case we have deal with very unstable samples and must to do everything to decrease (not increase as you recommended) electron dose. Drift is a second big problem for such application: to increase signal-noise ratio we have to use very thin support films. Images obtained at such conditions are noisy and in most cases we have to use image analysis tools to extract the data. It means that we have to digitize our images anyway. In such situation digital camera may help. As you, probably, remember I was a person who initiates this discussion. I think this discussion was very useful for many of us who are not friendly with digital camera's techniques. We understand the limitations of the modern digital cameras better now. I would like to say thank you everybody who was involved in this discussion. There is some conclusions I make for myself from discussion:
- Film is still cheap and universal material for recording and storage EM images, sorry CCD. - CCD TEM camera should not substitute film. Film and camera should work all together improving the flexibility of the TEM system. For this reason I will chose side-mount camera if will have money for it. - For cell-biology (thin sections) where the resolution of the sample is about 3 nm CCD camera may do a good job allowing users to make a huge number of pictures (cell-biology guys love it), instantly view and catalogize them. - Sometime the digital camera may help in area of high-resolution (relatively high, guys) EM when image will be digitized anyway. The major limitation here is small area of view (we need a lot of particles for image analysis sometime), but you could make the set of overlapping pictures and digitally combine it. I love, also, Mike Bode idea to make a few very short exposure pictures and combine it digitally later to reduce noise. The relatively big size of CCD's pixels is a real problem too. - CCD camera is expensive "toy". I am not sure that the benefits from using it will compensate astronomical price, actually the third of the electron microscope value ($70000 is it 1/3 of microscope's price on current market?). Currently, I would recognize the CCD TEM camera as funny "attachment" which may be useful if you rich enough to spend money on it (it mean, that you have everything else in your EM lab plus some extra $70000 for the fun playing with digital "toy"). - TEM CCD cameras are under extensive development now. Today's camera will be replaced on the new model (read better, faster, what else?) next year. Next year's camera will be easily forgotten next after the next year and so on... Each new camera will be better that previous one... CCD TEM camera it is not a good investment of money, I think. - We should keep in mind that many companies charged extra 3-4K$ for the installation and training (it is mandatory for Gatan for instance) and you, probably, have to buy service contract on it even if you have service contract on the microscope (JEOL's service contract on microscope do not cover the CCD camera even if you buy it from JEOL).
Best regard, Sergey
} Date: Fri, 02 Jun 2000 21:14:44 +1000 } From: jim {jim-at-proscitech.com.au} } Subject: RE: Film vs Digital } To: 'Mike Bode' {mb-at-Soft-Imaging.com} , } "'Microscopy-at-MSA.Microscopy.Com'" {Microscopy-at-sparc5.microscopy.com} } Cc: "'jim-at-proscitech.com.au'" {jim-at-proscitech.com.au} } Reply-to: "jim-at-proscitech.com.au" {jim-at-proscitech.com.au} } Organization: ProSciTech } X-Mailer: Microsoft Internet E-mail/MAPI - 8.0.0.4211 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
I agree with most of your statements, and I don't think that anybody would argue the point, that a raw CCD chip has a better resolution that film. As you pointed out, film can have a very small grain size ( {1 micron) and CCD chips usually have a few microns pixel size.
But that is not the end of the story. An optical system normally consists of more than a chip or a sheet of film. The question is, can I get the resolution I want or need. And here the situation is not as simple. For example: I used to do high-resolution TEM. What you do there is operate the microscope at optimum condition, then take a picture (on film). You then go to the darkroom and develop prints by blowing up the negative 10, 20 or even more times. When you then look at the images, you can usually see the grains of the film (especially if you then scan those into a copmputer). So, we are working at the resolution limit of the film, and according to your postings, we should not be able to see anything on a CCD. But that's not true. By using some geometrical properties (the camera sits further down in the column and sees an already enlarged image) and a tapered fiber-optic, you can acquire just as good and better images of the same structure. Both images are limited by the point resolution of the TEM and not by the Film or CCD resolution.
What I am trying to say, and what I have said before is, that film is definitely better when it comes to maximizing the product of resolution AND field of view. However, if we can trade one for the other, I believe in most cases you get better results from a CCD.
Having said that and looking at a print of Ansel Adams, I am glad there is film, though!!
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Chris Jeffree [mailto:cjeffree-at-srv0.bio.ed.ac.uk] Sent: Friday, June 02, 2000 10:16 AM To: joe fu Cc: microscopy-at-sparc5.microscopy.com
Date sent: Fri, 02 Jun 2000 11:03:03 -0400 To: c.jeffree-at-ed.ac.uk } From: joe fu {jofu-at-nist.gov}
Conventional wisdom said that when it came to using TEM for biological specimens (or beam sensitive specimens) it is always best to use a lower accelerating voltage. Many lower voltage microscopes (V { 120 kV) were sold on this assumption. However is this assumption true? At the present there are many research establishments, buying TEMs for biological use, who are using 200 to 300 kV beams. So obviously there has been a shift in the conventional way of thinking. Being materials based I am not sure what the status quo is in biological TEM. I know that the ratio of inelastic to elastic scattering cross sections is greater than one for the elements Z {12, but how does this change as the beam energy increases? What are your experiences? This is really just academic curiosity by the way, but I am sure many of you would appreciate the question. ******************************************************** Dr Jonathan Barnard
Analytical Materials Physics The Angstrom Laboratory, Uppsala University P O Box 534, SE-751 21 Uppsala, Sweden Phone: +46-(0)18-4716838 Fax: +46-(0)18-500131 Phone: Microscope room +46 18 471 6365 http://www.angstrom.uu.se/analytical/home.html ********************************************************
} } - CCD camera is expensive "toy". I am not sure that the } } benefits from using it will compensate astronomical price, actually } } the third of the electron microscope value ($70000 is it 1/3 of } } microscope's price on current market?). Currently, I would } } recognize the CCD TEM camera as funny "attachment" which may be } } useful if you rich enough to spend money on it (it mean, that you } } have everything else in your EM lab plus some extra $70000 for the } } fun playing with digital "toy"). - TEM CCD cameras are under extensive development now. Today's camera will be replaced on the new model (read better, faster, what else?) next year. Next year's camera will be easily forgotten next after the next year and so on... Each new camera will be better that previous one... CCD TEM camera it is not a good investment of money, I think. { {
Sergey:
I appreciate your post, but...
1. Remember that "time is money"...there is no question that there is a value in the nearly instantaneous result that derives from the use of digital imaging systems, particular on TEMs. Also, the ability to rapidly process and analyze your images to determine if they are "keepers" is priceless, IMO.
2. In our large, national multi-user facility we have been digital-only since about 1993 or so. We have no darkrooms for plate loading or enlarging. The only "chemicals" we handle in the entire photographic process are toner cartridges for our laser printers, or equivalent media for dye sub printers etc. In most instances, we process our images electronically all the way through to the final presentation. This includes preparation of PowerPoint slides for digital projection for talks, to sending full papers out for publication on disks or via e-mail attachments. No user has *ever* complained about the non-availability of film, or about the "lack of resolution of CCD images" compared to film. On the contrary, we have users that travel to our laboratory specifically to do work on our instruments because of the availability of digital imaging, when the same instruments in their own laboratories are not equipped with digital cameras.
3. Digital camera systems also provide the capability to conduct research with outside colleagues live-time via Telepresence Microscopy methods. This is a rapidly developing capability in our field that just cannot be done (on a TEM at least) if your microsocope only uses film for imaging.
4. The 1k x 1k Gatan CCD camera on our Hitachi HF-2000 was purchased for about $100K in 1993, and was upgraded about 5 years ago to a multi-scan capability. It is still functioning perfectly today, as is a similar camera on our JEOL 4000EX TEM. The very newest CCD cameras might offer faster read-out times, but there has been no order-of-magnitude improvement in capabilities to make our older camera so obsolete that we desire to purchase a new one. The point is that, unlike desk-top computers, the *expensive* digital camera you purchase today will definitely *not* be obsolete next year...
5. A CCD TEM camera is probably the *best* investment anyone could make to advance the research throughput in their microscopy facility.
All just MHO...
Larry
Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
} From: "Mike Bode" {mb-at-Soft-Imaging.com} To: "'Microscopy-at-MSA.Microscopy.Com'" {Microscopy-at-sparc5.microscopy.com} } Chris, } } I agree with most of your statements, and I don't think that anybody } would argue the point, that a raw CCD chip has a better resolution that } film. As you pointed out, film can have a very small grain size ( {1 } micron) and CCD chips usually have a few microns pixel size. } } But that is not the end of the story. An optical system normally } consists of more than a chip or a sheet of film. The question is, can I } get the resolution I want or need. And here the situation is not as } simple. For example: I used to do high-resolution TEM. What you do there } is operate the microscope at optimum condition, then take a picture (on } film). You then go to the darkroom and develop prints by blowing up the } negative 10, 20 or even more times. When you then look at the images, } you can usually see the grains of the film (especially if you then scan } those into a computer). So, we are working at the resolution limit of } the film, and according to your postings, we should not be able to see } anything on a CCD. But that's not true. By using some geometrical } properties (the camera sits further down in the column and sees an } already enlarged image) and a tapered fiber-optic, you can acquire just } as good and better images of the same structure. Both images are limited } by the point resolution of the TEM and not by the Film or CCD } resolution.c
For a scanning device be it light, electrons, x-rays or gamma rays is a CCD array the best way to capture the image. My experience has been with gamma rays and to some extent x rays. We have not found a good way to focus gamma rays so we use a crystal that emitted light and a photomultiplier tube and we got good images. The resolution depended on the aperture of the gamma ray source and was pretty large.
If you are scanning a sample wiht an electron beam the same principle should work. You would not need long dwell times but build the images out of multiple scans.
If you are scanning the sample the array of pixels seems redundant to me. Also a photomultiplier tube and crystal are a great deal more sensitive than CCD arrays and have a good deal more dynamic range.
The time to make a film image is allows going to be less than making a digital image. The ability to immediately see the digital image is a very handy thing. There are ways to develop B&W film that you can see your image in 5 minutes. For 4 X 5 images I am using BZT tubes that are tubs with a inter circumference of a little over 4 inches. You put the file and developer in the tube and put the tub in a pan of water and spin the tube in the water. The stop bath and fixing can be carried out in room light and it results in the most even development I have ever seen.
For archival storage I don't think 35 mm film can be beat for cost and resolution. Unfortunately you can't see the results until later occasional making it necessary to reshoot the session if you rely on film alone.
If you really need long term archival storage you might consider using a slide printer for a computer to print the digital images. We know the life of silver film is longer then 100 years. For the LM folks that are using color images you could use PhotoShop to do a color separation and print out three negatives on black and white for storage.
Hard drives And CD ROMS have come a long way but I loose about 20% of my hard drive storage a year and loose an occasional CDROM to scratches. Film would survive the scratches with a minimal loss of information insted loosing the whole CDROM.
Gordon
Gordon Couger gcouger-at-couger.com
Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
Please be aware that this is a commercial post that may be of interest to some of the list members.
Digital light microscope cameras are now available that use a highly touted CCD detector: Color or monochrome; fast interline transfer, high sensitivity, low noise; 6.7 micron square pixel; 2/3 inch 1.3 Million chip; PCI bus frame transfer. This detector is ideal to use in light microscopy applications. Now precision technology and fast high capacity cheap computers allow this detector to be "Micro Stepped" providing digital images of up to 12 million pixels to be captured very quickly and providing file sizes of up to 35 MB in 24 bit color images. "Inter Pixel Stepping" technology will allow a considerably less expensive approach to digital imaging that can now approach film resolving capability in a light microscope.
Hats of to Dr. Jeffree's explanation on optical resolution as it relates to light microscopy applications. With high pixel density capable digital cameras, now lower magnification, low NA (lower resolving power) objectives can be used to acquire matching digital resolution images of wide fields of view.
Information on Nikon's Instrument Division "Digital Eclipse" family of digital cameras for microscopy including the new DXM1200 digital color camera will soon be available on our web site www.nikonusa.com or you can go to our Dealer Locator at http://www.nikonusa.com/corpinfo/dealers/dealerSearch.cfm and ask your local Nikon authorized microscope dealer for further information.
Best Regards,
Stan Schwartz Manager, BioSciences Dept. Nikon Inc Instrument Division 1300 Walt Whitman Rd. Melville, NY 11747 631-547-8500 631-547-4033 Fax Schwartz-at-nikonincmail.com www.nikonusa.com
Earlier post ============================================================= Geoff Analysing this a little further: A 25 Mbyte image = 8.3 M pixels to cover 24x36mm or 864mm^2 this represents 98.2 pixels per mm, or 49 line pairs per mm. Pixel size 10.2 um. Resolution about 50 line pairs per mm at best.
A 25M pixels image used to capture a 24x36mm Kodachrome slide represents 28,935 pixels per mm^2 or 170 pixels per mm. This is equivalent to a maximum resolution of 85 line pairs per mm, which may be on the conservative side for Kodachrome. pixel size = 5.88um. The image will be approx. 6120x4082 pixels, generating a file size of approx 75Mb for a 24-bit (8+8+8bit) RGB image.
To record 120 line pairs per mm, which many top 35mm camera lenses can achieve, a minimum of 240 pixels per mm are required, each 4.2 um wide. This equates to 8640x5760 pixels for a 24x36mm frame = 50M-pixels or 50Mb in 8-bit greyscale, or 150Mb in 24-bit RGB.
At 320 line pairs per mm (Technical Pan) the minimum required 640 pixels per mm is a pixel size of 1.56 um
Presumably for a light image the diffraction limited resolution is approx 1/2 lambda which at 540nm is 0.27um. So looking to the future of ultimate-performance CCDs, direct recording of a diffraction limited light image projected onto the sensor requires at the very least 3703 pixels per image mm or 13,717,421 pixels per mm^2 (greyscale 8-bit)
However, if we are doing light microscopy with an NA 1.4 x100 objective, how much resolving power do we need on CCD or film?
Data is at 0.27um resolution (lambda = 540nm). Let's round this to 0.3um. Magnification at 24x36mm film image is x100, so pixels must be an absolute maximum of 30um wide to record the significant data = 33.3 pixels per mm, equivalent to 800x1200 pixels to record the whole 35mm negative area. However, most CCDs are much smaller than 35 mm frames, typically 1/3 inch. So an 800x1200 pixel CCD at 8x12 mm, 1/3 of the linear dimensions of a 35mm frame would use 9 pixels to record the smallest image details. This is about right from the point of view of resolution, but to record the whole 35mm frame we need about 2400x3600 pixels on our CCD.
Note also that Technical Pan has (depending on the criterion used to assess its performance) up to 10 times the resolving power required to record all there is to see in a diffraction-limited LM image made with a 100x NA 1.4 lens. So you can comfortably afford to use a 60x NA 1.4 lens, thereby getting the same resolution with a bigger field of view.
Many years ago, I took a photograph of a street scene using a Canon 35mm SLR loaded with Kodak Recordak (I think this was a single layer microfilm emulsion). Examined in a light microscope, the image clearly, legibly recorded the brand-name of a child's push chair. I tried to print this brand name using a DeVere point- source enlarger with an image size of 20x30 inches produced with a Schneider Componon lens, but was completely unable to produce a legible image. The point I am making here is that the combination of some high performance films, with high quality lenses of the standard produced by the leading camera manufacturers can record more detail on the film than you can easily get back out by conventional printing. I suspect the same is true of EM exposures.
So there is no contest - film beats CCDs for resolution hands down. And you can process the image on the cheap. No money goes to Intel or Microscoft, Adobe or Epson. But resolution is not primarily what we buy CCDs for. We buy them primarily for instant image capture in a format suitable for digital storage, digital transmission quantitative data recording and image processing.
Chris
===================================================================== DR CHRIS JEFFREE BIOSEM - BIOLOGICAL SCIENCES EM FACILITY UNIVERSITY OF EDINBURGH Daniel Rutherford Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JH, Scotland, UK Tel. #44 131 650 5345 FAX. #44 131 650 6563 Mobile 0410 585 401 email c.jeffree-at-ed.ac.uk SEM / TEM bookings sem-at-ed.ac.uk =====================================================================
I work with EPON sections of biological specimens. It seems clear to me that 60KV introduces more dammage than 80Kv. The sections are more unstable and tend to break more easily at the lower voltage.
Dr. A.P. Alves de Matos Pathology Department Curry Cabral Hospital Lisbon
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Conventional wisdom said that when it came to using TEM for biological specimens (or beam sensitive specimens) it is always best to use a lower accelerating voltage. Many lower voltage microscopes (V { 120 kV) were sold on this assumption. However is this assumption true? At the present there are many research establishments, buying TEMs for biological use, who are using 200 to 300 kV beams. So obviously there has been a shift in the conventional way of thinking. Being materials based I am not sure what the status quo is in biological TEM. I know that the ratio of inelastic to elastic scattering cross sections is greater than one for the elements Z {12, but how does this change as the beam energy increases? What are your experiences? This is really just academic curiosity by the way, but I am sure many of you would appreciate the question. ******************************************************** Dr Jonathan Barnard
Analytical Materials Physics The Angstrom Laboratory, Uppsala University P O Box 534, SE-751 21 Uppsala, Sweden Phone: +46-(0)18-4716838 Fax: +46-(0)18-500131 Phone: Microscope room +46 18 471 6365 http://www.angstrom.uu.se/analytical/home.html ********************************************************
At 02:21 PM 6/3/00, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I hope not.
} Digital light microscope cameras are now available that use a highly touted } CCD detector: Color or monochrome; fast interline transfer, high sensitivity, } low noise; 6.7 micron square pixel; 2/3 inch 1.3 Million chip; PCI bus frame } transfer. This detector is ideal to use in light microscopy applications. } Now precision technology and fast high capacity cheap computers allow this } detector to be "Micro Stepped" providing digital images of up to 12 million } pixels to be captured very quickly and providing file sizes of up to 35 MB in } 24 bit color images. "Inter Pixel Stepping" technology will allow a } considerably less expensive approach to digital imaging that can now approach } film resolving capability in a light microscope. } } Hats of to Dr. Jeffree's explanation on optical resolution as it relates to } light microscopy applications. With high pixel density capable digital } cameras, now lower magnification, low NA (lower resolving power) objectives } can be used to acquire matching digital resolution images of wide fields of } view. } } Information on Nikon's Instrument Division "Digital Eclipse" family of } digital cameras for microscopy including the new DXM1200 digital color camera } will soon be available on our web site www.nikonusa.com or you can go to } our Dealer Locator at } http://www.nikonusa.com/corpinfo/dealers/dealerSearch.cfm and ask your } local Nikon authorized microscope dealer for further information. } } Best Regards, } } Stan Schwartz } Manager, BioSciences Dept. } Nikon Inc Instrument Division } 1300 Walt Whitman Rd. } Melville, NY 11747 } 631-547-8500 } 631-547-4033 Fax } Schwartz-at-nikonincmail.com } www.nikonusa.com
As a long term past user of Nikon equipment, it is a tale of sorrow. Most recently, the digital E1 and E2 are dismal failures. Consumers are sending back 990's in droves (see rec.photo.marketplace.digital). I really think that Nikon blew it in regards to digicams. How Nikon can claim to get a 1.3M pixel imager to produce realistic 12M pixel images is rather absurd....if not offensive. The most recent D1 is a joke. Albeit, an expensive one.
Nikon blew it in the scanner arena and made huge blunders in high end pro digicams (E1 & E2). Total junk from my personal experiences. I am not at all prone to spend a dime on any new Nikon digital things. In fact, I have dumped my Nikon so-called pro lenses and bodies for Contax. But this is another story.
My advice is to be very wary....be very wary of Nikon. I do not use Nikon cameras anymore and I do not use Nikon microscopes anymore. But it is your money and your decision. As they say, "Caveat emptor."
gg
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Modern surfers use PC boards. You can too at http://photoweb.net ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Rubbish! With all due respect this is a complete distortion and mis- use of the point I made. To realise this objective with low NA lenses you would have to find some way to defeat the laws of physics. No CCD imager, however clever will make it possible to correct the shortcomings of cheap, low-performance optics. You clearly misunderstood the point, which is that microscope lens resoltuion is not fundamentally dependent on the magnification factor but on the numerical aperture, which also determines resolution. A x100 NA 1.4 lens resolves no more detail than a x60 NA 1.4, but simply magnifies the image further. This is "empty magnification", which is only useful if your image sensor (film or CCD) has limited resolving power. With a very high resolution film like Technical Pan, you can take advantage of this fact to capture a wide-field diffraction-limited image with a x60 1.4 lens. That is not an option with a low NA lens irrespective of the properties of the sensor, CCD or otherwise.
} Hats of to Dr. Jeffree's explanation on optical resolution as it relates to } light microscopy applications. With high pixel density capable digital } cameras, now lower magnification, low NA (lower resolving power) objectives } can be used to acquire matching digital resolution images of wide fields of } view. } } Information on Nikon's Instrument Division "Digital Eclipse" family of } digital cameras for microscopy including the new DXM1200 digital color camera } will soon be available on our web site www.nikonusa.com or you can go to } our Dealer Locator at } http://www.nikonusa.com/corpinfo/dealers/dealerSearch.cfm and ask your } local Nikon authorized microscope dealer for further information. } } Best Regards, } } Stan Schwartz } Manager, BioSciences Dept. } Nikon Inc Instrument Division } 1300 Walt Whitman Rd. } Melville, NY 11747 } 631-547-8500 } 631-547-4033 Fax } Schwartz-at-nikonincmail.com } www.nikonusa.com } } } Earlier post } ============================================================= } Geoff } Analysing this a little further: } A 25 Mbyte image = 8.3 M pixels to cover 24x36mm or 864mm^2 } this represents 98.2 pixels per mm, or 49 line pairs per mm. Pixel } size 10.2 um. Resolution about 50 line pairs per mm at best. } } A 25M pixels image used to capture a 24x36mm Kodachrome } slide represents 28,935 pixels per mm^2 or 170 pixels per mm. } This is equivalent to a maximum resolution of 85 line pairs per mm, } which may be on the conservative side for Kodachrome. pixel size } = 5.88um. The image will be approx. 6120x4082 pixels, generating } a file size of approx 75Mb for a 24-bit (8+8+8bit) RGB image. } } To record 120 line pairs per mm, which many top 35mm camera } lenses can achieve, a minimum of 240 pixels per mm are required, } each 4.2 um wide. This equates to 8640x5760 pixels for a } 24x36mm frame = 50M-pixels or 50Mb in 8-bit greyscale, or 150Mb } in 24-bit RGB. } } At 320 line pairs per mm (Technical Pan) the minimum required } 640 pixels per mm is a pixel size of 1.56 um } } Presumably for a light image the diffraction limited resolution is } approx 1/2 lambda which at 540nm is 0.27um. } So looking to the future of ultimate-performance CCDs, direct } recording of a diffraction limited light image projected onto the } sensor requires at the very least 3703 pixels per image mm or } 13,717,421 pixels per mm^2 (greyscale 8-bit) } } However, if we are doing light microscopy with an NA 1.4 x100 } objective, how much resolving power do we need on CCD or film? } } Data is at 0.27um resolution (lambda = 540nm). Let's round this to } 0.3um. Magnification at 24x36mm film image is x100, so pixels } must be an absolute maximum of 30um wide to record the } significant data = 33.3 pixels per mm, equivalent to 800x1200 } pixels to record the whole 35mm negative area. However, most } CCDs are much smaller than 35 mm frames, typically 1/3 inch. So } an 800x1200 pixel CCD at 8x12 mm, 1/3 of the linear dimensions } of a 35mm frame would use 9 pixels to record the smallest image } details. This is about right from the point of view of resolution, but } to record the whole 35mm frame we need about 2400x3600 pixels } on our CCD. } } Note also that Technical Pan has (depending on the criterion used } to assess its performance) up to 10 times the resolving power } required to record all there is to see in a diffraction-limited LM } image made with a 100x NA 1.4 lens. So you can comfortably } afford to use a 60x NA 1.4 lens, thereby getting the same } resolution with a bigger field of view. } } Many years ago, I took a photograph of a street scene using a } Canon 35mm SLR loaded with Kodak Recordak (I think this was a } single layer microfilm emulsion). Examined in a light microscope, } the image clearly, legibly recorded the brand-name of a child's } push chair. I tried to print this brand name using a DeVere point- } source enlarger with an image size of 20x30 inches produced with } a Schneider Componon lens, but was completely unable to } produce a legible image. The point I am making here is that the } combination of some high performance films, with high quality } lenses of the standard produced by the leading camera } manufacturers can record more detail on the film than you can } easily get back out by conventional printing. I suspect the same is } true of EM exposures. } } So there is no contest - film beats CCDs for resolution hands } down. And you can process the image on the cheap. No money } goes to Intel or Microscoft, Adobe or Epson. But resolution is not } primarily what we buy CCDs for. We buy them primarily for instant } image capture in a format suitable for digital storage, digital } transmission quantitative data recording and image processing. } } Chris } } ===================================================================== } DR CHRIS JEFFREE } BIOSEM - BIOLOGICAL SCIENCES EM FACILITY } UNIVERSITY OF EDINBURGH } Daniel Rutherford Building } King's Buildings, Mayfield Road } EDINBURGH, EH9 3JH, Scotland, UK } Tel. #44 131 650 5345 } FAX. #44 131 650 6563 } Mobile 0410 585 401 } email c.jeffree-at-ed.ac.uk } SEM / TEM bookings sem-at-ed.ac.uk } ===================================================================== }
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Dr Chris Jeffree University of Edinburgh Biological Sciences EM Facility Daniel Rutherford Building King's Buildings EDINBURGH EH9 3JH Tel: +44 (0) 131 650 5345 FAX: +44 (0) 131 650 6563
Lets place the exhausted and exhausting Digital topic aside. Sergey, you do have a challenging project and one that is worth discussing. Just maybe somebody has an idea that will help you. I think that we discuss in this forum pixels too much and microscopy too little.
You probably have tried most of my following suggestions, but just incase, here are a couple of my thoughts:
Dark field contrast is enhanced most when the density between specimen and background is greatest. So its most effective with no specimen support. You could try your luck with the superfine mesh grids now available; these thin bar grids can be purchased down to 2000 mesh and these grids have a 7.5um hole size. If that is not a fine enough support, then try holey films with a net-like structure.
Carbon coating will stabilize samples dramatically. A little loss of contrast is inevitable, but a least carbon on the grid or holey plastic film does not matter.
Don't see why you want to render the gold nano particles with detail (I assume some greys). In TEM I would expect gold above about 5nm to be black. In STEM or FESEM you could get greys easily, but you may not get the resolution required.
Increasing if available to 200 or more the kV, will give more brightness, less contrast, better resolution (specimen dependent). Most importantly, specimen damage frequently is less at higher kV, that depends on the specimen again. I think that damage is less in specimens with greater electron transparency, but one of our "physical" gurus may explain the whys and wherefores.
Without switching film-types, you can develop the TEM films in things like Microdol-X, Microphen or Rodinal (hope those developers still exist). You could also use D-19 more dilute than normal. For most EM purposes all of these would give too softer negatives, but they just may suit you. Over developing in light photography yields more contrast - not so in TEM, where more electrons are the main contrast mechanism. Hence slower film and denser negatives are preferred, especially by most biologists.
I think that we started out with too much contrast, but a grainy image because of insufficient electron exposure. Hmmm, I think that we should take a holiday. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Saturday, June 03, 2000 7:44 AM, Sergey Ryazantsev [SMTP:sryazant-at-ucla.edu] wrote: } } } Dear Jim } } I spent about two years trying to make good pictures of my NanoGold labeled } protein-DNA complexes. Doing this job I find two main problems: the } sample is unstable under the beam as any biological sample; NanoGold is } much more bright than protein core in dark-field. I find that it is } impossible to record equally perfect signals from NanoGold and protein core } because of short dynamic range for SO-163 film, I believe. I was trying to } make two pictures with different exposure, but it is tricky: in dark-field } mode the automatic exposure meter usually does not work and we have to set } exposure manually, in this case it is difficult to get "right" exposure } time in the right moment, you know. Again, because of sample's short life } under the beam, it is impossible to make a couple pictures at the different } conditions sometime. Keep in mind, please, that to change the film in the } microscope it takes about 10 seconds. Your idea about increasing } signal-noise ratio by collecting more electrons is bright but not } practical. For biological samples (I am talking about non-fixed, } non-stained samples of proteins, DNA or RNA-protein complexes etc) the } electron damage is a huge problem. People are trying to solve it in } different ways. Some using cryo temperature (to stabilize the biological } structure). I was using freeze-drying (I find that freeze-dried samples } are more stable under the beam). But in any case we have deal with very } unstable samples and must to do everything to decrease (not increase as you } recommended) electron dose. Drift is a second big problem for such } application: to increase signal-noise ratio we have to use very thin } support films. Images obtained at such conditions are noisy and in most } cases we have to use image analysis tools to extract the data. It means } that we have to digitize our images anyway. In such situation digital } camera may help. As you, probably, remember I was a person who initiates } this discussion. I think this discussion was very useful for many of us } who are not friendly with digital camera's techniques. We understand the } limitations of the modern digital cameras better now. I would like to say } thank you everybody who was involved in this discussion. There is some } conclusions I make for myself from discussion: } } } - Film is still cheap and universal material for recording and } storage EM images, sorry CCD. } - CCD TEM camera should not substitute film. Film and camera should } work all together improving the flexibility of the TEM system. For this } reason I will chose side-mount camera if will have money for it. } - For cell-biology (thin sections) where the resolution of the sample } is about 3 nm CCD camera may do a good job allowing users to make a huge } number of pictures (cell-biology guys love it), instantly view and } catalogize them. } - Sometime the digital camera may help in area of high-resolution } (relatively high, guys) EM when image will be digitized anyway. The major } limitation here is small area of view (we need a lot of particles for image } analysis sometime), but you could make the set of overlapping pictures and } digitally combine it. I love, also, Mike Bode idea to make a few very } short exposure pictures and combine it digitally later to reduce noise. The } relatively big size of CCD's pixels is a real problem too. } - CCD camera is expensive "toy". I am not sure that the benefits from } using it will compensate astronomical price, actually the third of the } electron microscope value ($70000 is it 1/3 of microscope's price on } current market?). Currently, I would recognize the CCD TEM camera as funny } "attachment" which may be useful if you rich enough to spend money on it } (it mean, that you have everything else in your EM lab plus some extra } $70000 for the fun playing with digital "toy"). } - TEM CCD cameras are under extensive development now. Today's } camera will be replaced on the new model (read better, faster, what else?) } next year. Next year's camera will be easily forgotten next after the next } year and so on... Each new camera will be better that previous one... CCD } TEM camera it is not a good investment of money, I think. } - We should keep in mind that many companies charged extra 3-4K$ for } the installation and training (it is mandatory for Gatan for instance) and } you, probably, have to buy service contract on it even if you have service } contract on the microscope (JEOL's service contract on microscope do not } cover the CCD camera even if you buy it from JEOL). } } Best regard, Sergey } } } } } } Date: Fri, 02 Jun 2000 21:14:44 +1000 } } From: jim {jim-at-proscitech.com.au} } } Subject: RE: Film vs Digital } } To: 'Mike Bode' {mb-at-Soft-Imaging.com} , } } "'Microscopy-at-MSA.Microscopy.Com'" {Microscopy-at-sparc5.microscopy.com} } } Cc: "'jim-at-proscitech.com.au'" {jim-at-proscitech.com.au} } } Reply-to: "jim-at-proscitech.com.au" {jim-at-proscitech.com.au} } } Organization: ProSciTech } } X-Mailer: Microsoft Internet E-mail/MAPI - 8.0.0.4211 } } } } ------------------------------------------------------------------------ } } 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 } } -----------------------------------------------------------------------. } } } } } } The world is full of possible solutions, but are they practical.? } } To produce high-resolution, dark-field or any others TEM images that require } } more electrons to form a clear image, Mike Bode would use multiple digital } } exposures. The exposures could be layered and combined into one superior } } image. } } This image would be made up of more pixel and is formed by more electrons } } and } } so would be noise-free and hence could be further enlarged then otherwise } } possible. Perhaps. } } Beam blanking would largely save the specimen from beam damage and drift } } could } } be compensated for by matching up the digitals. Great. } } How much time is required between exposures to transfer a minimum 10mb image } } per exposure? What would be the total time from focusing to the last } } exposure? } } What about Z-drift in the interim requiring objective changes and what about } } the total cost of this additional get-up. The mind boggles at a through } } focus } } series. } } When pushing the limits a piece of film seems more effective, cheaper and fa } } ster. } } Again, I don't doubt that there is now a large place for digital in TEM, but } } its no panacea. } } Cheers } } Jim Darley } } ProSciTech Microscopy PLUS } } PO Box 111, Thuringowa QLD 4817 Australia } } Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com } } Great microscopy catalogue, 500 Links, MSDS, User Notes } } www.proscitech.com } } } } On Friday, June 02, 2000 1:24 AM, Mike Bode [SMTP:mb-at-soft-imaging.com] wrote: } } } } } } } } No, it's not really a problem. It's been done with low density } } } microscopy all the time. Granted, there are some technical aspects to be } } } overcome, but (and I can only speak for ourselves) we have done that on } } } a number of microscopes. You are of course correct, that 10 images at } } } 0.8 seconds take longer than 8 seconds as the image has to be } } } transferred, etc. BUT: that's what beam blankers are for. It is pretty } } } straightforward to take an image at 0.8 seconds, then blank the beam } } } very quickly before taking the next image. That way you get pretty close } } } to the 8 sec total exposure. If there is no beam blanker on the } } } microscope, in most cases it can be added. } } } } } } I am not sure what you mean by "too sensitive". The cameras are usually } } } constructed so that 1 electron from the beam creates between a few tenth } } } to a few counts (these are all statistical data, of course). The well } } } width divided by this sensitivity then determines, how many primary } } } electrons are needed to fully expose one pixel. For example, if the well } } } width is 50,000 electrons and the sensitivity is 1 count/electron, one } } } needs 50,000 primary electrons to fill the well. This translates into } } } roughly a 0.4% statistical error. } } } } } } } From a practical standpoint: You can take images with most cameras when } } } the exposure meter on the microscope reads a couple of seconds without } } } overexposing the camera. On the other hand, you can reduce the intensity } } } of the beam until you see single electron events. } } } } } } The one area where CCD cameras may be too sensitive is diffraction. The } } } normally huge intensity in the transmitted beam often leads to } } } saturation. In CCDs this can lead to blooming (the intensity spills over } } } into neighboring pixels). This can be taken care of with special chips } } } that have anti-blooming features, but this usually has some other } } } drawbacks. Again, this can also be overcome somewhat with multiple } } } exposures. Film behaves more civilized here, as it simply stops } } } responding to the electrons, but this makes film more or less useless } } } for quantitative measurements of diffraction patterns. I have done } } } diffraction with CCDs many times and though it does require some } } } tweaking, one can get very good results from them. } } } } } } Michael Bode, Ph.D. } } } Soft Imaging System Corp. } } } 1675 Carr St., #105N } } } Lakewood, CO 80215 } } } =================================== } } } 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: jim [mailto:jim-at-proscitech.com.au] } } } Sent: Wednesday, May 31, 2000 9:37 PM } } } To: 'Mike Bode'; 'Microscopy-at-MSA.Microscopy.Com' } } } Subject: RE: Film vs Digital } } } } } } } } } ------------------------------------------------------------------------ } } } 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 } } } -----------------------------------------------------------------------. } } } } } } } } } Yes, it could be done "in theory". Somebody would need to figure out the } } } } } } software and perhaps modify the hardware. Then we would find that the } } } total } } } exposure of the specimen to the electron beam maybe a muliple of the } } } film's } } } exposure. Afterall, an 8 sec film exposure would not amount in digital } } } to } } } 10x0.8, but we would require considerable time in between exposures. } } } Since the } } } problems in the discussed circumstances are specimen movement and beam } } } damage, } } } it seems that taking multiple exposures is a poor option. } } } } } } Digital cameras are for some situation too sensitive to electron } } } exposure. } } } Cutting back on electrons is no option since its the electrons that form } } } the } } } image in the first instance. } } } Much easier in light microscopy . . . insert a neutral density filter. } } } Cheers } } } Jim Darley } } } ProSciTech Microscopy PLUS } } } PO Box 111, Thuringowa QLD 4817 Australia } } } Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com } } } Great microscopy catalogue, 500 Links, MSDS, User Notes } } } www.proscitech.com } } } } } _____________________________________ } } Sergey Ryazantsev Ph. D. } Electron Microscopy } UCLA School of Medicine } Department of Biological Chemistry } Box 951737 } Los Angeles, CA 90095-1737 } } Phone: (310) 825-1144 } Pager: (310) 845-0248 } FAX (departmental): (310) 206-5272 } mailto:sryazant-at-ucla.edu } http://www.bol.ucla.edu/~sryazant } }
I have two replies and Mike my well have four replies to these. 1. There has been a parallel discussion concerning resolution of film versus digital images. Clearly in raw power digital cannot compete since film has multi gigabyte capacity. I added to that thread that what matters is: does digital have enough power and that frequently it would. Mike reinforced and strengthened that argument, finishing with the note that he is glad for film when looking at prints by Ansel Adams. (Ansel Adams until about 30 years ago carted for decades large format cameras through US National Parks, especially Yosemite, producing fantastic landscape photographs and books) Adams' limited edition prints were contacts of 5x7 and 8x10" inch sheet film, probably rated at 400 ISO. The line resolution of such prints much exceeds our eyes' resolution, but still results in superior gradation and detail. TEM film, even when much enlarged has such details too. Why Mike, should we accept 2.5 Mb? That is a splendid file size for SEM, and because of the limited enlargability of light microscopy (concrete ceiling due to wavelengths) it's reasonable, but minimal for light microscopy. TEM can do and deserves better.
Incidentally, from the outset I cited increased "enlargability" to obtain high resolution TEM, as one of films major advantages, since greater depths of field at moderate powers makes high powers through photo enlarging a desirable technique. The small additional magnification yielded by placing a digital camera lower in the column does not compensate. So greater "enlargability" of digitals would be desirable, but is limited by pixel size.
2 The thread was initiated by Sergey. He had problems visualising certain specimens in dark field. The use of his "beaut" digital TEM camera made things worse. I pointed out that the shorter exposure reduced the number of electrons forming the image, hence more noise. I believe that a good part of Mike's case will be settled in his favour when we hear "Eureka" from Sergey's lab. I don't doubt that much more can be done with digital now and that further improvements are on the way. Mike's "solution" may well be possible, but I don't believe its a snap; in any case: Show Sergey! Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Saturday, June 03, 2000 2:58 AM, Mike Bode [SMTP:mb-at-Soft-Imaging.com] wrote: } } Well, let's see: } } Jim wrote: } } Mike Bode would use multiple digital } exposures. The exposures could be layered and combined into one superior } image. } This image would be made up of more pixel and is formed by more } electrons and } so would be noise-free and hence could be further enlarged then } otherwise } possible. Perhaps. } } No, I did not talk about further enlargements. All I wanted to say is, } that a more noise-free image can be achieved by adding multiple images, } and that this also to some extent helps with drift of the sample during } acquisition. } } Jim wrote: } } How much time is required between exposures to transfer a minimum 10mb } image } per exposure? } } How did you arrive at 10 MB? A 1280x1024 image with 16 bit pixel } information is about 2.5 MB (uncompressed). We acquire about 10 of those } per second and transfer them across the PC bus to the display. Putting } them on them into Memory might add a few tenth of a second. Writing to } HD can be done after all images are acquired. } } Jim wrote: } } What would be the total time from focusing to the last exposure? } What about Z-drift in the interim requiring objective changes } } Why would we have to worry about that, if we don't have to worry about } that when taking the image on film? In fact, we could take care of this } by looking at the image between exposures and correct for z-drift. } However, as you said, that would add to the overall time and exposure. I } was comparing a normal dark field image taken on film at 8 seconds with } acquiring the same image on a "too sensitive" CCD camera by adding up 10 } consecutive .8 second images. Why would the sample drift (in x, y or z) } substantially more in 8+delta seconds than in 8? } } Jim wrote: } } what about the total cost of this additional get-up } } That of course depends on the microscope and there is no general answer. } For example on a LEO 912 I believe the blanker is standard. The } additional cost to use an acquisition scheme like this with our software } is $0 plus perhaps a bit of time to write a small macro. On other } microscopes one might have to add a beam blanker and perhaps a control } mechanism for the beam blanker. But I would guess, that this cost is not } very high. All modern microscopes are computer controller anyway, so it } is most likely just a control command that needs to be sent to the } microscope over a serial port if the beam blanker is installed. Piece of } cake. } } Jim wrote: } } The mind boggles at a through focus series. } } You're right here. But I don't think we were talking about through-focus } series. Incidentally, we do through-focus series on light microscopes } and reconstruction routinely. Takes a few images at different focus (or } for a light microscope: stage) settings. The rest is done off-line. } Takes maybe a couple of minutes for about 20 images of about 1kx1k. I } agree that TEM is different here and much more complicated due to the } complicated Contrast Transfer Function. However, this could in principle } be sorted out. } } Jim wrote: } } Again, I don't doubt that there is now a large place for digital in TEM, } but } its no panacea. } } I also agree with you on that one. But using the additional computer } possibilities of digital imaging might take you further than expected. } } Michael } } } Michael Bode, Ph.D. } Soft Imaging System Corp. } 1675 Carr St., #105N } Lakewood, CO 80215 } =================================== } 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: jim [mailto:jim-at-proscitech.com.au] } Sent: Friday, June 02, 2000 5:15 AM } To: 'Mike Bode'; 'Microscopy-at-MSA.Microscopy.Com' } Cc: 'jim-at-proscitech.com.au' } Subject: RE: Film vs Digital } } } The world is full of possible solutions, but are they practical.? } To produce high-resolution, dark-field or any others TEM images that } require } more electrons to form a clear image, Mike Bode would use multiple } digital } exposures. The exposures could be layered and combined into one superior } image. } This image would be made up of more pixel and is formed by more } electrons and } so would be noise-free and hence could be further enlarged then } otherwise } possible. Perhaps. } Beam blanking would largely save the specimen from beam damage and drift } could } be compensated for by matching up the digitals. Great. } How much time is required between exposures to transfer a minimum 10mb } image } per exposure? What would be the total time from focusing to the last } exposure? } What about Z-drift in the interim requiring objective changes and what } about } the total cost of this additional get-up. The mind boggles at a through } focus } series. } When pushing the limits a piece of film seems more effective, cheaper } and fa } ster. } Again, I don't doubt that there is now a large place for digital in TEM, } but } its no panacea. } Cheers } Jim Darley } ProSciTech Microscopy PLUS } PO Box 111, Thuringowa QLD 4817 Australia } Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com } Great microscopy catalogue, 500 Links, MSDS, User Notes } www.proscitech.com } } On Friday, June 02, 2000 1:24 AM, Mike Bode [SMTP:mb-at-soft-imaging.com] } wrote: } } } } No, it's not really a problem. It's been done with low density } } microscopy all the time. Granted, there are some technical aspects to } be } } overcome, but (and I can only speak for ourselves) we have done that } on } } a number of microscopes. You are of course correct, that 10 images at } } 0.8 seconds take longer than 8 seconds as the image has to be } } transferred, etc. BUT: that's what beam blankers are for. It is pretty } } straightforward to take an image at 0.8 seconds, then blank the beam } } very quickly before taking the next image. That way you get pretty } close } } to the 8 sec total exposure. If there is no beam blanker on the } } microscope, in most cases it can be added. } } } } I am not sure what you mean by "too sensitive". The cameras are } usually } } constructed so that 1 electron from the beam creates between a few } tenth } } to a few counts (these are all statistical data, of course). The well } } width divided by this sensitivity then determines, how many primary } } electrons are needed to fully expose one pixel. For example, if the } well } } width is 50,000 electrons and the sensitivity is 1 count/electron, one } } needs 50,000 primary electrons to fill the well. This translates into } } roughly a 0.4% statistical error. } } } } } From a practical standpoint: You can take images with most cameras } when } } the exposure meter on the microscope reads a couple of seconds without } } overexposing the camera. On the other hand, you can reduce the } intensity } } of the beam until you see single electron events. } } } } The one area where CCD cameras may be too sensitive is diffraction. } The } } normally huge intensity in the transmitted beam often leads to } } saturation. In CCDs this can lead to blooming (the intensity spills } over } } into neighboring pixels). This can be taken care of with special chips } } that have anti-blooming features, but this usually has some other } } drawbacks. Again, this can also be overcome somewhat with multiple } } exposures. Film behaves more civilized here, as it simply stops } } responding to the electrons, but this makes film more or less useless } } for quantitative measurements of diffraction patterns. I have done } } diffraction with CCDs many times and though it does require some } } tweaking, one can get very good results from them. } } } } Michael Bode, Ph.D. } } Soft Imaging System Corp. } } 1675 Carr St., #105N } } Lakewood, CO 80215 } } =================================== } } 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: jim [mailto:jim-at-proscitech.com.au] } } Sent: Wednesday, May 31, 2000 9:37 PM } } To: 'Mike Bode'; 'Microscopy-at-MSA.Microscopy.Com' } } Subject: RE: Film vs Digital } } } } } } } ------------------------------------------------------------------------ } } 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 } } } -----------------------------------------------------------------------. } } } } } } Yes, it could be done "in theory". Somebody would need to figure out } the } } } } software and perhaps modify the hardware. Then we would find that the } } total } } exposure of the specimen to the electron beam maybe a muliple of the } } film's } } exposure. Afterall, an 8 sec film exposure would not amount in digital } } to } } 10x0.8, but we would require considerable time in between exposures. } } Since the } } problems in the discussed circumstances are specimen movement and beam } } damage, } } it seems that taking multiple exposures is a poor option. } } } } Digital cameras are for some situation too sensitive to electron } } exposure. } } Cutting back on electrons is no option since its the electrons that } form } } the } } image in the first instance. } } Much easier in light microscopy . . . insert a neutral density } filter. } } Cheers } } Jim Darley } } ProSciTech Microscopy PLUS } } PO Box 111, Thuringowa QLD 4817 Australia } } Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com } } Great microscopy catalogue, 500 Links, MSDS, User Notes } } www.proscitech.com } }
****This is a commercial response from a Vendor****
Gary and List,
I am sorry you have had a bad experience with Nikon products. However, you are entitled to your "opinion" and therefore; I am entitled to a response. First, the new Digital Eclipse Camera capable of 12M pixels using a stepper mode is exactly the same technology that the new Zeiss Axiocam and an Olympus model uses which has been very well received by the microscope community.
Next, the successor to E1, E2 (technically, made by Fuji) is the current Nikon D1 (Digital SLR) was NOT intended for microscopes. It CAN go on one, but is limited to Brightfield applications and has no NTSC (video) out for focusing. However, it is by far the standard in Digital Cameras! It has won every Journal Award in it's field and is the definitive choice for Professional Photo-Journalists, including the massive market share Nikon has and the majority of Pulitzer Prize winners. As for Film Scanner products, the Nikon Coolscan line is still the standard in the industry. Especially, high end units with Auto Feeders. All Nikon digital products are not perfect. However, these are technical products that are evolving 6 months at a time and are truly in their infantile state from where they will be in just a few years from now! I would prefer to address your specific problems, but you gave none and just called it "junk". With that mentality, I feel my Pentium 266 computer is "junk", but I do not blame the manufacturer because I own an older model.
As for the Nikon Coolpix line (990), pardon my sarcasm, but where are "all the returns" because we need them to fulfill the 38,000 unit Backorders! This camera is so wildly successful EVERY Dealer (Photo and Microscope) is begging for them. Check out Ebay for example; the cameras are selling for more than List Price. Sound to me like a success if you understand the basics of Economics and Supply and Demand. Is it the perfect microscope camera? No, I don't even think so...but it is the ONLY high resolution (3.34M) digital camera, with live video out, goes on any brand or model microscope for under $1000. Oh, did I forget to mention it also won almost every Magazine Award in its class (not all, it actually received a tie with the Olympus 3030 in one, which is a very nice camera, but does not mount a microscope).
Finally, I have been on this List for many years and have respected the use and rules of this forum. We Vendors for the most part are respectful of the opinions of our users. However, making a "blanket statement" like "stay away from Nikon", does not serve the public well; NOR YOURSELF! If you or any customer has specific problems; we want to know about it; as does any reputable manufacturer for future product improvement. The fact is Nikon has the largest market share in microscopes and professional camera equipment and is climbing very quickly on the digital camera list too. So, in short you are entitled to your opinion, but the rest of the world by far does NOT agree with it............
I personally apologize to any members this correspondence offends. Believe me, my preference is to serve the list and answer technical microscopy questions as I have for many years, but this unfair and libel attack required a response.
Regretfully,
Lawrence Kordon Nikon, Inc. Senior Bioscience Specialist nikon-at-jagunet.com
"Dr. Gary Gaugler" 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 } -----------------------------------------------------------------------. } } At 02:21 PM 6/3/00, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Hello all, } } } } Please be aware that this is a commercial post that may be of interest to } } some of the list members. } } I hope not. } } } Digital light microscope cameras are now available that use a highly touted } } CCD detector: Color or monochrome; fast interline transfer, high sensitivity, } } low noise; 6.7 micron square pixel; 2/3 inch 1.3 Million chip; PCI bus frame } } transfer. This detector is ideal to use in light microscopy applications. } } Now precision technology and fast high capacity cheap computers allow this } } detector to be "Micro Stepped" providing digital images of up to 12 million } } pixels to be captured very quickly and providing file sizes of up to 35 MB in } } 24 bit color images. "Inter Pixel Stepping" technology will allow a } } considerably less expensive approach to digital imaging that can now approach } } film resolving capability in a light microscope. } } } } Hats of to Dr. Jeffree's explanation on optical resolution as it relates to } } light microscopy applications. With high pixel density capable digital } } cameras, now lower magnification, low NA (lower resolving power) objectives } } can be used to acquire matching digital resolution images of wide fields of } } view. } } } } Information on Nikon's Instrument Division "Digital Eclipse" family of } } digital cameras for microscopy including the new DXM1200 digital color camera } } will soon be available on our web site www.nikonusa.com or you can go to } } our Dealer Locator at } } http://www.nikonusa.com/corpinfo/dealers/dealerSearch.cfm and ask your } } local Nikon authorized microscope dealer for further information. } } } } Best Regards, } } } } Stan Schwartz } } Manager, BioSciences Dept. } } Nikon Inc Instrument Division } } 1300 Walt Whitman Rd. } } Melville, NY 11747 } } 631-547-8500 } } 631-547-4033 Fax } } Schwartz-at-nikonincmail.com } } www.nikonusa.com } } As a long term past user of Nikon equipment, it is a tale of sorrow. } Most recently, the digital E1 and E2 are dismal failures. Consumers } are sending back 990's in droves (see rec.photo.marketplace.digital). } I really think that Nikon blew it in regards to digicams. How Nikon can } claim to get a 1.3M pixel imager to produce realistic 12M pixel images } is rather absurd....if not offensive. The most recent D1 is a joke. Albeit, } an expensive one. } } Nikon blew it in the scanner arena and made huge blunders in high end } pro digicams (E1 & E2). Total junk from my personal experiences. I am } not at all prone to spend a dime on any new Nikon digital things. In fact, } I have } dumped my Nikon so-called pro lenses and bodies for Contax. But this } is another story. } } My advice is to be very wary....be very wary of Nikon. I do not use Nikon } cameras anymore and I do not use Nikon microscopes anymore. } But it is your money and your decision. As they say, "Caveat emptor." } } gg } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } Modern surfers use PC boards. You can too at } http://photoweb.net } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
It's been some time since I posted news about Project MICRO, MSA's middle school educational outreach program. I'm happy to report that Nestor, wearing his Webmaster hat (yes, he owns more than one), has just posted a substantial revision of the MICRO website (URL below). You'll find new information on several pages and a LOT of new entries in the bibliography. Don't miss the "Cyclops" videos and the new CD-ROMs; the website hotlinks are much expanded also.
Caroline
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
I want to stain Procure-Araldite embedded material using the periodic-acid schiff procedure. One method I have is to hydrolyze in 1N HCl at 60 degrees C for 10 mins, water wash, stain in schiffs, metabisulphate rinses, counterstain, water wash (O'Brien and McCully, 1981). Does anyone use a method routinely which varies from this??
A fellow colleague has bought a JEOL 100s and wants and needs EDS X-ray Analysis. He's mounted his horizintal EDS detector , but can not get sample X-rays from the speciman. Contacting JEOL he found the problem to be a tilt problem. The 10 degree tilt from the normal SEG only tilts 10 degrees and a 30 to 60 degree tilt is necessary. At one time I was told that special sample holders and or double gap pole pieces were availabe. He wishs to find any one with a 100s for parts needed or information which would allow X-ray analysis. Please reply to mgmanders-at-aol.com or the list server.
I reacall some one looking for a desktop incubator. I came a across one on ebay. http://cgi.ebay.com/aw-cgi/eBayISAPI.dll?ViewItem&item=344913999 It is a little pricy by ebay standards. You can find a bunch of incubators by going to www.ebay.com and search for incubators you can do the same a http://www.labx.com
I don't have any connection with anyone involved in this.
Gordon
Gordon Couger gcouger-at-couger.com
Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
Toby, I used this technique years ago when I worked for Terry O'Brien at Monash Uni. Depending on the type of tissue you may want to do an aldehyde blockade before you begin the staining procedure. This technique is also in O'Brien and McCully. The blockade removes any aldehyde grouping that will react with the Schiffs reagent. You then generate and stain aldehyde groups during the procedure. Regards JVN
Toby Knight 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 want to stain Procure-Araldite embedded material using the periodic-acid } schiff procedure. One method I have is to hydrolyze in 1N HCl at } 60 degrees C for 10 mins, water wash, stain in schiffs, metabisulphate } rinses, counterstain, water wash (O'Brien and McCully, 1981). Does anyone } use a method routinely which varies from this?? } } thanks in advance, Toby Knight. } } -------------------------------------------------------- } Toby Knight } PhD student } } Department of Horticulture, Viticulture and Oenology } The University of Adelaide } Plant Research Centre } Waite Campus, PMB 1 } Glen Osmond, SA 5064 } } Tel: +61 8 8303 7224 or 8303 6668 } HVO: +61 8 8303 7242 } Fax: +61 8 8303 7116 } Email: tknight-at-waite.adelaide.edu.au } --------------------------------------------------------
-- **************************************************** John V Nailon Operations Manager Centre for Microscopy and Microanalysis The University of Queensland St. Lucia Queensland 4072 Phone: +61-7-3365-4214 Fax: +61-7-3365-4422 WWW: http://www.uq.edu.au/nanoworld/allstaff.html#Nailon ****************************************************
Toby- I've worked with the PAS stain with methyl methacrylate embedded tissue cut at 2 microns. I used a kit from Polyscientific (NY, USA) which supplied me with all of the necessary reagents (minus the ethanol and xylene). The protocol that was supplied with the kit is for paraffin embedded material but I made modifications to the protocol to accommodate methyl methacrylate. If you are interested in the modified protocol, please contact me. I should also mention that I automated this stain to save time and to maximize quality.
Good luck! Michelle Taurino Aventis Pharmaceuticals Bioimaging and Molecular Histology Tel-908-231-3357 Fax-908-231-3962 e-mail: Michelle.Taurino-at-aventis.com
-----Original Message----- } From: Toby Knight [mailto:tknight-at-waite.adelaide.edu.au] Sent: Sunday, June 04, 2000 10:59 PM To: Histonet; Microscopy list
I want to stain Procure-Araldite embedded material using the periodic-acid schiff procedure. One method I have is to hydrolyze in 1N HCl at 60 degrees C for 10 mins, water wash, stain in schiffs, metabisulphate rinses, counterstain, water wash (O'Brien and McCully, 1981). Does anyone use a method routinely which varies from this??
Just like to thank all those who responded to my squeal for help on BS detector resolution. Would like to add that the South African microscopy community is looking into establishing QA procedures and some of the suggestions maybe very helpful in this regard. Thanks, Malc.
-- Dr MP Roberts Phone: [+27](0)46 603 8313 Dept of Geology Fax: [+27](0)46 622 9715 Rhodes University Cell: 083 4060 262 (usually off) 6140 Grahamstown e-mail: malc-at-rock.ru.ac.za SOUTH AFRICA
I've been trying to look at Mycoplasms with SEM. Untill now, the results are not what we hoped for. We've used polycarbonate filters to prepare the Mycoplasms. Another way was preparing the Mycoplasms on agar. We have done this already for enterococcus and we know that this technique works well. But with the Mycoplasms we see almost nothing. It's like the Mycoplasms are IN the Agar. Has anyone have experience with MYCOPLASMS and SEM ???
De Pauw Bart Ghent University Faculty of Veterinary Medicine Morphology Salisburylaan 133 9820 Merelbeke Belgium Phone : 0032(0)9 264.77.19 Fax : 0032(0)9 264.77.90
Toby The procedure you describe is not in fact Periodic Acid-Schiff but the Feulgen reaction, for visualization of DNA in nuclei and mitochondria.
The PAS reaction uses periodic acid or sodium meta-periodate (typically 1% solution, ~10min) to oxidise the vicinal diols of some polysaccharides which are then stained with pararosaniline Schiff's reagent. It is not usually necessary to use metabisulphite in the wash water. The detailed procedure is also described in O'Brien & McCully 1981.
PAS (and Feulgen) may be viewed in a brightfield microscope or by fluorescence microscopy with a "rhodamine" filter set. An alternative fluorescent dye which works well in a fluorescence pseudo Schiff procedure is Lucifer Yellow CH, which needs blue excitation (FITC filter set).
Chris
Date sent: Mon, 5 Jun 2000 12:29:02 +0930 (CST) } From: Toby Knight {tknight-at-waite.adelaide.edu.au} Send reply to: Toby Knight {tknight-at-waite.adelaide.edu.au}
I just came across the following pearl in a 1982 Kodak publication, number PDS 61 "Selecting film from Kodak for photomicrography".
"The (Technical Pan) 2415 film has very low granularity and is capable of very great enlargement. In some cases the resolving power may be beyond that of the microscope image"
How do they know! Answers on a postcard please ....
Chris ===================================================================== DR CHRIS JEFFREE BIOSEM - BIOLOGICAL SCIENCES EM FACILITY UNIVERSITY OF EDINBURGH Daniel Rutherford Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JH, Scotland, UK Tel. #44 131 650 5345 FAX. #44 131 650 6563 Mobile 0410 585 401 email c.jeffree-at-ed.ac.uk SEM / TEM bookings sem-at-ed.ac.uk =====================================================================
The Electron Microscopy Core Facility at Mayo Clinic in Rochester, MN has an opening for a Biomedical EM technologist to support both clinical and research projects. The laboratory offers expertise to collaborative projects that involve transmission and scanning electron microscopy. The laboratory is well equipped and has a history of excellent productivity and adequate funding.
The successful candidate for this position will possess at least a bachelor's degree in Biology with experience in histology and/or electron microscopy. Additional courses or experience in Immunology, Cell Biology, and Digital Imaging is desirable. Operating knowledge of transmission and scanning electron microscopes is preferred. The applicant must have excellent communicative skills and the ability to work well with a variety of personalities.
The EM Technologist interacts with all laboratory users in order to accomplish specific research and clinical goals with respect to electron microscopy procedures. Duties include: All aspects of specimen preparation for a variety of biomedical samples for TEM and SEM, operation of TEM and SEM, darkroom developing and printing, digital image capture, and reporting. The technologist will also perform advanced research procedures including immunoelectron microscopy, x-ray microanalysis, and microwave processing.
Mayo offers a competitive salary and benefits package. If interested, please submit a cover letter and resume referencing job posting #00-0002365 to:
Jill Kelly Mayo Medical Center Human Resources-OE 1 Rochester, MN 55905 Fax: 507-284-1445 Email: kelly.jill-at-mayo.edu
Jon Charlesworth Coordinator Electron Microscopy Core Facility 1426 Gugg X4-3148
'I have two replies and Mike my well have four replies to these.'
I'll try. But I agree with you that we should let this thread die. jim, it seems you're a bit angy at me. If I unintentionally stepped on your toe I apologize. I did not expect that we would agree 100%, as you are selling film and I am involved in digital systems. I just hope that some other readers found some of this useful. I don't consider myself a "digital fanatic". This film vs. digital issue has many more facets, some of which we did not even touch, and which can be just as entertaining.
Jim wrote:
'Mike reinforced and strengthened that argument, finishing with the note that he is glad for film when looking at prints by Ansel Adams.'
Just for the record: I usually do not take a magnifying glass to photos. I mentioned Adams because I like his pictures. If he had taken them with a digital camera I would have liked them just as much. I also like some modern art paintings. That does not mean we should start drawing what we see in the microscopes rather than taking pictures ;-)
Jim wrote:
'Incidentally, from the outset I cited increased "enlargability" to obtain high resolution TEM'
Well, you said 'When great enlargements are required film is superior'. Perhaps I misunderstood. I thought, the term great enlargements referred to the microscope and meant 'small details', which you can take easier with a digital camera (no time delay between seeing something and taking an image, no mechanical vibration due to film movement, etc.). I have said many times before that film may be better if high resolution AND large field of view is required.
Jim wrote:
'in any case: Show Sergey!'
I'm in back-channel correspondence with him.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== 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: jim [mailto:jim-at-proscitech.com.au] Sent: Sunday, June 04, 2000 2:34 AM To: 'Mike Bode'; 'Microscopy-at-MSA.Microscopy.Com'
I have two replies and Mike my well have four replies to these. 1. There has been a parallel discussion concerning resolution of film versus digital images. Clearly in raw power digital cannot compete since film has multi gigabyte capacity. I added to that thread that what matters is: does digital have enough power and that frequently it would. Mike reinforced and strengthened that argument, finishing with the note that he is glad for film when looking at prints by Ansel Adams. (Ansel Adams until about 30 years ago carted for decades large format cameras through US National Parks, especially Yosemite, producing fantastic landscape photographs and books) Adams' limited edition prints were contacts of 5x7 and 8x10" inch sheet film, probably rated at 400 ISO. The line resolution of such prints much exceeds our eyes' resolution, but still results in superior gradation and detail. TEM film, even when much enlarged has such details too. Why Mike, should we accept 2.5 Mb? That is a splendid file size for SEM, and because of the limited enlargability of light microscopy (concrete ceiling due to wavelengths) it's reasonable, but minimal for light microscopy. TEM can do and deserves better.
Incidentally, from the outset I cited increased "enlargability" to obtain high resolution TEM, as one of films major advantages, since greater depths of field at moderate powers makes high powers through photo enlarging a desirable
technique. The small additional magnification yielded by placing a digital camera lower in the column does not compensate. So greater "enlargability" of digitals would be desirable, but is limited by pixel size.
2 The thread was initiated by Sergey. He had problems visualising certain specimens in dark field. The use of his "beaut" digital TEM camera made things worse. I pointed out that the shorter exposure reduced the number of electrons forming the image, hence more noise. I believe that a good part of Mike's case will be settled in his favour when we hear "Eureka" from Sergey's lab. I don't doubt that much more can be done with digital now and that further improvements are on the way. Mike's "solution" may well be possible, but I don't believe its a snap; in any case: Show Sergey! Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Saturday, June 03, 2000 2:58 AM, Mike Bode [SMTP:mb-at-Soft-Imaging.com] wrote: } } Well, let's see: } } Jim wrote: } } Mike Bode would use multiple digital } exposures. The exposures could be layered and combined into one superior } image. } This image would be made up of more pixel and is formed by more } electrons and } so would be noise-free and hence could be further enlarged then } otherwise } possible. Perhaps. } } No, I did not talk about further enlargements. All I wanted to say is, } that a more noise-free image can be achieved by adding multiple images, } and that this also to some extent helps with drift of the sample during } acquisition. } } Jim wrote: } } How much time is required between exposures to transfer a minimum 10mb } image } per exposure? } } How did you arrive at 10 MB? A 1280x1024 image with 16 bit pixel } information is about 2.5 MB (uncompressed). We acquire about 10 of those } per second and transfer them across the PC bus to the display. Putting } them on them into Memory might add a few tenth of a second. Writing to } HD can be done after all images are acquired. } } Jim wrote: } } What would be the total time from focusing to the last exposure? } What about Z-drift in the interim requiring objective changes } } Why would we have to worry about that, if we don't have to worry about } that when taking the image on film? In fact, we could take care of this } by looking at the image between exposures and correct for z-drift. } However, as you said, that would add to the overall time and exposure. I } was comparing a normal dark field image taken on film at 8 seconds with } acquiring the same image on a "too sensitive" CCD camera by adding up 10 } consecutive .8 second images. Why would the sample drift (in x, y or z) } substantially more in 8+delta seconds than in 8? } } Jim wrote: } } what about the total cost of this additional get-up } } That of course depends on the microscope and there is no general answer. } For example on a LEO 912 I believe the blanker is standard. The } additional cost to use an acquisition scheme like this with our software } is $0 plus perhaps a bit of time to write a small macro. On other } microscopes one might have to add a beam blanker and perhaps a control } mechanism for the beam blanker. But I would guess, that this cost is not } very high. All modern microscopes are computer controller anyway, so it } is most likely just a control command that needs to be sent to the } microscope over a serial port if the beam blanker is installed. Piece of } cake. } } Jim wrote: } } The mind boggles at a through focus series. } } You're right here. But I don't think we were talking about through-focus } series. Incidentally, we do through-focus series on light microscopes } and reconstruction routinely. Takes a few images at different focus (or } for a light microscope: stage) settings. The rest is done off-line. } Takes maybe a couple of minutes for about 20 images of about 1kx1k. I } agree that TEM is different here and much more complicated due to the } complicated Contrast Transfer Function. However, this could in principle } be sorted out. } } Jim wrote: } } Again, I don't doubt that there is now a large place for digital in TEM, } but } its no panacea. } } I also agree with you on that one. But using the additional computer } possibilities of digital imaging might take you further than expected. } } Michael } } } Michael Bode, Ph.D. } Soft Imaging System Corp. } 1675 Carr St., #105N } Lakewood, CO 80215 } =================================== } 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: jim [mailto:jim-at-proscitech.com.au] } Sent: Friday, June 02, 2000 5:15 AM } To: 'Mike Bode'; 'Microscopy-at-MSA.Microscopy.Com' } Cc: 'jim-at-proscitech.com.au' } Subject: RE: Film vs Digital } } } The world is full of possible solutions, but are they practical.? } To produce high-resolution, dark-field or any others TEM images that } require } more electrons to form a clear image, Mike Bode would use multiple } digital } exposures. The exposures could be layered and combined into one superior } image. } This image would be made up of more pixel and is formed by more } electrons and } so would be noise-free and hence could be further enlarged then } otherwise } possible. Perhaps. } Beam blanking would largely save the specimen from beam damage and drift } could } be compensated for by matching up the digitals. Great. } How much time is required between exposures to transfer a minimum 10mb } image } per exposure? What would be the total time from focusing to the last } exposure? } What about Z-drift in the interim requiring objective changes and what } about } the total cost of this additional get-up. The mind boggles at a through } focus } series. } When pushing the limits a piece of film seems more effective, cheaper } and fa } ster. } Again, I don't doubt that there is now a large place for digital in TEM, } but } its no panacea. } Cheers } Jim Darley } ProSciTech Microscopy PLUS } PO Box 111, Thuringowa QLD 4817 Australia } Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com } Great microscopy catalogue, 500 Links, MSDS, User Notes } www.proscitech.com } } On Friday, June 02, 2000 1:24 AM, Mike Bode [SMTP:mb-at-soft-imaging.com] } wrote: } } } } No, it's not really a problem. It's been done with low density } } microscopy all the time. Granted, there are some technical aspects to } be } } overcome, but (and I can only speak for ourselves) we have done that } on } } a number of microscopes. You are of course correct, that 10 images at } } 0.8 seconds take longer than 8 seconds as the image has to be } } transferred, etc. BUT: that's what beam blankers are for. It is pretty } } straightforward to take an image at 0.8 seconds, then blank the beam } } very quickly before taking the next image. That way you get pretty } close } } to the 8 sec total exposure. If there is no beam blanker on the } } microscope, in most cases it can be added. } } } } I am not sure what you mean by "too sensitive". The cameras are } usually } } constructed so that 1 electron from the beam creates between a few } tenth } } to a few counts (these are all statistical data, of course). The well } } width divided by this sensitivity then determines, how many primary } } electrons are needed to fully expose one pixel. For example, if the } well } } width is 50,000 electrons and the sensitivity is 1 count/electron, one } } needs 50,000 primary electrons to fill the well. This translates into } } roughly a 0.4% statistical error. } } } } } From a practical standpoint: You can take images with most cameras } when } } the exposure meter on the microscope reads a couple of seconds without } } overexposing the camera. On the other hand, you can reduce the } intensity } } of the beam until you see single electron events. } } } } The one area where CCD cameras may be too sensitive is diffraction. } The } } normally huge intensity in the transmitted beam often leads to } } saturation. In CCDs this can lead to blooming (the intensity spills } over } } into neighboring pixels). This can be taken care of with special chips } } that have anti-blooming features, but this usually has some other } } drawbacks. Again, this can also be overcome somewhat with multiple } } exposures. Film behaves more civilized here, as it simply stops } } responding to the electrons, but this makes film more or less useless } } for quantitative measurements of diffraction patterns. I have done } } diffraction with CCDs many times and though it does require some } } tweaking, one can get very good results from them. } } } } Michael Bode, Ph.D. } } Soft Imaging System Corp. } } 1675 Carr St., #105N } } Lakewood, CO 80215 } } =================================== } } 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: jim [mailto:jim-at-proscitech.com.au] } } Sent: Wednesday, May 31, 2000 9:37 PM } } To: 'Mike Bode'; 'Microscopy-at-MSA.Microscopy.Com' } } Subject: RE: Film vs Digital } } } } } } } ------------------------------------------------------------------------ } } 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 } } } -----------------------------------------------------------------------. } } } } } } Yes, it could be done "in theory". Somebody would need to figure out } the } } } } software and perhaps modify the hardware. Then we would find that the } } total } } exposure of the specimen to the electron beam maybe a muliple of the } } film's } } exposure. Afterall, an 8 sec film exposure would not amount in digital } } to } } 10x0.8, but we would require considerable time in between exposures. } } Since the } } problems in the discussed circumstances are specimen movement and beam } } damage, } } it seems that taking multiple exposures is a poor option. } } } } Digital cameras are for some situation too sensitive to electron } } exposure. } } Cutting back on electrons is no option since its the electrons that } form } } the } } image in the first instance. } } Much easier in light microscopy . . . insert a neutral density } filter. } } Cheers } } Jim Darley } } ProSciTech Microscopy PLUS } } PO Box 111, Thuringowa QLD 4817 Australia } } Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com } } Great microscopy catalogue, 500 Links, MSDS, User Notes } } www.proscitech.com } }
Jonathon Low kV is preferred for some biological applications chiefly because biologists experience difficulty getting enough contrast out of their specimens. As Dr Alves de Matos rightly says, low kV is more damaging to specimens than high kV, because the penetrating power of low-kV electrons is lower and therefore more beam energy is lost to the specimen. (We think of this exactly the other way round for SEM, but that's a story for another day!).
The main reason for the apparent shift in biologist's attitudes is that many are now less concerned with cell and tissue-level ultrastructure and increasingly interested in imaging macromolecules. 200kV or 300kV machines are preferred for cryo- microscopy of macromolecules in vitrified ice because the resolution is a lot better than at 80 or 120kV, and also because 200kV causes less specimen damage.
However, the cost of resolution either from higher kVs or the shorter working distance objective lenses favoured by materials scientists is poorer image contrast. Philips therefore supply "Biotwin" versions of some of their TEM range which use a long focal length objective which trading a little resolution for a little more contrast. This optical configuration is coincidentally very good for thickish specimens such as ultrathin biological sections where the ultimate resolution of the machine is pretty much academic anyway.
Images of thick (#100nm) specimens such as FIB sections of microelectronic devices seem crisper and more contrasty in our CM120 Biotwin than in a 200KV machine. Chris
} From: "A.P.Alves de Matos" {apmatos-at-ip.pt} To: {Microscopy-at-sparc5.microscopy.com}
Colleagues....
Some of you have misinterpreted my earlier message. Please feel free to carry on the discussion on film/digitization etc..
I only asked that the "tangental thread" centered about complaints on a specific product be taken off-line before any fire and brimstone starts up between a manufacturer and clients. This is not the forum for that type of feedback.
The previous dicussion should certainly continue as long as necessary.
Nestor Your Friendly Neighborhood SysOp
} } Nestor, } } It would be very unfortunate to squelch this important and central debate } concerning not only the corresponding resolution comparisons but also } differances in performance due to noise and greyscale depth. New methods of } enhansing resolution need to be critiqued as well. } } Dave Barnard } } Wadsworth Center HVEM } NYS Dept. Health } Albany NY
We are looking for the "T-tool" - a kind of fixture for TEM sample polishing. It is from T&T group in USA. We need to know the email address or telephone number of T&T group.
Any information about the T&T group or the T-tool will be highly appreciated!
We are looking to purchase a used JEOL 2010 TEM (LaB6, not FE) in good working condition. I would appreciate replies from anyone who knows if a JEOL 2010 is available or will become available in the next 6 months or so. A STEM unit would increase our interest, but isn't strictly necessary.
Also, we will be looking for a buyer for our JEOL 1200EX STEM which has been under service contract from day 1. It has been a highly reliable microscope and has given years of satisfactory performance.
Dave Joswiak Research Scientist Dept. of Astronomy, 351580 University of Washington Seattle, WA 98195 (206)543-7702 joswiak-at-astro.washington.edu
I am forwarding this request on behalf of Mr. Edoardo Nolfo. If anyone could be of assistance please send the information to him directly or (if you prefer) I will forward it to him.
} I am a student at the University of Oxford and am writing to ask for some } advice. I am currently working on multilayer reflectors in beetles; I plan to } use S.E.M. and T.E.M. to elucidate the fine structure of these reflectors. } Apparently the elytra of the beetles must be sliced very thinly for S.E.M. } analysis. My problem lies in the fact that the elytra are extremely } hard, which } means they cannot be sliced very thinly. Do you have any advice to } offer on how } to soften the elytra for this purpose? Perhaps there is a chemical } that is used } to make samples softer, or a standard technique used to soften very hard } samples. I would be extremely grateful to you if you could help me! Please do } not hesitate to contact me if you require any more information. } } Let me thank you in advance for your help; I look forward to hearing from you } soon.
#################################################################### John J. Bozzola, Ph.D., Director Micro-Imaging and Analysis Center 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 ####################################################################
Chris, This is because Kodak use EMs to measure the grain size of their silver halide grains. Tech Pan has a very small grain size (slow film, low ISO/ASA number) and is therefore capable of greater enlargement than film with a larger grains. The larger the grain the faster the film (higher ISO/ASA number). Regards JVN JVN
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 } -----------------------------------------------------------------------. } } I just came across the following pearl in a 1982 Kodak publication, } number PDS 61 "Selecting film from Kodak for photomicrography". } } "The (Technical Pan) 2415 film has very low granularity and is } capable of very great enlargement. In some cases the resolving } power may be beyond that of the microscope image" } } How do they know! } Answers on a postcard please .... } } Chris } ===================================================================== } DR CHRIS JEFFREE } BIOSEM - BIOLOGICAL SCIENCES EM FACILITY } UNIVERSITY OF EDINBURGH } Daniel Rutherford Building } King's Buildings, Mayfield Road } EDINBURGH, EH9 3JH, Scotland, UK } Tel. #44 131 650 5345 } FAX. #44 131 650 6563 } Mobile 0410 585 401 } email c.jeffree-at-ed.ac.uk } SEM / TEM bookings sem-at-ed.ac.uk } =====================================================================
-- **************************************************** John V Nailon Operations Manager Centre for Microscopy and Microanalysis The University of Queensland St. Lucia Queensland 4072 Phone: +61-7-3365-4214 Fax: +61-7-3365-4422 WWW: http://www.uq.edu.au/nanoworld/allstaff.html#Nailon ****************************************************
The "T" tool is a variation of the well known Tripod Polisherš. We are the manufacturers of the Tripod Polisherš and also manufacture the more compact BiPod Polisher. The BiPod polisher offers the smaller size as found on the T-tool while still incorporating our many years of experience in SEM and TEM cross section polishing as well as our expertise in precision machining. I know this doesn't help much in locating the T-tool, but it does offer you the information you need to acquire a tool that will effectively meet your requirements. If you would like additional information on these tools, please feel free to contact me.
David Henriks Vice President 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
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by Sha Zhu } ----------------------------------------------------------------------- The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We are looking for the "T-tool" - a kind of fixture for TEM sample polishing. It is from T&T group in USA. We need to know the email address or telephone number of T&T group.
Any information about the T&T group or the T-tool will be highly appreciated!
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Jim wrote:
'I have two replies and Mike my well have four replies to these.'
I'll try. But I agree with you that we should let this thread die. jim, it seems you're a bit angy at me. If I unintentionally stepped on your toe I apologize. I did not expect that we would agree 100%, as you are selling film and I am involved in digital systems. I just hope that some other readers found some of this useful. I don't consider myself a "digital fanatic". This film vs. digital issue has many more facets, some of which we did not even touch, and which can be just as entertaining.
Jim wrote:
'Mike reinforced and strengthened that argument, finishing with the note that he is glad for film when looking at prints by Ansel Adams.'
Just for the record: I usually do not take a magnifying glass to photos. I mentioned Adams because I like his pictures. If he had taken them with a digital camera I would have liked them just as much. I also like some modern art paintings. That does not mean we should start drawing what we see in the microscopes rather than taking pictures ;-)
Jim wrote:
'Incidentally, from the outset I cited increased "enlargability" to obtain high resolution TEM'
Well, you said 'When great enlargements are required film is superior'. Perhaps I misunderstood. I thought, the term great enlargements referred to the microscope and meant 'small details', which you can take easier with a digital camera (no time delay between seeing something and taking an image, no mechanical vibration due to film movement, etc.). I have said many times before that film may be better if high resolution AND large field of view is required.
Jim wrote:
'in any case: Show Sergey!'
I'm in back-channel correspondence with him.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== 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: jim [mailto:jim-at-proscitech.com.au] Sent: Sunday, June 04, 2000 2:34 AM To: 'Mike Bode'; 'Microscopy-at-MSA.Microscopy.Com'
I have two replies and Mike my well have four replies to these. 1. There has been a parallel discussion concerning resolution of film versus digital images. Clearly in raw power digital cannot compete since film has multi gigabyte capacity. I added to that thread that what matters is: does digital have enough power and that frequently it would. Mike reinforced and strengthened that argument, finishing with the note that he is glad for film when looking at prints by Ansel Adams. (Ansel Adams until about 30 years ago carted for decades large format cameras through US National Parks, especially Yosemite, producing fantastic landscape photographs and books) Adams' limited edition prints were contacts of 5x7 and 8x10" inch sheet film, probably rated at 400 ISO. The line resolution of such prints much exceeds our eyes' resolution, but still results in superior gradation and detail. TEM film, even when much enlarged has such details too. Why Mike, should we accept 2.5 Mb? That is a splendid file size for SEM, and because of the limited enlargability of light microscopy (concrete ceiling due to wavelengths) it's reasonable, but minimal for light microscopy. TEM can do and deserves better.
Incidentally, from the outset I cited increased "enlargability" to obtain high resolution TEM, as one of films major advantages, since greater depths of field at moderate powers makes high powers through photo enlarging a desirable
technique. The small additional magnification yielded by placing a digital camera lower in the column does not compensate. So greater "enlargability" of digitals would be desirable, but is limited by pixel size.
2 The thread was initiated by Sergey. He had problems visualising certain specimens in dark field. The use of his "beaut" digital TEM camera made things worse. I pointed out that the shorter exposure reduced the number of electrons forming the image, hence more noise. I believe that a good part of Mike's case will be settled in his favour when we hear "Eureka" from Sergey's lab. I don't doubt that much more can be done with digital now and that further improvements are on the way. Mike's "solution" may well be possible, but I don't believe its a snap; in any case: Show Sergey! Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Saturday, June 03, 2000 2:58 AM, Mike Bode [SMTP:mb-at-Soft-Imaging.com] wrote: } } Well, let's see: } } Jim wrote: } } Mike Bode would use multiple digital } exposures. The exposures could be layered and combined into one superior } image. } This image would be made up of more pixel and is formed by more } electrons and } so would be noise-free and hence could be further enlarged then } otherwise } possible. Perhaps. } } No, I did not talk about further enlargements. All I wanted to say is, } that a more noise-free image can be achieved by adding multiple images, } and that this also to some extent helps with drift of the sample during } acquisition. } } Jim wrote: } } How much time is required between exposures to transfer a minimum 10mb } image } per exposure? } } How did you arrive at 10 MB? A 1280x1024 image with 16 bit pixel } information is about 2.5 MB (uncompressed). We acquire about 10 of those } per second and transfer them across the PC bus to the display. Putting } them on them into Memory might add a few tenth of a second. Writing to } HD can be done after all images are acquired. } } Jim wrote: } } What would be the total time from focusing to the last exposure? } What about Z-drift in the interim requiring objective changes } } Why would we have to worry about that, if we don't have to worry about } that when taking the image on film? In fact, we could take care of this } by looking at the image between exposures and correct for z-drift. } However, as you said, that would add to the overall time and exposure. I } was comparing a normal dark field image taken on film at 8 seconds with } acquiring the same image on a "too sensitive" CCD camera by adding up 10 } consecutive .8 second images. Why would the sample drift (in x, y or z) } substantially more in 8+delta seconds than in 8? } } Jim wrote: } } what about the total cost of this additional get-up } } That of course depends on the microscope and there is no general answer. } For example on a LEO 912 I believe the blanker is standard. The } additional cost to use an acquisition scheme like this with our software } is $0 plus perhaps a bit of time to write a small macro. On other } microscopes one might have to add a beam blanker and perhaps a control } mechanism for the beam blanker. But I would guess, that this cost is not } very high. All modern microscopes are computer controller anyway, so it } is most likely just a control command that needs to be sent to the } microscope over a serial port if the beam blanker is installed. Piece of } cake. } } Jim wrote: } } The mind boggles at a through focus series. } } You're right here. But I don't think we were talking about through-focus } series. Incidentally, we do through-focus series on light microscopes } and reconstruction routinely. Takes a few images at different focus (or } for a light microscope: stage) settings. The rest is done off-line. } Takes maybe a couple of minutes for about 20 images of about 1kx1k. I } agree that TEM is different here and much more complicated due to the } complicated Contrast Transfer Function. However, this could in principle } be sorted out. } } Jim wrote: } } Again, I don't doubt that there is now a large place for digital in TEM, } but } its no panacea. } } I also agree with you on that one. But using the additional computer } possibilities of digital imaging might take you further than expected. } } Michael } } } Michael Bode, Ph.D. } Soft Imaging System Corp. } 1675 Carr St., #105N } Lakewood, CO 80215 } =================================== } 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: jim [mailto:jim-at-proscitech.com.au] } Sent: Friday, June 02, 2000 5:15 AM } To: 'Mike Bode'; 'Microscopy-at-MSA.Microscopy.Com' } Cc: 'jim-at-proscitech.com.au' } Subject: RE: Film vs Digital } } } The world is full of possible solutions, but are they practical.? } To produce high-resolution, dark-field or any others TEM images that } require } more electrons to form a clear image, Mike Bode would use multiple } digital } exposures. The exposures could be layered and combined into one superior } image. } This image would be made up of more pixel and is formed by more } electrons and } so would be noise-free and hence could be further enlarged then } otherwise } possible. Perhaps. } Beam blanking would largely save the specimen from beam damage and drift } could } be compensated for by matching up the digitals. Great. } How much time is required between exposures to transfer a minimum 10mb } image } per exposure? What would be the total time from focusing to the last } exposure? } What about Z-drift in the interim requiring objective changes and what } about } the total cost of this additional get-up. The mind boggles at a through } focus } series. } When pushing the limits a piece of film seems more effective, cheaper } and fa } ster. } Again, I don't doubt that there is now a large place for digital in TEM, } but } its no panacea. } Cheers } Jim Darley } ProSciTech Microscopy PLUS } PO Box 111, Thuringowa QLD 4817 Australia } Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com } Great microscopy catalogue, 500 Links, MSDS, User Notes } www.proscitech.com } } On Friday, June 02, 2000 1:24 AM, Mike Bode [SMTP:mb-at-soft-imaging.com] } wrote: } } } } No, it's not really a problem. It's been done with low density } } microscopy all the time. Granted, there are some technical aspects to } be } } overcome, but (and I can only speak for ourselves) we have done that } on } } a number of microscopes. You are of course correct, that 10 images at } } 0.8 seconds take longer than 8 seconds as the image has to be } } transferred, etc. BUT: that's what beam blankers are for. It is pretty } } straightforward to take an image at 0.8 seconds, then blank the beam } } very quickly before taking the next image. That way you get pretty } close } } to the 8 sec total exposure. If there is no beam blanker on the } } microscope, in most cases it can be added. } } } } I am not sure what you mean by "too sensitive". The cameras are } usually } } constructed so that 1 electron from the beam creates between a few } tenth } } to a few counts (these are all statistical data, of course). The well } } width divided by this sensitivity then determines, how many primary } } electrons are needed to fully expose one pixel. For example, if the } well } } width is 50,000 electrons and the sensitivity is 1 count/electron, one } } needs 50,000 primary electrons to fill the well. This translates into } } roughly a 0.4% statistical error. } } } } } From a practical standpoint: You can take images with most cameras } when } } the exposure meter on the microscope reads a couple of seconds without } } overexposing the camera. On the other hand, you can reduce the } intensity } } of the beam until you see single electron events. } } } } The one area where CCD cameras may be too sensitive is diffraction. } The } } normally huge intensity in the transmitted beam often leads to } } saturation. In CCDs this can lead to blooming (the intensity spills } over } } into neighboring pixels). This can be taken care of with special chips } } that have anti-blooming features, but this usually has some other } } drawbacks. Again, this can also be overcome somewhat with multiple } } exposures. Film behaves more civilized here, as it simply stops } } responding to the electrons, but this makes film more or less useless } } for quantitative measurements of diffraction patterns. I have done } } diffraction with CCDs many times and though it does require some } } tweaking, one can get very good results from them. } } } } Michael Bode, Ph.D. } } Soft Imaging System Corp. } } 1675 Carr St., #105N } } Lakewood, CO 80215 } } =================================== } } 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: jim [mailto:jim-at-proscitech.com.au] } } Sent: Wednesday, May 31, 2000 9:37 PM } } To: 'Mike Bode'; 'Microscopy-at-MSA.Microscopy.Com' } } Subject: RE: Film vs Digital } } } } } } } ------------------------------------------------------------------------ } } 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 } } } -----------------------------------------------------------------------. } } } } } } Yes, it could be done "in theory". Somebody would need to figure out } the } } } } software and perhaps modify the hardware. Then we would find that the } } total } } exposure of the specimen to the electron beam maybe a muliple of the } } film's } } exposure. Afterall, an 8 sec film exposure would not amount in digital } } to } } 10x0.8, but we would require considerable time in between exposures. } } Since the } } problems in the discussed circumstances are specimen movement and beam } } damage, } } it seems that taking multiple exposures is a poor option. } } } } Digital cameras are for some situation too sensitive to electron } } exposure. } } Cutting back on electrons is no option since its the electrons that } form } } the } } image in the first instance. } } Much easier in light microscopy . . . insert a neutral density } filter. } } Cheers } } Jim Darley } } ProSciTech Microscopy PLUS } } PO Box 111, Thuringowa QLD 4817 Australia } } Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com } } Great microscopy catalogue, 500 Links, MSDS, User Notes } } www.proscitech.com } }
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Jonathon Low kV is preferred for some biological applications chiefly because biologists experience difficulty getting enough contrast out of their specimens. As Dr Alves de Matos rightly says, low kV is more damaging to specimens than high kV, because the penetrating power of low-kV electrons is lower and therefore more beam energy is lost to the specimen. (We think of this exactly the other way round for SEM, but that's a story for another day!).
The main reason for the apparent shift in biologist's attitudes is that many are now less concerned with cell and tissue-level ultrastructure and increasingly interested in imaging macromolecules. 200kV or 300kV machines are preferred for cryo- microscopy of macromolecules in vitrified ice because the resolution is a lot better than at 80 or 120kV, and also because 200kV causes less specimen damage.
However, the cost of resolution either from higher kVs or the shorter working distance objective lenses favoured by materials scientists is poorer image contrast. Philips therefore supply "Biotwin" versions of some of their TEM range which use a long focal length objective which trading a little resolution for a little more contrast. This optical configuration is coincidentally very good for thickish specimens such as ultrathin biological sections where the ultimate resolution of the machine is pretty much academic anyway.
Images of thick (#100nm) specimens such as FIB sections of microelectronic devices seem crisper and more contrasty in our CM120 Biotwin than in a 200KV machine. Chris
} From: "A.P.Alves de Matos" {apmatos-at-ip.pt} To: {Microscopy-at-sparc5.microscopy.com}
Dear all,
1) I am working on oak trees, and I would like to describe the ultrastucture of leaves; also I am looking for a protocol (very detailed) to do it. 2) On oak leaves I would like to localize the superoxyde dismutase(s) enzymes by immunocytochemistry, also I am looking for a protocol (very detailed) to do it.
Thanks in advances. Dr. Didier Le Thiec
-------------------------------------------- Didier Le Thiec I.N.R.A. Centre de Recherches Forestieres Unite d'Ecophysiologie Forestiere Laboratoire de Pollution Atmospherique 54280 Champenoux - France
Following on from the very successful meeting held in 1999, this will review the current state-of-the-art of the UK's various FEG/TEM installations in operation.
The meeting will begin with a special keynote lecture on
Advanced TEM Instrumentation for Solving Critical Problems in Materials Science given by Professor Manfred Rhle Max-Planck Institute fr Metallforschung, Stuttgart
This lecture will include some news on the new German FEG/TEM project.
There are now ten FEG/TEM facilities in the UK (some with more than one instrument) and the main programme will consist of a series of invited presentations from each of the sites, reviewing the latest developments in instrumentation and covering specialised techniques such as holography, energy-filtered imaging, spectroscopic imaging, nano-scale analysis, etc.
With JIF, JREI and other special equipment funding now becoming available it is likely that there will be other sophisticated FEG/TEM installations in place in the near future. With this in view the meeting will also include a forward look: what will the next generation of instruments be like?
Speakers:
John Hutchison (University of Oxford)
Key-note Lecture - Professor Manfred RÄhle (Stuttgart) Per Bullough (University of Sheffield) Marin van Heel (Imperial College) John Berriman (MRC, Cambridge) Tony Cullis (University of Sheffield) Ian Jones (University of Birmingham) Rik Brydson (University of Leeds) Jeremy Sloan (University of Oxford) David Cherns (University of Bristol) Stephen McVitie (Glasgow University) Stephen Lloyd (University of Cambridge) John Titchmarsh (University of Oxford) Paul Midgley (University of Cambridge)
The meeting will conclude with the RMS AGM and Presidential Address.
For further details contact: Dr Paul Midgley (EMAG): pam33-at-cam.ac.uk or Dr John Hutchison (RMS): john.hutchison-at-materials.ox.ac.uk
Greetings, Just for the record, Tech Pan is interesting in that you can vary the grain size over an amzingly wide range by choice of developer and exposure conditions. You can technidol LC (I think) and get the teeeny weeeeny grains previously mentioned, or use D19 (I think) and get really high contrast big grains.
Just my few exposed halides, Tobias
} Chris, } This is because Kodak use EMs to measure the grain size of their silver } halide grains. Tech Pan has a very small grain size (slow film, low } ISO/ASA number) and is therefore capable of greater enlargement than } film with a larger grains. The larger the grain the faster the film } (higher ISO/ASA number). } Regards } JVN } JVN } } 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 } } -----------------------------------------------------------------------. } } } } I just came across the following pearl in a 1982 Kodak publication, } } number PDS 61 "Selecting film from Kodak for photomicrography". } } } } "The (Technical Pan) 2415 film has very low granularity and is } } capable of very great enlargement. In some cases the resolving } } power may be beyond that of the microscope image" } } } } How do they know! } } Answers on a postcard please .... } } } } Chris } } ===================================================================== } } DR CHRIS JEFFREE } } BIOSEM - BIOLOGICAL SCIENCES EM FACILITY } } UNIVERSITY OF EDINBURGH } } Daniel Rutherford Building } } King's Buildings, Mayfield Road } } EDINBURGH, EH9 3JH, Scotland, UK } } Tel. #44 131 650 5345 } } FAX. #44 131 650 6563 } } Mobile 0410 585 401 } } email c.jeffree-at-ed.ac.uk } } SEM / TEM bookings sem-at-ed.ac.uk } } ===================================================================== } } -- } **************************************************** } John V Nailon } Operations Manager } Centre for Microscopy and Microanalysis } The University of Queensland } St. Lucia Queensland 4072 } Phone: +61-7-3365-4214 } Fax: +61-7-3365-4422 } WWW: http://www.uq.edu.au/nanoworld/allstaff.html#Nailon } ****************************************************
Two weeks ago I posted a short message about the Ocean Optics spectrometer I had just received for doing Plasma diagnostics. I tried it and have found it very useful for examining a plasma cleaning process and optimizing operating conditions. I purchased a USB 2000 model that plugs into a USB port of a PC for less than $3000. Ocean Optics, 727-733-2447, www.OceanOptics.com
Ronald Vane XEI Scientific NEW WEB SITE! www.SEMCLEAN.com
-----Original Message----- } From: Ronald Vane {RVaneXEI-at-concentric.net} To: MSA listserver {Microscopy-at-sparc5.microscopy.com} ; Dr. Gary Gaugler {gary-at-gaugler.com}
My SEM facility has just been approached, and in the interest in providing what they want for a price based on an average, can I query the SEM community as to what they charge for commercial release of their SEM imagery. I'm looking for $/image, but if you charge on any type of sliding scale, I'd be interested in that too. I believe as well I shouldn't be undercutting commercial facilities, so I'm especially interested in these numbers. Reply to me direct, and I'll respond back to the list with the average, max and min, and no names.
cheerios, =shAf= :o)
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - mshaf-at-darkwing.uoregon.edu Geological Science's Electron Probe Facility - University of Oregon http://epmalab.uoregon.edu/
I work for North Carolina State University and we are looking in to using Fluorescence in-situ Hybridization (FISH) for identification of bacteria in biofilm. Do you know of any short courses in FISH here in the USA? I found some out of the country but would prefer to take a course in USA.
Thanks for your time
Tracey L. Daly {tldaly-at-unity.ncsu.edu}
Ron Anderson, IBM, Hopewell Jct., New York, USA. anderron-at-us.ibm.com
IBM Analytical Services; http://www.chips.ibm.com/services/asg
I have an associate who is doing cathodoluminescence studies on materials. He would like to extend his observations into the UV for a limited number of tests on a a few samples. He is presently limited by the optics of the microscope and the fiber optics cable connecting the microscope ocular to the input of the spectrometer. Does anyone have any suggestions for optical microscopes with UV optics that might be available for short term lease or loan that he could consider? Objective magnifications of 5X to 10 X and a 5X or 10X ocular would probably suffice.
Thanks,
Don Marshall
Donald J. Marshall Relion Industries P.O. Box 12 Bedford, MA 01730 Ph: 781-275-4695 FAX: 781-271-0252 email dmrelion-at-world.std.com
Cathodoluminescence, mass spectroscopy, electron beam technology
"A weed is a flower out of place."
(Please note: Do not send email with attachments to this address. Instead, send it to donbarlen-at-aol.com. Thank you.)
The Department of Microscopy and Microanalysis at Abbott Laboratories is recruiting an Electron Microscopist for its Biological Microscopy group. This group provides ultrastructural pathology support for Nonclinical Drug Safety studies, as well as for other biological microscopy projects, such as cell screening, virus identification and counting, and immunolabeling.
Requirements for this position include: a Masters' degree in a biological field, such as cell biology, anatomy, or zoology a thorough understanding of mammalian histology and ultrastructure excellent technical skills including tissue collection at necropsy, tissue and cell processing for TEM, sectioning, staining, operation of electron microscopes and related equipment, darkroom procedures ability to communicate information in technical reports and in oral presentations
Highly desirable, but not essential, are: experience in ultrastructural pathology or toxicologic pathology working knowledge of SEM specimen preparation and instrumentation working knowledge of immunocytochemistry, in situ hybridization, and other labeling methods at light or electron microscopic level familiarity with Good Laboratory Practices
We are looking for a team player with outstanding interpersonal skills and the ability to adjust readily to rapidly changing priorities and shifting deadlines. The ability to communicate clearly, both verbally and in writing, is essential. Careful attention to detail and accuracy are required.
The Department of Microscopy and Microanalysis provides corporate-wide support in Biological and Materials Microscopy to all divisions of Abbott Laboratories. The facility houses two TEMs (a Philips CM12 STEM and a LEO 910), two SEMs (a Philips XL30-FEG and AMRAY 1830i), three EDXS systems, a BioRad confocal scanning laser microscope, several fluorescent and light microscopes (polarized light, DIC), and a Quantimet Image Analysis system. We also have an Arcturus PixCell laser capture microdissection system and a Becton Dickinson FACSCalibur flow cytometer. Microtomes include Reichert Ultracut E and S ultramicrotomes, RMC 6000 XL cryoultrotome, Microm histological microtome, and Microm HM500 cryostat.
Please send letters of application and resumes to:
Jane A. Fagerland, Ph.D. Abbott Laboratories D45M/AP31 200 Abbott Park RD. Abbott Park IL 60064-6202
Dear Microscopists, I'm just curious if something is wrong on my end or I'm unsuscribed against my will. I did not receive any posting during the past two-three weeks, and this is impossible in view of previously received 10-20 postings/day. Is there any explanation? Kris Dr. Kristof Kovacs Associate Professor President, Hungarian Society for Microscopy Phone: +36-(88)-421-684 Fax: +36-(88)-328-643 Mailing Address: University of Veszprem, P.O.Box 158, Veszprem H-8201 Hungary
Dear all, I'm currently having a few problems preparing good cross-section specimens of ceramic thin films on LaAlO3 or NdGaO3 substrates.
I can cut them okay, but thinning them often results in cracking as the thickness goes below 100 microns. I have a Gatan model 623 disk grinder and currently have access to SiC abrasive papers to grit sizes of 1500 or 2000, and also have the Gatan diamond polishing disks and specimen lapping kit.
The problems are as follows: If we use the 1500 or 2000 grit SiC, we can often get the samples thin enough, but with poor surface finish which needs to be improved with dimpling (on both sides), we also risk cracking the sample as SiC papers often contain imperfections.
Using the Gatan specimen lapping kit, I find that as the disk grinder is altered to grind off a further 10 microns, the edge of the sample often catches on the diamond disk and tears some of the diamond coating off, leaving a lump on the surface which then risks catching the specimen and cracking it.
I have one possible alternative approach which I used in Sweden last year (with SiAlON ceramics) involving attaching the sample to a glass slide and polishing it using diamond spray on non-absorbent paper. I may consider trying this with these new materials.
Other ideas would be welcome, however, as would suggestions of how to improve my present method. Please note that I do not have the budget to buy any expensive new polishing equipment (such as a tripod polisher, for instance), so the most welcome suggestions would be those that involve improvements to current techniques, use of different types of polishing consumables, etc..
I hope to hear several suggestions, both from users and from the companies who producing polishing and lapping equipment. Why not post them with the list so we can all benefit from the sharing of experience?
Best wishes
===== Ian MacLaren Beijing Laboratory of Electron Microscopy Chinese Academy of Sciences, P.O. Box 2724 100080 Beijing China General Email: ian.maclaren-at-physics.org Work (esp. large attachments): maclaren-at-image.blem.ac.cn
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Since I am usually more involved in material analysis I am a little lost on the biological field. But now I am in the need of a good review article concerning the application of (analytical) TEM to biological materials. I am not looking for sophisticated latest developements but rather for basic applications with some examples, if possible including some examples of analytical TEM.
Hope to get some input :)
Petra
-------------------------------------------------------------- Dr. Petra Wahlbring Centre de Recherche Public - Gabriel Lippmann Laboratoire d'Analyse des Materiaux (LAM) 162a, av. de la Faiencerie L-1511 Luxembourg tel. +352-466644-402 fax +352-466644-400 e-mail: petra.wahlbring-at-crpgl.lu Visit our WWW site! http://www.crpgl.lu/~wahlbrin
ONLY 8 WEEKS TO GO UNTIL ICHC 2000 (3-8 Sept. York University, UK)
Abstracts are still welcome.
Register by the week or combinations of days.
For more details follow the links to the ICHC 2000 web-site below.
Best wishes
Gary Coulton Dr. Gary Coulton Molecular Pathology Division of Biomedical Sciences Imperial College School of Medicine The Sir Alexander Fleming Building South Kensington London SW7 2AZ
------------------------------------- Announcing the 11th International Congress of Histochemistry and Cytochemistry (ICHC 2000)
"Cell Biology and Imaging Tools for the New Century"
September 3-8, 2000, York, United Kingdom
ICHC 2000 comprises 27 symposia addressing latest developments and applications of histochemistry and cytochemistry in the life sciences including medicine.
Many leading experts to speak
8 weeks to go! Register for the week or by the day!
For further details go to http://www.med.ic.ac.uk/external/ichc_2000
I was happy to leave things at that, but there are a couple of things that Mike has raised that need an answer: Your actual quote concerning Ansel Adams was "Having said that and looking at a print of Ansel Adams, I am glad there is film, though!!" This is rather at odds with the new proclamation } " I mentioned Adams because I like his pictures. If he had taken them with } a digital camera I would have liked them just as much. Lots of people have made excellent compositions and many have make technically superior prints. Adam's has excelled by consistently winning on both account. Its a bit strange, but Mike I'll remind you why you his prints: It is not just Adam's great compositions but also the incredible sharpness and tonal range, which is only possible by huge data density and impossible with a 2.5mb enlarged print. Digital not only has problems with occasional great demands on print magnification (pixel size limitations) and lack of electron density (too short exposures), but because TEM negs, like Adam's prints, are capable of a terrific tonal range and resolution. An image appears better if theoretical data minima are exceeded.
I have been a stickler for showing possible conflict of interest. I realize now that I should have made a disclaimer along the way. I did not, simply because it never occurred to me that I could have a real or imagined gain. My aspirations are different, but since we are all subject to self-delusion I'll add the reality: We have very little mark-up on film and cannot export beyond New Zealand. For Australian research institutions money has been very tight and equipment sales have been low for some years. I doubt that at our exchange rate any digital TEM cameras will be installed here during the next couple of years at least. I think that my special friend Chuck is more likely to retain more film sales than ProSciTech may have lost - if anybody was influenced either way.
The word belief is fitting for all of this. Whatever the arguments, I expect people retain their "film versus digital" beliefs. Mike, we can lead a horse to water, but just try to make it float on its back. When you do, send a picture and I won't care if its digital. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Tuesday, June 06, 2000 1:10 AM, Mike Bode [SMTP:mb-at-soft-imaging.com] wrote: } Jim wrote: } } 'I have two replies and Mike my well have four replies to these.' } } I'll try. But I agree with you that we should let this thread die. jim, } it seems you're a bit angy at me. If I unintentionally stepped on your } toe I apologize. I did not expect that we would agree 100%, as you are } selling film and I am involved in digital systems. I just hope that some } other readers found some of this useful. I don't consider myself a } "digital fanatic". This film vs. digital issue has many more facets, } some of which we did not even touch, and which can be just as } entertaining. } } Jim wrote: } } 'Mike reinforced and strengthened that argument, finishing with the note } that he is glad for film when looking at prints by Ansel Adams.' } } Just for the record: I usually do not take a magnifying glass to photos. } I mentioned Adams because I like his pictures. If he had taken them with } a digital camera I would have liked them just as much. I also like some } modern art paintings. That does not mean we should start drawing what we } see in the microscopes rather than taking pictures ;-) } } Jim wrote: } } 'Incidentally, from the outset I cited increased "enlargability" to } obtain high } resolution TEM' } } Well, you said 'When great enlargements are required film is superior'. } Perhaps I misunderstood. I thought, the term great enlargements referred } to the microscope and meant 'small details', which you can take easier } with a digital camera (no time delay between seeing something and taking } an image, no mechanical vibration due to film movement, etc.). I have } said many times before that film may be better if high resolution AND } large field of view is required. } } Jim wrote: } } 'in any case: Show Sergey!' } } I'm in back-channel correspondence with him. } } } } Michael } } Michael Bode, Ph.D. } Soft Imaging System Corp. } 1675 Carr St., #105N } Lakewood, CO 80215 } =================================== } 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: jim [mailto:jim-at-proscitech.com.au] } Sent: Sunday, June 04, 2000 2:34 AM } To: 'Mike Bode'; 'Microscopy-at-MSA.Microscopy.Com' } Subject: RE: Film vs Digital } } } ------------------------------------------------------------------------ } 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 two replies and Mike my well have four replies to these. } 1. There has been a parallel discussion concerning resolution of } film versus } digital images. Clearly in raw power digital cannot compete since film } has } multi gigabyte capacity. I added to that thread that what matters is: } does } digital have enough power and that frequently it would. Mike reinforced } and } strengthened that argument, finishing with the note that he is glad for } film } when looking at prints by Ansel Adams. } (Ansel Adams until about 30 years ago carted for decades large format } cameras } through US National Parks, especially Yosemite, producing fantastic } landscape } photographs and books) } Adams' limited edition prints were contacts of 5x7 and 8x10" inch sheet } film, } probably rated at 400 ISO. The line resolution of such prints much } exceeds our } eyes' resolution, but still results in superior gradation and detail. } TEM film, } even when much enlarged has such details too. } Why Mike, should we accept 2.5 Mb? That is a splendid file size for SEM, } and } because of the limited enlargability of light microscopy (concrete } ceiling due } to wavelengths) it's reasonable, but minimal for light microscopy. TEM } can do } and deserves better. } } Incidentally, from the outset I cited increased "enlargability" to } obtain high } resolution TEM, as one of films major advantages, since greater depths } of field } at moderate powers makes high powers through photo enlarging a desirable } } technique. The small additional magnification yielded by placing a } digital } camera lower in the column does not compensate. So greater } "enlargability" of } digitals would be desirable, but is limited by pixel size. } } 2 The thread was initiated by Sergey. He had problems visualising } certain } specimens in dark field. The use of his "beaut" digital TEM camera made } things } worse. I pointed out that the shorter exposure reduced the number of } electrons } forming the image, hence more noise. I believe that a good part of } Mike's case } will be settled in his favour when we hear "Eureka" from Sergey's lab. I } don't } doubt that much more can be done with digital now and that further } improvements } are on the way. Mike's "solution" may well be possible, but I don't } believe its } a snap; in any case: Show Sergey! } Cheers } Jim Darley } ProSciTech Microscopy PLUS } PO Box 111, Thuringowa QLD 4817 Australia } Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com } Great microscopy catalogue, 500 Links, MSDS, User Notes } www.proscitech.com } } On Saturday, June 03, 2000 2:58 AM, Mike Bode [SMTP:mb-at-Soft-Imaging.com] } wrote: } } } } Well, let's see: } } } } Jim wrote: } } } } Mike Bode would use multiple digital } } exposures. The exposures could be layered and combined into one } superior } } image. } } This image would be made up of more pixel and is formed by more } } electrons and } } so would be noise-free and hence could be further enlarged then } } otherwise } } possible. Perhaps. } } } } No, I did not talk about further enlargements. All I wanted to say is, } } that a more noise-free image can be achieved by adding multiple } images, } } and that this also to some extent helps with drift of the sample } during } } acquisition. } } } } Jim wrote: } } } } How much time is required between exposures to transfer a minimum 10mb } } image } } per exposure? } } } } How did you arrive at 10 MB? A 1280x1024 image with 16 bit pixel } } information is about 2.5 MB (uncompressed). We acquire about 10 of } those } } per second and transfer them across the PC bus to the display. Putting } } them on them into Memory might add a few tenth of a second. Writing to } } HD can be done after all images are acquired. } } } } Jim wrote: } } } } What would be the total time from focusing to the last exposure? } } What about Z-drift in the interim requiring objective changes } } } } Why would we have to worry about that, if we don't have to worry about } } that when taking the image on film? In fact, we could take care of } this } } by looking at the image between exposures and correct for z-drift. } } However, as you said, that would add to the overall time and exposure. } I } } was comparing a normal dark field image taken on film at 8 seconds } with } } acquiring the same image on a "too sensitive" CCD camera by adding up } 10 } } consecutive .8 second images. Why would the sample drift (in x, y or } z) } } substantially more in 8+delta seconds than in 8? } } } } Jim wrote: } } } } what about the total cost of this additional get-up } } } } That of course depends on the microscope and there is no general } answer. } } For example on a LEO 912 I believe the blanker is standard. The } } additional cost to use an acquisition scheme like this with our } software } } is $0 plus perhaps a bit of time to write a small macro. On other } } microscopes one might have to add a beam blanker and perhaps a control } } mechanism for the beam blanker. But I would guess, that this cost is } not } } very high. All modern microscopes are computer controller anyway, so } it } } is most likely just a control command that needs to be sent to the } } microscope over a serial port if the beam blanker is installed. Piece } of } } cake. } } } } Jim wrote: } } } } The mind boggles at a through focus series. } } } } You're right here. But I don't think we were talking about } through-focus } } series. Incidentally, we do through-focus series on light microscopes } } and reconstruction routinely. Takes a few images at different focus } (or } } for a light microscope: stage) settings. The rest is done off-line. } } Takes maybe a couple of minutes for about 20 images of about 1kx1k. I } } agree that TEM is different here and much more complicated due to the } } complicated Contrast Transfer Function. However, this could in } principle } } be sorted out. } } } } Jim wrote: } } } } Again, I don't doubt that there is now a large place for digital in } TEM, } } but } } its no panacea. } } } } I also agree with you on that one. But using the additional computer } } possibilities of digital imaging might take you further than expected. } } } } Michael } } } } } } Michael Bode, Ph.D. } } Soft Imaging System Corp. } } 1675 Carr St., #105N } } Lakewood, CO 80215 } } =================================== } } 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: jim [mailto:jim-at-proscitech.com.au] } } Sent: Friday, June 02, 2000 5:15 AM } } To: 'Mike Bode'; 'Microscopy-at-MSA.Microscopy.Com' } } Cc: 'jim-at-proscitech.com.au' } } Subject: RE: Film vs Digital } } } } } } The world is full of possible solutions, but are they practical.? } } To produce high-resolution, dark-field or any others TEM images that } } require } } more electrons to form a clear image, Mike Bode would use multiple } } digital } } exposures. The exposures could be layered and combined into one } superior } } image. } } This image would be made up of more pixel and is formed by more } } electrons and } } so would be noise-free and hence could be further enlarged then } } otherwise } } possible. Perhaps. } } Beam blanking would largely save the specimen from beam damage and } drift } } could } } be compensated for by matching up the digitals. Great. } } How much time is required between exposures to transfer a minimum 10mb } } image } } per exposure? What would be the total time from focusing to the last } } exposure? } } What about Z-drift in the interim requiring objective changes and what } } about } } the total cost of this additional get-up. The mind boggles at a } through } } focus } } series. } } When pushing the limits a piece of film seems more effective, cheaper } } and fa } } ster. } } Again, I don't doubt that there is now a large place for digital in } TEM, } } but } } its no panacea. } } Cheers } } Jim Darley } } ProSciTech Microscopy PLUS } } PO Box 111, Thuringowa QLD 4817 Australia } } Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com } } Great microscopy catalogue, 500 Links, MSDS, User Notes } } www.proscitech.com } } } } On Friday, June 02, 2000 1:24 AM, Mike Bode [SMTP:mb-at-soft-imaging.com] } } wrote: } } } } } } No, it's not really a problem. It's been done with low density } } } microscopy all the time. Granted, there are some technical aspects } to } } be } } } overcome, but (and I can only speak for ourselves) we have done that } } on } } } a number of microscopes. You are of course correct, that 10 images } at } } } 0.8 seconds take longer than 8 seconds as the image has to be } } } transferred, etc. BUT: that's what beam blankers are for. It is } pretty } } } straightforward to take an image at 0.8 seconds, then blank the beam } } } very quickly before taking the next image. That way you get pretty } } close } } } to the 8 sec total exposure. If there is no beam blanker on the } } } microscope, in most cases it can be added. } } } } } } I am not sure what you mean by "too sensitive". The cameras are } } usually } } } constructed so that 1 electron from the beam creates between a few } } tenth } } } to a few counts (these are all statistical data, of course). The } well } } } width divided by this sensitivity then determines, how many primary } } } electrons are needed to fully expose one pixel. For example, if the } } well } } } width is 50,000 electrons and the sensitivity is 1 count/electron, } one } } } needs 50,000 primary electrons to fill the well. This translates } into } } } roughly a 0.4% statistical error. } } } } } } } From a practical standpoint: You can take images with most cameras } } when } } } the exposure meter on the microscope reads a couple of seconds } without } } } overexposing the camera. On the other hand, you can reduce the } } intensity } } } of the beam until you see single electron events. } } } } } } The one area where CCD cameras may be too sensitive is diffraction. } } The } } } normally huge intensity in the transmitted beam often leads to } } } saturation. In CCDs this can lead to blooming (the intensity spills } } over } } } into neighboring pixels). This can be taken care of with special } chips } } } that have anti-blooming features, but this usually has some other } } } drawbacks. Again, this can also be overcome somewhat with multiple } } } exposures. Film behaves more civilized here, as it simply stops } } } responding to the electrons, but this makes film more or less } useless } } } for quantitative measurements of diffraction patterns. I have done } } } diffraction with CCDs many times and though it does require some } } } tweaking, one can get very good results from them. } } } } } } Michael Bode, Ph.D. } } } Soft Imaging System Corp. } } } 1675 Carr St., #105N } } } Lakewood, CO 80215 } } } =================================== } } } 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: jim [mailto:jim-at-proscitech.com.au] } } } Sent: Wednesday, May 31, 2000 9:37 PM } } } To: 'Mike Bode'; 'Microscopy-at-MSA.Microscopy.Com' } } } Subject: RE: Film vs Digital } } } } } } } } } } } } ------------------------------------------------------------------------ } } } 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 } } } } } } -----------------------------------------------------------------------. } } } } } } } } } Yes, it could be done "in theory". Somebody would need to figure out } } the } } } } } } software and perhaps modify the hardware. Then we would find that } the } } } total } } } exposure of the specimen to the electron beam maybe a muliple of the } } } film's } } } exposure. Afterall, an 8 sec film exposure would not amount in } digital } } } to } } } 10x0.8, but we would require considerable time in between exposures. } } } Since the } } } problems in the discussed circumstances are specimen movement and } beam } } } damage, } } } it seems that taking multiple exposures is a poor option. } } } } } } Digital cameras are for some situation too sensitive to electron } } } exposure. } } } Cutting back on electrons is no option since its the electrons that } } form } } } the } } } image in the first instance. } } } Much easier in light microscopy . . . insert a neutral density } } filter. } } } Cheers } } } Jim Darley } } } ProSciTech Microscopy PLUS } } } PO Box 111, Thuringowa QLD 4817 Australia } } } Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com } } } Great microscopy catalogue, 500 Links, MSDS, User Notes } } } www.proscitech.com } } }
This sounds like a perfect application of the small angle cleavage technique. You might want to contact Ray Tweston at Univ. of Illinois because I think that we might have successfully prepared one of these while I visited there. At any rate, the technique is very inexpensive and you can prepare sample of superior quality. Get your hands on John McCaffrey and my paper in the MRS TEM sample Prep IV book (vol 480). It has a detailed description on how to do it. South Bay Technology sells the Microcleave kit and you should contact them as well.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
} -----Original Message----- } From: Ian MacLaren [mailto:maclariz-at-yahoo.co.uk] } Sent: Wednesday, June 07, 2000 3:28 AM } To: Microscopy list } Subject: Ceramic cross-section preparation } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Dear all, } I'm currently having a few problems preparing good } cross-section specimens } of ceramic thin films on LaAlO3 or NdGaO3 substrates. } } I can cut them okay, but thinning them often results in } cracking as the } thickness goes below 100 microns. I have a Gatan model 623 } disk grinder and } currently have access to SiC abrasive papers to grit sizes of } 1500 or 2000, } and also have the Gatan diamond polishing disks and specimen } lapping kit. } } The problems are as follows: } If we use the 1500 or 2000 grit SiC, we can often get the } samples thin } enough, but with poor surface finish which needs to be improved with } dimpling (on both sides), we also risk cracking the sample as } SiC papers } often contain imperfections. } } Using the Gatan specimen lapping kit, I find that as the disk } grinder is } altered to grind off a further 10 microns, the edge of the } sample often } catches on the diamond disk and tears some of the diamond } coating off, } leaving a lump on the surface which then risks catching the } specimen and } cracking it. } } I have one possible alternative approach which I used in } Sweden last year } (with SiAlON ceramics) involving attaching the sample to a } glass slide and } polishing it using diamond spray on non-absorbent paper. I } may consider } trying this with these new materials. } } Other ideas would be welcome, however, as would suggestions of how to } improve my present method. Please note that I do not have } the budget to buy } any expensive new polishing equipment (such as a tripod polisher, for } instance), so the most welcome suggestions would be those } that involve } improvements to current techniques, use of different types of } polishing } consumables, etc.. } } I hope to hear several suggestions, both from users and from } the companies } who producing polishing and lapping equipment. Why not post } them with the } list so we can all benefit from the sharing of experience? } } Best wishes } } ===== } Ian MacLaren } Beijing Laboratory of Electron Microscopy } Chinese Academy of Sciences, P.O. Box 2724 } 100080 Beijing } China } General Email: ian.maclaren-at-physics.org } Work (esp. large attachments): maclaren-at-image.blem.ac.cn } } ____________________________________________________________ } Do You Yahoo!? } Get your free -at-yahoo.co.uk address at http://mail.yahoo.co.uk } or your free -at-yahoo.ie address at http://mail.yahoo.ie }
Postdoctoral Position Immediately Available In situ UHV-TEM of Nanoparticle Reactions and Metal Oxidation
Materials Research Laboratory, University of Illinois at Urbana-Champaign and University of Pittsburgh
A postdoctoral position is immediately available in the Materials Research Laboratory at the University of Illinois at Urbana-Champaign in the area of nano-reactions and in situ UHV-TEM. This position is jointly sponsored between Professor Robert Averback (University of Illinois at Urbana-Champaign) and Professor Judith Yang (University of Pittsburgh).
The research project is two-fold: 1. Surface oxidation kinetics of metals and 2. Nanoparticle reactions/sintering. The research project is to combine the unique experimental information obtainable from in situ UHV-TEM and compare with theoretical models of these nanoscale reactions. The position requires a PhD in physical sciences/engineering. Hands-on experience in TEM techniques is highly desirable. The position is open to all qualified candidates and has an anticipated duration of 2 years. If you are interested in this opportunity, please send a resume and names of three references to the address below.
Dr. Judith C. Yang Assistant Professor Dept. of Materials Science & Engineering 848 Benedum Hall University of Pittsburgh Pittsburgh, PA 15261
I can reccomend you the following books: 1. D.C.Joy, A.D.Romig and J.I.Goldstein, "Principles of analytical electron microscopy", Plenum Press, 1989; it contains three chapters dedicated to bilogy: chapter 6, 12 and 13 2. J.J.Hren, J.I.Goldstein and D.C.Joy, "Introduction to AEM", Plenum Press, 1979; it contains chapters dedicated to biology. I hope this helps.
Corneliu Sarbu Dept.MTM KULeuven Belgium
- - - - - - - - - - - - - - Original Message - - - - - - - - - - - - - - ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Since I am usually more involved in material analysis I am a little lost on the biological field. But now I am in the need of a good review article concerning the application of (analytical) TEM to biological materials. I am not looking for sophisticated latest developements but rather for basic applications with some examples, if possible including some examples of analytical TEM.
Hope to get some input :)
Petra
-------------------------------------------------------------- Dr. Petra Wahlbring Centre de Recherche Public - Gabriel Lippmann Laboratoire d'Analyse des Materiaux (LAM) 162a, av. de la Faiencerie L-1511 Luxembourg tel. +352-466644-402 fax +352-466644-400 e-mail: petra.wahlbring-at-crpgl.lu Visit our WWW site! http://www.crpgl.lu/~wahlbrin
- - - - - - - - - - - - End of Original Message - - - - - - - - - - - -
Question: What is the proper procedure for increasing the filament current on a 2010 with a LaB6? How fast can you go up? Should the current be increased at a linear rate or should the rate be tapered off as you reach saturation? The manual suggests 30 sec/graduation while the service techs suggest a much slower rate.
Thanks, Kim ************************************************************** Kim W. Pierson, Ph.D. Dept of Physics & Astronmoy University of Wisconsin-Eau Claire (715) 836-5009 FAX 836-3955
I have a few comments for you here that may help. This is a little touchy as I don't want to knock my competitor's equipment. The Model 623 Disk Grinder that you have is going to cause problems as you describe because of it's design. The way that grinder works is that you use the dial on the top to make your sample extend below the base of the grinder. When you begin polishing, the entire weight of the grinder plus whatever weight you are applying by hand is being transferred to your very thin 3mm diameter sample. In addition to the additional weight, you can tend to apply too much pressure on one side of the sample which would cause the sample to "catch" on the diamond film as you describe.
We manufacture a series of polishing fixtures that are designed to prevent these problems. They are gravity feed fixtures. On these fixtures, the dial gauge at the top simply sets the amount of material that will be fed into the abrasive film. The only weight your sample sees is the weight of the central piston (with some of our fixtures, this weight can be counterbalanced to approach zero). The weight of the rest of the fixture is never transferred to the sample. Also, the sample surface that is being polished is always co-planar with the base of the polishing fixture. This ensures that you cannot apply uneven pressure and that you won't experience the "catching" that you described.
Now the good news. There is one of these fixtures (Model 150) already in use in your facility. There is also the Tripod Polisherš, Model 920 Lapping & Polishing Machine, Model 850 Wire Saw and Model 650 Diamond Wheel Saw. By separate e-mail,I will give you the details on where these are located and who you should contact to access them.
I also saw Scott Walck's post on the Microcleave technique. I will include a PDF of the paper he was talking about by separate e-mail.
If you have any questions or if I can be of any additional assistance, please contact me directly off-line. I hope this helps.
DISCLAIMER: South Bay Technology produces equipment and supplies as described above and, therefore, has a vested interest in promoting their use.
David Henriks Vice President 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
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy. Message text written by INTERNET:ian.maclaren-at-physics.org } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear all, I'm currently having a few problems preparing good cross-section specimens of ceramic thin films on LaAlO3 or NdGaO3 substrates.
I can cut them okay, but thinning them often results in cracking as the thickness goes below 100 microns. I have a Gatan model 623 disk grinder and currently have access to SiC abrasive papers to grit sizes of 1500 or 2000,
and also have the Gatan diamond polishing disks and specimen lapping kit.
The problems are as follows: If we use the 1500 or 2000 grit SiC, we can often get the samples thin enough, but with poor surface finish which needs to be improved with dimpling (on both sides), we also risk cracking the sample as SiC papers often contain imperfections.
Using the Gatan specimen lapping kit, I find that as the disk grinder is altered to grind off a further 10 microns, the edge of the sample often catches on the diamond disk and tears some of the diamond coating off, leaving a lump on the surface which then risks catching the specimen and cracking it.
I have one possible alternative approach which I used in Sweden last year (with SiAlON ceramics) involving attaching the sample to a glass slide and polishing it using diamond spray on non-absorbent paper. I may consider trying this with these new materials.
Other ideas would be welcome, however, as would suggestions of how to improve my present method. Please note that I do not have the budget to buy any expensive new polishing equipment (such as a tripod polisher, for instance), so the most welcome suggestions would be those that involve improvements to current techniques, use of different types of polishing consumables, etc..
I hope to hear several suggestions, both from users and from the companies who producing polishing and lapping equipment. Why not post them with the list so we can all benefit from the sharing of experience?
I'm trying to do immunogold studies on a membrane protein in a preparation of plant vesicles (from minus 80ŒC storage). I did an Epon embedding with standard procedure (glut and osmium) to check the ultrastructure and got decent-looking vesicles, but side by side an LR White prep (0.5% glut, 2% paraformaldehyde, without osmium) looks like a disaster, with granular material, vague membranish-like structures and whorls of membranes. Does anyone have any tips for LR White preps of vesicles, or any other ideas for embedding (resins, fixing, dehydration, embedding) to favour the immunogold process.
Thanks,
Mark
******************************************** Mark West, Unidad de Microscopia Electronica, (Electron Microscopy Unit) Instituto de Fisiologia Celular, Universidad Nacional Autonoma de Mexico, 04510 Mexico D.F.
A book that I have found to be very resourceful is Biomedical Electron Microscopy by Arvid B. Maunsbach and Bjorn Afzelius.
Hope this is useful to you as well. Regards- Michelle Taurino Aventis Pharmaceuticals Bioimaging and Molecular Histology Tel-908-231-3357 Fax-908-231-3962 e-mail: Michelle.Taurino-at-aventis.com
-----Original Message----- } From: Petra Wahlbring [mailto:wahlbrin-at-crpgl.lu] Sent: Wednesday, June 07, 2000 3:49 AM To: microscopy-at-sparc5.microscopy.com
Since I am usually more involved in material analysis I am a little lost on the biological field. But now I am in the need of a good review article concerning the application of (analytical) TEM to biological materials. I am not looking for sophisticated latest developements but rather for basic applications with some examples, if possible including some examples of analytical TEM.
Hope to get some input :)
Petra
-------------------------------------------------------------- Dr. Petra Wahlbring Centre de Recherche Public - Gabriel Lippmann Laboratoire d'Analyse des Materiaux (LAM) 162a, av. de la Faiencerie L-1511 Luxembourg tel. +352-466644-402 fax +352-466644-400 e-mail: petra.wahlbring-at-crpgl.lu Visit our WWW site! http://www.crpgl.lu/~wahlbrin
} My SEM facility has just been approached, and in the interest in } providing what they want for a price based on an average, can I query } the SEM community as to what they charge for commercial release of } their SEM imagery. I'm looking for $/image, but if you charge on any } type of sliding scale, I'd be interested in that too. I believe as } well I shouldn't be undercutting commercial facilities, so I'm } especially interested in these numbers. } Reply to me direct, and I'll respond back to the list with the } average, max and min, and no names.
shAf-
Please start with an inquiry to your university administration; you might save yourself a lot of grief. When I was running an interdepartmental lab at U.C. Berkeley, there were very specific university-wide rules that I was required to follow for such usage.
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
We have a 2010 and our procedure is to bring the filament dial (labeled off-10) to position #3. Then we go for coffee. After 20 min. or so we raise it to # 4, after a couple of minutes to #5 etc. until we get to the stop ( in our case we have it at about 7 1/2). Even at this point we have filament drift for quite some time, so , we are not ready to take pictures for another 1/2 hr or so. By the way, we have the bias settings at 7 and 7 which is high. I would be interested in knowing what your settings are.
At 14:19 +0200 3/6/00, Jonathan Barnard wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
My understanding is as follows ....
There are different damage mechnisms at different voltages.
At lower voltages ( {200 kV) the main damage mechanism is, in effect, specimen heating. And, the lower the voltage, the greater the interaction (elastic scattering) with the specimen - so more damage but greater contrast, the main reason users looking at resin sections prefer lower voltages.
Increasing the voltage reduces specimen damage and tends to improve resolution but at the expense of contrast. As the interest in biological TEM moves to molecular biology, resolution is more important. But, at high resolution, phase contrast becomes the dominant factor in producing image constrast and phase constrast can be significantly improved - without loss of resolution - by using highly coherent FEG sources.
At voltages between 300kV and 400 kV, the energy of the electron becomes sufficient to cause "direct" radiation damage - atoms are displaced. At this point, specimen damage rates increase. The particular voltage depends on the amtomic number of your specimen.
Damage rates can also be reduced by cooling. in particular, cooling a specimen to liquid helium temperatures allows significantly greater electron exposure before damage occurs.
So, for optimum imaging of molecular specimens you need 300 kV and a FEG gun plus a liquid helium stage.
Regards, -- Larry Stoter JEOL (UK) Ltd Silver Court, Watchmead, Welwyn Garden City, AL7 1LT, United Kingdom tel: +44-(0)1707-377117, fax: +44-(0)1707-373254, e-mail: larrys-at-jeoleuro.com
For EDS checking you can use specimen from Cu or Al foil so that it will have any angle, including 30-60 degrees. But I am afraid that if there is no spectra at all with 10 degree tilt then a problem is more complicated.
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} -----Original Message----- } From: "MGMANDERS-at-aol.com"-at-sparc5.microscopy.com } [mailto:"MGMANDERS-at-aol.com"-at-sparc5.microscopy.com] } Sent: Sunday, June 04, 2000 10:41 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: NEED HELP WITH EDS ON A JEOL 100S } } } -------------------------------------------------------------- } ---------- } 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/MicroscopyListser} ver/FAQ.html } } } -------------------------------------------------------------- } ---------. } } } A fellow colleague has bought a JEOL 100s and wants and } needs EDS X-ray } Analysis. He's mounted his horizintal EDS detector , but can } not get sample } X-rays from the speciman. Contacting JEOL he found the } problem to be a tilt } problem. The 10 degree tilt from the normal SEG only tilts } 10 degrees and a } 30 to 60 degree tilt is necessary. At one time I was told } that special } sample holders and or double gap pole pieces were availabe. } He wishs to find } any one with a 100s for parts needed or information which } would allow X-ray } analysis. Please reply to mgmanders-at-aol.com or the list server. } } Mike Manders } } }
We have the opportunity to acquire a TEM, but budget constraints mean that have to consider the cost of acquiring and operating this instrument carefully against alternatives. We plan to use it principally to study the morphology, behavior and size distribution of colloidal gold particles and other types of metal nanoparticles.
Can anyone give any information on what the maintenance, operating and facilities requirements and costs would be for this instrument, or point me to a good source? What renovations would be necessary to accommodate it - darkroom, ancillary equipment, cooling, water and power supplies, darkroom facilities? How much should we budget for supplies and consumables? Since we have no-one currently trained to use it, how long would it take to learn, and how much maintenance (time, supplies, contracts) would it require?
Thanks in advance,
Rick Powell
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Both Zeiss and Leica have long-standing reputations for quartz optics with transmission into the UV (typically, down to about 220nm). You are going to need more than just the objective; the binoc also needs to have a UV transmitting prism. A tip: inquire about microscopes used for microspectrophotometry or in semiconductor applications. From the biological/biomed realm: microscopes used for Fura or Indo studies.
Try Tom Calahan at Zeiss (914-681-7733) and Jan Hinsch at Leica/Allentown (201-236-5905).
Nikon has also been agressive in lens development, but I haven't had direct experience with any UV optics. Call Stan Schwartz (516-547-8529). ... and at Olympus: Reinhard Enders (516-844-5000).
Good hunting! Barbara Foster Microscopy/Microscopy Education 125 Paridon Street, Suitee 102 Springfield, MA 01118
Customized, on-site short courses in all areas of microscopy, sample prep, and image analysis. "Why didn't they teach us that sooner?" Probably because no one called MME!
At 03:27 PM 6/6/00 -0400, donald j marshall wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Suggest you buy a good upright stand which you can upgrade as your needs develop. I strongly suggest a combination of both transmitted and reflected light options.
Your feelings re: investing in the microscope vs. a high end camera at this stage are well founded.
Re: contrast techniques... I think that you might find Hoffman Modulation Contrast a better bet for what you are doing than Phase. Hoffman can be used singly for looking at surfaces (ex: your coatings) or in combination with polarized light. It is a good complement to DIC, which won't work if your powders, etc. respond to Pol.
All of the "big 4" (Olympus, Nikon, Leica, and Zeiss) have good equipment. The issues I would add to your shopping list might include how comfortable you feel operating a specific microscope (like trying on a coat or test-driving a new car) and how supportive your local dealer or representative is.
Your comment re: EM was interesting.... I have been doing a lot of work over the past 6 months with commercial companies who look at many of the applications you cited. Universally, they were astounded at the information they could get from the Light Microscope, quickly and with minimal sample prep. I am currently on assignment teaching a group of very competent EM people about Light Microscopy... and they are equally amazed. All by themselves, they came to the conclusion that they should take a look first with the Light Microscope and then, if necessary, go the SEM. (I added that they should also take a quick look with the stereo first).
By the way, you may be interesting in "Optimizing Light Microscopy", both as a reference for your lab and a text for your students. Details are on our website.
Customized, on-site short courses in all areas of microscopy, sample prep, and image analysis.
"Why didn't they teach us that sooner?" Probably because no one called MME!
At 09:47 AM 6/7/00 +0200, Bemporad, Edoardo wrote:
} } } }
{excerpt}
{fontfamily} {param} Arial {/param} {smaller} I am going to buy one ore two OM for our EM lab (XL30 and CM120), trying to convince myself that one brand is better than the other (quality/price rate included in the evaluation!).
{/smaller} {/fontfamily}
{fontfamily} {param} Arial {/param} {smaller} In our lab we do mainly metallographic analysis, interface studies of wear resistant coatings, and catalytic powders characterization, but I guess that the OM will be used for a wider range of investigation (W/O and O/W emulsions, asbestos, ..) and for didactical scopes too.
{/smaller} {/fontfamily}
{fontfamily} {param} Arial {/param} {smaller} So EPI and DIA illuminations, BF, DF, NIC, phase contrast, pol, I don't think we will need fluorescence; forgotten something?
{/smaller} {/fontfamily}
{fontfamily} {param} Arial {/param} {smaller} What about an inverse microscope? or always better two standard ones? {/smaller} {/fontfamily}
{fontfamily} {param} Arial {/param} {smaller} I have about 35K$ budget to make everybody happy (research group and students), and my position is to prefer spending on the microscope (or microscopes) rather than on a high-level digital camera. I read some threads about it here and I think something like a Nikon coolpix 990 (if it will works! :-) ) will be enough. (other opinions or point of views?)
{/smaller} {/fontfamily}
{fontfamily} {param} Arial {/param} {smaller} I do not have a so deep experience in OM but I was wandering if there are some key feature to keep in mind for an equipment that will be used in a EM lab, considering that where I will not able to go with the OM (depth of field, resolution) I can use our EM!
Most of the lenses should come off, either directly or with the help of a small Allen wrench.
My favorite approach to lens cleaning: "Puff, Huff, and Swirl".... 1. Puff off any debris or rough material which might scratch the lens using either a puffer (available from photographic supply houses) or just "puffing" the dry air from your mouth with your cheeks (easier to demo than explain). 2. Huff on the lens, using the warm, moist air from deep in your lungs, to deposit a fog of moisture on the lens then 3. Quickly and gently, remove the fog with a clean Q-tip (100% cotton ) or lens tissue (NOT Kim-Wipe!), starting at the middle of the lens and continuing in a spiral to the outer edge. Do not use the same area on the swab or tissue more than once; they will collect debris which can scratch the delicate coating. 4. If the dirt is persistent (ex: mascara, fingerprint, oily residue), dip the tip of a cotton swab in a good lens cleaning solution (also available at photo supply houses), shake off any excess, then repeat the "swirl" step.
I recently had a client who had a neat moist towelette made by Uvex. It worked really well, even on oil. I will have to find the contact info, so if you are interested, please email me off line.
Best regards, Barbara Foster Microscopy/Microscopy Education 125 Paridon Street, Suitee 102 Springfield, MA 01118
Customized, on-site short courses in all areas of microscopy, sample prep, and image analysis. "Why didn't they teach us that sooner?" Probably because no one called MME!
At 11:43 PM 6/6/00 -0500, wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
The real question is why should you increase the LaB6 heating current slowly? As I understand it there are two reasons. Firstly if the tip is new, the gun has been serviced or the gun has not been run at 200kV for some time it is important to run up slowly to ensure that you do not get any outgassing from the tip or other gun components that might cause a flashover and possible damage. Secondly if the machine is regularly used at 200kV and the gun is is good condition (normal case) then you want to avoid any thermal shock to the LaB6 tip which could cause damage or misalignment. The same applies to cooling the tip down after use.
In the first case I would certainly take several 10s of minutes while carefully watching the vacuum gauge and the emission current meter (and HT stability if you are connected to an oscilloscope). Exact time would depend on the condition but maybe 30 minutes to heat a new tip and 2 minutes for a normal tip in good condition. I would take a minute to cool down a tip.
In both cases the greatest heating effect is a square of the control position (power is proportional to V^2 or I^2) and this should be taken into acount. I heat to about 3.5 in 25 to 30 seconds then decrease my heating speed as I approach the stop (set at 5 in my case, bias 5.5). The particular setting will depend on the type of LaB6 tip used, the wenhelt to tip distance and how you run the emission. We use 5uA emission with the tip slightly undersaturated as it seems to give good results and reasonable long life at the high mags that we use. For the smallest probes we may desaturate further to reduce the probe size.
I am aware from visitors that we have from other sites that this is quicker than many people but it seems impractical to spend 30 minutes to run up the tip every time you want to change a specimen when we want to look at several in a session. Our tip life des not seem to be any worse than those who take much longer.
Regards, Ron
} ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } Question: } What is the proper procedure for increasing the filament current on a 2010 } with a LaB6? } How fast can you go up? } Should the current be increased at a linear rate or should the rate be } tapered off as you reach saturation? } The manual suggests 30 sec/graduation while the service techs suggest a } much slower rate. } } Thanks, } Kim } ************************************************************** } Kim W. Pierson, Ph.D. } Dept of Physics & Astronmoy } University of Wisconsin-Eau Claire } (715) 836-5009 } FAX 836-3955 } } }
=========================================================================== Mr. Ron Doole e-mail ron.doole-at-materials.ox.ac.uk Department of Materials, phone +44 (0) 1865 273701 University of Oxford, fax +44 (0) 1865 283333 Parks Road. Oxford. OX1 3PH. UK. ============================================================================
LR White won't look as good as Spurr's, but membrane whorls are indicative of insufficient fixation. Os is the better lipid (therefore membrane) fixative, since you cannot use Os, you may need to increase the time and or concentration of GA. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Thursday, June 08, 2000 1:51 AM, Mark West [SMTP:mwest-at-ifcsun1.ifisiol.unam.mx] wrote:
} } Hi, } } I'm trying to do immunogold studies on a membrane protein in a preparation } of plant vesicles (from minus 80oC storage). I did an Epon embedding with } standard procedure (glut and osmium) to check the ultrastructure and got } decent-looking vesicles, but side by side an LR White prep (0.5% glut, 2% } paraformaldehyde, without osmium) looks like a disaster, with granular } material, vague membranish-like structures and whorls of membranes. Does } anyone have any tips for LR White preps of vesicles, or any other ideas } for embedding (resins, fixing, dehydration, embedding) to favour the } immunogold process. } } Thanks, } } Mark } } } } } } } ******************************************** } Mark West, } Unidad de Microscopia Electronica, } (Electron Microscopy Unit) } Instituto de Fisiologia Celular, } Universidad Nacional Autonoma de Mexico, } 04510 Mexico D.F. } } tel (unidad/lab) *(525) 622 5610* } (casa/home) (525) 619 3020 } Fax (525) 616 2282 } ******************************************** }
Hi Kris, same thing happened to me, but re-subscribing took care of the problem. Mailed to listserver, but no respons at all.
} Dear Microscopists, } I'm just curious if something is wrong on my end or I'm unsuscribed against } my will. I did not receive any posting during the past two-three weeks, and } this is impossible in view of previously received 10-20 postings/day. } Is there any explanation? } Kris } Dr. Kristof Kovacs } Associate Professor } President, Hungarian Society for Microscopy } Phone: +36-(88)-421-684 } Fax: +36-(88)-328-643 } Mailing Address: } University of Veszprem, P.O.Box 158, Veszprem } H-8201 Hungary
Yours sincerely
Per Hrstedt Department of Medical Biosciences Pathology Unit for Electron Microscopy University of Ume S-90187 Ume Sweden
Literature indicates that specimen damage due to heating decreases with increasing accelerating voltage; however, there is a trade off because at higher voltages materials fall victim to knock-on and sputtering damage. A very good overview (with references) of specimen/beam interactions and beam damage can be found on PP 49-55 "Transmission Electron Microscopy" by Williams and Carter 1996 Plenum Press ISBN: 0-306-45342-X
Brenda I. Prenitzer, Ph.D. Member of Technical Staff Cirent Semiconductor (Lucent Technologies) 9333 S. John Young Parkway 6D-Lab Orlando, FL 32819-8612
-----Original Message----- } From: Larry Stoter [mailto:LPS-at-teknesis.demon.co.uk] Sent: Wednesday, June 07, 2000 3:49 PM To: Jonathan Barnard; microscopy-at-sparc5.microscopy.com
At 14:19 +0200 3/6/00, Jonathan Barnard wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
My understanding is as follows ....
There are different damage mechnisms at different voltages.
At lower voltages ( {200 kV) the main damage mechanism is, in effect, specimen heating. And, the lower the voltage, the greater the interaction (elastic scattering) with the specimen - so more damage but greater contrast, the main reason users looking at resin sections prefer lower voltages.
Increasing the voltage reduces specimen damage and tends to improve resolution but at the expense of contrast. As the interest in biological TEM moves to molecular biology, resolution is more important. But, at high resolution, phase contrast becomes the dominant factor in producing image constrast and phase constrast can be significantly improved - without loss of resolution - by using highly coherent FEG sources.
At voltages between 300kV and 400 kV, the energy of the electron becomes sufficient to cause "direct" radiation damage - atoms are displaced. At this point, specimen damage rates increase. The particular voltage depends on the amtomic number of your specimen.
Damage rates can also be reduced by cooling. in particular, cooling a specimen to liquid helium temperatures allows significantly greater electron exposure before damage occurs.
So, for optimum imaging of molecular specimens you need 300 kV and a FEG gun plus a liquid helium stage.
Regards, -- Larry Stoter JEOL (UK) Ltd Silver Court, Watchmead, Welwyn Garden City, AL7 1LT, United Kingdom tel: +44-(0)1707-377117, fax: +44-(0)1707-373254, e-mail: larrys-at-jeoleuro.com
I remember a message some time ago, I think, suggesting a way to flat embed in silicon molds using LRWhite resin. How the top was sealed escapes me. Could anyone with a better filing system and/or memory please send me a message or a reference on this.
Thanks.
Pete -- Peter Bond Plymouth Electron Microscopy Unit University of Plymouth Drake Circus Plymouth Devon UK PL4 8AA Tel/Fax: 01752 233092 email: pbond-at-plymouth.ac.uk
If you are looking for an inexpensive TEM and/or SEM, The University of Portland has two such units you might be interested in.
1) Zeiss EM9s2, transmission electron microscopy, with additional high voltage tank, replacement for vacuum tubes, fuses, new filaments, lots of film cassettes, and film, original schematics, and operation manuals.
2) ETEC AutoScan, scanning electron microscope, with 60 and 90 degree stages, several reconditioned column liner tubes, new apertures, new YAG scintillator in original container, original schematics and operating manuals, plus, video tapes on operation and use. This instrument was original manufactured for INTEL and has a 50kV power supply.
Both units are operational!
Any interested parties or individuals can contact me off the server.
Ken Tiekotter, Adjunct Professor The University of Portland Department of Biology 5000 N. Willamette Blvd. Portland, OR 97203 USA
EMSL Analytical Inc. (Westmont, NJ) is interested in hiring a Transmission Electron Microscopy Analyst (TEM) for its NJ Corporate Office. EMSL is the world leader in asbestos analysis since 1981 with over 20 laboratory locations worldwide.
Responsibilities include the preparation and analysis of air, bulk, and water samples submitted by clients for asbestos content and other specialty asbestos projects. College Degree in Materials Science or related field and past experience with electron microscopy analysis preferred. . The ideal candidate would be a detail orientated individual able to follow lab protocols and procedures and able to work in a fast paced environment.
Full benefits package with salary commensurate with experience. Interested individuals send resume and salary requirements to Stephen Siegel, CIH (ssiegel-at-emsl.com) or fax to 856-858-4960.
Stephen Siegel Stephen Siegel, CIH Asbestos Lab Manager EMSL Analytical, INC 107 Haddon Avenue Westmont, NJ 08108 Phone:800-220-3675x1209 Fax:609-858-4960 email:ssiegel-at-emsl.com
Your main problem is with the use of SiC for grinding and thinning.
Silicon carbide paper should not be used to prepare any ceramic material, it is not hard enough to cut it efficiently without damaging the substructure. SiC becomes dull quite rapidly, with ceramic a dull abrasive actually cracks the structure and causes pullout and excessive chipping. You are compounding your problem with the weight of the tool being used as well. While the disc grinder is a well designed product and very useful, it is quite heavy for samples of certain thicknesses not to mention the additional pressure you are applying is not helping any.
The solution is to use Diamond Lapping Film and apply some Diamond Extender at the final stages of thinning to reduce the surface tension between the water and the sample. This reduces the sheer stress being applied to the sample and will drastically reduce the possibility of cracking.
Should you have interest, we offer a polishing machine "The MultiPrep System" that allows you to prepare the sample without the possibility of the tool being misaligned or mishandled (tilted on edge) during prep. Only the sample makes contact with the abrasive and the plane of polish remains in tact throughout the polishing/grinding process. A dial indicator (1 micron increments) allows you to preset a known amount of material to be removed and there is no operator intervention until the sample is done. The amount of force applied to the sample is constant regardless of the operator.
If you wish to get more information about the MultiPrep System, you may visit our website: www.alliedhightech.com or contact me in person at 310- 635-2466.
Sincerely,
Gary Liechty Manager, Technical Products Allied High Tech Products, Inc. } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear all, } I'm currently having a few problems preparing good cross-section specimens } of ceramic thin films on LaAlO3 or NdGaO3 substrates. } } I can cut them okay, but thinning them often results in cracking as the } thickness goes below 100 microns. I have a Gatan model 623 disk grinder and } currently have access to SiC abrasive papers to grit sizes of 1500 or 2000, } and also have the Gatan diamond polishing disks and specimen lapping kit. } } The problems are as follows: } If we use the 1500 or 2000 grit SiC, we can often get the samples thin } enough, but with poor surface finish which needs to be improved with } dimpling (on both sides), we also risk cracking the sample as SiC papers } often contain imperfections. } } Using the Gatan specimen lapping kit, I find that as the disk grinder is } altered to grind off a further 10 microns, the edge of the sample often } catches on the diamond disk and tears some of the diamond coating off, } leaving a lump on the surface which then risks catching the specimen and } cracking it. } } I have one possible alternative approach which I used in Sweden last year } (with SiAlON ceramics) involving attaching the sample to a glass slide and } polishing it using diamond spray on non-absorbent paper. I may consider } trying this with these new materials. } } Other ideas would be welcome, however, as would suggestions of how to } improve my present method. Please note that I do not have the budget to buy } any expensive new polishing equipment (such as a tripod polisher, for } instance), so the most welcome suggestions would be those that involve } improvements to current techniques, use of different types of polishing } consumables, etc.. } } I hope to hear several suggestions, both from users and from the companies } who producing polishing and lapping equipment. Why not post them with the } list so we can all benefit from the sharing of experience? } } Best wishes } } ===== } Ian MacLaren } Beijing Laboratory of Electron Microscopy } Chinese Academy of Sciences, P.O. Box 2724 } 100080 Beijing } China } General Email: ian.maclaren-at-physics.org } Work (esp. large attachments): maclaren-at-image.blem.ac.cn } } ____________________________________________________________ } Do You Yahoo!? } Get your free -at-yahoo.co.uk address at http://mail.yahoo.co.uk } or your free -at-yahoo.ie address at http://mail.yahoo.ie }
We have JEOL 840 SEM that has develop a leak at the specimen chamber door interface and will not hold a vacuum. We have changed the o-ring around the door, look for nicks, scratches any other damage around the door interface, none found. We have also check the roughing valve (V4, LV3 and the pressure relief valve) for leaks. Pressure holds in the gun area. The large door is only opened when we have to replace or add a new attachment to the chamber and thats about once a year. We have a vacuum specimen exchange port to enter and retrieve samples. We have also check that seal and it also checks out OK. We have had 2 JEOL engineers take a look at the system and at this point no luck finding the cause or solution. You can respond to me off-line at bruce.arey-at-pnl.gov Thanks
hello Kim, Jordi & interested parties, We have a filament ramp circuit on our LaB6 2010 that controls the filament current. It is an accessory offered by Jeol. Jeol has set it to take about 8 minutes to bring the filament to max current. The max. current is still controlled by the (preset) filament current knob on L1. Once the filament is hot, when we change samples or other wise have the filament current off for {5mi. we use the "quick timer" feature of this circuit which brings the filament up in ~90 seconds. The max. filament current is set to run just under saturation (so you can just barely see the X when the beam is converged). The bias control is used to set beam current to 10 uA. The setting changes with KV & filament wear. The initial bias setting is a function of the physical position of the filament relative to the Wehnelt cap. It changes every time the filament or cap are serviced. As the tip wears the difference in bias settings say between 100 & 200 KV increases & both move up. We are a multiple user facility. Our filament sees a of cycles & we always have novice users. Filament life times are 400-1000 hours. This tends to track with the novice user density. As far as the drift Jordi mentioned. This is a non issue here & may be in the filament design. We use the Gimble Phillips 60-06. I do a lot of carbon work & can pretty much nail 3.4A when I get a beam. Yes the images are sharper after we have been running a bit.
Bruce Brinson Rice U.
usual disclaimer... no financial interest in companies mentioned.
Marti, Jordi 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 } -----------------------------------------------------------------------. } } Kim: } } We have a 2010 and our procedure is to bring the filament dial (labeled } off-10) to position #3. Then we go for coffee. After 20 min. or so we raise } it to # 4, after a couple of minutes to #5 etc. until we get to the stop ( } in our case we have it at about 7 1/2). Even at this point we have filament } drift for quite some time, so , we are not ready to take pictures for } another 1/2 hr or so. By the way, we have the bias settings at 7 and 7 which } is high. I would be interested in knowing what your settings are. } } I hope this helps } } Jordi Marti }
A user of our facility is interested in making height measurements from specimens viewed in the SEM. I am aware of the conventional way of doing this: stereo pairs and optical viewer (with stereometer parallax corrections). Is there a more modern (computerized) way of doing this, say with anaglyphs?
Thank you.
John B.
#################################################################### John J. Bozzola, Ph.D., Director Micro-Imaging and Analysis Center 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 ####################################################################
In a message dated 6/8/00 12:38:48 PM, bozzola-at-siu.edu writes:
} A user of our facility is interested in making height measurements } from specimens viewed in the SEM. I am aware of the conventional way } of doing this: stereo pairs and optical viewer (with stereometer } parallax corrections). Is there a more modern (computerized) way of } doing this, say with anaglyphs?
There was a thread recently on measurement of stereo pairs by computer-matching of points. Several software packages (one of the is Fovea Pro - http://members.aol.com/FoveaPro) were mentioned as having this capability. Whether or not the two images are put together as an anaglyph is unimportant.
Just a couple of additional comments to Barbara's procedure.
A Zeiss axiom: If the lens is not dirty, then cleaning it will never improve it, it only risks damaging it. Do the least amount possible to return the surface to clean. If the surface is only dusty, & puffing it removes the problem, stop there. I look at the lens being cleaned, using the ocular turned upside down as a loupe, at each step of the process. When it's clean, you're done. Usually only external surfaces are contaminated and need frequent or extensive cleaning. Internal lenses, a dusting often suffices.
In dealing with a dissecting microscope with a zoom magnification capability as opposed to one with click stops or a fixed magnification, be very careful about disturbing the positional settings of the internal lenses. Any changes to these settings will alter magnification for the each of the two eyes and make an image that can't be justified in the brain. This will require a service engineer to remedy. For heavily filmed lenses, use short or broken cotton tipped applicator and get in the best you can. Varying the zoom control will move the lenses up and down and maybe give you enough room to work.
As Barbara wrote, removing loose dust is essential to preserving lens quality. Lenses are coated with various coatings and these are easily damaged. Puffers from camera stores are good. The red bulb ear syringe for babies is excellent and usually readily available. I do use dusters and compressed air although both are frowned upon, by some, as possibly damaging lens coatings. If you use a duster, don't shake or tip it while dusting the lens. This will expel liquid from the can and contaminate the lens worse.
A good lens cleaner that is easy to get is Sparkle Glass Cleaner available from Ace hardware, and grocery stores (No financial interests). Never use Windex. It contains oils that will coat the lens.
As much as possible use the cotton tipped applicators rather than lens tissues. Unless you wear latex or polyethylene gloves, finger oils will get onto the tissue and be transferred to the lens. Another reason to shy away from the tissue is the tendency to scrub the lens. I agree with Barbara, NEVER, NEVER Kimwipes, I've been told from many sources they contain many silica strands, and will scratch the delicate optical coatings. On expensive lenses, make a single pass, using minimal pressure straight across the lens, with the applicator, rolling the stick between your fingers to present a fresh cotton surface to the lens and picking up and getting the dust away from the lens. Alternate between wet (either with lens cleaner or condensed breath (open mouth, no spit please) and dry cotton. (Yes, it is often necessary to use a swirling motion on oculars because of the amount of oil contamination from the eye lashes.)
It is best not to try to clean filters, there are a few recommended procedures but all potentially damage the very precise and thin coatings of the filter compromising its performance. (If anyone disagrees or has a favorite procedure for filter cleaning, contact me off list, I'd like to hear about it.)
Use of solvents or disassembling objectives or multiple lens stacks is best left to the service engineer. Lenses use a variety of air and cement interfaces to achieve resolution and aberration correction. Altering these by dissolving the cement will degrade the lens. Getting the small lenses back in exactly the same position is also very, very difficult and not for the faint hearted.
There is a microscope repair workshop in the initial planning stages for the Long Beach MSA meeting next year. It will target K-12 school teachers, but will hopefully address many levels. If members of the list will be at the meeting and are interested in attending this workshop to: 1. learn basic repairs for their microscopes. 2. get ideas for their outreach program. or 3. assist with putting on the workshop. Please e-mail me telling me of your interest or asking for more information.
If you have any questions feel free to contact me off line.
Jim is absolutely right about LR white. But instead of increasing the GA I will probably increase paraformaldehyde conc. to 4%.
Good luck,
Soumitra
} } Hi, } } } } I'm trying to do immunogold studies on a membrane protein in a preparation } } of plant vesicles (from minus 80oC storage). I did an Epon embedding with } } standard procedure (glut and osmium) to check the ultrastructure and got } } decent-looking vesicles, but side by side an LR White prep (0.5% glut, 2% } } paraformaldehyde, without osmium) looks like a disaster, with granular } } material, vague membranish-like structures and whorls of membranes. Does } } anyone have any tips for LR White preps of vesicles, or any other ideas } } for embedding (resins, fixing, dehydration, embedding) to favour the } } immunogold process. } } } } Thanks, } } } } Mark } } } } } } } } } } } } } } ******************************************** } } Mark West, } } Unidad de Microscopia Electronica, } } (Electron Microscopy Unit) } } Instituto de Fisiologia Celular, } } Universidad Nacional Autonoma de Mexico, } } 04510 Mexico D.F. } } } } tel (unidad/lab) *(525) 622 5610* } } (casa/home) (525) 619 3020 } } Fax (525) 616 2282 } } ******************************************** } }
***************************************************************** Soumitra Ghoshroy Ph.D. Electron Microscopy Lab Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3600 Fax: 505-646-5665 e-mail: ghoshroy-at-nmsu.edu http://confocal.nmsu.edu/eml
} ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. text deleted } } It is best not to try to clean filters, there are a few recommended } procedures but all potentially damage the very precise and thin coatings of } the filter compromising its performance. (If anyone disagrees or has a } favorite procedure for filter cleaning, contact me off list, I'd like to } hear about it.) } Please respond on-line! I'm sure that I'm not the only other person who would be interested in this!
Try treat the glut- & Os-fixed sections you have with sodium metaperiodate before doing immunolabeling. It may work.
Reference: M. Bendayan 1989. Protein A-gold and Protein G-gold postembedding immunoelectron microscopy. In Colloidal Gold: Principles, Methods and Applications Vol.1 Academic Press.
Ann Fook Yang EM Unit, Eastern Cereal and Oilseed Research Centre, Rm 2091, K.W. Neatby Bldg., Central Experimental Farm, Ottawa, Ontario, Canada K1A 0C6
Phone: 613-759-1638 Fax; 613-759-1701
} } } Mark West {mwest-at-ifcsun1.ifisiol.unam.mx} 06/07 11:50 AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi,
I'm trying to do immunogold studies on a membrane protein in a preparation of plant vesicles (from minus 80ŒC storage). I did an Epon embedding with standard procedure (glut and osmium) to check the ultrastructure and got decent-looking vesicles, but side by side an LR White prep (0.5% glut, 2% paraformaldehyde, without osmium) looks like a disaster, with granular material, vague membranish-like structures and whorls of membranes. Does anyone have any tips for LR White preps of vesicles, or any other ideas for embedding (resins, fixing, dehydration, embedding) to favour the immunogold process.
Thanks,
Mark
******************************************** Mark West, Unidad de Microscopia Electronica, (Electron Microscopy Unit) Instituto de Fisiologia Celular, Universidad Nacional Autonoma de Mexico, 04510 Mexico D.F.
Sorry, I don't have any useful suggestions, and for that reason, I'd rather see responses posted to the list, if possible, as this gives me a wonderful and otherwise unavailable opportunity to learn from the experience of others.
cheers
rtch
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } We have JEOL 840 SEM that has develop a leak at the specimen chamber door } interface and will not hold a vacuum. We have changed the o-ring around the } door, look for nicks, scratches any other damage around the door interface, none } found. We have also check the roughing valve (V4, LV3 and the pressure relief } valve) for leaks. Pressure holds in the gun area. The large door is only opened } when we have to replace or add a new attachment to the chamber and thats about } once a year. We have a vacuum specimen exchange port to enter and retrieve } samples. We have also check that seal and it also checks out OK. } We have had 2 JEOL engineers take a look at the system and at this point no luck } finding the cause or solution. You can respond to me off-line at } bruce.arey-at-pnl.gov } Thanks } } Bruce W. Arey } } PNNL-Battelle } Associate Scientist } Microstructural Characterization } bruce.arey-at-pnl.gov } 509-376-3363 } fax 509-376-6308 } } } } Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Mark, you didn't say how you post-stained after you immunolabeled. This step can make all the difference in the world. For LRW, I prefer 3-4 secs in saturated UA in 50% etoh, wash, followed by about 15secs in lead citrate. I find a major difference (inferior) when I use aqueous UA. Also, going too long in any of the solutions will give poor results.
} ===== Original Message From Mark West {mwest-at-ifcsun1.ifisiol.unam.mx} ===== } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hank Adams Manager Integrated Microscopy Core Molecular and Cellular Biology Baylor College of Medicine Houston, Tx 77030
John - I routinely make height measurements of 5 - 10 um interplanetary dust particles using the objective focus knob on our JEOL 1200EX in STEM mode. The procedure is simply to focus on the top of the particle and note the number of steps (or count the number of clicks of the knob) required when refocussing on the substrate next to the particle. By calibrating the objective focus step size with a known standard, you can routinely measure particle heights. In principle, a similar procedure should be possible on an SEM, depending on how the objective focus is configured. I don't think you can expect high accuracy with this method, however, so it may not be applicable to your needs.
Dave Joswiak Dept. of Astronomy University of Washington Seattle, WA 98195 joswiak-at-astro.washington.edu
On Thu, 8 Jun 2000, John J. Bozzola 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 } -----------------------------------------------------------------------. } } } A user of our facility is interested in making height measurements } from specimens viewed in the SEM. I am aware of the conventional way } of doing this: stereo pairs and optical viewer (with stereometer } parallax corrections). Is there a more modern (computerized) way of } doing this, say with anaglyphs? } } Thank you. } } John B. } } } #################################################################### } John J. Bozzola, Ph.D., Director } Micro-Imaging and Analysis Center } 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 } #################################################################### } }
Tamara et al : One superior method for cleaning sensitive optical components such as filters and front surfaced telescope mirrors is as follows: 1) gently puff of extraneous dust. 2) gently float the surface with deionized water. 3) if necessary to remove dirt film use a q-tip soaked in a non-ionic surfactant like a 0.01% solution of triton-x 100 (IMPORTANT DO NOT RUB ON THE SURFACE WITH FORCE BUT SIMPLY LET THE WEIGHT OF THE WET Q-TIP(S) BE THE ONLY "DOWNWARD PRESSURE" ON THE SURFACE OF THE GLASS. Such gently techniques such as these should help mitigate damage.
Tamara et al : (excuse mispelling in first copy sent) One superior method for cleaning sensitive optical components such as filters and front surfaced telescope mirrors is as follows: 1) gently puff off extraneous dust. 2) gently float the surface with deionized water. 3) if necessary to remove dirt film use a q-tip soaked in a non-ionic surfactant like a 0.01% solution of triton-x 100 (IMPORTANT DO NOT RUB ON THE SURFACE WITH FORCE BUT SIMPLY LET THE WEIGHT OF THE WET Q-TIP(S) BE THE ONLY "DOWNWARD PRESSURE" ON THE SURFACE OF THE GLASS. Such gently techniques such as these should help mitigate damage.
} I remember a message some time ago, I think, suggesting a way to flat } embed in silicon molds using LRWhite resin. How the top was sealed } escapes me. Could anyone with a better filing system and/or memory } please send me a message or a reference on this.
Peter Bond {P.Bond-at-plymouth.ac.uk}
} Pete -
If you have a silicon mold that isn't permeable to LR White, I'd like to know who makes it! There is a teflon flat mold available from Ted Pella that works well with LR White; it's their own product. It's sealed with Thermanox coverslips, so you might try them with your mold.
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
Peter Bond wrote: ============================================================== I remember a message some time ago, I think, suggesting a way to flat embed in silicon molds using LRWhite resin. How the top was sealed escapes me. Could anyone with a better filing system and/or memory please send me a message or a reference on this. ============================================================== This might have been an old posting of mine. Since UV transparency is usually required, we are talking about clear UV transparent silicone embedding molds and we recommend a slight "overfilling" of the cavities. Then when all cavities are filled, take another mold, just like the first one, and place it on top, bottom side down, cavity side up. The capillary action will result in a sealing out of any oxygen.
When the UV curing is complete, the top mold can be easily separated and since the cavity side is still unused, no wear and tear has been put on it in terms of taking away any of its lifetime.
Information about these transparent-to-UV embedding molds, and their use, can be found on the SPI website given below.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.spi.cc ######################## ============================================
Dear colleagues, We have a problem with spectra recording in our EDAX DX4 system mounted on Philips CM12/STEM. There is a hole in the spectrum between 0.5keV and 3.5keV. With service engineer, we have borrowed all the boards in electronics and exchange our boards with the borrowed ones. The hole in the spectrum remains. We have also dismounted detector from the EM column and the Be window was checked for contamination. The window was clean and intact. Please, can anybody give us any hints how to solve our problem? Many thanks in advance for any comment. Oldrich Benada
Our system specification: Philips CM12/STEM EDAX DX4 system with 184 preamp. based on win3.11
+-----------------------------------+ Oldrich Benada Acad. Sci. CR Institute of Microbiology Laboratory of electron microscopy Videnska 1083 CZ - 142 20 Prague 4 - Krc Czech Republic +------------------------------------+ Phone: +420-2-4752399 Fax: +420-2-4715743 WEB:http://www.biomed.cas.cz/mbu/lem113/lem.htm
Balzers Union renamed to BAL-TEC AG in 1992. But the product line was maintained and new developments are carried out. For additional information have a look at our website www.bal-tec.com. There are several coating systems and freeze fracture systems of this older types in use.
G'day Folks, I acquired some carbonate standards a while ago from the Smithsonian Insitute. Namely, Calcite, Siderite, Dolomite and Strontianite. Interestingly, when admiring them with our ED system, there appeared an anomalous peak around about where B is supposed to be. This is not a detector artifact as it is specific only to these materials. I just wondered if anyone else had noticed the same thing, or whether anyone has any clues on why this should be occuring. The programme suggests that there is about 50 wt % B in these things which seems unbelievable considering the compositions supplied by Washington. Ideas? Cheers, Malc.
-- Dr MP Roberts Phone: [+27](0)46 603 8313 Dept of Geology Fax: [+27](0)46 622 9715 Rhodes University Cell: 083 4060 262 (usually off) 6140 Grahamstown e-mail: malc-at-rock.ru.ac.za SOUTH AFRICA
With regard to the vacuum problems encountered by Bruce , without being familiar with the instrument we encountered a ' similar ' problem and were misled into thinking a chamber door leak existed by an over zealous leak detection unit . The problem on our sem actually turned out to be a hairline crack in a metal bellows attached to a valve that appeared OK . The vaccum would not exceed a certain level in pumpdown in its final stages .
I believe these bellows were often a source of problems on older instruments .
M.HARRIS harrism-at-esm-semi.co.uk ESM LTD South Wales , U.K .
Hi All, I'm posting this for a friend who is not on-line.
Does anyone out there have a used RMC MT-7000 for sale? My friend just moved to a lab that had an MT-5000 which she was told was operational. A major exaggeration. She is an RMC loyalist, and desparately wants another, but has a limited budget.
Please contact: Linda Burg Friedman at Columbia P & S at (212)305-9047
Thanks, Lee
Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Confocal Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
The specimen chamber is a vast area with many potential leak sources.
The door is the most likely if it is being opened on a regular basis, however do not forget that the stage drives pass through the door and they are being used all of the time.
If you are sure the door "O" ring is OK check the stage drive feed throughs as they may be the source of your problem?
Steve Chapman Senior Consultant Protrain For consultancy and professional training in EM world wide Tel 44+ 1280 814774 Fax 814007 www.emcourses.com
Rick, YOu don't mention make or modle of TEM, but i can tell you that my service contract (2 preventive maintenance call, unlimited service calls, parts, labor) costs around $15,500/year. You will need a water supply for cooling and electrical work to bring in a dedicated line. You will need a dakrroom with running water and a temp. control valve to process the film. If you wish to make photographic prints of your negatives, you will also need a point source enlarger (Durst was the best, but they are hared to find, Omega used to make one, too) and either pans (slow and painful) or a rapid processor. Budgeting for supplies is a tough call...it depends on your usage. If you find someone with an EM background to run it, its should't take them long to learn the individual instrument. Training someone from ground zero could take months to get dthe person on his/her way. Ancillary euqipment: it depends on what you will be doing, biological or materials, embedding/sectioning or particulates, negataive staining or metal shadowing.
A really good source to check is Audrey Glauert's Practical Methods in Electron Microscopy Vol. 4: Design of the Electron Microscope Laboratory, by Ronald H. Alderson, North-Holland publishers, 1975.
If you wish to contact me off-line, I can go over my lab's operating budget with you. I run a basic EM Core Facility here at (what used to be called) Cornell Medical College.
Lee
Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Confocal Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
JEOL has been very helpful and they are somewhat limited on what they can do to this instrument. The 840 has radioactive particles inside the chamber so we have to be careful each time we open the system up plus there hands on is limited to just oversight. I am in the process of trying to find a leak detector on site that can be used on radioactive system. But we also cannot pump down the chamber enough to use a leak detector. So we are in the process of making a plate that will fit over the door. We have taken off all our accessories off the chamber (EDS, OIM, BSE) and have pressurized the system and have found some evidence of a leak on the left hand side on the door, we have changed the o-ring and still the seal leaks on this side of the door. We have polished the door seal to make sure there is no major scratches or marks. We are pretty confident that specimen exchange port is OK we can pump this down and we see no signs of a leak. Why are we focusing our attention on the large o-ring and chamber door. We can clean the o-ring with alcohol (methanol or ethanol) and we can rough pump the chamber out and put the chamber on the high vac system but after a period of time the alcohol dries out and the system shuts down too a poor vacuum (overnight). Then we try to rough pump the chamber and we cannot go more than 20-30 mamps difference on the pirani gauge. We have done this several times with the same results. We are going to try to pressurize the system with He and try He sniffer to help us maybe pin point the leak. Thanks to all who have responded with suggestion on finding the leak and if we are successful in finding the leak I will post the finding on the server. Again JEOL has been very responsive in trying to find the leak and they are somewhat limited on this instrument do to its environment. Any other suggestion are welcomed.
---------- From: "Dopeyee-at-aol.com"-at-sparc5.microscopy.com Sent: Thursday, June 8, 2000 4:28 PM To: Microscopy-at-sparc5.microscopy.com Subject: LEAK IN JEOL
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GET A HOLD OF A LEAK DETECTOR AND FIND THE TRUE AREA THAT IS LEAKING
I DO BELEIVE THE MANUFACTURER SHOULD BE OF MORE HELP
I recall many years ago we had a similar vacuum leak on our 840A that was related to the specimen exchange port mechanism. The moving parts in the sliding door mechanism had become slightly miss-aligned. We ended up disassembling that mechanism, installing new o-rings and lubricating with Apiezon L. Problem gone. Hope your leak is that easy! Good Luck Brad Huggins BP Amoco, Naperville
} ---------- } From: Arey, Bruce W[SMTP:bruce.arey-at-pnl.gov] } Sent: Thursday, June 08, 2000 9:24 AM } To: 'Microscopy-at-msa.microscopy.com' } Subject: JEOL 840 SEM Vacuum Problem } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } We have JEOL 840 SEM that has develop a leak at the specimen chamber door } interface and will not hold a vacuum. We have changed the o-ring around } the } door, look for nicks, scratches any other damage around the door } interface, none } found. We have also check the roughing valve (V4, LV3 and the pressure } relief } valve) for leaks. Pressure holds in the gun area. The large door is only } opened } when we have to replace or add a new attachment to the chamber and thats } about } once a year. We have a vacuum specimen exchange port to enter and retrieve } samples. We have also check that seal and it also checks out OK. } We have had 2 JEOL engineers take a look at the system and at this point } no luck } finding the cause or solution. You can respond to me off-line at } bruce.arey-at-pnl.gov } Thanks } } Bruce W. Arey } } PNNL-Battelle } Associate Scientist } Microstructural Characterization } bruce.arey-at-pnl.gov } 509-376-3363 } fax 509-376-6308 } } }
I have always understood that deionized water would etch metal, that being the reason for PVC pipe being used to deliver it. If this is true, wouldn't that damage the first surface mirrors?
David, We have a B&L StereoZoom (0.7X - 3X) microscope which we intend to donate to the local school system (Computer VoTech Dept) for use in examining the circuit patterns on silicon wafers that I have donated to them.
My problem is that the microscope is not functioning properly. The two optical paths are not synchronized during zooming and therefore the image tends to rotate or it falls out of focus. We have tried to repair it in-house but we were unsuccessful. Knowing that that we would create additional problems, we did not disturb the lens or prism sub-assembly.
Is there a set of maintenance instructions or a tech manual available to help us align this scope?
Thanks Mike Urbanik Commercial Crystal Labs Naples, FL www.crystalguru.com
you may want to try the method below if your samples show very large structures. One of the nice features of an SEM is it's large depth of focus, unfortunately in many cases this prevents you from using the technique mentioned below. You can use a stereo technique to calculate a surface profile (see for example the stereo module on our web site or other stereo applications). Typical results from stereo images have a height resolution of about 1/10 the of the lateral resolution (in other words: if your lateral resolution is 1 micron between pixels, the height resolution will be on the order of 10 microns). This can be improved by sub-pixel interpolation but gives you an order of magnitude for the resolution.
Michael
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-----Original Message----- } From: David Joswiak [mailto:joswiak-at-orca.astro.washington.edu] Sent: Thursday, June 08, 2000 4:44 PM To: John J. Bozzola Cc: Microscopy-at-sparc5.microscopy.com
John - I routinely make height measurements of 5 - 10 um interplanetary dust particles using the objective focus knob on our JEOL 1200EX in STEM mode. The procedure is simply to focus on the top of the particle and note the number of steps (or count the number of clicks of the knob) required when refocussing on the substrate next to the particle. By calibrating the objective focus step size with a known standard, you can routinely measure particle heights. In principle, a similar procedure should be possible on an SEM, depending on how the objective focus is configured. I don't think you can expect high accuracy with this method, however, so it may not be applicable to your needs.
Dave Joswiak Dept. of Astronomy University of Washington Seattle, WA 98195 joswiak-at-astro.washington.edu
On Thu, 8 Jun 2000, John J. Bozzola 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 } -----------------------------------------------------------------------. } } } A user of our facility is interested in making height measurements } from specimens viewed in the SEM. I am aware of the conventional way } of doing this: stereo pairs and optical viewer (with stereometer } parallax corrections). Is there a more modern (computerized) way of } doing this, say with anaglyphs? } } Thank you. } } John B. } } } #################################################################### } John J. Bozzola, Ph.D., Director } Micro-Imaging and Analysis Center } 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 } #################################################################### } }
This may sound like "bucket science" but we've used the disposable aluminum weighing dishes to flat embed material. If you fill the bottom tray about 1/2 full and set another tray inside it, press gently until a little LR white oozes up the sides, what remains in the middle will be protected from the air and should polymerize nicely. (Maybe we've just been lucky!) good luck!
Tracey Pepper Supervisor Bessey Microscopy Facility Iowa State University ph: 515-294-3872 fax: 515.294.1337
Some Chlorine-L lines, as well as Sr lines have nearly identical energy/wavelength as B Ka. I don't remember the chemistry of these standards offhand, but I know some of them were Sr-bearing and some may also have Cl. These are likely the peaks you are seeing at the low end.
Jim -- ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ James J. McGee (email: jmcgee-at-sc.edu) Department of Geological Sciences University of South Carolina Columbia, SC 29208
Tel: 803-777-6300 Fax: 803-777-6610
Dr Malcolm Roberts 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 } -----------------------------------------------------------------------. } } G'day Folks, } I acquired some carbonate standards a while ago from the Smithsonian } Insitute. Namely, Calcite, Siderite, Dolomite and Strontianite. } Interestingly, when admiring them with our ED system, there appeared an } anomalous peak around about where B is supposed to be. This is not a } detector artifact as it is specific only to these materials. I just } wondered if anyone else had noticed the same thing, or whether anyone } has any clues on why this should be occuring. The programme suggests } that there is about 50 wt % B in these things which seems unbelievable } considering the compositions supplied by Washington. Ideas? } Cheers, } Malc. } } -- } Dr MP Roberts Phone: [+27](0)46 603 8313 } Dept of Geology Fax: [+27](0)46 622 9715 } Rhodes University Cell: 083 4060 262 (usually off) } 6140 Grahamstown e-mail: malc-at-rock.ru.ac.za } SOUTH AFRICA
If the instrument has been physically moved recently, this can sometimes
hasten a crack in these stainless steel "flex" vacuum lines. I have found these to develop mostly at the end of the tube where it has been flared for the fitting connector. Check the roughing lines first.
Happy hunting...
Bob Roberts EM Lab Services, Inc 2409 S. rural Rd Suite C Tempe, Arizona 85282 480.967.3946
} Oldrich Need a little more information. Do you actually have spectral information in the spectrum at energies { 0.5, or is there possibly, only noise in this low energy range? If the signal present in your spectra at these very lowest energies is just noise-like signal. Then it is possible that you have a severe alignment problem with the EDS detector/specimen geometry. A combination of high noise (due to vibration or other sources) and poor line of sight with the detector window (resulting in detection of only the higher energy x-rays ) could give you this "hole in the EDS spectrum" effect. A miss-aligned detector, and/or a detector making contact with the internal parts of the microscope might create this situation. Does the information in the low energy region correlate with the specimen composition? Does the high energy signal correlate with the specimen composition?
} ---------- } From: Oldrich Benada[SMTP:benada-at-biomed.cas.cz] } Sent: Friday, June 09, 2000 2:19 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: A hole in the EDS spectrum } } ------------------------------------------------------------------------ } 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, } We have a problem with spectra recording in our EDAX DX4 system } mounted on Philips CM12/STEM. There is a hole in the spectrum between } 0.5keV and 3.5keV. } With service engineer, we have borrowed all the boards in electronics } and exchange our boards with the borrowed ones. The hole in the } spectrum remains. We have also dismounted detector from the EM column } and the Be window was checked for contamination. The window was clean } and intact. } Please, can anybody give us any hints how to solve our problem? Many } thanks in advance for any comment. } } Oldrich Benada } } Our system specification: } Philips CM12/STEM } EDAX DX4 system with 184 preamp. based on win3.11 } } } } } } +-----------------------------------+ } Oldrich Benada } Acad. Sci. CR } Institute of Microbiology } Laboratory of electron microscopy } Videnska 1083 } CZ - 142 20 Prague 4 - Krc } Czech Republic } +------------------------------------+ } Phone: +420-2-4752399 } Fax: +420-2-4715743 } WEB:http://www.biomed.cas.cz/mbu/lem113/lem.htm }
Title: Optical Microscopy and Imaging in the Biomedical Sciences
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Applications to live cells will be emphasized; other specimens will be covered as well.
Students will have direct hands-on experience with state-of-the-art microscopes, video cameras, recorders and image processing equipment provided by major optical and electronics companies. Instruction will be provided by experienced staff from universities and industry.
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Postdoctoral Research Assistant - Electron Microscopy of Crystalline Materials
Salary £16,286 - £24,479 p.a.
A three-year position is available in a research group being developed by Professor David Cockayne FRS for the study of a range of materials using electron diffraction, electron microscopy (EM) and modelling techniques. Quantitative microscopy and materials modelling will refine structural models of technologically important materials. Extensive expertise in EM, diffraction, the preparation of crystalline materials for microscopy, and strong computing skills, are essential. Expertise in microscopy of semiconductors including quantum dots and computational techniques for image simulation would be an advantage. The Department has outstanding EM and modelling facilities. Please quote ref. DJ00/12.
Applications including a curriculum vitae, list of publications and the names and addresses of three referees should be sent to The Administrator, Department of Materials, University of Oxford, Parks Road, Oxford, OX1 3PH, from whom further particulars are available. The closing date for applications is 14 June 2000.
I have found the use of clear silcone sealant (as used for windows etc) quite effective to temporally fix or help to isolate large and medium vacuum leaks. In many instance it can be applied externally over various fittings. Screw holes are best covered first with a little tape, to facilitate the later the removal of the dry silicone. Silicone outgases a fair bit for some hours, so only major leaks can be determined immediately after applying the silicone. The method seems crude but is effective to eliminate numerous fittings as the source of a leak. I once operated a TEM for several months with a split stainless bellow, patched with a smear of silicone sealant. I don't suggest the use of that sealant on a permanent basis or in ion gutter pumped parts of a column. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Saturday, June 10, 2000 12:05 AM, Arey, Bruce W [SMTP:bruce.arey-at-pnl.gov] wrote: } } } JEOL has been very helpful and they are somewhat limited on what they can do } to } this instrument. The 840 has radioactive particles inside the chamber so we } have } to be careful each time we open the system up plus there hands on is limited } to } just oversight. I am in the process of trying to find a leak detector on site } that can be used on radioactive system. But we also cannot pump down the } chamber } enough to use a leak detector. So we are in the process of making a plate } that } will fit over the door. We have taken off all our accessories off the chamber } (EDS, OIM, BSE) and have pressurized the system and have found some evidence } of } a leak on the left hand side on the door, we have changed the o-ring and } still } the seal leaks on this side of the door. We have polished the door seal to } make } sure there is no major scratches or marks. We are pretty confident that } specimen } exchange port is OK we can pump this down and we see no signs of a leak. Why } are } we focusing our attention on the large o-ring and chamber door. We can clean } the o-ring with alcohol (methanol or ethanol) and we can rough pump the } chamber } out and put the chamber on the high vac system but after a period of time the } alcohol dries out and the system shuts down too a poor vacuum (overnight). } Then } we try to rough pump the chamber and we cannot go more than 20-30 mamps } difference on the pirani gauge. We have done this several times with the same } results. We are going to try to pressurize the system with He and try He } sniffer } to help us maybe pin point the leak. Thanks to all who have responded with } suggestion on finding the leak and if we are successful in finding the leak I } will post the finding on the server. Again JEOL has been very responsive in } trying to find the leak and they are somewhat limited on this instrument do } to } its environment. Any other suggestion are welcomed. } } } Bruce W. Arey } } PNNL-Battelle } Associate Scientist } Microstructural Characterization } bruce.arey-at-pnl.gov } 509-376-3363 } fax 509-376-6308 } } } ---------- } From: "Dopeyee-at-aol.com"-at-sparc5.microscopy.com } Sent: Thursday, June 8, 2000 4:28 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: LEAK IN JEOL } } ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } GET A HOLD OF A LEAK DETECTOR AND FIND THE TRUE AREA THAT IS LEAKING } } I DO BELEIVE THE MANUFACTURER SHOULD BE OF MORE HELP } } THEY HAVE TO FIX IT!!!! }
Hmmm, and "I didn't even know that", but believed that deionised water had metal ions removed from it and so in that respect its purer than 2x glass distilled water. Then I was taught and believed! that metal pipes would re-introduce metal ion back into the water! Being deionised the water has no buffering capacity and therefore is neither acid nor alkaline, they told me and I believed. Education is expensive; must ask for my money back. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Saturday, June 10, 2000 12:24 AM, "milesd-at-us.ibm.com"-at-sparc5.microscopy.com [SMTP:"milesd-at-us.ibm.com"-at-sparc5.microscopy.com] wrote: } } } I have always understood that deionized water would etch metal, } that being the reason for PVC pipe being used to deliver it. If this } is true, wouldn't that damage the first surface mirrors? } } Darrell }
Ahhh, but I have now been educated! Re-contamination of the painstakingly purified water is the concern, and there is no threat to the durability of the pipes. I had been mislead.
Hi Vacuum leaks, what a pleasure! Our tried and tested methods include using Petroleum Ether or Ethanol and then Bostick Prestic or Blue tac as it is known in Australia. Spraying alcohol around the suspected areas should show up the leak. If you ant to try and stop a leak use the Prestic. Its that stuff you buy at the stationary shop that is used to stick posters and pictures to a wall. That is pliable, removable and really handy at sealing off a few suspect areas. We are currently working on a JEOL840 here with a leak. After many hours we found that it was the gauge head that was leaking.
Good luck Luc Harmsen Anaspec, South Africa Technical support on microscopy. Tel + 27 (0) 11 476 3455 www.anaspec.co.za
ICEM 15 will be in Durban, South Africa, 2002. www.icem15.com
-----Original Message----- } From: Arey, Bruce W [mailto:bruce.arey-at-pnl.gov] Sent: Friday, June 09, 2000 4:05 PM To: 'microscopy-at-msa.microscopy.com'
JEOL has been very helpful and they are somewhat limited on what they can do to this instrument. The 840 has radioactive particles inside the chamber so we have to be careful each time we open the system up plus there hands on is limited to just oversight. I am in the process of trying to find a leak detector on site that can be used on radioactive system. But we also cannot pump down the chamber enough to use a leak detector. So we are in the process of making a plate that will fit over the door. We have taken off all our accessories off the chamber (EDS, OIM, BSE) and have pressurized the system and have found some evidence of a leak on the left hand side on the door, we have changed the o-ring and still the seal leaks on this side of the door. We have polished the door seal to make sure there is no major scratches or marks. We are pretty confident that specimen exchange port is OK we can pump this down and we see no signs of a leak. Why are we focusing our attention on the large o-ring and chamber door. We can clean the o-ring with alcohol (methanol or ethanol) and we can rough pump the chamber out and put the chamber on the high vac system but after a period of time the alcohol dries out and the system shuts down too a poor vacuum (overnight). Then we try to rough pump the chamber and we cannot go more than 20-30 mamps difference on the pirani gauge. We have done this several times with the same results. We are going to try to pressurize the system with He and try He sniffer to help us maybe pin point the leak. Thanks to all who have responded with suggestion on finding the leak and if we are successful in finding the leak I will post the finding on the server. Again JEOL has been very responsive in trying to find the leak and they are somewhat limited on this instrument do to its environment. Any other suggestion are welcomed.
---------- From: "Dopeyee-at-aol.com"-at-sparc5.microscopy.com Sent: Thursday, June 8, 2000 4:28 PM To: Microscopy-at-sparc5.microscopy.com Subject: LEAK IN JEOL
------------------------------------------------------------------------ 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 -----------------------------------------------------------------------.
GET A HOLD OF A LEAK DETECTOR AND FIND THE TRUE AREA THAT IS LEAKING
I DO BELEIVE THE MANUFACTURER SHOULD BE OF MORE HELP
We are consolidating two electron microscopy labs in the Boston area and have some equipment which someone may want. 1) JEOL JEM 100CX electron microscope which has always been under service contract and is still being used. It is about 20 years old and we would like to see it in a new home rather than trashing it. You would need to have it moved. 2)JEOL JEE 4C vacuum evaporator for Carbon. Still under vacuum and yours for the taking. 3)Durst Laborator 138S floor model enlarger with many condensers and an Agfa Rapidoprint DD6400 processor, both in very good shape and for sale. Contact me directly with any questions.
Norman Michaud Director, Morphology Mass Eye and Ear Infirmary Ophthalmology-5th flr. 243 Charles St, Boston, MA 02114 norman_michaud-at-MEEI.Harvard.edu Tel:617-573-3316; Fax:617-573-4290
Bruce W. Arey referred in his request to find a vacuum leak in his JSM840 to 'radioactive particles' inside the chamber of his SEM. I wonder if he could be more specific, because I feel if we start calling a SEM as a 'radioactive system' many safety officers will have a field day. I have been working with various SEMs since 1968 and never felt that I might be bombarded by radioactive particles, even when I opened the chamber (e.g. ETEC) of the SEM.
Hans Brinkies Senior Lecturer Swinburne, University of Technology School of Engineering and Science P.O.Box 218 - Hawthorn - Vic -3122 - Australia Phone: +61 3 9214 8657 Fax: +61 3 9214 8264 Email: Hbrinkies-at-swin.edu.au
I have a student who has been trying to jet polish tungsten for TEM. She has tried various concentrations of sodium hydroxide in water, as well as 40g trisodium phosphate/250ml water, and 55.8g magnesium perchlorate/250ml methanol, at a range of voltages. We have a Fischione jet-polishing unit. So far, none of the samples has been close to good. Can anyone help us out here, with past experience or general suggestions?
Many thanks in advance,
Gill
Dr Gillian M. Bond Department of Materials & Metallurgical Engineering New Mexico Tech Socorro, NM 87801
Certainly the best reference you can have for any jet polishing inquiry is Bernie Kestel at Argonne National Laboratory. I will forward this to him to see if he can add anything else. In digging through my extensive "Bernie Archives" I did find a paper titled "A Jet Polishing Solution for Silicon Germanium, Tantalum, Niobium and Tungsten-Rhenium" Ultramicrscopy 9 (1982) 379-384.
He was able to get a good polish under the following conditions: Temperature: -50 C Jet Height: 4.5mm (Single vertical jet system) Pump setting: 6 Volts: 40 Current: 20mA
This was done using his BK-1 solution. BK-1 is prepared by mixing 500ml methanol, 100ml butyl cellosolve, 90ml H2SO4 and 30ml HF.
He also has another paper MRS Volume 199 "Improved Methods and Novel Techniques for Jet Electropolishing of TEM Foils" which lists a method for electropolishing a 0.010" tungsten wire.
He was able to get a good polish under the following conditions: Temperature: -50 C Pump setting: 6 Volts: 120V
Using 6% HF, 12% sulphuric acid, 68% methanol, and 14% butyl cellosolve.
Of course these were done with a South Bay Vertical Jet system so you will need to adjust the parameters for your system. Please get the reference papers or contact me and I will send them to you. I have a long list of Bernie's papers I could send you along with many other references on TEM sample preparation that may be of interest. If you'd like to see more, please contact me.
DISCLAIMER: South Bay Technology produces the Model 550 D Single Vertical Jet Electropolisher as described above and, therefore, has a vested interest in promoting its use.
David Henriks Vice President 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
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by Gillian Bond } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have a student who has been trying to jet polish tungsten for TEM. She has tried various concentrations of sodium hydroxide in water, as well as 40g trisodium phosphate/250ml water, and 55.8g magnesium perchlorate/250ml methanol, at a range of voltages. We have a Fischione jet-polishing unit. So far, none of the samples has been close to good. Can anyone help us out here, with past experience or general suggestions?
Many thanks in advance,
Gill
Dr Gillian M. Bond Department of Materials & Metallurgical Engineering New Mexico Tech Socorro, NM 87801 {
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Business continuity and disaster recovery planning has now been made easier than ever with Version 7.3 of the Disaster Recovery System (DRS) product. DRS provides a plan for inaccessibility or inoperability (a disaster situation).
DRS is an industry standard software product, used by thousands worldwide. DRS users are in large and small companies across a wide variety of industries.
DRS conforms to federal regulations and meets insurance, auditing and legal requirements. DRS runs under Windows, with stand-alone and network versions available. DRS provides the most complete, easy-to-use product available today.
To prove its value, a free trial is available. For more information, visit our web site at www.drsbytamp.com {http://www.drsbytamp.com} . Dealer and distributor inquiries are welcomed
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Hi, one of our TEM's , a Zeiss EM 109, is equipped with an ion getter hi-vac pump. We are now planning to install a turbopump including all the piping and valves that are necessary. The reason for this is that the high vacuum is not good enough ( only in low E-6 area ) and the ion pump has to work too hard and the life-time of the electrodes becomes very short. Also the housing is clogged with trapped gas molecules and has to be regenerated far too often. Technically and electronically this exchange is fairly easily done, but my question is if anyone has done it and if so, what's the experience? Yours sincerely
Per Hrstedt Department of Medical Biosciences Pathology Unit for Electron Microscopy University of Ume S-90187 Ume Sweden
Dear Friends, For a long time we used Fab fragments-HRP from BioSys (France) for the immuno EM labelling of proteins with subsequent preembedding. These fragments were the best. However, recently the company cancelled its activity and does not send any more these products. Would you be so kind to tell me what has happened (if you know) and what fragments have the same quality?
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I changed my former SEM (Etec w/D.P)to a a Leybold mag-lev turbo a number of years ago. The ball bearing pumps (of the day) caused too much vibration. The results were excellent! The Etec plumbing allowed the turbo to run continuously during vent cycles which was of great benefit.
Keep in mind that for any gas, turbos have a fixed compression ratio. Among other things, the foreline pressure will have a direct influence on the ultimate vacuum.
The ion pumos are better than turbos for the cleanest, highest vacuum, but you have observed one weakness. They do not excel at pumping large volumes of garbage-loaded gasses.
Good Luck,
Woody White McDermott Technology
Hi, one of our TEM's , a Zeiss EM 109, is equipped with an ion getter hi-vac pump. We are now planning to install a turbopump including all the piping SNIP
I believe The SEM to which Bruce refered was/is used to examine radioactive materials. The "zoomies" are not from the SEM itself, but contamination from his specimens. My (former) Etec was in a similar condition. Over the years, my work mix resulted in a stage/chamber activitly level in the thousands of cpm generated by radioactive products of nuclear fission. ...Kept the really loose stuff at a minimum, but certainly had to exercise the appropriate precautions when working on the system.
Woody White McDermott Technology ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Bruce W. Arey referred in his request to find a vacuum leak in his JSM840 to 'radioactive particles' inside the chamber of his SEM. I wonder if he could be more specific, because I feel if we start calling a SEM as a 'radioactive system' many safety officers will have a field day. I have been working with various SEMs since 1968 and never felt that I might be bombarded by radioactive particles, even when I opened the chamber (e.g. ETEC) of the SEM.
Hans Brinkies Senior Lecturer Swinburne, University of Technology School of Engineering and Science P.O.Box 218 - Hawthorn - Vic -3122 - Australia Phone: +61 3 9214 8657 Fax: +61 3 9214 8264 Email: Hbrinkies-at-swin.edu.au
Dear Friends, For a long time we used Fab fragments-HRP from BioSys (France) for the immuno EM labelling of proteins with subsequent preembedding. These fragments were the best. However, recently the company cancelled its activity and does not send any more these products. Would you be so kind to tell me what has happened (if you know) and what fragments have the same quality?
The Laboratory of Electron Microscopy is looking for an used critical point drier to obtain it in donation.... We can pay all the costs of shipping and handling For any questions, please contact to me....
best regards.... =================================================== Fernando D. Balducci Laboratorio de Microscopia Electrnica Facultad de Ingeniera - Bioingenieria Universidad Nacional de Entre Rios Argentina e-mail: microsc-at-fi.uner.edu.ar tel: 54 43 975100 fax: 54 43 975077
I have done this on a JEOL 4000 but not a Zeiss. I used a Maglev TMP (Seiko Seiki) and an antivibration bellows to couple the pump because I was afraid of vibration degrading the 0.25nm resolution. I need not have worried, even with the antivibration bellows shorted out I was OK. Check that the TMP you choose does not give out any magnetic fields when running. If vibration is a problem then the Balzers (Pfieiffer) antivibration bellows that has a large worm drive clip around them can be tuned (by tightening the clip) to avoid the instrument resonant frequency.
Good luck, Ron
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
On Tue, 13 Jun 2000 09:17:46 +0200 Per =?iso-8859-1?Q?H=F6rstedt?= {per.horstedt-at-pathol.umu.se} 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, } one of our TEM's , a Zeiss EM 109, is equipped with an ion getter hi-vac } pump. We are now planning to install a turbopump including all the piping } and valves that are necessary. The reason for this is that the high vacuum } is not good enough ( only in low E-6 area ) and the ion pump has to work } too hard and the life-time of the electrodes becomes very short. Also the } housing is clogged with trapped gas molecules and has to be regenerated far } too often. } Technically and electronically this exchange is fairly easily done, but my } question is if anyone has done it and if so, what's the experience? } Yours sincerely } } Per Hörstedt } Department of Medical Biosciences } Pathology } Unit for Electron Microscopy } University of Umeå } S-90187 Umeå } Sweden } } phone int-46-90-7851541 } fax int-46-90-7851215
After many years of staining grids with uranyl acetate and lead citrate, we have begun to see a needle like or shard precipitate, (about 1/2 inch long at 100,000x's; resembles the lead precipitate on page 469 of Electron Microscopy second edition John Bozzola and Lonnie Russell). We have been using a 2.5% aqueous uranyl acetate and Reynold's lead citrate (filtered through a 2 micron filter) for the past 4 years or so with no problems. I have stained the grids with just UA and can see no precipitate and have stained grids with just the lead citrate and still not see the precipitate. I have also checked the water and cannot still see the precipitate. However, when I stain with UA followed by lead citrate it mysteriously reappears much to my dissatisfaction. I have also tried the basic lead citrate and just recently tried Sato's lead stain and had the same problem. I have made up UA from a newly purchased bottle. I have also lessened the staining times from 30 minutes in UA to 7 minutes and from 20 minutes in lead citrate to 5 minutes and the precipitate is less but still there. I have also checked the grids before staining them and cannot see the precipitate. Please help, I grow more grey day by day.
Phoebe J. Doss Manager/Adjunct Instructor Electron Microscope Lab Oklahoma State University
Back in the olden days, when BioRad sold microscopy supplies, they had an epoxy cleaner (to remove epoxies from hands, benches, etc., not made from epoxies). It was blue gunk in a little jar. Does anyone know what happened to this stuff? Or have an alternate?
I've just been digging through catalogues and Ted Pella sells a liquid cleaner - any experience with it?
a student here needs to fix and dehydrate strep mutans on polystyrene petri dishes. Can anyone point me to a good (simple?) protocol? Since we are primarily a materials research lab, we don't have a lot of biological references.
I don't know the Bio-Rad product, but Ladd sold (still sells?) a product called Met-a-terge that gets rid of uncured resins. A little bit goes a long way. I've used it for years.
John Mansfield's post regarding surplus equipment prompts me to make the following observations regarding the transfer of University-owned equipment in the United States. I would imagine that many other countries might have similar policies.
It often seems to come as a surprise to people to discover that the US government won't buy the same piece of equipment twice. What I mean by this is that if a piece of equipment has originally been purchased using federal funds (regardless of who currently holds title), then another institution cannot use federal funds (regardless of the source) to buy the equipment from the first owner.
To use the specific example of John's equipment: suppose it was bought originally with, say, an NSF grant, and I find that I could make use of it now. In order to do that I would need to find a non-US-government source of money, as I could not use even the income from my facility operation (which is regarded by the accountants as government money, as it originates predominantly from government research grants).
The logic of this policy, of course, is quite inescapable, however unpalatable it may be to the present owners of the equipment.
The policy only covers the cost of purchasing the equipment, by the way. If, to use the same example, John were to give me his surplus equipment, it would be perfectly acceptable for me to use federal funds to pay for the packing and shipping (and, if appropriate, reinstallation)
Tony Garratt-Reed.
** Anthony J. Garratt-Reed ** MIT Room # 13-1027 ** 77 Massachusetts Avenue ** Cambridge, MA 02139-4307 ** USA ** ** Phone: (+) 1-617-253-4622 ** Fax: (+) 1-617-258-6479 **
I am using a Denton Desk II sputter coater with a Pt target. I have noticed recently that a dark spot has formed in the middle of the target. I have not seen this before. Is this due to impurities in the target, problems with the vacuum, contamination from outgassing samples, or something else? Everything appears to be running fine and sample types have not changed. Do I clean it or just leave it alone? What is it telling me (if the coater could talk)?
Thanks in advance for all your expertise.
David BG Rose WL Gore and Associates 297 BLue Ball Road Elkton, MD 21921 410-506-2958
We recently had a rash of horrible precipitate problems that we couldn't seem to trace. Our staining procedures are similar to yours. We made up fresh stains, changed all our syringe filters, used every precaution we could think of.
Then we had the water system checked. Our in-line reverse-osmosis, deionization system had become a mess, although we had assumed (there's that word!) that the company we leased it from was maintaining it properly. Turns out that they thought we owned the system, while in fact we only rented it and paid them to service it.
Anyway, to keep it short, we purchased a Millipore bench-top, low-volume water-polishing unit and used our old water to feed it (after getting the thing serviced properly). Our precipitates disappeared and have not yet reappeared.
For what it's worth.
(No financial interest in Millipore, etc.)
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: Phoebe J Doss [mailto:pjdoss-at-okstate.edu] Sent: Tuesday, June 13, 2000 9:33 AM To: microscopy-at-sparc5.microscopy.com
Hello:
After many years of staining grids with uranyl acetate and lead citrate, we have begun to see a needle like or shard precipitate, (about 1/2 inch long at 100,000x's; resembles the lead precipitate on page 469 of Electron Microscopy second edition John Bozzola and Lonnie Russell). We have been using a 2.5% aqueous uranyl acetate and Reynold's lead citrate (filtered through a 2 micron filter) for the past 4 years or so with no problems. I have stained the grids with just UA and can see no precipitate and have stained grids with just the lead citrate and still not see the precipitate. I have also checked the water and cannot still see the precipitate. However, when I stain with UA followed by lead citrate it mysteriously reappears much to my dissatisfaction. I have also tried the basic lead citrate and just recently tried Sato's lead stain and had the same problem. I have made up UA from a newly purchased bottle. I have also lessened the staining times from 30 minutes in UA to 7 minutes and from 20 minutes in lead citrate to 5 minutes and the precipitate is less but still there. I have also checked the grids before staining them and cannot see the precipitate. Please help, I grow more grey day by day.
Phoebe J. Doss Manager/Adjunct Instructor Electron Microscope Lab Oklahoma State University
Transmission Electron Microscopist (TEM) / Engineer
United Technologies Corporation is seeking an engineer to fill the TEM operator/engineer position at the United Technologies Research Center in East Hartford, CT. This position will provide support to the United Technologies Corporation Business Units including Pratt & Whitney, Carrier, Sikorsky Aircraft, Hamilton Sundstrand, Otis Elevator, and International Fuel Cells. The TEM operator will be responsible for the dailyoperations of the TEM laboratory; including preparation of TEM samples using various techniques such as dimpling, ion milling, jet polishing, microtoming, and replication. Project duties include conducting failure analyses, characterization of surface coatings, and analysis of advanced metal and ceramic materials. The candidate should have experience with both TEM sample preparation and conventional TEM operation. Good communication and interpersonal skills are essential. The ability to recognize fracture modes and origins of fractures is desired. Experience with Scanning Electron Microscopy (SEM) is a plus.
Qualified candidates will have a BS in Materials Science or equivalent, with a minimum of 2 years TEM experience. U. S. citizenship or permanent residency is required.
Please visit our web site at www.utrc.utc.com, and send your resume to Employment Opportunities, Code MATS-2050-9049, 411 Silver Lane, East Hartford, CT 06118 or e-mail employment-at-utrc.utc.com. United Technologies Corporation is an equal opportunity employer.
} } Back in the olden days, when BioRad sold microscopy supplies, they had an } epoxy cleaner (to remove epoxies from hands, benches, etc., not made from } epoxies). It was blue gunk in a little jar. Does anyone know what happened } to this stuff? Or have an alternate? } } I've just been digging through catalogues and Ted Pella sells a liquid } cleaner - any experience with it? }
I seem to remember that soaking in N,N-dimethyl formamide dissolves epoxy, you might care to beg a little from a freiendly chemistry dept and try it, it's not very expensive. It has a moderately offensive smeel, though.
rtch
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Yes, Ladd stills sells Met-A-Terge (catalog #13045). Please check our web site http://www.laddresearch.com for more information or contact me off line.
JD Arnott
Disclaimer: As stated above, Ladd sell Met-A-Terge and thus has a commercial interest in this thread.
LADD RESEARCH 131 Dorset Lane Williston, VT 05495 USA
TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net
ALL NEW WEB SITE UP NOW AT OUR NEW URL:
http://www.laddresearch.com
Beverly_E_Maleeff-at-sbphrd.com-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 } -----------------------------------------------------------------------.} } Tamara: } } I don't know the Bio-Rad product, but Ladd sold (still sells?) a product called } Met-a-terge that gets rid of uncured resins. A little bit goes a long way. } I've used it for years. } } Hope this helps. } } Regards, } Bev Maleeff } SmithKline Beecham Pharmaceuticals
Hi Tamara, I have used the Ted Pella Epoxy Hand Cleaner and it works well. It will remove epoxy from hands and also glassware. Jo Dee Fish
PS I am not affiliated with Ted Pella, just love their products!
Tamara Howard 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 } -----------------------------------------------------------------------. } } Back in the olden days, when BioRad sold microscopy supplies, they had an } epoxy cleaner (to remove epoxies from hands, benches, etc., not made from } epoxies). It was blue gunk in a little jar. Does anyone know what happened } to this stuff? Or have an alternate? } } I've just been digging through catalogues and Ted Pella sells a liquid } cleaner - any experience with it? } } Thanks! } } Tamara Howard } CSHL
-- Jo Dee Fish Electron Microscopy Assistant Cell Analysis Facility The Burnham Institute 10901 N. Torrey Pines Rd. La Jolla, CA 92037 (858)646-3100 ext. 3620
My husband uses epoxy on a sailboat he's building. He cleans everything up... hands, spills, etc... with plain old vinegar. We go through a LOT of vinegar. If it's dried, then soak acetone on it until it softens, then use vinegar (or 5% acetic acid) to mop up the residues.
connie m
At 10:31 AM 06/13/2000 -0400, Tamara Howard wrote: } Back in the olden days, when BioRad sold microscopy supplies, they had an } epoxy cleaner (to remove epoxies from hands, benches, etc., not made from } epoxies). It was blue gunk in a little jar. Does anyone know what happened } to this stuff? Or have an alternate? } } I've just been digging through catalogues and Ted Pella sells a liquid } cleaner - any experience with it? } } Thanks! } } Tamara Howard } CSHL } } } Connie McManus Veterinary Diagnostics Lab Utah State University Logan, UT USA
} To use the specific example of John's equipment: suppose } it was bought originally with, say, an NSF grant, and } I find that I could make use of it now. In order to do } that I would need to find a non-US-government source } of money, as I could not use even the income from my } facility operation (which is regarded by the accountants } as government money, as it originates } predominantly from government research grants).
I would assume it can even get messy if, at least some, of my facility's income had come from outside sources. I would imagine the accountants will first assume it is government $$ ... in which case I would have to show I had taken in an equivelent in outside $$ ... but over what time frame? ... this fiscal? ... last 10 years?
cheerios, =shAf= :o)
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - mshaf-at-darkwing.uoregon.edu Geological Science's Electron Probe Facility - University of Oregon http://epmalab.uoregon.edu/
I use methylene dichloride (dimethyl chloride) to dissolve most any epoxy.
I have 3-5 gallons of it. I do not know if it is still generally available.
gg
At 07:31 AM 6/13/00, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
The listserver is at MSA listserver {Microscopy-at-sparc5.microscopy.com} Just sent email say, "Please subscribe". It is fairly active but is only about 1/2 relevant as most data is biology oriented.
Earl JCNABITY-at-aol.com wrote:
} Dear Earl, } } Could you tell me how to subscribe to the MSA listserver? Greg talked highly } of it and I'm thinking I will set up another e-mail account to use for it. } Since it is pretty active, I didn't want all the messages going in with my } normal e-mail, but a using a separate account will resolve that issue. } } Joe
Our EM Unit has a Zeiss EM109 which uses 70mm roll film - Agfa Scientia 23D56. Manufacture of this film ceased quite a while back. We are trying to locate unused stock of this film for our usage. If you have any surplus stock for sale please contact me.
Thanks Mohamed
****************************** M. A. Jaffer Electron Microscope Unit R. W. James Building University of Cape Town Private Bag Rondebosch, 7701 South Africa
At 06:09 PM 06/13/2000 -0700, Don Hammer wrote: } Stuff is cheap too and if there is any left after the zillions of home uses, } great on salads!!!!
yeah, especially the used stuff........ eeeeuewwwwwwww! *G*
connie m } } Don Hammer, Retired Guy } ----- Original Message ----- } From: Connie McManus {conmac-at-cc.usu.edu} } To: Tamara Howard {howard-at-cshl.org} ; Microscopy Listserver } {Microscopy-at-sparc5.microscopy.com} ; Histology listserver } {histonet-at-pathology.swmed.edu} } Sent: Tuesday, June 13, 2000 2:29 PM } Subject: Re: Epoxy cleaner? } } } } My husband uses epoxy on a sailboat he's building. He cleans everything } } up... hands, spills, etc... with plain old vinegar. We go through a LOT } of } } vinegar. If it's dried, then soak acetone on it until it softens, then } use } } vinegar (or 5% acetic acid) to mop up the residues. } } } } connie m } } } } At 10:31 AM 06/13/2000 -0400, Tamara Howard wrote: } } } Back in the olden days, when BioRad sold microscopy supplies, they had an } } } epoxy cleaner (to remove epoxies from hands, benches, etc., not made from } } } epoxies). It was blue gunk in a little jar. Does anyone know what } happened } } } to this stuff? Or have an alternate? } } } } } } I've just been digging through catalogues and Ted Pella sells a liquid } } } cleaner - any experience with it? } } } } } } Thanks! } } } } } } Tamara Howard } } } CSHL } } } } } } } } } } } Connie McManus } } Veterinary Diagnostics Lab } } Utah State University } } Logan, UT } } USA } } } } } } Connie McManus Veterinary Diagnostics Lab Utah State University Logan, UT USA
Hi to all, Does anybody have some information on a possible relationship between lipid droplets and wax production in plant cells. Are there some evidences that lipid droplets could actually be storage sites of wax precursors ? I looked for that in literature but found nothing... Thus : HELP ! References will be welcome Thanks in advance for answering Bye Pascal
""""""""' ( O)(o ) --------------------0000--------------0000---------- Pascal VEYS Laboratory of Plant Biology Catholic University of Louvain Place Croix du Sud 5 (bte 14) B 1348 Louvain-la-Neuve Belgium Phone : 0032 10473004 Fax : 0032 10473471 Email : Veys-at-bota.ucl.ac.be ---------------------ooooO-----------Ooooo-------- ( ) (_)(_) ( ) ) ( ) ( (_) (_)
Gary Gaugler wrote: =============================================================== I use methylene dichloride (dimethyl chloride) to dissolve most any epoxy. I have 3-5 gallons of it. I do not know if it is still generally available. ================================================================ If we are talking about methylene dichloride, or dichloromethane, CAS # 75- 09-2, this is a pretty bad actor, and is on the list of Prop. 65 chemicals for the State of California as being cancer causing. Chemicals on the Prop . 65 list are so highly restricted that in some organizations, they are allowed in only with the approval of top management.
But I do have a question: I always thought that most epoxies, certainly the ones used in microscopy, ended up being three dimensionally crosslinked intractable solids. The only way such a material is going to be "dissolved" is for chemical bonds to be broken. And yet, I don't see how chemically, methylene dichloride is going to be breaking chemical bonds. Or the same comment for some of the other materials mentioned. These materials might plasticize (e.g. soften) an epoxy and aid in its removal from a surface, but do any of these really "dissolve" a three dimensionally crosslinked epoxy system?
I am very interested in this topic because we believe that at least in terms of getting epoxy out of the "nooks and crannies" of a non-smooth surface, an oxygen plasma is needed. But perhaps we are wrong about that, that is why I ask the question.
Disclaimer: SPI Supplies manufactures the Plasma Prep II plasma etcher and has an interest in seeing more applications for plasma etching.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.spi.cc ######################## ============================================
David, I had the same problem with a gold target-a dark spot in the middle. Also, what was being sputtered on my samples was not gold. I cleaned the target with acetone and it has been working fine since. Joyce Craig Chicago State University
We are working with Leishmania major, a parasite that is incorporated into macrophages. We have had good success with fixation of lymph nodes infected with Leishmania. We have not been as happy with the results of fixation of cell cultures of infected bone marrow macrophages. The membranes of the Leishmania and of the internal compartments within the macrophages that hold the Leishmania are well fixed, but the external cell membranes of the macrophages are somewhat discontiuous. We have not done cell cultures before. Is this a problem to be expected? We are fixing with 2+2 glutaraldehyde/paraformaldehyde in 0.1 M phosphate buffer, post-fixing with 2% buffered Osmium, dehydrating with ethanol and propylene oxide, then embedding in epoxy. We have shortened all times compared to those we use with tissue samples. Joyce Craig Chicago State University
I am looking for a non-fluorescent plastic coverslip that allows confocal laser microscopy and subsequent sectioning for Transmission Electron Microscopy. I will greatly appreciate your suggestions.
Sincerely, Karthi Subramanian Department of Microbiology University of Guelph Guelph Ontario N1G 2W1 Phone: (519)824-4120 ext.8904 Fax:(519)837-1802
I perform stereo measurements only occasionally, so I prefer to save money on specialized equipment or software. All I use is just freeware program ImageTool (good for on-screen stereo pair measurements) and Excel (not bad for calculations).
Of course, for a big project it's better to buy a software.
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} -----Original Message----- } From: John J. Bozzola [mailto:bozzola-at-siu.edu] } Sent: Thursday, June 08, 2000 11:13 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Stereometry by computer } } } -------------------------------------------------------------- } ---------- } 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 } } -------------------------------------------------------------- } ---------. } } } A user of our facility is interested in making height measurements } from specimens viewed in the SEM. I am aware of the conventional way } of doing this: stereo pairs and optical viewer (with stereometer } parallax corrections). Is there a more modern (computerized) way of } doing this, say with anaglyphs? } } Thank you. } } John B. } } } #################################################################### } John J. Bozzola, Ph.D., Director } Micro-Imaging and Analysis Center } 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 } #################################################################### }
I need to label Si-OH groups with something that will show up in TEM. If it were proteins or sugars, I'd immuno- or lectin label. Any ideas how I can get gold or ferritin or whatever onto these groups?
Mahalo! Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
} Hello, } } Can anyone recommend a supplier for uranyl acetate? } Also, does anyone out there have experience with Sigma's Lowicryl kit? } If possible, please respond to me directly at: mpilgrim-at-mendelbio.com } } Many thanks, } Marsha
Dear Marsha,
We at Ladd Research, and most of the other supply companies, can sell you this. In our case it is catalog # 23620 and more information can be found on our web site, http://www.laddresearch.com
John Arnott --
LADD RESEARCH 131 Dorset Lane Williston, VT 05495 USA
TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net
Help - A faculty member's plant tissue is autofluorescing. She is using aniline blue to look at pollen tubes (through the styles). She would like to reduce the background fluorescence. I gave her a copy of a borohyride reference from a '97 listserv posting. Can anyone recommend a fixation that will decrease or eliminate the autofluorescence? Any thoughts on this matter would be greatly appreciated. thanks, Beth
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
It took me a few days to find this protocol from my collegue Lucinda Swatzell. It sounded intriguing. Although I've not tried it myself yet, she's used it with success.
I've copied the following out of her e-mail. Things that need to be kept in mind are that she polymerizes in a vacuum oven, she's refering to plant seedlings, and that I don't have any financial interest that I know of (i.e. I haven't checked my mutual fund prospectus) in Rubbermaid, Inc.
Here it is:
"There is a way to flat embed with LR White: This works great for me when I am keeping them on their agar blocks and maintaining orientation, but it should also work for regular seedlings to keep them flat instead of crooked in the bottom of the capsules. Use rubbermaid ice cube trays. Place the specimens in the flat bottoms of the tray. cover with about 1/4 in of resin. In each well, place another well that has been cut from it's tray. The single loose wells will nestle down into the ice cub tray on top of the resin. Because rubbermaid is dishwasher safe it will take the heat, but get soft enough to snuggle in tightly and keep out oxygen. When you pump the vaccuum the extra resin also snakes up into the cracks, so that you get a seal. Now the thing that is scary: will loose seedlings just suck up into the cracks? I haven't tried them."
Heather A. Owen, Director Electron Microscope Laboratory Department of Biological Sciences University of Wisconsin - Milwaukee (414)229-6816
I've used it often to unpackage epoxy microchips. These epoxies are silicone/epoxy and not true epoxy. They are typically called plastic packaged ICs. Whatever. I've used MEC and DiMEC to do this. Some packages had to be oxygen ion blasted open. Over time, the plasma method has been much safer for the operator and the chip. And it can make the removal process much faster. Plasma is the current method of choice.
Whether the "epoxies" talked about here are the same as those for IC packages is likely not to be.
gary g.
At 09:40 AM 6/14/00, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I used to prepare W FIM emitter tips with a 5% NaOH solution and I think the voltage was around 5 to 10 volts ac. When I used to electropolish, the FIM solutions and conditions were frequently similar to the jet polishing solutions.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
} -----Original Message----- } From: Gillian Bond [mailto:gbond-at-nmt.edu] } Sent: Monday, June 12, 2000 7:43 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Jet polishing of tungsten } } } -------------------------------------------------------------- } ---------- } 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 student who has been trying to jet polish tungsten } for TEM. She } has tried various concentrations of sodium hydroxide in } water, as well as } 40g trisodium phosphate/250ml water, and 55.8g magnesium } perchlorate/250ml } methanol, at a range of voltages. We have a Fischione jet-polishing } unit. So far, none of the samples has been close to good. Can anyone } help us out here, with past experience or general suggestions? } } Many thanks in advance, } } Gill } } Dr Gillian M. Bond } Department of Materials & Metallurgical Engineering } New Mexico Tech } Socorro, NM 87801 } }
My name is Tim Strovas and I am a graduate student at the U of Washington bioengineering department. I am investigating the halo effect from single myofibrils (from bumblebee muscle tissue) as seen under a phase contrast microscope. Has anyone encountered a previous investigation into this effect and its possible relationship with structured water? Note: The Halo does not appear as ripples that are normally associated with optical microscope artifacts. The halo is single broad band that borders the tissue sample.
My name is Tim Strovas and I am a graduate student at the U of Washington bioengineering department. I am investigating the halo effect from single myofibrils (from bumblebee muscle tissue) as seen under a phase contrast microscope. Has anyone encountered a previous investigation into this effect and its possible relationship with structured water? Note: The Halo does not appear as ripples that are normally associated with optical microscope artifacts. The halo is single broad band that borders the tissue sample.
My name is Tim Strovas and I am a graduate student at the U of Washington bioengineering department. I am investigating the halo effect from single myofibrils (from bumblebee muscle tissue) as seen under a phase contrast microscope. Has anyone encountered a previous investigation into this effect and its possible relationship with structured water? Note: The Halo does not appear as ripples that are normally associated with optical microscope artifacts. The halo is single broad band that borders the tissue sample.
G'day folks, Be very wary of any Good Epoxy Solvent, do not use it to clean epoxy off your skin. Anything that is a good solvent for epoxy will probably be a good solvent for the oils and lipids in/on your skin. These oils and lipids are your protection against epoxy resins entering your body. Remember, all epoxy resins at carcinogenic, soap and water are probably the safest agents to remove epoxies from your skin. Regards JVN
Connie McManus 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 } -----------------------------------------------------------------------. } } At 06:09 PM 06/13/2000 -0700, Don Hammer wrote: } } Stuff is cheap too and if there is any left after the zillions of home uses, } } great on salads!!!! } } yeah, especially the used stuff........ eeeeuewwwwwwww! *G* } } connie m } } } } Don Hammer, Retired Guy } } ----- Original Message ----- } } From: Connie McManus {conmac-at-cc.usu.edu} } } To: Tamara Howard {howard-at-cshl.org} ; Microscopy Listserver } } {Microscopy-at-sparc5.microscopy.com} ; Histology listserver } } {histonet-at-pathology.swmed.edu} } } Sent: Tuesday, June 13, 2000 2:29 PM } } Subject: Re: Epoxy cleaner? } } } } } } } My husband uses epoxy on a sailboat he's building. He cleans everything } } } up... hands, spills, etc... with plain old vinegar. We go through a LOT } } of } } } vinegar. If it's dried, then soak acetone on it until it softens, then } } use } } } vinegar (or 5% acetic acid) to mop up the residues. } } } } } } connie m } } } } } } At 10:31 AM 06/13/2000 -0400, Tamara Howard wrote: } } } } Back in the olden days, when BioRad sold microscopy supplies, they had an } } } } epoxy cleaner (to remove epoxies from hands, benches, etc., not made from } } } } epoxies). It was blue gunk in a little jar. Does anyone know what } } happened } } } } to this stuff? Or have an alternate? } } } } } } } } I've just been digging through catalogues and Ted Pella sells a liquid } } } } cleaner - any experience with it? } } } } } } } } Thanks! } } } } } } } } Tamara Howard } } } } CSHL } } } } } } } } } } } } } } } Connie McManus } } } Veterinary Diagnostics Lab } } } Utah State University } } } Logan, UT } } } USA } } } } } } } } } } } Connie McManus } Veterinary Diagnostics Lab } Utah State University } Logan, UT } USA
-- **************************************************** John V Nailon Operations Manager Centre for Microscopy and Microanalysis The University of Queensland St. Lucia Queensland 4072 Phone: +61-7-3365-4214 Fax: +61-7-3365-4422 WWW: http://www.uq.edu.au/nanoworld/allstaff.html#Nailon ****************************************************
I suggest you copy this message to the plant surfaces mail list
To join, send the command join plant-surfaces firstname lastname to: mailbase-at-mailbase.ac.uk "Firstname" can be one or more names or initials. The last word in this command will be interpreted as the last name. The email address will be extracted automatically from the message.
Chris Jeffree
Date sent: Wed, 14 Jun 2000 16:18:08 +0100 To: microscopy-at-sparc5.microscopy.com } From: veys-at-bota.ucl.ac.be (Pascal Veys)
Hello Tina:
I would approach your problem by either modifying the Si-OH groups with a hapten and detecting with antibody- or streptavidin-gold, or converting them to amines or thiols then labeling with a gold labeling reagent (disclaimer - we make gold labeling reagents). You could introduce amino- groups at the Si-OH groups using a silylating reagent such as 3-{Tris[2-(2-methoxyethoxy)ethoxy]silyl}propylamine or 3-[Tris(trimethylsiloxy)silyl]propylamine (both from Fluka), then either biotinylate with NHS-biotin and detect with streptavidin-gold, or label the amines with Mono-Sulfo-NHS-Nanogold.
I have not actually tried this, and since I don't know what types of samples you are looking at, it's difficult to say what else in them might affect the reaction. If there are already other primary amines in your sample, they need to be blocked first.
If you would like other ideas, a text on solid-phase oligo- or peptide synthesis might be another good starting point - the chemistry used to functionalize the beads used in these systems may also be transferable to your situation.
Hope this is helpful,
Rick Powell
} } Oh wise and helpful microscopists- } } I need to label Si-OH groups with something that will show up in TEM. If } it were proteins or sugars, I'd immuno- or lectin label. Any ideas how I } can get gold or ferritin or whatever onto these groups? } } Mahalo! } Tina } } **************************************************************************** } * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * } * Biological Electron Microscope Facility * (808) 956-6251 * } * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* } ****************************************************************************
John Nailon is making a very good argument for using gloves or working very cleanly. All epoxies I understand are "somewhat" carcinogenic. The much quoted John Luft, years ago advised me that photographic fixer (sodium thiosulphate) solution, chemically changed epoxies so they would not be carcinogenic. If he was right, then first washing any body parts contaminated by epoxy resin in photographic fixer should avert the worse. Those fixers do not dissolve or clean epoxies. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com
On Thursday, June 15, 2000 11:36 AM, John Nailon [SMTP:mmjnailo-at-dingo.cc.uq.edu.au] wrote: } } G'day folks, } Be very wary of any Good Epoxy Solvent, do not use it to clean epoxy off } your skin. Anything that is a good solvent for epoxy will probably be a } good solvent for the oils and lipids in/on your skin. These oils and } lipids are your protection against epoxy resins entering your body. } Remember, all epoxy resins at carcinogenic, soap and water are probably } the safest agents to remove epoxies from your skin. } Regards } JVN } } Connie McManus 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 } } -----------------------------------------------------------------------. } } } } At 06:09 PM 06/13/2000 -0700, Don Hammer wrote: } } } Stuff is cheap too and if there is any left after the zillions of home } } } uses, } } } great on salads!!!! } } } } yeah, especially the used stuff........ eeeeuewwwwwwww! *G* } } } } connie m } } } } } } Don Hammer, Retired Guy } } } ----- Original Message ----- } } } From: Connie McManus {conmac-at-cc.usu.edu} } } } To: Tamara Howard {howard-at-cshl.org} ; Microscopy Listserver } } } {Microscopy-at-sparc5.microscopy.com} ; Histology listserver } } } {histonet-at-pathology.swmed.edu} } } } Sent: Tuesday, June 13, 2000 2:29 PM } } } Subject: Re: Epoxy cleaner? } } } } } } } } } } My husband uses epoxy on a sailboat he's building. He cleans everything } } } } up... hands, spills, etc... with plain old vinegar. We go through a LOT } } } of } } } } vinegar. If it's dried, then soak acetone on it until it softens, then } } } use } } } } vinegar (or 5% acetic acid) to mop up the residues. } } } } } } } } connie m } } } } } } } } At 10:31 AM 06/13/2000 -0400, Tamara Howard wrote: } } } } } Back in the olden days, when BioRad sold microscopy supplies, they had } } } } } an } } } } } epoxy cleaner (to remove epoxies from hands, benches, etc., not made } } } } } from } } } } } epoxies). It was blue gunk in a little jar. Does anyone know what } } } happened } } } } } to this stuff? Or have an alternate? } } } } } } } } } } I've just been digging through catalogues and Ted Pella sells a liquid } } } } } cleaner - any experience with it? } } } } } } } } } } Thanks! } } } } } } } } } } Tamara Howard } } } } } CSHL } } } } } } } } } } } } } } } } } } } Connie McManus } } } } Veterinary Diagnostics Lab } } } } Utah State University } } } } Logan, UT } } } } USA } } } } } } } } } } } } } } } } Connie McManus } } Veterinary Diagnostics Lab } } Utah State University } } Logan, UT } } USA } } -- } **************************************************** } John V Nailon } Operations Manager } Centre for Microscopy and Microanalysis } The University of Queensland } St. Lucia Queensland 4072 } Phone: +61-7-3365-4214 } Fax: +61-7-3365-4422 } WWW: http://www.uq.edu.au/nanoworld/allstaff.html#Nailon } ****************************************************
I needed to have a cross sectional view into an adhesive layer so that I could count the layers, if possible. Since the adhesive acts like a highly viscous liquid, polishing it is out of the question. Instead, I submerged the film into liquid Nitrogen and cut it quickly w/ a scissors. The sound of the cut was more like a breaking sound. When I examined the cross sectional surface I observed a brain like surface. There were columns with a highly consistent diameter and orientation. It looked crystalline. There was zero evidence of material smearing that one would expect if one cut a material. Is the proper interpretation that these columns existed before the fracture and that the fracture occurred along the boundaries? Or is the cross sectional surface generated by a rippled distortion of an highly viscous liquid? The regularity of the surface seems to make the latter interpretation unlikely.
I would be interested in any opinions on the interpretation of the image or alternative means of cross sectioning the adhesive layer.
I could e-mail an image to anyone who is interested.
We haven't had a need to unpackage 'plastic' encapsulated IC's for a while but may have a need to do so soon (preferrably not impairing functionality?!). I am told that the last person to do this here (now retired) dripped fuming sulfuric acid on the plastic and used frequent water rinses. We have a plasma etcher but I was afraid it would take forever to get through the plastic.
Any hints and suggestions would be greatly appreciated.
Diane Ciaburri Senior Materials Engineer General Dynamics 100 Plastics Ave. Pittsfield MA 01210
I've used it often to unpackage epoxy microchips. These epoxies are silicone/epoxy and not true epoxy. They are typically called plastic packaged ICs. Whatever. I've used MEC and DiMEC to do this. Some packages had to be oxygen ion blasted open. Over time, the plasma method has been much safer for the operator and the chip. And it can make the removal process much faster. Plasma is the current method of choice.
Whether the "epoxies" talked about here are the same as those for IC packages is likely not to be.
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Hi, All:
This one is not really related to microscopy. But since I am a TEM guy, I hope I can get some help here.
Does anybody have some information about MgAl2O4 as a substrate for perovskite films? I know it is not often used, so I am worndering if it has a major disadvantage so that nobody is using it. Any references would be welcome.
I have a control PC board for the E5200 sputter coater. This is the model with an Intel single chip MPU on one end and a 4-conductor socket on the other end. The board uses a VME connector for main interface.
Coater is trashed. Board is OK. If anybody can use the board, first request gets it.
Beth, more details are needed on how the tissue was processed before viewing. I have used aniline blue to view pollen tubes in style of Salicornia virginica which had ben fixed in Nawashin's fixative. Have also viewed tubes of Melilotus which had simply been preserved in 70% EtOH. If one uses glut as a fixative, it fluoresces so you won't be able to distinguish the PT from everything else. Mary Pfauth
Can't comment on sulfuric, but I have used red fuming nitric at near boiling temperature. Apply acid, let react. Flush witn more acid, let react, etc.
Woody White
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Gary and others,
We haven't had a need to unpackage 'plastic' encapsulated IC's for a while but may have a need to do so soon (preferrably not impairing functionality?!). I am told that the last person to do this here (now retired) dripped fuming sulfuric acid on the plastic and used frequent water rinses. We have a plasma etcher but I was afraid it would take forever to get through the plastic.
Any hints and suggestions would be greatly appreciated.
Diane Ciaburri Senior Materials Engineer General Dynamics 100 Plastics Ave. Pittsfield MA 01210
I've used it often to unpackage epoxy microchips. These epoxies are silicone/epoxy and not true epoxy. They are typically called plastic packaged ICs. Whatever. I've used MEC and DiMEC to do this. Some packages had to be oxygen ion blasted open. Over time, the plasma method has been much safer for the operator and the chip. And it can make the removal process much faster. Plasma is the current method of choice.
Whether the "epoxies" talked about here are the same as those for IC packages is likely not to be.
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Hello,
Does anyone know the shear modulus for LaAlO3?
Thanks
Yan Xin ======================================= Yan Xin Magnet Science & Technology National High Magnetic Field Laboratory Florida State University 1800 E. Paul Dirac Drive Tallahassee, FL 32310 Tel: (850) 644 1529 Fax: (850) 644 0867 ========================================
The ion beam approach works well. I have not used it recently on finer pitch ICs. With as-built feature sizes of 2-4u, it is fine. It will stop at the passivation and leave the Al bond wires intact. The resulting package looks like it has a V-shaped pit in it (which it does). The extent of the pit depends on the size of the die and if you want to blast down to the lead frame or substrate.
I have not done this on finer pitch devices. I would be a bit skeptical about these mostly because of the smaller bond pads. The etching would still stop at the passivation.
There are numerous places in Silicon Valley that do this on an outsource basis. Typical costs are about $75 per IC. I can get some contacts for you if you'd like.
gary g.
At 06:55 AM 6/15/00, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
MME is currently conducting research through Microscopy & Analysis regarding the impact of the internet on microscopy and imaging facilities. If you have not yet faxed back your responses, we'd appreciate your participation. The questionnaire is in the center of the May issue of M&A.
Results of this survey will be reported in a Fall issue of Microscopy & Analysis; specific information on the impact of the internet will be presented along with data collected from other recent MME surveys and reports from other meetings in the "Microbrew" column in the July issue of Advanced Imaging.
Many thanks.
Best regards, Barbara Foster Microscopy/Marketing & Education 125 Paridon Street, Suite 102 Springfield, MA 01118-2130 PH: 413-746-6931 FX: 413-746-9311 email:mme-at-map.com
To the wealth of knowledge on the Microscopy list server,
I have a question about Osmium fixation..Basically I was curious to know if there are any references to Osmium fixation at room temperature? Everything I have seen so far only talks about fixation for 2 hours in the refrigerator at 4 degrees...
Are there any drawback or problems that can occur if tissues specifically Kidney and muscle would or could have? i.e. precipitation etc... etc....
} } We haven't had a need to unpackage 'plastic' encapsulated IC's for a while but } may have a need to do so soon (preferrably not impairing functionality?!). I am } told that the last person to do this here (now retired) dripped fuming sulfuric } acid on the plastic and used frequent water rinses. We have a plasma etcher but } I was afraid it would take forever to get through the plastic. } } Any hints and suggestions would be greatly appreciated. } } Diane Ciaburri } Senior Materials Engineer } General Dynamics } 100 Plastics Ave. } Pittsfield MA 01210
Diane, Yes, hot fuming sulfuric and/or hot fuming nitric are used as standard procedures for removing plastic from IC's. The process is not quite that simple. For example, water rinses will almost certainly etch the bond pads on the IC and thus removing connection to the outside world. Additionally, the plastic contains fire retardants which some regions don't like being washed down the drain. There is more detailed help through EDFAS.org (one of ASM's branches). B&G International sells a very safe, effective etcher which performs decapsulation automatically in minutes.
I have no association with B&G International.
David Saxon Analytical Microscope Services 11826 Reservoir Rd. E. Puyallup, WA 98374 253-848-7701 voice & fax email: info-at-analyticalmicroscope.com website: www.analyticalmicroscope.com
I am looking for information about an analysis software package by the name of ONCOR. Does any have an adress for the company.... which may not exist anymore?? Thanks Blystone in Texas
Robert V. Blystone, PH.D. Professor of Biology Trinity University San Antonio, Texas 78212 rblyston-at-trinity.edu 210-999-7243 FAX 210-999-7229
Being unable to afford a digital camera for my TEM. I'm wondering if the next best option is to get a high end scanner to scan in negatives and then print them on a decent printer. Any advice regarding this idea and brands of scanners and printers that are useful? Also, what image analysis systems are user friendly?
Dr. Thomas P. Bonner Department of Biological Sciences SUNY at Brockport Brockport, NY 14420
I'm looking for advice on embedding bovine oocytes (~100 micron diameter). I'd like to embed them in araldite for probing with labelled lectins as this seems to be fairly well established in the literature. The hangup is this: we are using fairly large plastic cassettes and are having problems losing the oocytes within the volume of araldite. Does anyone know of a way around this? Is there something we can pre-embed the oocytes in to make a smaller chip that we can then embed in the larger block (that is, of course, compatible with polymerizing / clearing the araldite? Or does anyone know of an altogether different method for oocyte embedding that is more effective? Thanks in advance!
--Carrie Golash
Carrie Golash John O. Almquist Research Center Penn State University University Park, PA 16802 W: (814) 865-5896 H: (814) 692-7926 http://www.das.psu.edu/dbrc/dbrc.htm
Our materials science microscopy lab is in need of a used atomic force microscope. No specific model or make. All reasonable offers will be considered. Please respond directly to Don Kierstead at or call 330-794-6600. Any help in this effort would be greatly appreciated.
Dichloromethane and dimethylformamide are relatively effective disrupters of most epoxies but their action is accompanied by great swelling because the polymer becomes engorged with the liquid before any significant solvation takes place. This will destroy wire bonds on an IC.
Fuming (essentially anhydrous) sulfuric acid acts by the completely different process of sulfonating reactive groups that remain on the polymer. The depolymerized and sulfonated byproducts are quite soluble not only in the acid but usually in water as well. The worst thing that you could do in this relatively straightforward process is to wash with water at intervals because this would initiate almost instantaneous corrosion. It would be advisable for a chemist, as someone trained in the handling of reactive materials, to carry this out or at least to establish procedures and train others with less experience. The action of sulfuric acid in this regard is quite different than that of nitric. Nearly anhydrous nitric acid (completely anhydrous is extremely difficult to prepare) is a very powerful oxidizer and could lead to unstable, dangerous byproducts whereas the sulfonates resulting from the sulfuric acid reaction are relatively stable. Water must, of course, be prevented from splashing into any concentrated acid, especially sulfuric.
A very strong acid such as sulfuric behaves completely differently in the absence of water. Since most acids are highly hygroscopic and are sold as water solutions, most people do not observe this other side of their behavior. Without water to create an ionized electrolyte, corrosion of metals will not take place. I have de-encapsulated ICs for failure analysis in 200 degree sulfuric acid and been able to operate the IC without replacing the .001" aluminum wirebonds that it came with. I recall one instance where our company built prototype hybrid microelectronic circuits out of such de-encapsulated ICs when their supplier was late getting a new design on the market and the only ones available were already encapsulated.
The key is to realize that water must be excluded until the sulfonating acid has been completely rinsed away by a non-aqueous liquid. As Mr. Saxon said, there are simple and safe devices available for doing this operation. However, with proper care and protective gear it can be done in a beaker on a hot plate in a fume hood. A few ml.s of sulfuric acid are heated to drive off water until heavy vapors are observed over the liquid (which may darken during heating due to trace impurities). The IC is carefully lowered into the hot acid and a vigorous reaction ensues with the epoxy almost instantly washing into the solution. After a few seconds the IC is then quickly lifted out and held over a receiving vessel and flooded with a stream of ethanol. Only after this is a final rinse in deionized water carried out, followed by fresh electronic grade ethanol and forced drying in warm air.
The ready made devices which carry out the operation are typically a small bowl with a hinged lid from which air is withdrawn by a gentle vacuum. An inert metal feeder tube leads from a heated reservoir for the sulfuric acid and passes through the wall of the bowl to a position where the encapsulated device is secured. When the lid is closed and the slight vacuum applied, the hot acid is pulled into the bowl over the device. It is somewhat self-limiting in that, if the lid is opened, there is no driving force to bring more acid into the container. Naturally, the vacuum source needs to be protected by a trap and all waste products properly handled no matter how the procedure is carried out.
John Twilley Conservation Scientist (formerly, Manager of the Reliability Analysis Center, Teledyne Microelectronics)
DAVID I SAXON 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 haven't had a need to unpackage 'plastic' encapsulated IC's for a while but } } may have a need to do so soon (preferrably not impairing functionality?!). I am } } told that the last person to do this here (now retired) dripped fuming sulfuric } } acid on the plastic and used frequent water rinses. We have a plasma etcher but } } I was afraid it would take forever to get through the plastic. } } } } Any hints and suggestions would be greatly appreciated. } } } } Diane Ciaburri } } Senior Materials Engineer } } General Dynamics } } 100 Plastics Ave. } } Pittsfield MA 01210 } } Diane, } Yes, hot fuming sulfuric and/or hot fuming nitric are used as standard } procedures for removing plastic from IC's. The process is not quite that } simple. For example, water rinses will almost certainly etch the bond pads } on the IC and thus removing connection to the outside world. Additionally, } the plastic contains fire retardants which some regions don't like being } washed down the drain. There is more detailed help through EDFAS.org (one } of ASM's branches). B&G International sells a very safe, effective etcher } which performs decapsulation automatically in minutes. } } I have no association with B&G International. } } David Saxon } Analytical Microscope Services } 11826 Reservoir Rd. E. } Puyallup, WA 98374 } 253-848-7701 voice & fax } email: info-at-analyticalmicroscope.com } website: www.analyticalmicroscope.com
We often work with clients who wish to look inside materials. Firstly the SEM is very clever it will tell you if a material is cut with a blade or a knife or scissors!
The only way to see the true internal structure of a material is to fracture it. Drop the material into LN2 wait until the bubbles stop and then take it out and using heavy duty tweezers crack it.
If a material (like hair and some polymer fibres) will not crack you need to support them in some way to make them crack. We use a water based carbon solution and two SEM stubs. Glue the two stubs together with the water soluble adhesive (try Spi) and then drill two or three small holes through the stubs (about 1mm diameter). Glue the hairs together with the carbon solution and pass then through the holes (messy). When all is dry plunge into LN2. Tap a blade between the two stubs and ALL the material should fracture.
Alternatively, take a fine bore drinking straw and pass the hairs plus carbon solution into the straw. Wait until dry, dump in LN2 and flex the straw to crack it and its contents.
Such fractures of layered materials (e.g. paints) will be best viewed in BSE each "phase" will either be of a different contrast or fracture in a different way. Great fun, try it?
Steve Chapman Senior Consultant Protrain For consultancy and professional training in EM world wide Tel 44+ 1280 814774 Fax 814007 www.emcourses.com
Hi, I want to contact by Email Prof. N.D. Hallam (formerly at Melbourne and LA Tobe - Australia) Does anybody have his contact adress Thanks to all in advance Pascal
""""""""' ( O)(o ) --------------------0000--------------0000---------- Pascal VEYS Laboratory of Plant Biology Catholic University of Louvain Place Croix du Sud 5 (bte 14) B 1348 Louvain-la-Neuve Belgium Phone : 0032 10473004 Fax : 0032 10473471 Email : Veys-at-bota.ucl.ac.be ---------------------ooooO-----------Ooooo-------- ( ) (_)(_) ( ) ) ( ) ( (_) (_)
I«m thinking in buying a saphire knife for ultramicrotomy since they seem to be less expensive than the traditional diamond ones. I need to cut thin sections of sponges that are difficul to cut with glass knifes. Does anyone have experience with these knifes? How do they compare with diamond in terms of cutting properties and durability?
Thanks
Dr. A.P. Alves de Matos Dental Medical School Lisbon
Many of our customers are using Agfa Duoscan scanners with excellent results. These are "flatbed" type scanners but handle films in a separate drawer, similar to a negative carrier in an enlarger. The advantage of this system is not scanning through glass, eliminating the chance of Newton Rings. In addition, the Duoscan line offers high optical resolutions(up to 2500x2500ppi)and high dynamic ranges. The Umax Powerlook III is also an excellent scanner where budgets may be limited. It has 1200x2400 optical resolution and is a traditional "flatbed" design. Also new is the Linocolor 1400 with 1200x2400 resolution with a letter size scan bed.
Choosing a printer is more difficult, depending on your output needs. High end photographic printers such as the Fuji Pictrography or dyesub printers from Kodak and Sony offer top quality output but at a high price for both hardware and cost per print. For publication quality prints, these are the best. Ink jet printers continue to improve in image quality, and more importantly, long term image stability. The cost of these printers is very low although they are very slow, and still somewhat costly per print when used with the higher quality print materials. Most inkjets are also better at producing color prints than monochrome prints. Another favorite of ours is the Tektronix Phaser 850. This high quality plain paper printer uses a unique Solid Ink technology. Ink is supplied not in a liquid form but a solid blocks. Cost per print is very low and black ink is free for the life of the printer. The Phaser 850 will also handle any "office" type output such as letters and reports with the advantage of integrating images into pages instead of attaching all photos at the end.
We are currently working on a project involving blastocysts and since we aren't osmicating the samples, we also have the problem of seeing them in the resin. Embedding them in agar helps somewhat. Even though it's also relatively transparent, the larger size of the agar chunk makes it easier to see.
We perform our primary fixation, then buffer washes, then make a 2% agar solution on the hot plate. When the agar cools down enough to be quite warm to the touch (but before the gelling stage), we pipette our cells into it on a microscope slide or cover slip, then put it into the fridge to harden. It hardens almost immediately. Then we cut the piece of agar with the sample into a tiny cube and continue processing it normally.
Hope this helps.
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: Carrie Golash [mailto:cdg126-at-psu.edu] Sent: Thursday, June 15, 2000 9:14 PM To: Microscopy-at-sparc5.microscopy.com
Hello all -
I'm looking for advice on embedding bovine oocytes (~100 micron diameter). I'd like to embed them in araldite for probing with labelled lectins as this seems to be fairly well established in the literature. The hangup is this: we are using fairly large plastic cassettes and are having problems losing the oocytes within the volume of araldite. Does anyone know of a way around this? Is there something we can pre-embed the oocytes in to make a smaller chip that we can then embed in the larger block (that is, of course, compatible with polymerizing / clearing the araldite? Or does anyone know of an altogether different method for oocyte embedding that is more effective? Thanks in advance!
--Carrie Golash
Carrie Golash John O. Almquist Research Center Penn State University University Park, PA 16802 W: (814) 865-5896 H: (814) 692-7926 http://www.das.psu.edu/dbrc/dbrc.htm
At 11:44 AM +0100 6/16/0, A.P. Alves de Matos wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
********************* I had bought a saphire knife, years ago (early 1980's). Our lab bought it with the idea that it was a good half-way stop for a new tech who needed something better than glass. It had certain drawbacks....the edge seemed to collect debris and was more difficult to clean than a diamond, and of course it wore faster too. she used it for a while (6 months?) and then we were able to buy another diamond knife.
I haven't tried a saphire knife since, although I am partial to them in jewelry!
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
When we were working with porcine oocytes it was necessary to handle each individually so we enrobed them in agarose inside a cell of nylon net of a dark color, using a dissecting microscope. We saved enough of the excess nylon net to use as a "handle" to pick up the sample and moved it from solution to solution. This should be done after fixation, since glut fixed agarose is sometimes a problem. We also used the low temp gelling agarose, so that we had time to work. Then put it in the frig to solidify. It will then remain solid at room temp. You might get better lectin labeling using one of the acrylic resins rather than an epoxy
At 09:13 PM 06/15/2000 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Gregory W. Erdos, Ph.D. Assistant Director, Biotechnology Program PO Box 110580 University of Florida Gainesville, FL 32611
I see no reason why this type of cross sectional view cannot be achieved by sectioning with a cryostat, this is the type of use our cryostats are supplied for. If you would like more information please get back to me, I would be happy to section some samples for you to inspect.
Best Regards
Alan Bright
Bright Instrument Co.Ltd. St Margaret's Way Huntingdon PE18 6EB England
Tel No:+44 (0)1480 454528 Fax No:+44 (0)1480 456031 Email: AlanBright-at-brightinstruments.com Web Site: www.brightinstruments.com
-----Original Message----- } From: Smartech [mailto:smartech-at-javanet.com] Sent: 15 June 2000 15:03 To: To all on the list
I needed to have a cross sectional view into an adhesive layer so that I could count the layers, if possible. Since the adhesive acts like a highly viscous liquid, polishing it is out of the question. Instead, I submerged the film into liquid Nitrogen and cut it quickly w/ a scissors. The sound of the cut was more like a breaking sound. When I examined the cross sectional surface I observed a brain like surface. There were columns with a highly consistent diameter and orientation. It looked crystalline. There was zero evidence of material smearing that one would expect if one cut a material. Is the proper interpretation that these columns existed before the fracture and that the fracture occurred along the boundaries? Or is the cross sectional surface generated by a rippled distortion of an highly viscous liquid? The regularity of the surface seems to make the latter interpretation unlikely.
I would be interested in any opinions on the interpretation of the image or alternative means of cross sectioning the adhesive layer.
I could e-mail an image to anyone who is interested.
I'll ring in & say yes. I am quite happy with this combination. You get the digitized images with a much large field of view. Photo quality ink jets are cheap & in general do well. You will always find extremist in on the subjects of the infinitely best scanner & printer but here is what I bought for {9K$. AGFA Duoscan T2500 ~$4500 500MHz PC with 1/2 Gig memory & 19" hi res monitor, CD writer ~2.5K$ Epson Stylus 870 ~$300 Photo Shop, Fovea 1.0 IP software {1K$ with student ver. of PS Misc. supplies some $
For a MacPerson, my understanding is that in terms of image processing speed the Macs are 2-4x faster that PC but I don't have any benchmarks on the latest generations.
If your printing a lot, the ink jets will drain cartridges pretty quick. Down the road I will probably pick up one of the wax printers. They cost something like 3k$ but I think it is Tektronix that offers to supply all the black wax you can user for life, they are pretty fast & no secret papers are required (much cheap per BW page).
Just my thoughts before coffee.
Bruce Brinson
disclaimer... no financial interest in any companies mentioned.
tbonner 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 } -----------------------------------------------------------------------. } } Being unable to afford a digital camera for my TEM. I'm wondering if the next } best option is to get a high end scanner to scan in negatives and then print } them on a decent printer. Any advice regarding this idea and brands of } scanners and printers that are useful? Also, what image analysis systems are } user friendly? } } Dr. Thomas P. Bonner } Department of Biological Sciences } SUNY at Brockport } Brockport, NY 14420
I have been trying to replace the photocell on the Jetpolisher that I have been using. It is about 15 years old and is made by Struers. The Model is a Tenupol and the power supply is type is Polipower. Struers no longer makes replacement parts for these units. If anyone has any information concerning the photocells of this model (who I might contact to replace it or the sensitivity of the photocell) it would be greatly appreciated. Regards Kevin
} Being unable to afford a digital camera for my TEM. I'm } wondering if the next best option is to get a high end } scanner to scan in negatives and then print } them on a decent printer. ...
The next best option would be a 4x5 film scanner (~US$4k). The problem with typical film scanners is their anticipated dynamic range for photographic film, which is where TEM digital capture excels. I suggest you take a representative film and evaluate the Polaroid "4x5 Ultra". Althought I'm unfamiliar with this particular 4x5 scanner, it is the only one (I'm aware of) which is purported to scan an OD better than 3.5 (approximately 14 f/stops ... 4 f/stops per OD unit ... correct me if I'm wrong). Less expensive (~US$1.2k) would be a flat bed scanner designed for transparencies as well as hardcopy. This additional feature could be a "drawer" for film, or a optional "lid" which provides a lamp from above.
I need tips, descriptions, references regarding the preparation of cross-section specimens of the magnetic layer of magnetic recording tape for high magnfication SEM and conventional TEM observation. Thanks.
Actually the printing part has recently gotten much easier. ElectroImage (http://www.electroimage.com) is offering new technology that lets you print real grey scale images on simple inkjet printers. They have grey inks and new printer drivers.
Bill Miller
At 08:47 AM 6/16/00 -0700, George Laing wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Off hand I know I have information on the cross sectioning of hard disk media using the Tripod Polisherš. I'm not sure if I have anything on magnetic recording tape. If you send me your mailing address, I'll send you whatever I can find that comes close.
David Henriks Vice President 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
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by INTERNET:"EBMet-at-aol.com"-at-sparc5.microscopy.com } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
To All:
I need tips, descriptions, references regarding the preparation of cross-section specimens of the magnetic layer of magnetic recording tape for high magnfication SEM and conventional TEM observation. Thanks.
In response to Eric's question, I run a diagnostic Pathology lab at the University of South Florida, and have a one day processing schedule for kidney biopsies that calls for osmication in 1% buffered osmium at room temperature for 30 minutes. I have found that for tissue pieces in the order of 1/2 millimeter thick in one dimension the fixation is fine, and is equivalent to our routine processing osmium fixation of one hour at 4 degrees. I haven't tried this on muscle biopsies, but if they meet the thickness criteria they should be O.K. too,. Just make sure to rinse these extensively (3x 10 minutes, perhaps) in buffer to remove the excess osmium from the muscle tissue, as fluids enter and leave muscle slower because of the extensive connective tissue sheaths around the myocytes. Overosmication is a definite possibility, with subsequent tissue brittleness, if tissue is left in osmium too long, no matter what the temperature. If the tissue is too thick, uneven osmication can occur, where the outside of the tissue is well fixed and a fixation gradient is set up with poor fixation towards the center of the tissue. I observed this happening at a renal lab in Pittsburgh where I used to work. Our unstained thick sections were darker at the periphery than in the center. Another sign of this problem is lack of specimen contrast at the center of thin sections. So, Eric, as far as my experience goes, it is possible to start with 4 degree, buffered osmium, and to osmicate at room temperature for a half of an hour and get results equal to those from osmication at 4 degrees for one hour if your tissue is sufficiently thin in at least one dimension. I haven't noticed any precipitation problems with this technique, nor does the osmium discolor during fixation. Our lab has been using this technique for several years now. Take care! Ed Haller, U.S.F. Pathology
You may want to ask Struers for a users list. There may be someone out there with an older unit that is no longer being used who may be willing to give it to you for spare parts. If they can't give you a list, let me know - I think I can dig up an old list I put together of some previous Tenupol users you may be able to contact. If that doesn't work, you may want to consider upgrading to a South Bay Technology Model 550D Jet Polisher. If you have an interest in getting more information on that option, please contact me and I'll send you information.
David Henriks Vice President 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
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by INTERNET:"kklos-at-mail.mse.ufl.edu"-at-sparc5.microscopy.com } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have been trying to replace the photocell on the Jetpolisher that I have been using. It is about 15 years old and is made by Struers. The Model is a Tenupol and the power supply is type is Polipower. Struers no longer makes replacement parts for these units. If anyone has any information concerning the photocells of this model (who I might contact to replace it or the sensitivity of the photocell) it would be greatly appreciated. Regards Kevin {
To the person asking about the possibility of doing immunofluorescence microscopy on cells grown on plastic coverslips, someone has published a technique in BioTechniques that I saved in case I needed it. Volume24, number 6, pages 910-914, 1998 is the article titled "Mounting technique allows observation of immuno-labeled cells on plastic coverslips". The basic technique from M. F. Donohue et al involves using Thermanox coverslips on which cells are grown and immunolabeled. Following labeling, this group uses a drop of aqueous mounting medium to mount the side of the coverslip without cells on it to a glass slide. On top of this, the group then mounted a regular glass coverslip with an additional drop of aqueous mountant, and then could do their microscopy. The authors state that the inherent strong autofluorescence is greatly reduced by this technique, and the problem with the plastic not transmitting light well is overcome. Although I haven't tried the technique yet, it sounds like a simple fix for a sticky problem. I hope this is of help to you! Ed Haller, U.S.F. Pathology
Hi, We are currently in the market for a tissue processor for electron microscopy. It will mainly be used for biolgical specimens (some quite small). Although I have considerable experience with the processor sold by RMC (Ventana), I know virtually nothing about the Lynx (now being sold by EMS, I think). Any information on the advantages or disadvantages of either model (or any other one that might be out there) would be appreciated. Offline replies are welcome.
Tom Januszewski Senior Electron Microscopist Molecular and Cellular Imaging Facility UT Southwestern Medical Center at Dallas 5323 Harry Hines Blvd. Dallas, TX 75390 Email: tom.januszewski-at-email.swmed.edu
We are using the Agfa T2500 to scan TEM and SEM negatives, which also gives us the ability to scan prints. The 1200 dpi is sufficient for most needs, with 2500 dpi getting used less often and mostly with low mag images. It is currently connected to a 233 MHz Mac G3/160 Mb RAM, being replaced with a 400 MHz G4/320 Mb. The Agfa is driven by either a stand alone app. or through a plug-in that runs under most software, such as Photoshop or Object Image. You will want a lot RAM and drive space, CDR, Ord, Jaz or DVD-RAM drives. Zip drives fill far too quickly.
Photoshop is used for publication images, although some users prefer Canvas. Most of our image analysis uses the Object Image enhanced version of NIH Image. It is very easy to use. I've less experience with Image/J, but it is quickly adding capabilities and will display } 8 bits, whereas Object Image will process 16 but only displays 8 bits. Printing is either to an Epson 850, 3000, or our venerable Phaser IIsdx. BTW, Tektronix sold its printer division to Xerox.
tbonner 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 } -----------------------------------------------------------------------. } } Being unable to afford a digital camera for my TEM. I'm wondering if the next } best option is to get a high end scanner to scan in negatives and then print } them on a decent printer. Any advice regarding this idea and brands of } scanners and printers that are useful? Also, what image analysis systems are } user friendly? } } Dr. Thomas P. Bonner } Department of Biological Sciences } SUNY at Brockport } Brockport, NY 14420
--
Glen MacDonald Virginia Merrill Bloedel Hearing Research Center Box 357923 University of Washington Seattle, WA 98195-7923 glenmac-at-u.washington.edu (206) 616-4156 (206) 616-1828 fax *********************************************************** C:} The box said "Requires Windows95 or better". So I bought a Macintosh. ***********************************************************
} I«m thinking in buying a saphire knife for ultramicrotomy since they seem to } be less expensive than the traditional diamond ones. I need to cut thin } sections of sponges that are difficul to cut with glass knifes. Does anyone } have experience with these knifes? How do they compare with diamond in terms } of cutting properties and durability? } } Dr. A.P. Alves de Matos } Dental Medical School } Lisbon
Unfortunately, you get what you pay for. Sapphire does NOT have the durability of diamond and will be damaged by the sponge spicules. And, to my knowldge, they can't be resharpened.
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
It could be a solution if you mainly want an alternative to silver prints, or to acquire images for Powerpoint presentations. However, many of the really useful features of a digital camera on a TEM are unavailable if you adopt this approach, namely instant verification of image capture, greyscale expansion, image averaging, online analysis, and many more. Also, you still need to allocate some space to a darkroom.
I looked at large-format transparency scanners a couple of years ago, when there was little of this kind in the market, and concluded that their combinations of bit depth, pixels per inch and sensitivity and dynamic range at the high-density end of the negative (i.e. highlight detail) was close to what was required if the objective was merely to obtain publication quality images from a large proportion of negative area (these images require optimised contrast and crispness, but being small do not demand much resolution), but really inadequate for scanning of image details (organelles, molecules), for dense exposures and highlights, and for high contrast subjects like replicas. Most of these scanners appeared to be optimised for scanning positive images (large format colour transparencies) where discrimination of detail in the extreme shadows is not top priority. However, this becomes a major shortcoming when dealing with negatives.
I would certainly like to know whether anyone feels that there is an adequate solution available today.
I don't think user friendliness is the most useful criterion for discriminating between image analysis packages. This is in any case a fairly subjective property, depending very considerably on the computer - literacy of the user. Most IA packages (analySIS, Optimas, etc) are GUI-based systems, and therefore are reasonably intuitive. One of the features that differentiates them is the balance between the provision of off-the-peg analysis solutions and programmability. The range of tasks demanded of an IA package is potentially so great that there is little alternative but to evaluate them and see if they suit your needs. However, I warn you that your needs are likely to evolve. What seems like a simple and user- friendly solution today will probably feel like a very limited and inflexible one tomorrow if it has insufficient functionality and programmability, and these things unavoidably add complexity.
Chris Jeffree
Date sent: Thu, 15 Jun 2000 21:14:18 -0500 To: Microscopy-at-sparc5.microscopy.com } From: tbonner {tbonner-at-brockport.edu}
tbonner said: } Being unable to afford a digital camera for my TEM. I'm wondering } if the next best option is to get a high end scanner to scan in } negatives and then print them on a decent printer.
Using a flat bed scanner over a digital camera for TEM images is definately your best option budget wise. We use an Agfa Duoscan scanner with great success. it is very versitile, we use it to scan gels, scan TEM negatives and scan old prints. It can scan slides for powerpoint quality presentations also. So check with Agfa to see their latest lineup.
For printers inkjets work great when combined with photoquality papers. Any (Epson HP & Cannon) 300dpi or higher color ink jet will give decent images suitable for posters. For publications dye sublimation printers work well but get one that uses a cartridge (one piece) to replace empty media. For proofing look for laser printers that are at least 1200dpi with extra ram (64meg on the printer is a nice number to start with) and large toner cartridges, graphic images burn a lot of toner. Look for a printer that will print alot before replacement of the toner.
For software, Photoshop is a good choice. Before you spend alot on a venders image analysis package try some of the freeware out there like NIH image and Image J both from the NIH website. If you can't get the free stuff to work, then spend the extra money. We use Image J here and it works well.
For computers (Apple or PC) consider one scanner with a SCSI interface with need a SCSI card (avoid parallel port and universal serial bus (USB) scanners unless you like coffee breaks). So you need one computer to hook up to the scanner but Ideally you would also have three printers (inkjet, Laser & Dye sub or comparable) But Inkjets are cheap make your users get one and maintain it. I bet somewhere in your department is a networked laser printer so print to a networked printer elsewhere. Keep the high end printer (Dye sub/thermal printer close by) Invest in removable media that others can use like a CD-r (HP or Plextor) and a zip drive at the bare minimum.
Hope this helps Jon Ekman Associate Research Specialist Deptartment of Biological Sciences University of Wisconsin-Milwaukee phone W:414.229.6471 Web1 http://www.graffitimasters.com Web2 http://www.uwm.edu/~jekman
Embed it, Elliot and diamond knife section it in an ultramicrotome, using the block face for the FE-SEM examination and the ultrathin sections (20-200 nm thick) for examination in the TEM. A good reference is Ho et al, Specimen Preparation for TEM of Materials, Mat. Res. Soc. Symp. Proc., vol. 115, pp. 149-154 (MRS, Pittsburgh, 1988). You may have to search around for someone who can do this for you, but it is far and away the best technique for your needs.
Tom Malis Group Leader - Characterization Materials Technology Laboratory Natural Resources Canada (Govt. of Canada) 568 Booth St., Ottawa, Canada ph. 613-992-2310 FAX 613-992-8735 email: malis-at-nrcan.gc.ca
---------- From: "EBMet-at-aol.com"-at-sparc5.microscopy.com [SMTP:"EBMet-at-aol.com"-at-sparc5.microscopy.com] Sent: Friday, June 16, 2000 11:35 AM To: microscopy-at-sparc5.microscopy.com Subject: re: Cross-section Preparation of Magnetic Recording Tape
------------------------------------------------------------------------ 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 need tips, descriptions, references regarding the preparation of cross-section specimens of the magnetic layer of magnetic recording tape for high magnfication SEM and conventional TEM observation. Thanks.
I am ion milling gold. nearly 100 nm. I want to know how long it takes to make it thin enough to be transparent under TEM. Thanks.
Gen ****************************************************************** Gen Pei Department of Materials Science and Engineering Cornell University 328 Thurston Hall tele: (607)255-5177 fax:(607)255-2365 gp35-at-cornell.edu
a) So far as I know they are not made any more and have not been made for at least ten years, and
b) The economics have changed drastically from twenty years ago when the sapphire knife did enjoy a bit of popularity. In real terms, diamond knives, now because of the competition from Microstar have drop significantly from what they once were, perhaps 50%, so whatever pricing advantage there was at one time, did not exist any more. So the Japanese company that made them discontinued their production. It was called "Saphatome" or something like that. Ted Pella would probably know their history, perhaps better than I do.
Also, because of your interest in education, take a look at www.microscopy-advantage.com . Tell me what you think. Attendees at the coming meetings of APEM, EUREM and MSA will automatically receive a CD in their registration materials. If you would like a copy, send me your UPS address and I will make sure that one gets sent to you. But it will "work" exactly as if you were on line. --- Original Message --- Caroline Schooley {schooley-at-mcn.org} Wrote on Fri, 16 Jun 2000 11:47:54 -0700 ------------------ ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} IŽm thinking in buying a saphire knife for ultramicrotomy since they seem to } be less expensive than the traditional diamond ones. I need to cut thin } sections of sponges that are difficul to cut with glass knifes. Does anyone } have experience with these knifes? How do they compare with diamond in terms } of cutting properties and durability? } } Dr. A.P. Alves de Matos } Dental Medical School } Lisbon
Unfortunately, you get what you pay for. Sapphire does NOT have the durability of diamond and will be damaged by the sponge spicules. And, to my knowldge, they can't be resharpened.
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
----- Sent using MailStart.com ( http://MailStart.Com/welcome.html ) The FREE way to access your mailbox via any web browser, anywhere!
} It could be a solution if you mainly want an alternative to silver prints, } or to acquire images for Powerpoint presentations. However, many } of the really useful features of a digital camera on a TEM are } unavailable if you adopt this approach, namely instant verification of } image capture, greyscale expansion, image averaging, online } analysis, and many more. Also, you still need to allocate some } space to a darkroom.
Why not just outsource the processing of the film? Depending on where one resides/operates, there are typically numerous professional and non-professional labs which will do same day development of b/w film. I do this for 4x5 cut sheet film and 120/220 roll film from a regular camera and from the SEM recording camera. Unless there is some overriding need or requirement for an on-site darkroom, why not just send the film out whenever it is needed? I could see the rationale for an on-site facility if the TEM was producing hundreds of negs per day or perhaps per week. Then it is a make-buy decision regarding in-house or out-house processing.
If one is concerned about whether a shot will turn out (instant verification), just shoot a couple more sheets or frames bracketed around the "optimum/normal" exposure time. The cost of the film and processing is way too low to justify a high cost digicam for TEM. SEM imaging is of course a totally different matter.
I find that grey scale expansion is not the sole domain of the digicam. In a neg, additional information is there--but typically the eye cannot see it. This is where image analysis and image processing programs are very beneficial.
} I looked at large-format transparency scanners a couple of years } ago, when there was little of this kind in the market, and concluded } that their combinations of bit depth, pixels per inch and sensitivity } and dynamic range at the high-density end of the negative (i.e. } highlight detail) was close to what was required if the objective was } merely to obtain publication quality images from a large proportion } of negative area (these images require optimised contrast and } crispness, but being small do not demand much resolution), but } really inadequate for scanning of image details (organelles, } molecules), for dense exposures and highlights, and for high } contrast subjects like replicas. Most of these scanners appeared to } be optimised for scanning positive images (large format colour } transparencies) where discrimination of detail in the extreme } shadows is not top priority. However, this becomes a major } shortcoming when dealing with negatives.
Discrimination of detail in shadows is a major concern for users of transparencies. This is why they seek high D rated scanners. I typically scan negative and transparencies as transmitted RGB or greyscale. This is because I find that the scanner picks up more detail across the whole image when scanned as a tranny.
} [snip]
} and see if they suit your needs. However, I warn you that your } needs are likely to evolve. What seems like a simple and user- } friendly solution today will probably feel like a very limited and } inflexible one tomorrow if it has insufficient functionality and } programmability, and these things unavoidably add complexity. } } Chris Jeffree
I agree that needs may evolve. That means that when making the initial purchase of an image analysis program, it should be flexible enough to allow custom augmentation. Most of the higher end ones do. But it may turn out that one program alone is not as good as two different programs--each being good at different aspects of image analysis.
gary g.
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Modern surfers use PC boards. You can too at http://photoweb.net ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Tom Malis wrote: ====================================================== Embed it, Elliot and diamond knife section it in an ultramicrotome, using the block face for the FE-SEM examination and the ultrathin sections (20-200 nm thick) for examination in the TEM. A good reference is Ho et al, Specimen Preparation for TEM of Materials, Mat. Res. Soc. Symp. Proc., vol. 115, pp. 149-154 (MRS, Pittsburgh, 1988). You may have to search around for someone who can do this for you, but it is far and away the best technique for your needs. ======================================================
This has been our experience too, however we add the following to the preparation protocols:
a) We coat one side of the recording tape with, say, Pt, the other side with Al, because once in the TEM, it is important to validate that i] nothing has fractured off during the ultramicrotomy and ii] you can keep straight which side is which for the asymmetric cross-section. If the two metallization lines are present in the TEM, with embedding resin on the other side, you can be certain you are seeing the entire cross-section. If one is missing, you might not have the entire cross-section.
b) For looking at the "faced-off-piece", we suggest an ever so slight amount of oxygen plasma etching, in order to bring out a bit more contrast between the ferrite or other inorganics from the matrix polymer. The inorganics stand up like little "mesas" in the desert, giving greatly enhanced contrast. Since you now have an element of three dimensional nature to the same, you can gain some insight into orientation, something that would not otherwise be possible
Disclaimer: If you are looking for someone to do this kind of work, look no further, we have been doing this kind of sample preparation for clients on a contract basis since the early 1970's! Our own Plasma Prep II plasma etcher would do the described etching on the faced-off-piece after about 120 seconds of exposure.
Chuck
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And if you're satisfied with prints at 720x1440 dpi, Epson has just made a major leap in ink and paper longevity; read about it at http://www.epson.com/whatsnew/ygtsi/lightfast.html http://www.wilhelm-research.com/ . Unfortunately, the new ink cartridge won't fit old (as in last year's) printers. Oh well - what else might I spend $370 on?
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
List Recipients: I am posting this message at the request of Joe Lester and all correspondence should be sent to him. Dave Audette david.audette-at-sylvania.com
JOB OPENING Scanning Electron Microscopist in Analytical Laboratory ( Beverly, MA)
OSRAM Sylvania, Inc. 71 Cherry Hill Drive Beverly MA 01915
DESCRIPTION: Structural and elemental characterization of materials used in incandescent, fluorescent and discharge lamps, especially by optical and electron microscopy. Failure analyses of lamps and lighting components. Technical problem solving as a member of a team. Oral and written communication of results and conclusions with client population.
POSITION REQUIREMENTS: Competence in optical and electron microscopy of materials including EDS. An understanding of failure analysis. Ability to work independently and/or in a team and to communicate effectively.
EDUCATION AND EXPERIENCE REQUIREMENTS: B.S., or higher, in Materials Science, Chemistry, or Physics. 2-5 years experience in SEM/EDS. Experience with lamp components is desirable.
Please send a resume to
Dr. Joe Lester Technical Assistance Lab OSRAM Sylvania Inc. 71 Cherry Hill Drive Beverly, MA 01915 e-mail: joe.lester-at-sylvania.com
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G'day All, In my experience Sapphire Knives are an excellent replacement for glass knives when working with soft materials. Sapphires are much softer than diamond and are more easily damaged than diamond. Sponges are NOT soft tissue, they contain very hard inorganic salt spicules that damage both glass and sapphire knives. Regards JVN
Leona Cohen-Gould 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 } -----------------------------------------------------------------------. } } At 11:44 AM +0100 6/16/0, A.P. Alves de Matos wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Dear All } } } } } } I«m thinking in buying a saphire knife for ultramicrotomy since they seem to } } be less expensive than the traditional diamond ones. I need to cut thin } } sections of sponges that are difficul to cut with glass knifes. Does anyone } } have experience with these knifes? How do they compare with diamond in terms } } of cutting properties and durability? } } } } } } Thanks } } } } Dr. A.P. Alves de Matos } } Dental Medical School } } Lisbon } } ********************* } I had bought a saphire knife, years ago (early 1980's). Our lab bought it } with the idea that it was a good half-way stop for a new tech who needed } something better than glass. It had certain drawbacks....the edge seemed } to collect debris and was more difficult to clean than a diamond, and of } course it wore faster too. she used it for a while (6 months?) and then } we were able to buy another diamond knife. } } I haven't tried a saphire knife since, although I am partial to them in } jewelry! } } 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
-- **************************************************** John V Nailon Operations Manager Centre for Microscopy and Microanalysis The University of Queensland St. Lucia Queensland 4072 Phone: +61-7-3365-4214 Fax: +61-7-3365-4422 WWW: http://www.uq.edu.au/nanoworld/allstaff.html#Nailon ****************************************************
I had a argument with my husband, he says he is going to the Kunming for an EM meeting,
http://www.iphy.ac.cn/microsc/IKSM.html
He says there are many good scientists going which I believe with doubt. But I think he is going for a sight seeing. There is a Chinese saying: The mountains and waters in Guilin are the most beautiful ones under the sky. I have asked him to bring me, he agreed but unable to get the same airline ticket (UA fully booked). Any body is going and knows alternative airlines, please contact me. Thanks a lot.
I have two DVD-RAM drives and 15 media (Type I and II). I have found that these are riddled with write & read errors. Be careful when using this storage media.
My main unit is a Panasonic LF-D101 (SCSI) and the second one is the same. The third is a Matshushita ID unit.
Scandisk will report either many errors that are fixed or no errors. Either way, the media/drive will write faulty file contents.
Hi Hao, We have in stock (100), (110) and (111) MgAl2O4 substrates with 7 angstrom finish ready for laser ablation or other kinds of epitaxial film deposition. Contact me for info regarding perovskite lattice matching. Best regards, Mike Urbanik www.crystalguru.com
{ { Subj: information about MgAl2O4 Date: 6/15/00 3:33:44 PM Eastern Daylight Time From: haoli-at-glue.umd.edu (Hao Li) To: Microscopy-at-sparc5.microscopy.com
------------------------------------------------------------------------ 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, All:
This one is not really related to microscopy. But since I am a TEM guy, I hope I can get some help here.
Does anybody have some information about MgAl2O4 as a substrate for perovskite films? I know it is not often used, so I am worndering if it has a major disadvantage so that nobody is using it. Any references would be welcome.
Hi All, I processed a series of pellets of yeast for a client using a glut-pfa fix, osmium, dehydration through ethanols, and a day & a half step-wise infiltration into Spurr's resin (1:1 with Ethanol for 4 hr, pure resin overnight under light vacuum, then fresh for 3 more hours, then embed in fresh). Polymerization was overnight at 60C. About half the blocks were soft and had to be returned to the oven for a prolonged polym. (over the weekend). I was able to get sections from each of the 10 samples, but in the 'scope, many of these looked a bit like swiss cheese. those that were not lacy exhibited areas where the resin pulled away from the cell coats of the yeasts. Clearly something went wrong with the infiltration/polymerization. I used the same batch of resin for other things, and its fine.
Does anyone out there have experience with yeast? Any suggestions? I'd like to give this peson some usable data!
Thanks in advance, 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
My only thought is that your infiltration/dehydration were too short, or your 100% ethanol had absorbed too much water. My understanding is that Spurr's is very sensitive to small quantities of water, with soft blocks and holes being symptoms of incomplete dehydration. Sometimes we extend the dehydration through 3 changes of 100% ETOH with molecular sieves, then on through 2-3 changes of propylene oxide. Infiltration is usually 1:2 PO:Resin, followed by 1:1, 2:1, then a couple changes of pure resin overnight or for 4-8 hours, then final embedding in another change of pure resin.
I don't know if yeast is more problematic than other things in this respect, however.
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: Leona Cohen-Gould [mailto:lcgould-at-mail.med.cornell.edu] Sent: Monday, June 19, 2000 8:47 AM To: Microscopy-at-sparc5.microscopy.com
Hi All, I processed a series of pellets of yeast for a client using a glut-pfa fix, osmium, dehydration through ethanols, and a day & a half step-wise infiltration into Spurr's resin (1:1 with Ethanol for 4 hr, pure resin overnight under light vacuum, then fresh for 3 more hours, then embed in fresh). Polymerization was overnight at 60C. About half the blocks were soft and had to be returned to the oven for a prolonged polym. (over the weekend). I was able to get sections from each of the 10 samples, but in the 'scope, many of these looked a bit like swiss cheese. those that were not lacy exhibited areas where the resin pulled away from the cell coats of the yeasts. Clearly something went wrong with the infiltration/polymerization. I used the same batch of resin for other things, and its fine.
Does anyone out there have experience with yeast? Any suggestions? I'd like to give this peson some usable data!
Thanks in advance, 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
Dear Gen Pei, I have looked at gold contacts, lifted from an electronic device, that were supposed to be 100 nm thick. I could see the structure clearly at 200 kV. At 10:57 PM 6/16/00 -0400, you wrote:
} HI, } } I am ion milling gold. nearly 100 nm. I want to know how long it } takes to make it thin enough to be transparent under TEM. Thanks. } } Gen
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
I was just visited by one of our EH&S folks who wanted to know why I had 1,2 dichloroethane.
Seems they track purchases and I bought some last year to make formvar films.
1,2 dichloroethane is on their bad list as a carcinogen. He was actually here to figure out how much might be going up the fume hood so he could make a report to the local air quality agency. But as we talked, it seemed like it would be better to not have the stuff around.
Anyone have experience with formvar in chloroform? I read it works but have never tried it. According to our EH&S guys, chloroform would be better than 1,2, dichloroethane.
BTW we are in California and must abide by some pretty strict rules, it may seem like they are going overboard, but they are just trying to do their job.
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
{P} Good day to all on the listserver, {/P} {P} {/P} {P} I have a person in my department who is interested in processing horse sperm for TEM. Does anyone who does this routinely be willing to give me some pointers as to how to process them? I've only worked with muscle and brain tissue so this is kinda new - I have processed cells from cell culture for TEM would it be the same procedure? {/P} {P} {/P} {P} Thanks so much, {/P} {P} Connie Cummings, DVM {/P} {P} Instructor Anatomic Pathology {/P} {P} Department VBP {/P} {P} Oklahoma State University {/P} {P} {/P}
Hi Y'all: We are looking for a for-hire independent FIB company that has experience in preparing TEM cross-sections of semiconductors. Please contact me if you do this, or know of a lab that does. Regards, Michael Coviello Lab Manager Materials Science University of Texas at Arlington
Thanks to everyone for your helpful comments. I'm taking another stab at it with smaller pellets, longer times and the addition of prop. ox. steps after the ethanol. I had pretty much decided to do all that anyway, but its nice to gets confirmation ofone's ideas!
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
Try FIBICS in Ottawa, Canada. Contacts are Mike Phaneuf or Louise Weaver. Their telephone number is 613-860-0861. email: mphaneuf-at-fibics.com or lweaver-at-fibics.com
-----Original Message----- } From: Mike Coviello [mailto:coviello-at-mae.uta.edu] Sent: Monday, June 19, 2000 4:12 PM To: listserver
Hi Y'all: We are looking for a for-hire independent FIB company that has experience in preparing TEM cross-sections of semiconductors. Please contact me if you do this, or know of a lab that does. Regards, Michael Coviello Lab Manager Materials Science University of Texas at Arlington
Hi Lee, We follow a very similar protocol to yours with the exception that we use propylene oxide rather than ethanol for infiltration. PO : Spurrs 1:1 1 hour PO : Spurrs 1:3 1- 2 hours Spurrs 1 - 2 hours Spurrs overnight (we do not infiltrate under vacuum) fresh Spurrs 1 - 2 hours polymerization at 60C 48 hours
We had the problem you describe when we tried to embed yeast in EPON equivalents. Switching to Spurrs was the fix for us. Frank
At 09:46 AM 6/19/00 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi Leona, The standard way for dealing with yeast cells is to remove the cell wall. This is done with glusulase (fancy name for snail guts) and or lyticase (both from Sigma). If the wall cannot be removed for experimental reasons (i.e. study of the plasma membrane/cell wall interface) then you need to modify the carbohydrate linkages of the cell wall to make the wall more permeable. This can be done by treating the cells, after fixation, with 1% sodium metaperiodate for about 15 minutes. However, most researchers simply wishing to examine yeast morphology remove the wall because this not only improves inflitration of the resin it also allows more extraction of the cytoplasm and thus makes it easier to resolve structures and membranes within the ribosome rich yeast cell. A classic protocol, by Byers and Goetsch, can be found in Vol 194, Methods in Enzymology (AKA Guthrie and Fink) pg 602. I strongly encourage you to read this article as yeast can be very problematic. You also want to use the Hard Spurrs formulation and use 100% acetone (or propylene oxide) as the last dehydration step before going into 1:1 resin. I have not observed any difference in the ultrastructure of cells embedded in Spurrs vs. polybed 812 (when the wall is removed). If you want to do immuno-EM you might find our protocol useful; checkout: http://genome-www.stanford.edu/group/botlab/protocols/EM_protocol.pdf.
Jon Mulholland Genetics Dept Stanford University School of Medicine Stanford, CA 94305-5120
On Mon, 19 Jun 2000, Leona Cohen-Gould 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 All, } I processed a series of pellets of yeast for a client using a glut-pfa fix, } osmium, dehydration through ethanols, and a day & a half step-wise } infiltration into Spurr's resin (1:1 with Ethanol for 4 hr, pure resin } overnight under light vacuum, then fresh for 3 more hours, then embed in } fresh). Polymerization was overnight at 60C. About half the blocks were } soft and had to be returned to the oven for a prolonged polym. (over the } weekend). I was able to get sections from each of the 10 samples, but in } the 'scope, many of these looked a bit like swiss cheese. those that were } not lacy exhibited areas where the resin pulled away from the cell coats of } the yeasts. Clearly something went wrong with the } infiltration/polymerization. I used the same batch of resin for other } things, and its fine. } } Does anyone out there have experience with yeast? Any suggestions? } I'd like to give this peson some usable data! } } Thanks in advance, } 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 } } }
Formvar in chloroform works well. I don't think we have ever tried dochloroethane.
I have had to do the job myself recently and made a minor discovery - the film seems to stick very well to acid washed slides! So, not so cleverly clean. Then the second trick - float the film soon after it has dried, within a minute. Then it seems to work better.
Keith
_______________________ Keith Ryan (Dr) Marine Biological Association Citadel Hill Plymouth Devon PL1 2PB England
Dear all, Thanks to all who responded to my enquiry and gave me useful advice concerning the preparation of cross-sections of ceramic thin films.
Hopefully, I should now be able to prepare some better thin film specimens using one or more of the suggestions that I received.
Thanks again
Best wishes
===== Ian MacLaren Beijing Laboratory of Electron Microscopy Chinese Academy of Sciences, P.O. Box 2724 100080 Beijing China General Email: ian.maclaren-at-physics.org Work (esp. large attachments): maclaren-at-image.blem.ac.cn
____________________________________________________________ Do You Yahoo!? Get your free -at-yahoo.co.uk address at http://mail.yahoo.co.uk or your free -at-yahoo.ie address at http://mail.yahoo.ie
12th Scandinavian Conference on Image Analysis SCIA 2001
June 11-14, 2001 in Bergen, Norway
Sponsored by: IAPR (The International Association for Pattern Recognition) http://www.iapr.org
First Announcement and Call for Papers:
http://www.ux.his.no/scia2001/
Invitation to the 12th SCIA. Following the previous conferences in Greenland, SCIA 2001 - the 12th Scandinavian Conference on Image Analysis - will be held in Bergen on the west coast of Norway. The conference is arranged by the Norwegian Society for Image Processing and Pattern Recognition (NOBIM) and sponsored by the International Association for Pattern Recognition (IAPR). The conference venue is Grieghallen, located in the city centre.
Scientific Program: The conference will offer internationally acclaimed speakers in plenary talks and parallel sessions with selected oral presentations and posters. The conference language is English. The different presentations will cover unpublished theoretical or applied research results.
Invited Speakers: Professor Theo Pavilidis, State University of New York at Stony Brook, USA: "History of Image Analysis" Professor Josef Begün, University of Halmstad, Sweden: "Biometric Person Authentication" Professor Matti Pietikäinen, University of Oulu, Finland: "Machine Vision and Media Processing" Associate Professor Torbjørn Eltoft, University of Tromsø, Norway: "Neural Network approaches to Cluster-Detection-and-Labelling"
In addition, we are working to find an invited speaker for the subject: "Images in the future mobile terminals".
Pre-conference Workshop/Tutorial: A set of pre-conference half-day workshops/tutorials will be held on June 11, 2001: 1. ICA (Independent Component Analysis): Professor Erkki Oja, Helsinki University of Technology, Finland. 2. Data fusion: Professor Jon Atli Benediktsson, University of Iceland.
Paper submission and registration for presenting authors: Only full papers in English will be accepted, and the length should not exceed eight pages. All papers will be refereed by two reviewers for publication in the conference proceedings. Please send four copies of your paper to:
SCIA2001, Department of Electrical and Computer Engineering, Stavanger University College, P.O.Box 2557 Ullandhaug, N-4091 Stavanger, Norway
Important dates:
Paper submission deadline: November 6, 2000 Notification of acceptance: January 19, 2001 *Camera-ready copy: March 19, 2001
*Camera-ready copy must be accompanied by registration and payment by presenting author. The cover page must contain: * Title of the paper * Name(s), complete address and e-mail for the author(s) * Brief abstract (150-200 words) * Keywords describing the main subject of the paper (3-5 words)
* Author's opinion on whether the paper is most suitable for oral or poster presentation * Name and address for correspondence
Papers considerably longer than the final size, risk being rejected. The fee for one or two extra pages is NOK 500 per page. The fee for colour illustrations is NOK 4000 per page. The decision on oral or poster presentation will be taken solely on suitability, not on paper quality. Paper submission information is available at http://www.ux.his.no/scia2001/, where the LaTeX style file and an example file in the recommended two-column LaTeX format is available.
Enquires: If you have scientific questions, please contact Ivar.Austvoll-at-tn.his.no. The program with further information about registration and payment will be send to you February 1, 2001. If you occasionally have seen this announcement, and want to have the program sent to you, please contact scia-at-plus-convention.no.
Program Committee: Dr. Ivar Austvoll (Chairman), Norway Prof. Jussi Parkkinen, Finland Prof. Fritz Albregtsen, Norway Dr. Alfred Hanssen, Norway Prof. Gunilla Borgefors, Sweden Dr. Anne Solberg, Norway Dr. Bjarne Ersbøll, Danmark
Organized by: Norsk forening for bildebehandling og mønstergjenkjenning (NOBIM) (Norwegian Society for Image Processing and Pattern Recognition) http://www.nobim.no/
SCIA2001 web site:
http://www.his.no/scia2001/ or http://www.ux.his.no/scia2001/
At 12:42 PM -0500 6/19/0, Connie A Cummings wrote:
} Good day to all on the listserver, } } I have a person in my department who is interested in processing horse } sperm for TEM. Does anyone who does this routinely be willing to give me } some pointers as to how to process them? I've only worked with muscle and } brain tissue so this is kinda new - I have processed cells from cell } culture for TEM would it be the same procedure? } } Thanks so much, } Connie Cummings, DVM } Instructor Anatomic Pathology } Department VBP } Oklahoma State University **************************** Connie, Assuming that your colleague will bring you a semen sample, and that orientation is not critical, you can spin the sperm to a pellet and treat it like any other cell pellet. I've done various rodent, marsupial and human sperm samples this way. Once at the microscope, you will have to hunt around a bit to fine the appropriate views (head, mid-piece, tails,etc), but since there are so many cells in the pellet, I've always found what we wre looking for. Its the easiest way to go. Good luck, Lee
Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
I am posting this question to see if we can get some help for our building engineer to deal with problems we are having with our SEM (JEOL T330A).
We suspect that we are having a problem with the building power supply to our SEM. Other folks in our building using other types of equipment have found it necessary to use power conditioners or UPS systems to run their equipment - these systems seem to do the trick for them.
We are trying this approach with the power supply to our SEM, but find that there are some problems. When using the UPS system, we have to connect it up, put it on bypass, start up the SEM, then switch the UPS over after that - a real nuisance. If we don't do this, the SEM won't start.
Does anyone else have this problem, and if so, has found an easy solution? Please contact me offline if you have any suggestions and other words of wisdom. Thanks in advance. Also, thanks for all the help you as a group have given me when I posted previous questions - sometimes I forget to say thank you.
Paula.
Paula Allan-Wojtas Research Scientist - Food Microstructure Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
This is a preliminary message to all Gatan users who will be attending the Microscopy and Microanalysis Meeting in Philadelphia this August (13th to the 17th). I think it has become time to form a User's Group to discuss the level of service and support that we are receiving from Gatan. We should determine where the company should be focusing its efforts and lobby them to correct problems that are most important to us. Please let me know if you are interested in attending such a meeting so I can gauge whether it should be held, and how large a conference room I would need to reserve in Philly.
Note, Gatan representatives are encouraged to attend this meeting, but it will be a user meeting run by the users.
I will not rant and rave here in a completely open forum, as I believe it would be unfair. If you have concerns or comments on this subject, whether or not you are attending M&M2000, please contact me directly. Do NOT reply to the list, check the "To:" header before sending your message, it should say "jfmjfm-at-engin.umich.edu" only.
Thank You.
John Mansfield.
Disclaimer: Opinions expressed in this message are my own personal ones and do not represent necessarily those of my employers.
--
Dr. John Mansfield CPhys MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 763-2282 Cellular Phone: (734) 358-7555 (Leaving a phone message at 936-3352 is preferable to 358-7555) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42¡ 16' 48" Long. 83¡ 43' 48"
Does anyone have experience taking specimens from acetone into LR Gold resin? Some of the brochures on LR White seem to recommend against it, but we are hoping it may be OK for LR Gold. We are doing freeze substitution through acetone for EM-immunocytochemistry.
Thanks in advance for your help. David H. Hall Center for C. elegans Anatomy Department of Neuroscience 1410 Pelham Parkway Albert Einstein College of Medicine Bronx, NY 10461
Sounds like the UPS capacity (VA) is sufficient to run the system, but under rated for the start-up surge. One solution would be a higher capacity UPS. One sized to handle the surge load, however, could be quite large and expensive.
If the nature of your problems is related to line noise, but not voltage levels, you might consider an "ultra isolation transformer" and experiment with various grounding options to minimize interference.
If voltage flucuations are the problem, investigate a "ferro resonant" transformer to stabilize the line. This transformer *should* be somewhat more economical than a similar capacity UPS. BEWARE: These devices produce a loud hum. You don't want it in the same room without some sort of noise attenuation.
.Haven't checked relative pricing, but here is a typical link: http://www.sola-hevi-duty.com/
Another posibility... Can you seperate the vacuum pump(s) supply from the electronics, powering the pumps directly and using your UPS for only the electronics? That may lessen the start-up load enough for the UPS to funcion normally.
Woody White McDermott Technology, Inc.
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi, all,
I am posting this question to see if we can get some help for our building engineer to deal with problems we are having with our SEM (JEOL T330A).
We suspect that we are having a problem with the building power supply to our SEM. Other folks in our building using other types of equipment have found it necessary to use power conditioners or UPS systems to run their equipment - these systems seem to do the trick for them.
We are trying this approach with the power supply to our SEM, but find that there are some problems. When using the UPS system, we have to connect it up, put it on bypass, start up the SEM, then switch the UPS over after that - a real nuisance. If we don't do this, the SEM won't start.
Does anyone else have this problem, and if so, has found an easy solution? Please contact me offline if you have any suggestions and other words of wisdom. Thanks in advance. Also, thanks for all the help you as a group have given me when I posted previous questions - sometimes I forget to say thank you.
Paula.
Paula Allan-Wojtas Research Scientist - Food Microstructure Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
HI: For the second year in a row I am having to have our thin window (low element type-brand name with held) replaced. We have the EDS connected to a TOPCON SM-500 environmental SEM. I look at a lot industrial dusts for particle size and composition and the manuf. of the EDS system says they poke holes in the polymer window. Has anyone else have this problem? I have elected to have a thin beryllium installed this time since my boss is upset about spending $7000 every year plus the downtime and since low element detection is not critical . Thanks Terry Ellis Hallmark Cards Inc.
Does anyone have suggestions for encapsulating cell pellets in agarose?
I am working with flatfish cells but the pellets don't look cohesive enough to withstand washing, dehydration etc. We have some Type I agarose (Sigma, gel temp 36C, melting temp. 86C). I plan to post-fix in 1% osmium tetroxide followed by 1% aqueous uranyl acetate then embed in Spurr's resin.
As this is my first time working with cells, any help will be greatly appreciated.
Thank you very much, Carla Aiwohi Western Fisheries Research Center Seattle, WA
The particulate (have also heard) can "shoot" holes in a thin window. My former SEM/EDS was an Etec with a LARGE roughing system and a turret detector with Be, UTW, and "open" positions. Both windows lasted the 15 odd years it was in use before replacement. An important point is to evacuate and vent slowly so as to not accelerate the particles into the window.
I added a manual valve to the Etec between the roughing system and the chamber. With the automatic valves closed, I could slowly open the manual valve to rough the chamber down to a point where any particulate is not disturbed, close the valve, then switch to automatic to finish the evacuation. Venting gas was from my cryo of liquid nitrogen (pretty dry!) which I pressure/flow controlled for similar results on venting.
My modification was driven by the need to avoid damage to fluffy ceramic specimens, but worked well to protect the detector also.
Have not added such a feature to the new SEM/Thin Window EDS system, but (for now) the chamber is pristine and the fluffy specimens are not part of the work mix.
Woody White McDermott Technology, Inc
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HI: For the second year in a row I am having to have our thin window (low element type-brand name with held) replaced. We have the EDS connected to a TOPCON SM-500 environmental SEM. I look at a lot industrial dusts for particle size and composition and the manuf. of the EDS system says they poke holes in the polymer window. Has anyone else have this problem? I have elected to have a thin beryllium installed this time since my boss is upset about spending $7000 every year plus the downtime and since low element detection is not critical . Thanks Terry Ellis Hallmark Cards Inc.
My guess is that the SEM inititally draws power that exceeds the ratings of the UPS. It is probably due to the rotary pumps that can take up to ten amps upon start up.
The alternatives are to increase the power rating of the UPS or rewire the rotary pump so it draws it's power from the line and not through the UPS (the pumps is not that sensitive anyway).
Good Luck,
Earl weltmer
Paula Allan-Wojtas 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, all, } } I am posting this question to see if we can get some help for our building engineer to deal with problems we are having with our SEM (JEOL T330A). } } We suspect that we are having a problem with the building power supply to our SEM. Other folks in our building using other types of equipment have found it necessary to use power conditioners or UPS systems to run their equipment - these systems seem to do the trick for them. } } We are trying this approach with the power supply to our SEM, but find that there are some problems. When using the UPS system, we have to connect it up, put it on bypass, start up the SEM, then switch the UPS over after that - a real nuisance. If we don't do this, the SEM won't start. } } Does anyone else have this problem, and if so, has found an easy solution? Please contact me offline if you have any suggestions and other words of wisdom. Thanks in advance. Also, thanks for all the help you as a group have given me when I posted previous questions - sometimes I forget to say thank you. } } Paula. } } Paula Allan-Wojtas } Research Scientist - Food Microstructure } Agriculture and Agri-Food Canada } Atlantic Food and Horticulture Research Centre } Kentville, Nova Scotia Canada B4N 1J5 } } Tel: (902) 679-5566 } FAX: (902) 679-2311 } } email: allanwojtasp-at-em.agr.ca
John Mansfield's reminded me that I would like to have some sort of users' group meeting for Emispec users for mutual support and information exchange. I have discussed this with the Emispec folks, but I don't think that they have followed up on it. Could I seed some level of support for a get-together at M&M MM so that I can send it to Emispec. Please respond to me offline and I will send them the number and names of the people that would desire something like that. I would like to see something in a positive and instructive type of meeting.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
this is a message for EM people in Sydney or Woolongong, Australia.
The rotory pump on my JEOL 2020F TEM is short of oil and making a bit of noise so I'm trying to find some quickly. Our JEOL service guys have ordered some "MR100" but it may take a couple of weeks to arrive. Can any one lend me a few hundred mls in the meantime?
Can anyone suggest a local supplier of this rotory pump oil?
Cheers,
Mark Blackford TEM Group Materials Division, ANSTO PMB 1, Menai, N.S.W. Australia 2234
Phone 61 2 9717 3027 Fax 61 2 9543 7179
Disclaimer: The views expressed in this E-mail message do not necessarily represent the official views of ANSTO from which this message was conveyed.
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My guess would be the current loading at start up. When you turn on an SEM from cold, everything starts and draws current. In particular, the rotary pump kicks in and draws a high current as it starts. You will need to be able to set up the UPS so that it can handle this - or set it on a short timer so that it automatically switches in, say, 10 mins after start up.
Regards, -- Larry Stoter JEOL (UK) Ltd Silver Court, Watchmead, Welwyn Garden City, AL7 1LT, United Kingdom tel: +44-(0)1707-377117, fax: +44-(0)1707-373254, e-mail: larrys-at-jeoleuro.com
Probably you are having problems with the high switch on current the sem draws. You could try switching on different parts of the SEM system not at the same time (if possible). You could also consider to only power the electronics of the SEM via the UPS, in that way you reduce the required current drastically, while there is little use for a pump to sit on the UPS power. Alternatively you could look for a more powerfull UPS that can handle the larger currents (for short moment is enough- see data sheets of different UPS's).
Hope this helps...
Jo
************************************************************* * Jo Verbeeck * * University of Antwerp * * Dept. EMAT (Electron Microscopy for Materials Research) * * e-mail: joverbee-at-ruca.ua.ac.be * * tel: +32(0)3 218 02 49 * * fax: +32(0)3 218 02 57 * *************************************************************
On Tue, 20 Jun 2000, Paula Allan-Wojtas 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, all, } } I am posting this question to see if we can get some help for our building engineer to deal with problems we are having with our SEM (JEOL T330A). } } We suspect that we are having a problem with the building power supply to our SEM. Other folks in our building using other types of equipment have found it necessary to use power conditioners or UPS systems to run their equipment - these systems seem to do the trick for them. } } We are trying this approach with the power supply to our SEM, but find that there are some problems. When using the UPS system, we have to connect it up, put it on bypass, start up the SEM, then switch the UPS over after that - a real nuisance. If we don't do this, the SEM won't start. } } Does anyone else have this problem, and if so, has found an easy solution? Please contact me offline if you have any suggestions and other words of wisdom. Thanks in advance. Also, thanks for all the help you as a group have given me when I posted previous questions - sometimes I forget to say thank you. } } Paula. } } Paula Allan-Wojtas } Research Scientist - Food Microstructure } Agriculture and Agri-Food Canada } Atlantic Food and Horticulture Research Centre } Kentville, Nova Scotia Canada B4N 1J5 } } Tel: (902) 679-5566 } FAX: (902) 679-2311 } } email: allanwojtasp-at-em.agr.ca } } }
If you are using water vapour in your environmental SEM you may want to check the suitability of a Be window. I have a vague memory that water will make holes in the thin (6-8um) Be, check it out with your supplier.
I have used two polymer windows on an EDX detector in a TEM gas reaction cell. The front window is on a replaceable mount in front of the detector. This is easily exchangeable in case I cover it with reaction products from the in-situ experiment; I don't want to see them in all subsequent spectra. Much cheaper then a window replacement and can be carried out by me on site. Contact me for further details if you want to.
Reducing the disturbance to the vacuum system gasses during pumpdown and venting by limiting the speed may help prevent dust particles damaging the window in the first place.
Regards, Ron
On Tue, 20 Jun 2000 tellis2-at-hallmark.com-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 } -----------------------------------------------------------------------. } } } HI: } For the second year in a row I am having to have our thin window (low } element type-brand name with held) replaced. } We have the EDS connected to a TOPCON SM-500 environmental SEM. I look at a } lot industrial dusts for particle size and composition and the manuf. of } the EDS system says they poke holes in the polymer window. } Has anyone else have this problem? } I have elected to have a thin beryllium installed this time since my } boss is upset about spending $7000 every year plus the downtime and since } low element detection is not critical . } Thanks } Terry Ellis } Hallmark Cards Inc. } } }
=========================================================================== Mr. Ron Doole e-mail ron.doole-at-materials.ox.ac.uk Department of Materials, phone +44 (0) 1865 273701 University of Oxford, fax +44 (0) 1865 283333 Parks Road. Oxford. OX1 3PH. UK. ============================================================================
I am currently studying for an MSc in Textile studies, at Bolton Institute, England.
For part of my studies I have been analysing some polyester film that has been treated with 10% sodium hydroxide under imposed load under polarized light on an optical microscope. Vivid colours have been noted, Reds, greens, and I am struggling to find information to outline what these colours are actually indicating. I therefore write and ask for any information you may deem relevant, I would be extremely grateful for.
Hi, I am working on powders of mixed WC and Co and I have some problems with EDS. EDS analysis on grains whose diffractions patterns are unambiguously indexed in the Co structure give a majority of W, and EDS analysis on grains whose diffractions patterns are unambiguously indexed in the WC structure (without distortion and superstructures) always indicate the presence of Co (with a majority of W). So I was wondering if there could be any problem with EDS on Cobalt (because of magnetism????)
I have an Electroscan 2010 and routinely have problems with the window. I have ascribed it to particles flying around from the surface of the sample. This generally occurs at startup when there may be some charging. We solved the problem by retrofiting the detector so that it can be retracted from the chamber when changing specimens. Its a pain in the butt, but less so than having to replace windows.
Thanks Robert Carlton Aventis Pharmaceuticals robert.carlton-at-aventis.com
Greetings, Yes, those colors are beautiful, arn't they? The colors indicate the magnitude of birefringent retardation in your sample. In essence, you are seeing subtraction colors resulting from the diminution of intensity at certain wavelengths. Say your sample has a retardation of 550 nm. When linearly polarized light of that wavelength passes through the sample, it will be retarded by exactly one wavelength, which is the same as zero retardation. Thus, it will not be affected by the sample and will be blocked by the analyzer. But light of longer or shorter wavelengths will be retarded by more or less than a wave and so will emerge as elliptically polarized light and thus will have a component transmitted through the analyzer. So for any actual sample retardation, the color will result from exactly how much of each wavelength gets through. Because our eyes are very sensitive to color, this has been used for more than 100 years to measure/estimate retardation. There is a chart that reproduces the colors as a function of retardation. I have no idea if this chart has made it on line but I would be careful. The colors are very tricky to print and great care was used to get them right. You would do far better to look this one up in your library. I am sure the Bolton Textile Institute will have excellent books on polarized light microscopy, and while they might be dusty, this particular corner of science has been well understood for years and years. Hope this helps, Tobias Baskin
} } } Dear Sir/Madam } } I am currently studying for an MSc in Textile studies, at Bolton } Institute, England. } } For part of my studies I have been analysing some polyester film that } has been treated with 10% sodium hydroxide under imposed load under } polarized light on an optical microscope. Vivid colours have been noted, } Reds, greens, and I am struggling to find information to outline what } these colours are actually indicating. I therefore write and ask for any } information you may deem relevant, I would be extremely grateful for. } } I thank you for your time. } } Yours Faithfully } } Kelly Goodman } } EMAIL: suite666-at-netscapeonline.co.uk
A colleage of mine here at the Medical College of Wisconsin is working on a new teaching manual for our first-year medical students' cells & tissues course. We are currently putting together a portion of the manual which shows students electron micrographs of cells and organelles. We are looking for high quality TEM images (non-copyrighted) of structures such as the following:
We would be grateful to anyone who feels they have something useful to contribute. The plan is to scan appropriate images for the lab manual and return them as soon as possible to the owner. There will be a list at the end of the manual acknowledging all contributors.
Thank you everyone,
Susan K. Danielson, MS Neuromuscular Lab Coordinator Dept. Neurology, Medical College of Wisconsin ph: 414.259.3836 email: sdaniels-at-mcw.edu
We are looking for qualified professionals to provide applications-based sales and technical support for our complete line of optical microscopes, sample preparation equipment, consumables, and digital imaging systems in the Dallas/Ft. Worth and Austin, TX areas. Must have at least 2 years experience in failure analysis or materials analysis using microscopes and sample preparation equipment. Prior sales experience is not necessary. Understanding of dimensional measurement and image analysis systems a plus.
Please respond by e-mail to mms-at-micrometsys.com
We are looking for qualified professionals to provide applications-based sales and technical support for our complete line of optical microscopes, sample preparation equipment, consumables, and digital imaging systems in the Dallas/Ft. Worth and Austin, TX areas. Must have at least 2 years experience in failure analysis or materials analysis using microscopes and sample preparation equipment. Prior sales experience is not necessary. Understanding of dimensional measurement and image analysis systems a plus.
Please respond by e-mail to mms-at-micrometsys.com
Responding to the message of {3.0.5.32.20000620115226.008c7a80-at-mailserver.aecom.yu.edu} from "David H. Hall" {hall-at-aecom.yu.edu} :
David,
I've done it, but the thing to watch out for is that if the temperature of your acetone/LR Gold mixtures - say 1:1 acetone:LR Gold - drops below about -35 C, some components of the LR Gold will start to freeze out, solutions gets cloudy. For methanol sub-solution's, LR Gold starts to freeze out at even higher temps, about -27 C.
Just experiment with your sub mixtures at various temperatures to see if you get any freeze-out like this happening, before you do an actual sub run. If so just make sure you stay warmer than freezing points.
I'm curious why this happens, anyone else got any info on this?
Gib Ahlstrand
} Does anyone have experience taking specimens from acetone into LR Gold } resin? Some of the brochures on LR White seem to recommend against it, but } we are hoping it may be OK for LR Gold. We are doing freeze substitution } through acetone for EM-immunocytochemistry. } } Thanks in advance for your help. } David H. Hall } Center for C. elegans Anatomy } Department of Neuroscience } 1410 Pelham Parkway } Albert Einstein College of Medicine } Bronx, NY 10461 } } phone (718) 430-2195 FAX (718) 430-8821 } hall-at-aecom.yu.edu } website: www.aecom.yu.edu/wormem
Gib Ahlstrand Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN. USA. 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.cdl.umn.edu http://biosci.umn.edu/MIC/consortium.html