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
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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 can’t 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 prŽcis 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 - Universitˆ 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 UniversitŠt Erlangen-NŸrnberg, 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 Universitˆ 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 InouŽ 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 InouŽ 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 Electr—nica Facultad de Ingenier’a - 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 – 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 (1Ž0 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
Jesœs Ricote ------------------------------------------------------------- Dr. Jesœs Ricote Departamento de Materiales FerroelŽctricos 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 LŸšND [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 LŸšND 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 Electr—nica Facultad de Ingenier’a - 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
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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: VŠisŠlŠ M, SevŽus 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).
SevŽus L, Kuusisto A, VŠisŠlŠ M, HemmilŠ I., and Soini E: Lanthanide chelates as labels in in situ hybridization and immunohistochemistry, Micron and Microscopica Acta 21; 3, (1990).
SevŽus L, VŠisŠlŠ M, SyrjŠnen S, Sandberg M, Kuusisto A, Harju R, Salo J, HemmilŠ I, Kojola H, and Soini E: Time-Resolved Fluorescence Imaging of Europium Chelate Label in Immunohistochemistry and In Situ Hybridization, Cytometry 13; 329, (1992).
SevŽus L, Kuusisto A, VŠisŠlŠ M, HemmilŠ 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 FacultŽ de MŽdecine 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 Magalh‹es 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 Polisher¨ 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
********************************************************************** * - PLEASE NOTE OUR NEW ADDRESS - EFFECTIVE FEBRUARY 15, 2000 - * * * * NANOPROBES, Incorporated | US Toll-free: (877) 447-6266 * * 95 Horse Block Road | Tel: (919) 510-0590 * * Yaphank, NY 11980-9710, | Fax: (919) 510-0590 * * USA | rpowell-at-nanoprobes.com * * * * NOW EASY TO FIND ON THE WEB: http://www.nanoprobes.com * **********************************************************************
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
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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 60¼C 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 fŸr Botanik (210), UniversitŠt 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 PolitŽcnica 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 PolitŽcnica 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 PolitŽcnica 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 Electr—nica Facultad de Ingenier’a - 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 PolitŽcnica 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-Tak¨ 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 Fisiolog’a de Insectos Dpto. Cs. Biol—gicas, 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 fŸr Botanik (210), UniversitŠt 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 fŸr Botanik (210), UniversitŠt 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 fŸr Botanik (210), UniversitŠt 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, QuŽbec } 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
AndrŽ 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 Žlectronique
INRS-Institut Armand-Frappier Centre de Microbiologie et Biotechnologie 531 boul. des Prairies Laval, QuŽbec 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 Electr—nica Facultad de Ingenier’a - 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–›”Š’”Œ –””‡Œ’Œ----- ë˜: Dr Malcolm Roberts {malc-at-rock.ru.ac.za} 攓—: Microprobe discussion group {Microscopy-at-sparc5.microscopy.com} €ˆ˜ˆ: 19 “ˆØ 2000 ‹. 16:13 ñŒ“ˆ: 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.
JOB SUMMARY
Provide the principal conduit for technical information and feedback between the end user/sales network and Nikon Factory engineers on: product applications, product acceptance, competitive changes, required product improvements, new technologies and methodologies that impact Nikon's present and future business. Provide the sales force with technical assistance to successfully satisfy customer's needs and complete the sale. Create and conduct training curriculum on advanced microscopy and new technologies to Nikon distribution and end users. Attend trade shows, workshops and national dealer / sales meetings. Write technical and applications bulletins to end users and distribution channels as well articles for publication in Bio journals highlighting Nikon technology or applications.
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.
Mail or fax your resume, which MUST include salary requirements, to: HR Department, Nikon, 1300 Walt Whitman Road, Melville, NY 11747. Fax: 631-547-4025.
Or send a digital cover letter and resume / CV in (plain text, MS Word, or .pdf file) to biosales-at-nikonincmail.com
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Nikon is an Equal Opportunity Employer =============================================================
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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, BUEHLER¨ 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 BUEHLER¨ , 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 Polisher¨. We are the manufacturers of the Tripod Polisher¨ 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 RŸhle Max-Planck Institute fŸr 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 Polisher¨, 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 80¼C 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 Hšrstedt Department of Medical Biosciences Pathology Unit for Electron Microscopy University of UmeŒ S-90187 UmeŒ 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 80¼C 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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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 Hšrstedt Department of Medical Biosciences Pathology Unit for Electron Microscopy University of UmeŒ S-90187 UmeŒ 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 Electr—nica Facultad de Ingenier’a - 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 Polisher¨. 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
} 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
----- 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.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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
Even though we have not yet had our Microscopy and Microanalysis 2000 meeting in Philadelphia the Program Committee for the M&M 2001 meeting is preparing the symposia topics for the meeting in Long Beach. If anyone has suggestions for topics that they would like to see included in the Long Beach meeting please contact one of the Program Committee Officers listed below. Please note that we may not be able to respond to all suggestions for this year, but voicing your opinion now may get a topic of interest on the program list for upcoming years.
Thanks.
Bob Price
Program Officers:
Bob Price, M&M 2001 Program Chair Price-at-med.sc.edu
Inga Holl Musselman, M&M 2001 MAS Program Co-chair imusselm-at-utdallas.edu
Edgar Voelkl, M&M 2001 Program Vice-chair/M&M 2002 Program Chair vog-at-ornl.gov
Robert L. Price Director, Instrumentation Resource Facility USC School of Medicine Garner's Ferry Road Columbia, SC 29208 Phone: 803-733-3393 Fax:803-733-1533
I couldn't find this in the archives but I cannot imagine someone has not asked this already. If so, please direct me to the approximate month, year. If not, here goes:
We are decomissioning a laboratory refrigerator. Over the 30+ years of use it has accumulated osmium black on the walls. It has been determined by our EH&S people (after consultation with a hazardous waste shipping compnay) that disposal requires a sealed, separate container for the refrigerator, several licenses, and a hazardous waste truck. The estimate is not in yet, but everyone is talking thousands of dollars. If that is the only alternative for safe disposal, so be it. However, I cannot imagine that there is not a safe alternative to decontaminate before shipping. After all we clean up spills with corn oil and reduce osmium tetroxide with 5% sodium bisulfite solution or sodium metabisulfite. Any definitive advice or references will be appreciated.
Thanks-
-- 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 - ----------------------------------------------
W. Gray Jerome wrote: =========================================================== I couldn't find this in the archives but I cannot imagine someone has not asked this already. If so, please direct me to the approximate month, year. If not, here goes:
We are decomissioning a laboratory refrigerator. Over the 30+ years of use it has accumulated osmium black on the walls. It has been determined by our EH&S people (after consultation with a hazardous waste shipping compnay) that disposal requires a sealed, separate container for the refrigerator, several licenses, and a hazardous waste truck. The estimate is not in yet, but everyone is talking thousands of dollars. If that is the only alternative for safe disposal, so be it. However, I cannot imagine that there is not a safe alternative to decontaminate before shipping. After all we clean up spills with corn oil and reduce osmium tetroxide with 5% sodium bisulfite solution or sodium metabisulfite. Any definitive advice or references will be appreciated. ============================================================= I will risk showing what maybe I don't know, but this is really an important question. Laboratories should not be wasting thousands of dollars needlessly, when other alternatives are available.
I thought that Os (IV) oxide, that is, the tetroxide form of osmium, was clear, after all, crystals in ampoules are fairly clear or translucent, and 4% aqueous osmium tetroxide is water clear. And I thought that in the reduced form, that is, the dioxide or Os (II) form, the color was black. Putting it another way, if it is in the reduced form (e.g. black), it is not in its "hazardous" form, and indeed might even be fairly innocuous. A **quick** survey of shipping regulations does not even indicate that osmium (II) oxide is a controlled material from the standpoint of shipping. I want to emphasize the word "quick", but I could not find a UN number asigned for it.
If that is the case, and if I am right, why could not the black deposit be collected, perhaps by way of some rubbing and scrubbing, and disposed of (or better yet, recycled) and the refrigerator disposed of as any ordinary refrigerator might be dealt with?
Remember, I am not saying that this could be done, legally or otherwise, I am just asking what it is that I might be missing here.
Disclaimer: SPI Supplies is a major supplier of osmium tetroxide to EM laboratories worldwide.
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 need chemical reference standards for the analysis of a Ni-base alloy.
-We intend to use the energy-dispersive X-ray analysis, coupled to a Scanning Electron Microscope(SEM).
-The composition of the alloy is going to vary between 18 to 28 wt% Aluminium ; Al is the most important element to be tested, the remainder of the alloy being mostly Ni of course.
-Secondary main elements are : Cr from 4 to 10 wt% ; Co from 10 to 15 wt% ; Elements in low concentration (below 3%) are Ti and Mo.
-Accuracy : 0.5 % in Al is required.
Questions are :
-How many different standards would we need ?
-What should be the chemical variations from standard to standard to reach the required accuracy ?
-What would be the price for such standards ? Time for delivery ? Where to order this ?
-Can you send us information (reprints from technical journals, articles) on the topic of chemical analysis by means of energy-dispersive (or wavelength-dispersive) X-ray systems ?
If you need any further information, please contact me.
In anticipation, many thanks for a prompt answer.
Yours faithfully,
PAUL-LAURENT CAPRON JR. S.A. LABORIMPEX N.V. RUE DES ALLIES 78-80 BONDGENOTENSTRAAT 78-80 1190 BRUSSEL/BRUXELLES BELGIUM TEL.: 00 32 2 345.99.94 FAX.: 00 32 2 347.39.63
Fellow EM Users, Just a short query ----- would anyone installed a modern day alternative to the 1920's !!! style glass ionization vacuum gauge ("electron current") + electronics box for the JEOL 4B coater? Thankyou for your time, Barry EM UNIT UNSW
Very funny, in every meaning of that word. Of course, the waste company would make a meal of it, why ask?
Its not Os04 that is coating the inside of the fridge, but a tiny amount of osmium metal. The metal is not particularly toxic. Os and Ir are the heaviest metals, with a specific gravity over 22, but nonetheless, its hard to see that you would have 2g of Os in any refrigerator coating. Finely dispersed Os produces tiny amounts of Os04 - if that is a problem, than that would have been so in the confined lab space. If that fridge was dumped conventionally in a land-fill, the tiny amount of OsO4 would be reduced before ever reaching the surface. If it was smelted for scrap, arguably it would, however, immeasurably improve the quality of the scrap. There is an argument for smearing a bit of vegetable oil over the inside of the fridge prior to disposal. Have those safety people given their fully developed reasons? I love to know them.
The bigger picture is interesting too. Safety people are not selected for, nor seem to get taught much logic or a sense of proportion. So we seem to get more and more hassles about things that hardly matter at all, but there is no money and often no objection to the really large environmental issues. Its been said that the old Roman civilasation's collapse was partially due to the difficult Roman numerals, which made the efficient administration of the empire impossible. Our "Western Society" has seen a rapid increase of useless, if not counterproductive rules and regulations, I fear if that trend continues that these "well-meaning" rules will undermine our society. 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 22, 2000 7:07 AM, Jay Jerome [SMTP:jjerome-at-wfubmc.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 couldn't find this in the archives but I cannot imagine someone has } not asked this already. If so, please direct me to the approximate } month, year. If not, here goes: } } We are decomissioning a laboratory refrigerator. Over the 30+ years of } use it has accumulated osmium black on the walls. It has been determined } by our EH&S people (after consultation with a hazardous waste shipping } compnay) that disposal requires a sealed, separate container for the } refrigerator, several licenses, and a hazardous waste truck. The } estimate is not in yet, but everyone is talking thousands of dollars. If } that is the only alternative for safe disposal, so be it. However, I } cannot imagine that there is not a safe alternative to decontaminate } before shipping. After all we clean up spills with corn oil and reduce } osmium tetroxide with 5% sodium bisulfite solution or sodium } metabisulfite. Any definitive advice or references will be } appreciated. } } Thanks- } } -- } 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 - } ---------------------------------------------- } }
Rotary pump oil is a refined mineral oil, which needs to have low vapour pressure, low viscosity and good lubricating properties. such oils are made by "oil companies". In my lab days I never had the luxury to purchase oil from a microscope manufacturer, because their on-cost for such items are horrific and real and my budgets were small.
Disclaimer: ProSciTech sell Rotary Pump Oil. Don't even ask to ship it to USA; the material is cheap, but expensive to ship. Our price includes shipping within Australia only. 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, June 21, 2000 2:33 PM, Mark Blackford [SMTP:mgb-at-ansto.gov.au] 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, } } 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. } }
I remember using the old Geissler tube on an evaporator 32 years ago and very likely they existed long before then. They were actually a bit of an ongoing maintenance expense, but I loved the pretty colours. Cannot see any reasons why other type gauges could not be used, except the manufacturer would find the Geissler tube a cheaper alternative.
Disclaimer: ProSciTech supplies vacuum gauges, circa year 2000
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 22, 2000 10:24 AM, Barry Searle [SMTP:B.Searle-at-unsw.edu.au] wrote: } } Fellow EM Users, Just a short query ----- would anyone installed a } modern day alternative to the 1920's !!! style glass ionization vacuum } gauge ("electron current") + electronics box for the JEOL 4B coater? } Thankyou for your time, Barry EM UNIT UNSW }
Regarding the colours in the PET film, the optical part is as Tobias Baskin says. In regard to the polymer science side of it, sretched films or fibres show optical anisotropy, because the polarizability along the polymer chain is different from that transverse to it. The more highly stretched the fibre, the more the chains will be oriented, and this will increase the optical anisotropy of the fibre. Multiply this by the thickness of the fibre and that will give you the birefringence. Most likely you will be seeing coloured fringes parallel to the fibre direction, which indicate the different viewing thickness through the different parts of the fibre, just like the colours in an oil slick under a car indicate different thicknesses of an oil film.
+------------------------------------------------------------------------+ | 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 am not familiar with microscopy. The most I know is how SEM/EDX can examine a cross sectional sample.
Currently, I am looking for some methodology to analyse interface structure, elementry and chemical compound especially between the solder and metal, i.e. the IMC layer. The solder height will be ranging from 2-4 mil (SMT process). Wondering anybody can advice the best method to analyse the IMC layer. EDX is capable of detecting the element contain insde the IMC but I am seeing more and more alloy up to ternary or quartenary alloy at the layer. Problem is I do not have any idea about where and how it come from.
I'm looking for a manual for the JVG-N2 ionization vacuum gauge. I've already tried JEOL and they could find JVG-N1 manual (thanks!), but not -N2, and -N1 and -N2 are different enough (different connectors, apparently EB grid heating in -N2 etc.) that the -N1 manual is not enough to get the gauge operational.
So, anyone with manual for JVG-N2 ??
btw: The JVG-N1 manual mentions "Forgel tube" which looks in drawing just like normal hot-filament ionization gauge tube.. Is "Forgel tube" just an old term for one?
For some idea of what can be done, check out the soldering iron tip images and x-ray data at my web site.
Keep in mind that I did not not take time to achieve the best possible polish and most of the work was at rather low magnification.
Site: http://woody.white.home.att.net
Woody White
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Hello,
I am not familiar with microscopy. The most I know is how SEM/EDX can examine a cross sectional sample.
Currently, I am looking for some methodology to analyse interface structure, elementry and chemical compound especially between the solder and metal, i.e. the IMC layer. The solder height will be ranging from 2-4 mil (SMT process). Wondering anybody can advice the best method to analyse the IMC layer. EDX is capable of detecting the element contain insde the IMC but I am seeing more and more alloy up to ternary or quartenary alloy at the layer. Problem is I do not have any idea about where and how it come from.
} question. Laboratories should not be wasting thousands of dollars } needlessly, when other alternatives are available. } } I thought that Os (IV) oxide, that is, the tetroxide form of osmium, was } clear, after all, crystals in ampoules are fairly clear or translucent, and } 4% aqueous osmium tetroxide is water clear. And I thought that in the } reduced form, that is, the dioxide or Os (II) form, the color was black. } Putting it another way, if it is in the reduced form (e.g. black), it is not } in its "hazardous" form, and indeed might even be fairly innocuous. A } **quick** survey of shipping regulations does not even indicate that osmium } (II) oxide is a controlled material from the standpoint of shipping. I want } to emphasize the word "quick", but I could not find a UN number asigned for } it. } } If that is the case, and if I am right, why could not the black deposit be } collected, perhaps by way of some rubbing and scrubbing, and disposed of } (or better yet, recycled) and the refrigerator disposed of as any ordinary } refrigerator might be dealt with? }
The problem is that any form of osmium is a heavy metal, and therefore should be treated as one would cadmium or arsenic. It is not innocuous.
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
We have made magnetic nanodots and metastable magnetic phases. The physical structures are well revealed by TEM. However, we would like to relate the magnetic properties with magnetic structure and physical structure. Are there other methods such as neutron diffraction, X-ray or spin polarized electrons that can characterize the magnetic structure/ordering. Please contact me if you could offer such info/collaboration. Thanks a lot.
Yi
******************************************************************* Yi Liu Department of Mechanical Eng. and CMRA 104 N Walter Scott Engineering Center University of Nebraska-Lincoln Lincoln, NE 68588-0656 Tel. (402) 472-7759 (Office) Tel. (402) 472-8762 (EM lab) Fax (402) 472-1465 Email: yliu-at-unlserve.unl.edu *******************************************************************
This is a very complex problem. You don't mention the thickness of the intermetallic layers. EDX analysis has relatively poor spatial resolution in the SEM (of the order of microns) so you can clearly see features that you cannot analyze well. People often use backscatter electron imaging (which can be calibrated) in cases such as yours. You should read a good book on SEM - the subject is too complex to cover in an e-mail like this.
If you can make thin foils of the sample, AEM (analytical electron microscopy) analysis would overcome most of the problems. A LaB6 instrument will give you better than 10 nm analytical resolution, while a field-emitter will approach 1 nm resolution. The problem, though, will be specimen preparation. I would be tempted to suggest using a microtome to cut thin slices, but I don't know the form of your material.
Good luck!
Tony Garratt-Reed
At 06:37 PM 06/22/2000 +0800, 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 **
Dear Paul-Laurent, The usual standards that are used for alloy analysis in the SEM+EDX are pure element standards. You can prepare pure (three or four nines, e.g. 99.9 or 99.99%) metal standards yourself, or any of the EM catalog houses will list standards consisting of pure metals or minerals covering the range of most common elements. They are fairly expensive (~$2000US) and I am not sure of delivery time. Tousimis is one standard supplier that I know of. The same standards are used for EDX or WDX analysis. You collect the spectra from these standards under carefully reproducible conditions in your SEM and follow the instructions in your EDX software to do fully standard analysis with matrix corrections. You can store the standard spectra in the EDX computer, so you only need to run them once for each set of conditions. It is quite difficult to get an accurate and precise analysis of Al in Ni alloys, because there is a large correction factor for absorption of the Al x-rays by the Ni matrix, and because the best SEM conditions for Ni analysis, 20 kV, is a large overvoltage for Al. It is very helpful if you can make or obtain a well-characterized, homogeneous alloy in the range of the alloy you will be testing, so you can check the results of your EDX analysis. The last time I did a careful phase analysis of an alloy consisting of 5% Al in Zn, I eventually went to analysing the Al at 10 kV and doing the Zn by difference. This was the only way to get the Al results with the accuracy required and agreeing with the phase diagram. With the other elements present in your system, you cannot use the element-by-difference method, but you could check to see if you get the same Al results at 10 kV, doing the Ni by difference (forgetting the other elements), as you get in your full, standard analysis at 20 kV. Most of the EDX supplier companies have excellent instruction books that cover the topics of chemical analysis by EDX and the exact conditions required for accurate analysis. Some offer courses in EDX analysis and they are a very good way to get familiar with the strengths and weaknesses of the system. Please contact me if you need any more information. At 07:07 PM 6/21/00 -0500, you wrote:
} CAN SOMEBODY HELP ME?: } } -We need chemical reference standards for the analysis of a Ni-base alloy. } } -We intend to use the energy-dispersive X-ray analysis, coupled to a } Scanning } Electron Microscope(SEM). } } -The composition of the alloy is going to vary between 18 to 28 wt% } Aluminium ; } Al is the most important element to be tested, the remainder of the alloy } being } mostly Ni of course. } } -Secondary main elements are : Cr from 4 to 10 wt% ; Co from 10 to 15 } wt% ; Elements in low concentration (below 3%) are Ti and Mo. } } -Accuracy : 0.5 % in Al is required. } } } Questions are : } } -How many different standards would we need ? } } -What should be the chemical variations from standard to standard to reach } the } required accuracy ? } } -What would be the price for such standards ? Time for delivery ? Where to } order this ? } } -Can you send us information (reprints from technical journals, articles) on } the } topic of chemical analysis by means of energy-dispersive (or } wavelength-dispersive) X-ray systems ? } } If you need any further information, please contact me. } } In anticipation, many thanks for a prompt answer. } } Yours faithfully, } } PAUL-LAURENT CAPRON JR.
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 Listservers, We are in the process of replacing our old critical point dryer and have narrowed our selection to the following choices: Tousimis Samdri 795 semi automatic and the EMS 850 critical point dryers. If anyone out there is familiar with either of these instruments and would care to comment on their advantages and disadvantages, I would appreciate hearing from you. Offline replies are welcome. I will report back with summary of results.
Tom Januszewski Senior 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 Fax: 214-648-6408 Email: tom.januszewski-at-email.swmed.edu
Dear Kristian, I also have an old JEOL evaporator (JEE-4B) and, yes, the Forgel tube was just a hot-wire ionization gauge. I got tired of buying new ones, so I successfullly replaced that with a cold cathode gauge attached to a tube of the right diameter. At 01:54 PM 6/22/00 +0300, you wrote: } Hello, } } I'm looking for a manual for the JVG-N2 ionization } vacuum gauge. I've already tried JEOL and they could } find JVG-N1 manual (thanks!), but not -N2, and -N1 and } -N2 are different enough (different connectors, apparently } EB grid heating in -N2 etc.) that the -N1 manual is not } enough to get the gauge operational. } } So, anyone with manual for JVG-N2 ?? } } btw: The JVG-N1 manual mentions "Forgel tube" which looks } in drawing just like normal hot-filament ionization } gauge tube.. Is "Forgel tube" just an old term for one? } } Thanks, } } Kristian Ukkonen. 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
This doesn't seem to cover it. Gold and platinum are heavy metals. So are we to treat them as we treat osmium tetroxide? Where does light end and heavy begin anyway? There is a real need here for a proper definition of the risk.
} } The problem is that any form of osmium is a heavy metal, and therefore } should be treated as one would cadmium or arsenic. It is not innocuous. } } 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 } } } }
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 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
I received a request for EM autoradiography of C14 labeled polymers. The idea is to trace the distribution of the polymer once it is internalized in the cell. It has been a long while since I have done autoradiography and I was wondering if there are new books and information that is available out there. Is anyone still using this technique. What are the alternatives? I am not too keen on handling radioactive materials again. I would appreciate comments.
******************************************************* 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
Greetings: I am a new subscriber to this list and am looking for input/advice regarding the use of SEM's in undergraduate research. I teach at a liberal arts university with only undergraduate programs in the natural sciences. A geology colleague and myself (I'm a biologist) are exploring possible funding sources which would allow us to purchase an SEM that would be used by biology and geology undergrads in their independent and senior thesis projects. (We currently have a non-functioning ETEC that we would like to replace -- we've been advised that further repair and head-banging would not be cost effective). If anyone is currently engaged in SEM-based research with undergrads, I would like to get your feedback on your experiences (both the pluses and minuses). Although we don't have a TEM, input from those using TEM with undergrads would also be helpful.
Specific Questions:
1) What types of projects are your students pursuing?
2) Are you conducting interdisciplinary projects? e.g., micropaleontology or environmental applications.
3) Are there particular models of SEMs which would be more user friendly since we would be training undergrads and we would also be the primary trainers and technicians?
4) Have you developed an undergrad EM course?
Thanks, Ken
----------------------------------------------------------------------- Kenneth Long, PhD email: long-at-clunet.edu Associate Professor, Biology Office: (805) 493-3346 California Lutheran University Fax: (805) 493-3392 60 West Olsen Rd. #3700 http://www.clunet.edu/Biology/Anatomy Thousand Oaks, CA 91360
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Fellow Microscopists: Stuart Mckernan,M&M'00 Program Chair, has arranged for an additional "EXPERTS" session for the topic of Facility Management. This special session will be held on Wednesday, August 16 at 9:00Am in Room 106. Advertising will be done by flyers, daily news bulletin, and word of mouth.
The room will be available all morning so that we can be somewhat flexible in timing. However, in order to try to make the session as constructive as possible, I would like to identify 2-3 main topics of interest to lab managers from both academic and industrial facilities. These topics will be covered first. Additional topics can then be discussed based on interest and time.
I would appreciate it if those who are interested in attending this session would provide the following: a) 2-3 topics of primary interest to you ranked in order of that interest. b) suggestions of individuals who might be asked to introduce a specific topic so as to give a framework for further discussion.
I am heading out for vacation on July 1 and will unsubscribe from the list a few days prior to then. Please send the above information directly to me rather than to the list so I am sure to get it. We will announce the topics to be discussed and approximate times they will be discussed so that you can choose when you want to attend the session.
If the session is well attended, then it may be added to the agendas of future meeting. Hope to hear from many of you regarding the topics. 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
by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id SAA15867 for dist-Microscopy; Thu, 22 Jun 2000 18:00:09 -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 RAA15863 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Thu, 22 Jun 2000 17:59:39 -0500 (CDT) Received: from pdx.edu (tuttle.oit.pdx.edu [131.252.120.29]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id RAA15856 for {microscopy-at-sparc5.microscopy.com} ; Thu, 22 Jun 2000 17:59:28 -0500 (CDT) Received: from [131.252.192.235] (host-192-235.dhcp.pdx.edu [131.252.192.235]) by pdx.edu (8.9.3/8.9.3) with ESMTP id PAA26977 for {microscopy-at-sparc5.microscopy.com} ; Thu, 22 Jun 2000 15:53:53 -0700 (PDT) Message-Id: {200006222253.PAA26977-at-pdx.edu} X-Mailer: Microsoft Outlook Express Macintosh Edition - 4.5 (0410)
Hi everyone,
I'm currently making 1.5 litre batches of 4% formaldehyde using paraformaldehyde, sodium hydroxide and distilled water and not having much success. The mixture immediately begins to polymerize. The recipe I used is simply an amplified version of Karnovsky's recipe for 25ml of 4% (which works perfectly). I am wondering whether my assumption of a simple linear relationship between the proportions of NaOH and paraformaldehyde is incorrect. Making stock solutions of higher concentrations does not work either.
Does anyone know a more successful method of preparing large volumes of formaldehyde? We do not want to use phosphates or preservatives.
regards Elizabeth McKenzie -------------------------------------------------- Geomicrobiology and Electron Microscopy Laboratory Room S9 Cramer Hall 1721 SW Broadway Portland State University Portland OR97201
Fellow EM Users, Would anyone have a list of the O rings for use on the JEOL JEE4B/4C vacuum evaporator. The photocopy of pages from the manual that I have does not list any o-rings. Thankyou Barry EM UNIT UNSW
One response. No follow through. Board is going into the circular file.
gg
} Date: Thu, 15 Jun 2000 07:55:38 -0700 } To: MSA listserver } From: "Dr. Gary Gaugler" {gary-at-gaugler.com} } Subject: Polaron E5200 control board } } 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. } } gary g.
I had a look at the excellent site http://www.webelements.com/index.html
clicking on osmium and then asking for "biology" this info is given: "Osmium metal does not normally cause problems as it is relatively unreactive but all osmium compounds should be regarded as highly toxic. The metal dust is an irritant and presents a fire and explosion hazard. Osmium oxide, OsO4, is highly toxic, and boils at 130?C (760 mm). Concentrations in air as low as 10-7 g m-3 can cause lung congestion, skin damage, and severe eye damage. The oxide, in particular, should only ever be handled by a properly qualified chemist." We all know about the tetroxide, but the point I made in my previous email was that the fridge contamination is Osmium metal and this has little reactivity. Osmium when finely dispersed would produce a tiny amount of tetroxide. Possibly not enough to worry while in the lab, certainly not if its in landfill or smelted. 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 23, 2000 6:58 AM, Chris Jeffree [SMTP:cjeffree-at-srv0.bio.ed.ac.uk] wrote:} } } This doesn't seem to cover it. Gold and platinum are heavy metals. } So are we to treat them as we treat osmium tetroxide? } Where does light end and heavy begin anyway? There is a real } need here for a proper definition of the risk. } } } } } The problem is that any form of osmium is a heavy metal, and therefore } } should be treated as one would cadmium or arsenic. It is not innocuous. } } } } 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 } } } } } } } } } } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } 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 } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Dear colleagues, Many thanks for all responses to my question considering the hole in the EDX spectrum. We had a service engineer in our lab and he tried to solve the problem but he was not successful. I have prepared WEB page with the images of spectra, that were recorded from following samples: Cu_Al sample (usually used for calibration) Ti grid AL grid K standard
The url of that page is following: http://www.biomed.cas.cz/~benada/EDX_page/index.htm
Please, if anyone would be so kind and could comment the spectra, we will be very happy. Best regards from Prague Oldrich 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 WEB:http://www.biomed.cas.cz/mbu/lem113/lem.htm
There's nothing mysterious about this. It should be possible to make any volume of formaldehyde solution over a wide range of concentrations. We routinely make 10% solutions without problems in volumes much greater than 25ml. There is no reason to suppose that the volume is relevant, and the concentration is not critical. The temperature needs to be } 60oC and the solution needs to be slightly alkaline. Otherwise it is extremely straightforward. Could you tell us your procedure, step by step? Then maybe it will be possible to diagnose the problem. Chris
Date sent: Thu, 22 Jun 2000 15:50:11 -0700
Commerially available formaldehyde contains formic acid and methanol and is therefore unsuitable for EM. Instead, one usually prepares a solution of formaldehyde from paraformaldehyde. An aqueous solution (40%, w/v) can be made by warming for 1 h at 65¡V (constant stirring). Milkyness can be overcome by adding a few drops of a 40% (w/v) NaOH solution. This solution is stable for several weeks at 4¡c (in the dark). Hopes this helps.
De Pauw Bart Ghent University Faculty of Veterinary Medicine Department Morphology
"E. J. McKenzie" schreef:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 currently making 1.5 litre batches of 4% formaldehyde } using paraformaldehyde, sodium hydroxide and distilled water and not } having much success. The mixture immediately begins to polymerize. The } recipe I used is simply an amplified version of Karnovsky's recipe for 25ml } of 4% (which works perfectly). I am wondering whether my assumption of a } simple linear relationship between the proportions of NaOH and } paraformaldehyde is incorrect. Making stock solutions of higher } concentrations } does not work either. } } Does anyone know a more successful method of preparing large } volumes of formaldehyde? We do not want to use phosphates or } preservatives. } } regards } Elizabeth McKenzie } -------------------------------------------------- } Geomicrobiology and Electron Microscopy Laboratory } Room S9 Cramer Hall } 1721 SW Broadway } Portland State University } Portland } OR97201 } } ph:503 725 3362 } fax:503 725 3025
Here's an interesting problem. The other day I was examining some plankton trap samples in our ESEM, and found a few bits of what I took to be pieces of foraminiferal shell - they were slightly curved, crystalline-looking shards with closely spaced fine openings (the foramen, for which forams are named) and a few large holes (possible spine attachment localities). I ran EDS on them to make sure that is indeed, what they were. Planktic foram shells should be fairly pure calcium carbonate. What I got instead were spectra each showing a fairly large strontium peak, a smaller sulfur peak and a little oxygen. There were also very high aluminum peaks on each one, but since the sample was simply air dried on an aluminum stub, I assume that was the source of the Al. In 20-some years of micropaleontology I've never heard of any marine organism that uses strontium to build its shell. Sr is just below Ca in the periodic table, so I suppose it shares some of the same reactability characteristics (but I'm no chemist), so perhaps it could substitute, if no Ca was available (which would be darn funny in a surface marine sample). Let me add that all the other peaks were in their proper places, and the instrument had been calibrated quite recently, and I can think of no source of strontium contamination which could have gotten in there. We're planning on sending some of these bits out for analysis by some other means just to make sure, but for the time being, it's quite a mystery. Any ideas from the marine biologists? The chemists? The EDS gurus? Psychics?
F.C. Thomas MicroAnalysis Facility Geological Survey of Canada (Atlantic) Bedford Institute of Oceanography Dartmouth, Nova Scotia Canada B2Y 4A2
We use the Agfa DD3700 paper processor. However, the wonderful graded papers are no longer available and have not been for some time now. Our back-up system is the Mohr Pro-8. Most of our older trained users are unwilling to use the AGFA multi-contrast graded papers, stating that the iamge is "not as crisp, not as white, harder to work with", etc. If anyone has used AGFA graded papers in the past, and has found a resin-coated graded paper alternative that works to give sharp, crisp, clean, white, micrographs from the DURST Point source, I would like to hear about your solutions. Some un-tried recommendations have been: FOMA papers, FORTE papers, ILFORD papers, and then there is the rest of the list, EFKE, Kentmere, Kodak, MACO, Oriental, adn ECCO. Does anyone know if there is a graded paper comparable to the AGFA RAPITONE GRADES P1,P2, P 3, and P4. AGFA only suggests to use their polycontrast resin papers. I have swithced to digital, but the older folks with all the great publications refuse to lower their standards. I would lke to know more about how I can get the wanted details back onto my micrographs. ANY IDEAS ? ....-TOM
Thomas A Baginski Technical Coordinator for Microscopy Uniformed Services University of the Health Sciences Bethesda, MD 20814-4799
Depending on the polymorph of CaCO3 that foram shells are made of, a few hundred (calcite) to a few thousand (aragonite) ppm of Sr would not be unusual in carbonate precipitated from standard sea-water. SO4 ion is also a fairly significant component of sea-water and S can be incorporated in the structures of carbonate minerals in small amounts. It could also be present as fluid inclusions. Another possibility is that the seawater present in the pores of the forams on recovery from the ocean has precipitated celestite (SrSO4) on dessication, in which case the Sr might have a very patchy distribution.
Roger Mason
Frank Thomas 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 } -----------------------------------------------------------------------. } } Here's an interesting problem. The other day I was examining some plankton } trap samples in our ESEM, and found a few bits of what I took to be pieces } of foraminiferal shell - they were slightly curved, crystalline-looking } shards with closely spaced fine openings (the foramen, for which forams are } named) and a few large holes (possible spine attachment localities). I ran } EDS on them to make sure that is indeed, what they were. Planktic foram } shells should be fairly pure calcium carbonate. What I got instead were } spectra each showing a fairly large strontium peak, a smaller sulfur peak } and a little oxygen. There were also very high aluminum peaks on each one, } but since the sample was simply air dried on an aluminum stub, I assume } that was the source of the Al. }
} We use the Agfa DD3700 paper processor. However, the wonderful } graded papers are no longer available and have not been for some time now. } Our back-up system is the Mohr Pro-8. Most of our older trained users are } unwilling to use the AGFA multi-contrast graded papers, stating that the } iamge is "not as crisp, not as white, harder to work with", etc. } If anyone has used AGFA graded papers in the past, and has found a } resin-coated graded paper alternative that works to give sharp, crisp, clean, } white, micrographs from the DURST Point source, I would like to hear about } your solutions. } Some un-tried recommendations have been: FOMA papers, FORTE papers, } ILFORD papers, and then there is the rest of the list, EFKE, Kentmere, } Kodak, MACO, Oriental, adn ECCO. Does anyone know if there is a graded } paper comparable to the AGFA RAPITONE GRADES P1,P2, P 3, and P4. AGFA } only suggests to use their polycontrast resin papers. } I have swithced to digital, but the older folks with all the great } publications refuse to lower their standards. } I would lke to know more about how I can get the wanted details } back onto my micrographs. ANY IDEAS ? ....-TOM } } Thomas A Baginski } Technical Coordinator for Microscopy } Uniformed Services University of the Health Sciences } Bethesda, MD 20814-4799 } } Voice Phone: 301 295 5691 } Fax: 301 319 8218 } Email: tbaginski-at-usuhs.mil
Dear Tom:
I have had good results with both Kodak and Ilford multigrade resin coated papers in my DD-3700. I am surprised that you can see a difference in sharpness between modern. glossy papers. Is Kodabrom RC still available in grades?
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 **********************************************
Frank, I am not a microbiologist or even close - I work in cathodoluminescence instrumentation. I recalled that Sheldon Sommer made some comments on Sr in shells in his work at Penn State. On page 282 of his paper, he talks about 70 mole percent strontianite in pelecypods, for what it is worth. Sommer, S. E. CL of carbonates,2. Geological Applications. Chemical geology, 1972, pp. 275-284.
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.)
There is a group of marine protistans that uses strontium sulphate to make their tests. Amoeboid critters closely related to ... wait a bit while I dredge through my memory here, my texts are at home ... Actinaria? Relatives of the Radiolarians and Heliozoa IF I'm remembering correctly. They're mentioned inter alia in the book "Synoptic Classification of Living Organisms" and any good invertebrate zoology text or Protistology text.
Phil The Al is most likely from the stub ... try a carbon mount.
} Here's an interesting problem. The other day I was examining } some plankton } trap samples in our ESEM, and found a few bits of what I took to be pieces } of foraminiferal shell - they were slightly curved, crystalline-looking } shards with closely spaced fine openings (the foramen, for which forams are } named) and a few large holes (possible spine attachment localities). I ran } EDS on them to make sure that is indeed, what they were. Planktic foram } shells should be fairly pure calcium carbonate. What I got instead were } spectra each showing a fairly large strontium peak, a smaller sulfur peak } and a little oxygen. There were also very high aluminum peaks on each one, } but since the sample was simply air dried on an aluminum stub, I assume } that was the source of the Al. } In 20-some years of micropaleontology I've never heard of any marine } organism that uses strontium to build its shell. Sr is just below Ca in the } periodic table, so I suppose it shares some of the same reactability } characteristics (but I'm no chemist), so perhaps it could substitute, if no } Ca was available (which would be darn funny in a surface marine sample). } Let me add that all the other peaks were in their proper } places, and the } instrument had been calibrated quite recently, and I can think of no source } of strontium contamination which could have gotten in there. We're planning } on sending some of these bits out for analysis by some other means just to } make sure, but for the time being, it's quite a mystery. } Any ideas from the marine biologists? The chemists? The EDS gurus? } Psychics? } } } F.C. Thomas } MicroAnalysis Facility } Geological Survey of Canada (Atlantic) } Bedford Institute of Oceanography } Dartmouth, Nova Scotia } Canada B2Y 4A2
-- }}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{ 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)
Below is the result of your feedback form. It was submitted by (barbarac-at-biols.susx.ac.uk) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Friday, June 23, 2000 at 08:00:37 ---------------------------------------------------------------------------
Email: barbarac-at-biols.susx.ac.uk Name: Barbara Ciani School: University of Sussex Question: Hi everyone,
I need to do some congo red staining on my peptides and i need to make sure they stick on the microscope slides.
I have been suggested to use gelatine with chrome alum but my slides don't seem to get 'sticky'.
Any suggestion?
I use 0.5% gelatine (300 bloom) and 0.05% chrome alum.
Date sent: Mon, 19 Jun 2000 09:46:49 -0400 To: {Microscopy-at-sparc5.microscopy.com} } From: Leona Cohen-Gould {lcgould-at-mail.med.cornell.edu}
Tom,
Although we do very little photographic printing anymore, I printed extensively in the past and have gotten excellent results from Kodak and Ilford multi-contrast resin-coated papers, using a Durst professional enlarger, as well as Beseler enlargers of various types. What I have never done, though, is make much use of a paper processor. Force of habit, maybe, but I always preferred tray processing, even when I had to make lots of prints in short amounts of time.
Even resin-coated papers respond somewhat to variations in developing times/temperatures, etc., allowing minor adjustments during processing. Often it is possible to watch the image come up in the tray and know within 15-30 seconds whether it will acceptable or what changes will be necessary. I can only speak for myself, but quality-wise I believe that tray processing is superior, in terms of tonality, grey-scale range, etc. I also believe it's as fast or faster.
That said, I know that people accustomed to processors are just as stubborn as we dinosaurs who prefer tray processing, and nobody is likely to change long-established habits. Any mainstream resin-coated, glossy paper should capture any needed detail that's present in the negative. I'm not saying that photo paper is as "sharp" in terms of lines/mm as a TEM negative, but only that the detail it won't capture is too small to see in normal viewing, and certainly won't survive the reproduction process in a journal. If you're visibly losing detail in the print at a standard, or even close, viewing distance without a lens, I would be checking the negatives and enlarger.
I won't even get into the debate about journals considering digital imaging as a "lowering of standards"! Not going there... :-)
Just my very subjective two cents.
All the best, 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: Thomas A. Baginski [mailto:tombg-at-bictom.usuf1.usuhs.mil] Sent: Friday, June 23, 2000 7:09 AM To: Microscopy-at-sparc5.microscopy.com
We use the Agfa DD3700 paper processor. However, the wonderful graded papers are no longer available and have not been for some time now.
Our back-up system is the Mohr Pro-8. Most of our older trained users are
unwilling to use the AGFA multi-contrast graded papers, stating that the iamge is "not as crisp, not as white, harder to work with", etc. If anyone has used AGFA graded papers in the past, and has found a resin-coated graded paper alternative that works to give sharp, crisp, clean, white, micrographs from the DURST Point source, I would like to hear about
your solutions. Some un-tried recommendations have been: FOMA papers, FORTE papers,
ILFORD papers, and then there is the rest of the list, EFKE, Kentmere, Kodak, MACO, Oriental, adn ECCO. Does anyone know if there is a graded paper comparable to the AGFA RAPITONE GRADES P1,P2, P 3, and P4. AGFA only suggests to use their polycontrast resin papers. I have swithced to digital, but the older folks with all the great publications refuse to lower their standards. I would lke to know more about how I can get the wanted details back onto my micrographs. ANY IDEAS ? ....-TOM
Thomas A Baginski Technical Coordinator for Microscopy Uniformed Services University of the Health Sciences Bethesda, MD 20814-4799
} Dear all } } I just found some cells with granules resembling neurosecretory granules in } invertebrate tissues, and I need to confirm their nature. } } Can anyone recomend a straightforward and reliable method to identify } neurosecretory cells (at the light or/and electron microscopic levels)? } } Thanks to all } Dr. A.P. Alves de Matos
Chrome-alum hematoxylin or Falhmi's aldehyde fuchsin work well with mammalian tissues. Check almost any edition of Humason's "Animal Tissue Techniques" for specifics. Modifications for invert tissues are out there but I don't have references handy.
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 **********************************************
Hi Y'all: Thanks for your overwelming response to my request for an independent FIB lab that can do TEM x-sections. Over the years, this forum has been an invaluable resource for me. Regards, Mike Coviello Lab Manager University of Texas -at- Arlington
Hello everyone, I'm studying microstructures and intergrowths of fibrous minerals by TEM. I'm looking for a sample of amosite and a sample of antophillite fibres in a rock matrix in order to continue this study. I'm gratefully if someone will send my these samples and will be interested to collaborate.
Thank you everyone, Elena Belluso
---------------------------------------------------- Elena BELLUSO Dipartimento di Scienze Mineralogiche e Petrologiche Via Valperga Caluso, 35 I-10125 TORINO - ITALIA tel:(39) 011.670.71.35 - fax: (39) 011.670.71.28 e-mail: belluso-at-dsmp.unito.it http://www.dsmp.unito.it ----------------------------------------------------
The deadline for contributions to the "LATE-BREAKING POSTER SESSION" at MICROSCOPY AND MICROANALYSIS 2000 is very close at hand.
This poster session will be 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. The abstracts will obviously not be published in the proceedings, since it is already being printed, but will be available at the meeting. The posters will be advertized in the daily meeting newsletters to alert attendees to their presence.
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 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).
It is not too early to register for the meeting, the pre-meeting congress, or the short courses (or any combination of these events). The early registration deadline is also fast approaching, and can be done using the online registration site at: http://www.peregrine.net/mm2000/
Looking forward to a great meeting in Philadelphia,
__________________ 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
Hello everyone, I'm studying microstructures and intergrowths of fibrous minerals by TEM. I'm looking for a sample of amosite and a sample of antophillite fibres in a rock matrix in order to continue this study. I'm gratefully if someone will send my these samples and will be interested to collaborate.
Thank you everyone, Elena Belluso
---------------------------------------------------- Elena BELLUSO Dipartimento di Scienze Mineralogiche e Petrologiche Via Valperga Caluso, 35 I-10125 TORINO - ITALIA tel:(39) 011.670.71.35 - fax: (39) 011.670.71.28 e-mail: belluso-at-dsmp.unito.it http://www.dsmp.unito.it ----------------------------------------------------
Yes...very interesting result! My guess is that you have happened upon a very rare group of microplankton which make a celestite test (SrSO4). If I remember my micropaleo, look for a group called Acantharia (?) Try Haq & Boersma's "Marine Micropaleontology" for a start. The reason you have not come across them in 20-odd years is that the celestite tests are very unstable...I saw a few beautiful examples in filtered water from the Arabian Sea, never in sediment samples. So not much use for paleoclimatology, but an interesting diversion. If you can post some pics to a website, I'd like to take a look at them.
Matt
Matthew J. Lynn, Ph.D. Center for Advanced Microscopy University of Miami (305)284-4736 mlynn-at-miami.edu
On Friday, June 23, 2000 6:59 AM, Frank Thomas [SMTP:thomasf-at-AGC.BIO.NS.CA] 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 } -----------------------------------------------------------------------. } } } Here's an interesting problem. The other day I was examining some plankton } trap samples in our ESEM, and found a few bits of what I took to be pieces } of foraminiferal shell - they were slightly curved, crystalline-looking } shards with closely spaced fine openings (the foramen, for which forams are } named) and a few large holes (possible spine attachment localities). I ran } EDS on them to make sure that is indeed, what they were. Planktic foram } shells should be fairly pure calcium carbonate. What I got instead were } spectra each showing a fairly large strontium peak, a smaller sulfur peak } and a little oxygen. There were also very high aluminum peaks on each one, } but since the sample was simply air dried on an aluminum stub, I assume } that was the source of the Al. } In 20-some years of micropaleontology I've never heard of any marine } organism that uses strontium to build its shell. Sr is just below Ca in the } periodic table, so I suppose it shares some of the same reactability } characteristics (but I'm no chemist), so perhaps it could substitute, if no } Ca was available (which would be darn funny in a surface marine sample). } Let me add that all the other peaks were in their proper places, and the } instrument had been calibrated quite recently, and I can think of no source } of strontium contamination which could have gotten in there. We're planning } on sending some of these bits out for analysis by some other means just to } make sure, but for the time being, it's quite a mystery. } Any ideas from the marine biologists? The chemists? The EDS gurus? } Psychics? } } } F.C. Thomas } MicroAnalysis Facility } Geological Survey of Canada (Atlantic) } Bedford Institute of Oceanography } Dartmouth, Nova Scotia } Canada B2Y 4A2
I think you have massive icing of your detector crystal, or equally massive contamination of your window. I have modelled your Al spectrum (using DTSA) and can get quite good agreement with your spectrum by putting in 300 microns of ice in the model. The low energy rise is probably caused simply by the tail of the electronic noise which is amplified so much by the scale at which you display your spectrum. Because of the way x-ray absorption works, I can't really tell the difference between absorption in ice and absorption in oil. Anyway, fixing an icing problem is far easier than fixing an oil problem - you simply have to warm up the detector while pumping on it. This will need an adaptor to fit on the pumping port of your detector - your service engineer should have, or be able to get hold of, one of these. So I would try warming the detector and seeing of it solved the problem.
I wouldn't stake my professional reputation on this diagnosis, but I am certain that if the problem is not contamination, then it is a geometrical problem - the collimator may have moved, or some other mechanical derangement taken place.
Good luck,
Tony Garratt-Reed.
At 11:11 AM 06/23/2000 +0200, 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 **
Dear Ken, I have always run undergraduate courses on the SEM/EDX and we currently run a course that puts groups of second year engineering students doing independent research projects. They research the composition of many common household and commercial appliances, taking them apart to see what metals, plastics and ceramics are used for the various applications such as small electric motors, heating elements and temperature control. The students are very appreciative of being allowed to sit down and use the instrument with minimal supervision and I find they are very careful with it. I run courses for Anthropology and Electrical Engineering and all disciplines of Engineering, Physics and Chemistry use the instrument for both undergraduate and graduate research. I would suggest you get an EDX and a low vacuum ("environmental") SEM for full versatility in both physical and biological EM. All of my EM instruments (two SEMs with EDX, one 200 kV TEM and SEM with EBSP) are Hitachi, as I have found them the most robust. They run for years of student use and nothing ever seems to go wrong with them. I had an ETEC before my first Hitachi SEM and I liked it too, but they have been out of business for a long time. We also have a fourth year course that covers XRD, SEM, EDX and TEM/EDX. The professor in charge has developed the lab course to touch all of the important principles and demonstrate them in the lab. We cover changing kV, EDX excitation volume, EDX matrix corrections, TEM use and TEM/EDX issues. Please contact me if I can give you any other information. At 03:05 PM 6/22/00 -0700, you wrote: } } Greetings: } I am a new subscriber to this list and am looking for input/advice } regarding the use of SEM's in undergraduate research. I teach at a liberal } arts university with only undergraduate programs in the natural sciences. } A geology colleague and myself (I'm a biologist) are exploring possible } funding sources which would allow us to purchase an SEM that would be used } by biology and geology undergrads in their independent and senior thesis } projects. (We currently have a non-functioning ETEC that we would like to } replace -- we've been advised that further repair and head-banging would } not be cost effective). If anyone is currently engaged in SEM-based } research with undergrads, I would like to get your feedback on your } experiences (both the pluses and minuses). Although we don't have a TEM, } input from those using TEM with undergrads would also be helpful. } } Specific Questions: } } 1) What types of projects are your students pursuing? } } 2) Are you conducting interdisciplinary projects? e.g., micropaleontology } or environmental applications. } } 3) Are there particular models of SEMs which would be more user friendly } since we would be training undergrads and we would also be the primary } trainers and technicians? } } 4) Have you developed an undergrad EM course? } } Thanks, } Ken
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
Date sent: Fri, 23 Jun 2000 11:57:42 -0400 (EDT) } From: Donald Lovett {lovett-at-TCNJ.EDU} To: Chris Jeffree {cjeffree-at-srv0.bio.ed.ac.uk}
When AGFA (Bayer) pulled the plug on the graded papers, we switched to Kodak Polycontrast RCIII. So I pulled the plug on their activator and fixer and now use Kodak Polymax developer 1:4 dilution and Rapid Fixer in my DD3700. Still use my Durst S45EM with point light source and Kodak Polycontrast Filter Kit. Bob Santoianni Emory University Hospital Atlanta, GA
} Here's an interesting problem. The other day I was examining some } plankton } trap samples in our ESEM, and found a few bits of what I took to be pieces } of foraminiferal shell - they were slightly curved, crystalline-looking } shards with closely spaced fine openings (the foramen, for which forams are } named) and a few large holes (possible spine attachment localities). I ran } EDS on them to make sure that is indeed, what they were. Planktic foram } shells should be fairly pure calcium carbonate. What I got instead were } spectra each showing a fairly large strontium peak, a smaller sulfur peak } and a little oxygen. There were also very high aluminum peaks on each one, } but since the sample was simply air dried on an aluminum stub, I assume } that was the source of the Al. } In 20-some years of micropaleontology I've never heard of any marine } organism that uses strontium to build its shell. Sr is just below Ca in the } periodic table, so I suppose it shares some of the same reactability } characteristics (but I'm no chemist), so perhaps it could substitute, if no } Ca was available (which would be darn funny in a surface marine sample). } Let me add that all the other peaks were in their proper places, } and the } instrument had been calibrated quite recently, and I can think of no source } of strontium contamination which could have gotten in there. We're planning } on sending some of these bits out for analysis by some other means just to } make sure, but for the time being, it's quite a mystery. } Any ideas from the marine biologists? The chemists? The EDS gurus? } Psychics?
Frank -
You've just rediscovered the strontianate Radiolaria. They DO selectively concentrate strontium. I almost selected them as my thesis topic 'way back in the dark ages; I dropped the idea when I realised that there was no way to culture them in the lab. I don't think there is, yet. } 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
This may be a bit of a shameless plug for a paper I contributed to, but here's the reference:
Payne CM, Cromey DW (1987) Ultrastructural similarities between Chlorohydra viridissima and human neurosecretory granules: A cytochemical study using the uranaffin reaction. Cytobios 50:191-203.
Back when I was doing clinical TEM, we used the Uranaffin Method as the "gold standard" for diagnostic identification of neurosecretory granules in tumors. It was more reliable than immunomarkers and its fairly simple to do. The reference section from this particular article should include Dr. Payne's original publication on the technique. The most important thing is to follow the protocol, failure to use the specified buffers usually causes problems.
Yours, Doug Cromey
At 09:46 AM 6/23/2000 -0400, you wrote: } "A.P. Alves de Matos" wrote: } } I just found some cells with granules resembling neurosecretory granules in } } invertebrate tissues, and I need to confirm their nature. } } } } Can anyone recomend a straightforward and reliable method to identify } } neurosecretory cells (at the light or/and electron microscopic levels)? } } Chrome-alum hematoxylin or Falhmi's aldehyde fuchsin work well with mammalian } tissues. Check almost any edition of Humason's "Animal Tissue Techniques" for } specifics. Modifications for invert tissues are out there but I don't have } references handy.
.................................................................... : 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"
Sorry if this is double-posted....server troubles!
Frank,
Yes...very interesting result! My guess is that you have happened upon a very rare group of microplankton which make a celestite test (SrSO4). If I remember my micropaleo, look for a group called Acantharia (?) Try Haq & Boersma's "Marine Micropaleontology" for a start. The reason you have not come across them in 20-odd years is that the celestite tests are very unstable...I saw a few beautiful examples in filtered water from the Arabian Sea, never in sediment samples. So not much use for paleoclimatology, but an interesting diversion. If you can post some pics to a website, I'd like to take a look at them.
Matt
On Friday, June 23, 2000 6:59 AM, Frank Thomas [SMTP:thomasf-at-AGC.BIO.NS.CA] 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 } -----------------------------------------------------------------------. } } } Here's an interesting problem. The other day I was examining some plankton } trap samples in our ESEM, and found a few bits of what I took to be pieces } of foraminiferal shell - they were slightly curved, crystalline-looking } shards with closely spaced fine openings (the foramen, for which forams are } named) and a few large holes (possible spine attachment localities). I ran } EDS on them to make sure that is indeed, what they were. Planktic foram } shells should be fairly pure calcium carbonate. What I got instead were } spectra each showing a fairly large strontium peak, a smaller sulfur peak } and a little oxygen. There were also very high aluminum peaks on each one, } but since the sample was simply air dried on an aluminum stub, I assume } that was the source of the Al. } In 20-some years of micropaleontology I've never heard of any marine } organism that uses strontium to build its shell. Sr is just below Ca in the } periodic table, so I suppose it shares some of the same reactability } characteristics (but I'm no chemist), so perhaps it could substitute, if no } Ca was available (which would be darn funny in a surface marine sample). } Let me add that all the other peaks were in their proper places, and the } instrument had been calibrated quite recently, and I can think of no source } of strontium contamination which could have gotten in there. We're planning } on sending some of these bits out for analysis by some other means just to } make sure, but for the time being, it's quite a mystery. } Any ideas from the marine biologists? The chemists? The EDS gurus? } Psychics? }
Matthew J. Lynn, Ph.D. Center for Advanced Microscopy University of Miami (305)284-4736 mlynn-at-miami.edu
Oldrich- I am sorry, but I have forgotten all of the details of your problem. Have you thermally cycled your detector? letting the detector be a room temperature for at least 24hrs can remove ice build-up in the detector. It can also be useful to purge the dewar with nitrogen at that time to remove any ice that may be in the dewar thus reducing cooling efficiency. I would also suggest cleaning your detector window (I don't know what kind you have.) Be very sure that you are using the proper technique and chemicals for cleaning the window as the polymer windows are extremely easy to damage (i.e., don't blow a dust particle off of it, see the current thread on particulate damage to EDX windows.) In my case, as my window gets coated with pump oil from the vacuum, etc. the Cu L line reduces in intensity relative to the K line. For my super thin window the ratio should be L:K } =2:1. If I put a thin graphite sheet between the detector and the sample, I get spectra very similar to the ones you have posted with the low energy x-rays blocked out (you'll still have the electronic noise peak at the lowest energy) and then a normal looking higher energy range. Perhaps some of this will help. I'll be on vacation the next couple of weeks, but give me a call after 7 July if you have any questions I can help with. Matthew Ervin MErvin-at-ARL.mil 301-394-0017
Oldrich Benada {benada-at-biomed.cas.cz} on 06/23/2000 05:11:15 AM
To: Microscopy-at-sparc5.microscopy.com cc:
Dear colleagues, Many thanks for all responses to my question considering the hole in the EDX spectrum. We had a service engineer in our lab and he tried to solve the problem but he was not successful. I have prepared WEB page with the images of spectra, that were recorded from following samples: Cu_Al sample (usually used for calibration) Ti grid AL grid K standard
The url of that page is following: http://www.biomed.cas.cz/~benada/EDX_page/index.htm
Please, if anyone would be so kind and could comment the spectra, we will be very happy. Best regards from Prague Oldrich 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 WEB:http://www.biomed.cas.cz/mbu/lem113/lem.htm
To all: We have been using Kodabrome II RC paper F1-F4 for years with our Durst Laborator 1200 and Rapidoprint DD3700. As of the end of 1999, the paper is still available and gives good results.
Peggy Sherwood Photopathology Wellman Labs of Photomedicine-W224 70 Blossom Street Boston, MA 02114
Does anyone know how a really good counterfeiter would make a change in the date field of a rare coin look as though it weren't altered? I've looked for markings associated with moving the metal around, a joint between the numbers and the base coin, and differing metallurgy, with no "smoking gun"....
Any thoughts??
Brian ---------------------------------------------- Brian McIntyre Electron Microscopy Lab- River Campus Univ of Rochester Rochester, NY 14627 716-275-3058/4875
Sorry if this is double-posted....server troubles!
Frank,
Yes...very interesting result! My guess is that you have happened upon a very rare group of microplankton which make a celestite test (SrSO4). If I remember my micropaleo, look for a group called Acantharia (?) Try Haq & Boersma's "Marine Micropaleontology" for a start. The reason you have not come across them in 20-odd years is that the celestite tests are very unstable...I saw a few beautiful examples in filtered water from the Arabian Sea, never in sediment samples. So not much use for paleoclimatology, but an interesting diversion. If you can post some pics to a website, I'd like to take a look at them.
Matt
On Friday, June 23, 2000 6:59 AM, Frank Thomas [SMTP:thomasf-at-AGC.BIO.NS.CA] 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 } -----------------------------------------------------------------------. } } } Here's an interesting problem. The other day I was examining some plankton } trap samples in our ESEM, and found a few bits of what I took to be pieces } of foraminiferal shell - they were slightly curved, crystalline-looking } shards with closely spaced fine openings (the foramen, for which forams are } named) and a few large holes (possible spine attachment localities). I ran } EDS on them to make sure that is indeed, what they were. Planktic foram } shells should be fairly pure calcium carbonate. What I got instead were } spectra each showing a fairly large strontium peak, a smaller sulfur peak } and a little oxygen. There were also very high aluminum peaks on each one, } but since the sample was simply air dried on an aluminum stub, I assume } that was the source of the Al. } In 20-some years of micropaleontology I've never heard of any marine } organism that uses strontium to build its shell. Sr is just below Ca in the } periodic table, so I suppose it shares some of the same reactability } characteristics (but I'm no chemist), so perhaps it could substitute, if no } Ca was available (which would be darn funny in a surface marine sample). } Let me add that all the other peaks were in their proper places, and the } instrument had been calibrated quite recently, and I can think of no source } of strontium contamination which could have gotten in there. We're planning } on sending some of these bits out for analysis by some other means just to } make sure, but for the time being, it's quite a mystery. } Any ideas from the marine biologists? The chemists? The EDS gurus? } Psychics? }
Matthew J. Lynn, Ph.D. Center for Advanced Microscopy University of Miami (305)284-4736 mlynn-at-miami.edu
A really good counterfeiter could do this at the molecular level by using and FIB or equivalent.
I haven't really explored exactly how this could be done but it has crossed my mind.
regards,
Earl Weltmer
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 all- } } Does anyone know how a really good counterfeiter would make a change in the } date field of a rare coin look as though it weren't altered? I've looked } for markings associated with moving the metal around, a joint between the } numbers and the base coin, and differing metallurgy, with no "smoking gun".... } } Any thoughts?? } } Brian } ---------------------------------------------- } Brian McIntyre } Electron Microscopy Lab- River Campus } Univ of Rochester } Rochester, NY 14627 } 716-275-3058/4875
There was a recent posting about UA precipitating during staining.
Well wouldn't you know I too have experinecing similar problem. It took me a long time to figure out but I narrowed it down to our rinsing water that is used to rinse the grid in after it is stained.
After chaging the filters on our water purification system the problem was gone.
Rajesh Patel Robert Wood Johnson Medical School Dept. of Pathology Electron Microscopy Lab 675 Hoes Lane Piscataway, NJ 08854
how about looking at sub-surface damage (using some channeling techniques or similar)? I could imagine, that the damage from stamping the coin is different from Ion bombarding the coin.
Or, look for implanted atoms. The FIB process uses metal atoms to bombard the sample. Some of them get implanted into the material. So you might find higher concentrations where the sample was altered.
Finally, how about a surface layer like an oxide or patina. There may be differences between the very old layer and the new layer.
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: Brian McIntyre [mailto:mcintyre-at-optics.rochester.edu] Sent: Friday, June 23, 2000 12:14 PM To: Microscopy-at-sparc5.microscopy.com
Hi all-
Does anyone know how a really good counterfeiter would make a change in the date field of a rare coin look as though it weren't altered? I've looked for markings associated with moving the metal around, a joint between the numbers and the base coin, and differing metallurgy, with no "smoking gun"....
Any thoughts??
Brian ---------------------------------------------- Brian McIntyre Electron Microscopy Lab- River Campus Univ of Rochester Rochester, NY 14627 716-275-3058/4875
Over time I saw some questions regarding the installation of a UPS or some power conditioner to improve the stability of all sorts of microscopy equipment. I understand that quite a few instruments are sensitive to the quality of the line power they get. As I was working on EMC (electro magnetic compatibility) in a former life, this seems strange to me. In Europe we have very strict EC-rules that guarantee that equipment meets its specifications even under severe conditions (including HF noise on the power, fluctuations in the power, high EM fields, temperature variations etc)
I agree that microscopes are far more sensitive devices than, say, videogames but does anyone know of the EMC rules for microscopes and related equipment? It seems strange that some people are using UPS's while especially power fluctuations can be easily overcome with a good design of the electronics...
Regards,
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 * *************************************************************
I've got my references now: The protists with SrSO4 tests are the class *Acantherea*, members of the phylum Sarcodina (amoebae), subphylum Actinopoda (used to be known as the Radiolaria).
By my somewhat outdated reference ("A Synoptic Classification of Living Organisms", R.S.K. Barnes, ed.). The names have likely been changed, and the Actinopoda broken up to protect the careers of systematists.
Forget I ever mentioned Actinaria or Acanthella. The long-term storage in my wetware is getting flakey (I misremembered).
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 peoshel-at-facstaff.wisc.edu fax: (608) 262-7420 (dept. fax)
Could anyone recommend an independent U.K based company specialising in the repair and maintenance of electron microscopes ?
Without mentioning names I feel that the service we receive re our SEM maintenance contract is particularly poor and am beginning to wonder is there any alternative to complaint without improvement and excuse after excuse .
I have noticed that little repair is carried out at component level during service engineer visits but am not aware of how ' specific ' most parts are to a particular make of instrument . If parts always have to be ordered through the instrument supplier there would be no advantage in independent maintenance . Another alternative perhaps would be in-house maintenance but do courses exist to train the novice type like myself to a reasonably competent level or is this not advised ?
Keen to hear comments ...
M.Harris Email harrism-at-esm-semi.co.uk ESM LTD , South Wales , U.K
} Date: Mon, 26 Jun 2000 09:56:43 +0200 } To: Microscopy-at-sparc5.microscopy.com } From: Rudi Lurz {Lurz-at-molgen.mpg.de} } Subject: Re: ***TEM Darkroom Users*** AGFA Paper has become extinct ! } } To all: } The identical technology (developer in emulsion + activator) as with } Rapidoprint papers is used with Agfa Brovira-Speed papers. I have replaced } years ago the Rapitone paper by Brovira-Speed 310 RC because I prefered } more the blue-black of Brovira-Speed to Rapitone which showed more brown } tendency. Brovira-Speed is available from soft to extra hard and should be } still produced (hopefully) as I was told from my dealer this morning. } This is for Germany - I do not know the situation in the US. } Agfa seems to reduce drastically the B/W programme. The Agfa Scientia } 23D56 films are no longer produced too and we have to switch to Kodak SO-163. } Best regards } Rudi Lurz
_________________________________ Rudi Lurz Phone: X - 30-8413-1271 MPI fŸr Molekulare Genetik Fax: X - 30-8413-1385 Ihnestrasse 73 D-14195 Berlin E-mail to: Lurz-at-molgen.mpg.de
Teaching microscopy at both the undergrad and grad student levels would be an excellent symposium or discussion group at the Long Beach meeting. Especially since the life of EM facilities depends on having users, and getting students interested in microscopy is one of the best ways to create users.
Phil
} } Greetings: } } I am a new subscriber to this list and am looking for input/advice } } regarding the use of SEM's in undergraduate research. I teach at a liberal } } arts university with only undergraduate programs in the natural sciences. } } A geology colleague and myself (I'm a biologist) are exploring possible } } funding sources which would allow us to purchase an SEM that would be used } } by biology and geology undergrads in their independent and senior thesis } } projects. (We currently have a non-functioning ETEC that we would like to } } replace -- we've been advised that further repair and head-banging would } } not be cost effective). If anyone is currently engaged in SEM-based } } research with undergrads, I would like to get your feedback on your } } experiences (both the pluses and minuses). Although we don't have a TEM, } } input from those using TEM with undergrads would also be helpful. } } } } Specific Questions: } } } } 1) What types of projects are your students pursuing? } } } } 2) Are you conducting interdisciplinary projects? e.g., micropaleontology } } or environmental applications. } } } } 3) Are there particular models of SEMs which would be more user friendly } } since we would be training undergrads and we would also be the primary } } trainers and technicians? } } } } 4) Have you developed an undergrad EM course? } } } } Thanks, } } Ken -- }}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{ 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 should clarify that the coin in question could not have been altered after 1900 so modern techniques are obviously out of the question. the issue becomes one of... if you wanted to alter a coin's date how would you have done it 100 years ago, and how could it be uncovered using modern analysis techniques.
thanks! b- ---------------------------------------------- Brian McIntyre Electron Microscopy Lab- River Campus Univ of Rochester Rochester, NY 14627 716-275-3058/4875
Use of an x-ray microprobe should be rather definitive, I would think. While there are exotic (and expensive) techniques that could be used to remove a raised date and place a new one, it would be extremely difficult to exactly match the elemental contents, particlarly those of the trace elements. For example, gold, even when refined to 99.999% pure, contains enough impurities for any batch to be matched to a particular mine that it came from.
The only other method I could imagine is a close characterization of the grain structures. I don't know if the stamping process would have much of an affect on the surface grain, but I imagine any implantation method would leave a much different structure as completely different conditions were used to form the metal.
Allen R. Sampson, Owner Advanced Research Systems, St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
On Friday, June 23, 2000 1:14 PM, Brian McIntyre [SMTP:mcintyre-at-optics.rochester.edu] wrote:
} Hi all- } } Does anyone know how a really good counterfeiter would make a change in the } date field of a rare coin look as though it weren't altered? I've looked } for markings associated with moving the metal around, a joint between the } numbers and the base coin, and differing metallurgy, with no "smoking gun".... } } Any thoughts?? } } Brian } ---------------------------------------------- } Brian McIntyre } Electron Microscopy Lab- River Campus } Univ of Rochester } Rochester, NY 14627 } 716-275-3058/4875 } }
A couple of weeks ago I wrote in witha problem trying to fix plant vesicle preps for LR White inclusion and immunogold studies, without osmicating. The pellets were disappearing during dehydration. I included an osmium step (5, 30 or 60 minutes) and the pellets stopped disappearing, so I guess there is so little protein that a formaldehyde/gluteraldehyde fix doesn't manage to stabilize them. I'll have to check whether these osmicated vesicles work in the immunogold reaction and check the effect of a metaperiodate treatment to expose the proteins. Thanks for the advice - another problem solved.
Someone brought us some frogs' eggs to fix. It looks like they have a hard skin. The researcher is interested in extracting the nuclei (huge) which also look like they're enclosed in a hard skin, and he wants to know whether the extracted nuclei still retain some endoplasmic reticulum, so I thought the best way would be to compare a whole egg with an extracted nucleus. Any ideas/experience on how to fix this thing?
Thanks,
Mark
******************************************** Mark West, Unidad de Microscopia Electronica, (Electron Microscopy Unit) Instituto de Fisiologia Celular, Universidad Nacional Autonoma de Mexico, 04510 Mexico D.F.
I just want some opinions about the problem I got recently with our SEM.
We use this machine for electron beam lithography. This application is a little more sensitive than imaging. The problem I have now is, when I image at very low magnification, I can see the circular rim of the final section of the column. But from a couple of weeks, it changed suddenly, now the circular image is distorted like an oval shape and at second saturation point, normally only one position of gun alignment gives the maximam beam current, but I see there are clearly two distinct, well separated positions. We think something is blocking in the middle of the beam path, but want to know whether there is any other one who experienced similar thing and know the reason. I appreciate if anyone can give me his/her similar experince.
Coins have been faked outright and updated using modern dies, and assembled from two separate halves using solder. Coins whose value depends on date have been "enriched" by removing the original date and soldering in its place numbers of a more valuable date. This latter case might be discerned using scanning electron microscopy to examine surface texture and degradation and adjunct energy-dispersive analysis to detect the presence of solder.
James Martin Principal/Research Scientist Orion Analytical, LLC P.O. Box 550 Williamstown, MA 01267 www.orionanalytical.com
} i should clarify that the coin in question could not have been altered } after 1900 so modern techniques are obviously out of the question. the } issue becomes one of... if you wanted to alter a coin's date how would you } have done it 100 years ago, and how could it be uncovered using modern } analysis techniques. } } thanks! } b- } ---------------------------------------------- } Brian McIntyre } Electron Microscopy Lab- River Campus } Univ of Rochester } Rochester, NY 14627 } 716-275-3058/4875 }
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You don't have to wait that long! Go to the Philadelphia meeting web page search engine http://www.msa.microscopy.com/cgi-bin/M&M00Program.pl and enter "teaching microscopy". Steve Barlow has organized an excellent all-day session.
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
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Clearly, I have several vested interests here! As I am answering this from home, I don't actually know as I write if you are specifically concerned about a JEOL SEM, although I will check when I get to the office tomorrow.
First, I would suggest you complain, with specifics, to the service manager of the relevant company. If that is ineffective, direct your complaint to the Salesperson covering your area - sales people, perhaps, have a much stronger vested interest in happy customers. A happy customer is possibly going to buy again in the future, an unhappy customer makes their feelings known to others and can influence future sales.
I hope you are not a JEOL user - if you are, please contact me directly and I will attempt to resolve your problems.
If the SEM in question is an older instrument, independent service companies will do a good job on all of the more routine problems. You should also be able to resolve these yourself, in principle, with a reasonable knowledge of vacuum technology and electronics.
I don't know where you might get training on SEM service - possibly Protrain?
The two main independent service companies that I am aware of operating in the UK are ISS and EOS, both based in the Manchester area but with engineers throughout most of the UK. Contact details from sources on the web and Microscopy & Analysis. If you contect me directly, I will forward you details.
Problems arise with newer instruments, especially PC-controlled ones. Self-service and independent service will cover the basics but without detailed training, circuit diagrams, software tools, it gets very difficult to handle anything more than the basics.
Incidentally, this is one of the disadvantages for customers of computer controlled EMs - it makes it easier for ALL manufacturers to restrict service to approved service engineers. Not that this happens just with scientific instruments - a similar issue exists with cars/automobiles. I believe there is/has been a court case in the US by independent auto-repair companies against the manufacturers to force them to release service software.
Even on older instruments, major problems can be difficult to resolve for anybody other than the manufacturer.
You will probably find that service via independents is cheaper than via the manufacturers. Specialist parts will have to be purchased from the manufacturer but even then, the overall cost will probably be lower. However, I would add that I don't believe any EM manufacturer is interested in providing anything other than value-for-money service and, to a great extent, you do get what you pay for - at least, you should! If you are paying less, overall, the service you get is probably less - it may not be obvious, and you may never see the difference but, in the long run, you will get what you pay for.
You also need to keep in mind that the more comprehensive service contracts provided by manufactuers contain a significant 'insurance' element. Major components are expensive - with a comprehensive service contract from the maufacturer, you will never know how expensive but with service via an independent, you may find out!
Hope that is useful.
Best 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
It would very much help to know what sort of an SEM you are using and at what kV you are operating. I suspect you may have had a minor vacuum accident which caused one of your "spray apertures" to be blown out of position by inrushing air. Pull the liner tube (if your instrument has one) and reposition all the apertures. It might not be a bad idea to do a good cleaning job while inside the column.
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 REPAIR -----Original Message----- } From: Heejun Jeong {" hjjeong"-at-physics.purdue.edu} {Heejun Jeong {" hjjeong"-at-physics.purdue.edu} } To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
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There are essentially two types of UPS systems. One acts in a passive mode and switches to battery backup when power fails. The other is an active mode which converts AC input to DC and uses this to generate stable AC output.
Power conditioners are typically microprocessor-controlled units which adjust taps on a transformer to maintain reasonably stable output voltage. These can also include traps for spikes and overvoltage conditions (Topaz for example).
The issue here is the stability of the basic potentials and currents which are fed to or are delivered to the electron gun. In this regard, I think that we are talking about very narrow margins of input voltage variation and output voltages. I think that one will have line sag no matter what--on occasion. Thus, the active UPS units bypass these conditions and provide a constant, stable AC supply to the SEM. Line conditioners are rated at +/- 6% down to +/- 3%. Active UPS can do better.... but their surge current capabilities are less than those of the conditioner units.
Consequently, there is not one, singularly perfect solution. But there are options which the user should consider.
gg
At 12:29 AM 6/26/00, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} From a practical point of view my experience with computer controlled UPS systems is that cause more problems than they solve. They provide a computer with good enough power in most cases. This power is not good enough for equipment that is not designed for considerable voltage changes and micro power losses.
Most of my experience comes from computer back up power supplies and a project that was measuring sunlight. Since it was sunlight and we needed fast response we did not have any low pass filter. When working on it in the lab I used filtered DC since I was having problems with the 120 Hz flicker of incandescent bulbs. I could see sags and glitches in the AC power through the UPS systems even with the filters. It was good enough for what I was doing but I could see a good deal difference in the stability of the UPS and a battery.
I have worked with a few radio systems that use AC to charge a battery bank and then power the radios from the batteries. These are far more reliable than UPS systems.
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
} From: "Dr. Gary Gaugler" {gary-at-gaugler.com} } } There are essentially two types of UPS systems. One acts in } a passive mode and switches to battery backup when power } fails. The other is an active mode which converts AC input } to DC and uses this to generate stable AC output. } } Power conditioners are typically microprocessor-controlled units which } adjust taps on a transformer to maintain reasonably stable } output voltage. These can also include traps for spikes and } overvoltage conditions (Topaz for example). } } The issue here is the stability of the basic potentials and currents } which are fed to or are delivered to the electron gun. In this } regard, I think that we are talking about very narrow margins of } input voltage variation and output voltages. I think that one will } have line sag no matter what--on occasion. Thus, the active } UPS units bypass these conditions and provide a constant, } stable AC supply to the SEM. Line conditioners are rated at } +/- 6% down to +/- 3%. Active UPS can do better.... but their } surge current capabilities are less than those of the conditioner units. } } Consequently, there is not one, singularly perfect solution. But there are } options which the user should consider. } } gg }
} } } } Hello all, } } } } Over time I saw some questions regarding the installation of a UPS or some } } power conditioner to improve the stability of all sorts of microscopy } } equipment. } } I understand that quite a few instruments are sensitive to the quality of } } the line power they get. } } As I was working on EMC (electro magnetic compatibility) in a former life, } } this seems strange to me. } } In Europe we have very strict EC-rules that guarantee that equipment meets } } its specifications even under severe conditions (including HF noise on the } } power, fluctuations in the power, high EM fields, temperature variations } } etc) } } } } I agree that microscopes are far more sensitive devices than, say, } } videogames but does anyone know of the EMC rules for microscopes and } } related equipment? } } It seems strange that some people are using UPS's while especially power } } fluctuations can be easily overcome with a good design of the } } electronics... } } } } Regards, } } } } Jo
I am trying to locate a copy of a graph that shows the thicknesses of (glass) windows that can support atmospheric pressure against a vacuum for various radii of O ring support. Kaufman glass has one for high pressures for several glass types but it doesn't cover the range of atmospheric pressure and relatively small radii(1 to 3 cm). Pyrex, leaded glass, and quartz are the three principal materials of interest. Appreciate any leads.
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.)
You may want to contact David Gard, Ph.D., Biology Department, University of Utah, Salt Lake City, UT. He has done a great deal of work with frog eggs.
good luck
Ramin Rahbari Pfizer Global Research & Development 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: Mark West [mailto:mwest-at-ifcsun1.ifisiol.unam.mx] Sent: Monday, June 26, 2000 3:37 PM To: Microscopy-at-sparc5.microscopy.com
Hi,
A couple of weeks ago I wrote in witha problem trying to fix plant vesicle preps for LR White inclusion and immunogold studies, without osmicating. The pellets were disappearing during dehydration. I included an osmium step (5, 30 or 60 minutes) and the pellets stopped disappearing, so I guess there is so little protein that a formaldehyde/gluteraldehyde fix doesn't manage to stabilize them. I'll have to check whether these osmicated vesicles work in the immunogold reaction and check the effect of a metaperiodate treatment to expose the proteins. Thanks for the advice - another problem solved.
Someone brought us some frogs' eggs to fix. It looks like they have a hard skin. The researcher is interested in extracting the nuclei (huge) which also look like they're enclosed in a hard skin, and he wants to know whether the extracted nuclei still retain some endoplasmic reticulum, so I thought the best way would be to compare a whole egg with an extracted nucleus. Any ideas/experience on how to fix this thing?
Thanks,
Mark
******************************************** Mark West, Unidad de Microscopia Electronica, (Electron Microscopy Unit) Instituto de Fisiologia Celular, Universidad Nacional Autonoma de Mexico, 04510 Mexico D.F.
We're in the process of acquiring an EDS system for our JEOL 5600 and I'm curious about the experience of others with EDS systems on scopes without specimen airlocks (column vents to exchange specimens). Any thoughts/recommendations? I'm particularly concerned with contamination,
window integrity and the like. Are there any advantages/disadvantages of
Be vs. thin window detectors here?
Thanks,
Jim Ehrman
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
I've received a query from a climatologist who wants to photograph fine varves and/or laminations (~0.5 mm- 1 mm each layer) in carbonate (calcite) rich silt (or marl) through a binocular light microscope. The samples are flat, finely cut or polished sections that can be etched if necessary, but he's really looking for a staining method that would enhance the difference between the carbonate-rich layers and the clay-rich layers to make them more usefully photogenic. Can anyone advise us on how to proceed to best advantage?
Many thanks, Dee
*************************************************************** Dee Breger Mgr. SEM/EDX Facility Lamont-Doherty Earth Observatory 61 Route 9W Palisades, NY 10964 USA T: 914/365-8640 F: 914/365-8155
http://www.ldeo.columbia.edu/micro http://www.discovery.com/area/science/micro/micro1.html http://www.lsc.org/antarctica/front.html Journeys in Microspace (Columbia University Press, 1995)
We have two different models of thin window detectors on two different scopes. Our Hitachi 2460N does not have an airlock and is vented for every sample change. Our JEOL 840A has an airlock, but does get the chamber vented fairly often. We have only had one pin-hole develop in the detector on our 840 in the 15-20 instrument-years that we have had the detectors.
Therefore, I don't think you would be gaining much, if anything, by switching to a Be window. And you would be giving up that wonderful light element capability. I would be hard pressed not to be able to detect C and O. I would go for the thin window and be reasonably careful.
Warren S.
At 10:59 AM 6/27/2000 -0300, you wrote:
} Hi all, } } We're in the process of acquiring an EDS system for our JEOL 5600 and } I'm curious about the experience of others with EDS systems on scopes } without specimen airlocks (column vents to exchange specimens). Any } thoughts/recommendations? I'm particularly concerned with contamination, } } window integrity and the like. Are there any advantages/disadvantages of } } Be vs. thin window detectors here? } } Thanks, } } Jim Ehrman } } -- } } James M. Ehrman } Digital Microscopy Facility } Mount Allison University } Sackville, NB E4L 1G7 } CANADA } } phone: 506-364-2519 } fax: 506-364-2505 } email: jehrman-at-mta.ca } www: http://www.mta.ca/~jehrman }
} Could anyone recommend an independent U.K based company } specialising in the repair and maintenance of electron microscopes ? } } Keen to hear comments ...
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An investigator just brought me 6 samples - heads of flies. He was instructed to bring the entire body but he cuts the heads off when he does TEM. The investigator knows nothing about SEM. I have done alot of scanning of fly eyes but the body is attached. My question is: how do I mount the heads on the stubs without the graphite or silver paint causing capillary action over the entire head? We will scan on a JEOL 840A after Au/Pd coating.
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 Fax 214-648-6408 eMail: George.Lawton-at-email.swmed.edu
I would use one of the double-sided carbon sticky tabs available from several vendors. They are good for small samples that would be buried in liquids. If they don't provide enough contact area, you could very carefully dab a conductive paint around the base of the mounted heads with a pointy stick.
Cheers, 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: George Lawton [mailto:George.Lawton-at-email.swmed.edu] Sent: Tuesday, June 27, 2000 10:03 AM To: microscopy-at-sparc5.microscopy.com
An investigator just brought me 6 samples - heads of flies. He was instructed to bring the entire body but he cuts the heads off when he does TEM. The investigator knows nothing about SEM. I have done alot of scanning of fly eyes but the body is attached. My question is: how do I mount the heads on the stubs without the graphite or silver paint causing capillary action over the entire head? We will scan on a JEOL 840A after Au/Pd coating.
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 Fax 214-648-6408 eMail: George.Lawton-at-email.swmed.edu
I have dug through the archives of this fine list and found a bunch of information on vibration and microscopy. I work for an engineering consulting company, and part of our work is related to vibration of various sorts (implosions, blasting, construction, pile driving, vehicular vibration). A current project is for a medical facility that is expanding, including operating rooms that have surgical microscopes. The facility is very close to a freight line, and we are working with them to provide mitigation measures before the facility is constructed.
My question to the list is, does anybody know of actual vibration criteria in terms of amplitude (peak or RMS) and frequency for ANY microscopes. The microscope manufacturers we have contacted indicate no research in the area, which is surprising to me.
Our previous work with both human perception and machine sensitivity has usually involved some specific vibration criterion, whether it be for damage, annoyance, or whatever purpose. The discussion of the various isolation tables on the list is interesting, including the innertubes and tennis balls. They appear to be a "One isolator fits all" approach, which, if it works, is also surprising to me. Any help or leads in this regard would be greatly appreciated. Thanks
Doug Anderson
Douglas A. Anderson, PhD Senior Consultant Schnabel Engineering Associates (http://www.schnabel-eng.com) 510 East Gay Street West Chester, PA 19380 Phone: 610 696-6066, Fax: 610 696-7771
Brian, that's a very intriguing problem. What is the metal or alloy of the coin? What is the particular date you are dealing with? Could a counterfeiter have done something so simple as cutting out segments of a pre-existing numeral to alter the date? If so, perhaps channelling would reveal the coining deformation pattern below the missing segments.
Also, is it clear that the WHOLE COIN is not a counterfeit? Ordinarily, the effort and cost to create a die set for counterfeiting a whole coin would deter anyone from the effort, unless the coin is spectacularly valuable. On the other hand, if someone had a genuine coin (or a carefully modified one) to use as a pattern, it would not be too dreadfully difficult (even a century ago) to make a ceramic mold directly, or a metal die indirectly, from the pattern. Then you could produce "knock-offs" from that die. And your "real McCoy" could be undamaged. This would take a clever, patient, and very meticulous craftsman to pull it off, however.
==================================== Roy Arrowood, Associate Professor Metallurgical and Materials Engineering UTEP, El Paso, TX 79968-0520 (915)747-6934 NEW E-MAIL ADDRESS: arrowood-at-miners.utep.edu
This worked for amphipod maxilla, which are minute or smaller and setose, so it should work for fly heads: 1) mount on double-sticky carbon-conductive tabs; cut these from a sheet, or buy as stub-sized tabs 2) sharpen toothpicks or other sticks, some to a point and others to a pen nib (with or without the slot found in a fountain pen nib) 3) place a small blob of Ag paint at the edge of the stub connecting the surface of the sticky tab to the metal of the stub; I prefer Ag paint dissoved in methylethylketone 4) use the stick to draw the paint from the blob to the fly head (etc.); connect this to a part of the head that you don't care about; draw a circle around the head just far enough not to touch the head -- this creates a shorter conductive path from the specimen to the Ag paint
Careful use of the sticks and paint will allow you to use Ag paint on very small, hairy specimens. The paint can be allowed to partially dry (solvent to evaporate) to thicken it, but this may just form a skin on the surface, and does lead to stringing of the partly dry paint which can be very annoying.
Phil
} An investigator just brought me 6 samples - heads of flies. He was } instructed to bring the entire body but he cuts the heads off when } he does TEM. The investigator knows nothing about SEM. I have done } alot of scanning of fly eyes but the body is attached. } My question is: how do I mount the heads on the stubs without the } graphite or silver paint causing capillary action over the entire } head? We will scan on a JEOL 840A after Au/Pd coating. } } 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 } Fax 214-648-6408 } eMail: George.Lawton-at-email.swmed.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)
{bold} {bigger} {bigger} Microscopy and Molecular Biology Postdoctoral Position
Department of Zoology
Miami University
Oxford, OH {/bold} {FontFamily} {param} TIMES {/param}
{FontFamily} {param} Arial {/param} A two-year postdoctoral position is available for investigating the role of inner arm dyneins in ciliary motility. We have seven different inner arm dynein heavy chain genes cloned from {underline} Tetrahymena {/underline} {underline} thermophila {/underline} and are in the process of creating knockout mutations in those genes. The successful applicant will be involved in all aspects of the work, including creation of the knockout mutants and analysis of the phenotypes. The successful candidate should be experienced in light and electron microscopy and willing to learn molecular biology and some protein biochemistry (preferred) or experienced in molecular biology and protein biochemistry and willing to learn light and electron microscopy. Send letter, curriculum vitae, and reference letters to:
David Pennock
Department of Zoology
Miami University, Oxford, OH 45056
Email: pennocdg-at-muohio.edu {FontFamily} {param} TIMES {/param} {smaller} {smaller}
Drill small holes in stubs and glue a dress making pin in each hole spike upwards. Coat this in a sputter coater.
Push each insect head onto a pin and sputter coat at a specimen-target distance of 5cms.
This technique is used by a number of our clients.
Steve Chapman Senior Consultant Protrain For consultancy and professional training in EM world wide. www.emcourses.com Tel 44+ 1280 814774 Fax 44+ 1280 814007
I've done a little work with varves...let me start by saying that typically this imaging is done using X-rays or by making conventional thin sections for petrographic microscopy. That said, I vaguely recall staining carbonate thin sections with Alizarin Red S. There is a different stain for dolomite. I'm not sure how well the technique can be applied to large sections....are they still wet? Dehydrated? Vacuum impregnated? Let me know if this is on the right track and I will hunt for some references.
Matt
Matthew J. Lynn, Ph.D. Center for Advanced Microscopy University of Miami (305)284-4736 mlynn-at-miami.edu
On Tuesday, June 27, 2000 10:21 AM, Dee Breger [SMTP:micro-at-ldeo.columbia.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, } } I've received a query from a climatologist who wants to photograph fine } varves and/or laminations (~0.5 mm- 1 mm each layer) in carbonate (calcite) } rich silt (or marl) through a binocular light microscope. The samples are } flat, finely cut or polished sections that can be etched if necessary, but } he's really looking for a staining method that would enhance the difference } between the carbonate-rich layers and the clay-rich layers to make them } more usefully photogenic. Can anyone advise us on how to proceed to best } advantage? } } Many thanks, } Dee } } } } } *************************************************************** } Dee Breger } Mgr. SEM/EDX Facility } Lamont-Doherty Earth Observatory } 61 Route 9W } Palisades, NY 10964 USA } T: 914/365-8640 } F: 914/365-8155 } } http://www.ldeo.columbia.edu/micro } http://www.discovery.com/area/science/micro/micro1.html } http://www.lsc.org/antarctica/front.html } Journeys in Microspace (Columbia University Press, 1995) }
Dear Jim, We have a Kevex Quantum light element EDX on a Hitachi that vents the whole chamber and it has been no problem, but I am told that the rate of venting can make quite a difference. I bought our EDX detector used and the previous owner claimed to have replaced the window every year for the five years that he had it. I have never had to replace the window and that is in a student lab. This particulars SEM takes over a minute to vent and makes no discernable sound, whereas my other SEM vents in ten seconds with an audible hiss. It, fortunately, has a Be-window detector, which is definitely tougher. If your SEM vents quickly, you might try to put some sort of restricter or filter on the vent to clean and slow it down. At 10:59 AM 6/27/00 -0300, you wrote: } } Hi all, } } We're in the process of acquiring an EDS system for our JEOL 5600 and } I'm curious about the experience of others with EDS systems on scopes } without specimen airlocks (column vents to exchange specimens). Any } thoughts/recommendations? I'm particularly concerned with contamination, } } window integrity and the like. Are there any advantages/disadvantages of } } Be vs. thin window detectors here? } } Thanks, } } Jim Ehrman 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
A colleague of mine has asked me to post the follwoing job announcement for an opening in our Cancer Research area. Please send resumes to the address at the bottom of the ad, or visit the website, which is also at the bottom of the ad. Please do not contact me directly concerning this position. Thanks!
Jane A. Fagerland, Ph.D. Dept. Microscopy and Microanalysis Abbott Laboratories
ABBOTT LABORATORIES PHARMACEUTICAL PRODUCTS DIVISION Abbott Park, Illinois
Abbott Laboratories is a global diversified company dedicated to the discovery, development, manufacture and marketing of health care products and services. Around the world, the company,s 54,000+ employees are committed to improving people,s lives by providing cost-effective health care technologies. We are interested in interviewing candidates for the following opportunity:
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Please send resumes to: 100 Abbott Park Road, D-583, AP9A, Abbott Park, Illinois 60064 Attn. Job Code 2K-KDA3422. Or visit our web site at www.abbott.com
Before staining, try using a UV light source to see if the calcite fluoresces - can fluoresce red to pink, orange, white, yellow or blue.
If you need to stain, use Alizarin Red S for staining. If it turns a deep red color, it's most likely calcite. Procedures for this can be found in "Laboratory Handbook of Petrographic Techniques" by Charles S. Hutchinson, 1974, John Wiley and Sons. Or you can have a thin section preparation lab do the staining for you - this will be much easier since they are set up to do this routinely.
Cheers, James Talbot
K/T GeoServices, Inc. X-ray diffraction petrologic studies visit my web site at http://www.ktgeo.com Argyle, TX, USA, (214) 403-6342
} ----- Original Message ----- } From: Dee Breger {micro-at-ldeo.columbia.edu} } To: {microscopy-at-sparc5.microscopy.com} } Sent: Tuesday, June 27, 2000 9:20 AM } Subject: LM geological stain question } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society 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've received a query from a climatologist who wants to photograph fine } } varves and/or laminations (~0.5 mm- 1 mm each layer) in carbonate } (calcite) } } rich silt (or marl) through a binocular light microscope. The samples are } } flat, finely cut or polished sections that can be etched if necessary, but } } he's really looking for a staining method that would enhance the } difference } } between the carbonate-rich layers and the clay-rich layers to make them } } more usefully photogenic. Can anyone advise us on how to proceed to best } } advantage? } } } } Many thanks, } } Dee } } } } } } } } } } *************************************************************** } } Dee Breger } } Mgr. SEM/EDX Facility } } Lamont-Doherty Earth Observatory } } 61 Route 9W } } Palisades, NY 10964 USA } } T: 914/365-8640 } } F: 914/365-8155 } } } } http://www.ldeo.columbia.edu/micro } } http://www.discovery.com/area/science/micro/micro1.html } } http://www.lsc.org/antarctica/front.html } } Journeys in Microspace (Columbia University Press, 1995) } } } } } } } } }
I've mounted small insects similar to what you now have using the following method:
1. Get some dissecting pins, preferably, or any very thin, rigid pin or wire and cut about 1/4 inch long.
2. Mount it vertically on an SEM stub using carbon paint, or any quick drying glue, and let it dry completely, put in warming oven to speed it up, or use hair dryer.
3. Position the fly heads upsidedown on clean surface, like lens tissue or glass petri dish.
4. Put a fresh drop of carbon paint, or glue, on a nearby surface, pick up the SEM stub with stub handling forceps, dip the tip of the pin into the fresh paint or glue just enough to get a tiny little blob on the pin head, then under a dissecting or low power scope touch the tip of the pin to the back surface of the fly head. It should stick and you can pick it up at that point, place right side up and let dry completely.
5. Coat in vacuum evaporator or sputter coater as usual. After coating run, vent gas into chamber slowly to not disturb delicate sample.
Its a bit of a prep, but you also get the fly head off the stub surface, little or no background junk in the view. Good luck!
Gib
Responding to the message of {s9587bef.020-at-mednet.swmed.edu} from "George Lawton" {George.Lawton-at-email.swmed.edu} : } } An investigator just brought me 6 samples - heads of flies. He was } instructed to bring the entire body but he cuts the heads off when he does } TEM. The investigator knows nothing about SEM. I have done alot of scanning } of fly eyes but the body is attached. } My question is: how do I mount the heads on the stubs without the graphite } or silver paint causing capillary action over the entire head? We will scan } on a JEOL 840A after Au/Pd coating. } } 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 } Fax 214-648-6408 } eMail: George.Lawton-at-email.swmed.edu } } } .
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
Just a reminder while we are on the subject of thin window detectors - as well as controlling the gas flow on venting also make sure that the chamber can not overpressure. On SEMs the chamber door should be able to open freely as soon as it reaches atmospheric pressure. On TEMs an aperture mechanism or similar should be freed so that it can release at atmospheric pressure.
Ron
On Tue, 27 Jun 2000, Mary Mager 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 Jim, } We have a Kevex Quantum light element EDX on a Hitachi that vents the whole } chamber and it has been no problem, but I am told that the rate of venting } can make quite a difference. I bought our EDX detector used and the previous } owner claimed to have replaced the window every year for the five years that } he had it. I have never had to replace the window and that is in a student } lab. This particulars SEM takes over a minute to vent and makes no } discernable sound, whereas my other SEM vents in ten seconds with an audible } hiss. It, fortunately, has a Be-window detector, which is definitely } tougher. If your SEM vents quickly, you might try to put some sort of } restricter or filter on the vent to clean and slow it down. } At 10:59 AM 6/27/00 -0300, you wrote: } } } } Hi all, } } } } We're in the process of acquiring an EDS system for our JEOL 5600 and } } I'm curious about the experience of others with EDS systems on scopes } } without specimen airlocks (column vents to exchange specimens). Any } } thoughts/recommendations? I'm particularly concerned with contamination, } } } } window integrity and the like. Are there any advantages/disadvantages of } } } } Be vs. thin window detectors here? } } } } Thanks, } } } } Jim Ehrman } 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 } } }
=========================================================================== 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. ============================================================================
OK thanks for the change of address, as usual we are looking for: books on microscopes catalogs of microscopes microscope instruction manuals books on application of microscopes either in biological or material sciences .. anything! Optical, em sem, afm, etc etc etc.
----- items and literature relating to any form of electrical communication and engineering of thereof (yes telephone, telegraph, early wireless telegraphy ---- artifacts and literature on radio and TV broadcasting --- Radar and radar countermeasures ----- cryptanalysis --------
thanks in advance as always Ed Sharpe archivist for SMECC
I have used my Umax Powerlook III flatbed scanner with transparency adapter many times to get high quality scans of my TEM negatives but today it is driving me crazy. I have some great gold labeling of pretty electron dense granules. On the negative I can clearly see the gold against the granule matrix but when I scan, the 256 grey levels compresses all that info into one level of black and therefore I can't discern the gold. any scan gurus out there who can help? I have tried playing with the gamma without much luck. thanks in advance, 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)
Tom, Have you tried telling the scanner that the image is a positive transparency rather than a negative? And then reverse the contrast in photoshop rather than with the scanner software. Greg
At 12:20 PM 06/28/2000 -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
we have a Epson 1600, sometimes this problem happens too. my solution is to scan the negative in positive mode first then do a inversion after that. Hope this helps.
Hao Li
----- Original Message ----- } From: "Tom Phillips" {PhillipsT-at-missouri.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, June 28, 2000 1:20 PM
Hi, Thomas
I find it best to scan a negative in as a positive transparency at 6 million colors (rather than the 256 greys you are using), and then reverse it. This gives me the best tonal range.
Aloha, Tina
} } I have used my Umax Powerlook III flatbed scanner with transparency } } adapter many times to get high quality scans of my TEM negatives but } } today it is driving me crazy. I have some great gold labeling of } } pretty electron dense granules. On the negative I can clearly see } } the gold against the granule matrix but when I scan, the 256 grey } } levels compresses all that info into one level of black and therefore } } I can't discern the gold. any scan gurus out there who can help? I } } have tried playing with the gamma without much luck. thanks in } } advance, tom } }
**************************************************************************** * 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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Oxford Instruments supply a wide range of CL detection systems, from low cost broad-band detectors to multiple-band, UV, IR and spectroscopic systems. As far as I am aware, they are the only company supplying such a wide range of CL systems - if there are others, sorry, and I would also like to know!
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
Thanks for all the suggestions. We did indeed find that scanning in the color mode (which is 14 bit) solved the problem. I don't know why UMax has their greyscale image only go to 256 colors. I will ultimately incorporate this in a lecture when i am teaching image analysis since it clearly shows the limitations of 8 bit images. once again, 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)
Stephan, I know that Oxford instruments can supply you with the necessary equipment to do CL in the SEM. I am not sure whether this is standard equipment or whether it is special order, but I have seen it done. The equipment usually involves a parabolic mirror and a light-pipe with the relevant detector on the end of this. Then there is the software .etc. There is a contact email address on their web page:
http://www.oxford-instruments.com/
give them a try and see what they say.
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 ********************************************************
Has anyone tried Kodak's MDS 100 imaging system? I have a user who needs a quick documentation system and this might work for her. She is using NIH Image on a compound scope in our lab and she wants to become independent and do the work in her own lab, and maybe at other locations.
As an alternative, anyone with a Nubus frame grabber, ie Scion LG3 or equiv., that would sell it to her to use with NIH Image?
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
} I have used my Umax Powerlook III flatbed scanner with transparency } adapter many times to get high quality scans of my TEM negatives but } today it is driving me crazy. I have some great gold labeling of } pretty electron dense granules. On the negative I can clearly see } the gold against the granule matrix but when I scan, the 256 grey } levels compresses all that info into one level of black and therefore } I can't discern the gold. any scan gurus out there who can help? I } have tried playing with the gamma without much luck. thanks in } advance, tom } } Thomas E. Phillips, Ph.D. } Associate Professor of Biological Sciences } Director, Molecular Cytology Core Facility } Tom -
You can get a really good book on scanner use at www.scantips.com or download it as a pdf,if you have the patience for 218 pp.
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
Can anyone advise what the water flows should be in the three parallel branches (OL; DPs; and power boards) of the 840A) The manual spec is } 5 l/min total, but I need to know particularly what the flow should be thru the OL.
TIA
rtch
ps replies welcome from JEOL
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 an Oxford MiniCL detector attached to a JEOL 6300 - same column as the 6400. This is an inexpensive detector which suits our needs but there are other suppliers. No matter who you obtain one from, give some thought to what ports are free and to your analytical working distance (presume you have EDS or WDS). On the 6400 this is most likely 15mm - some ingenuity from Oxford or other supplier will be needed to clear the pole-piece eg an inclined configuration. It would be worth it - rather than have to alter the sample height for quant analysis.
Regards
Stafford Stafford McKnight Geology & Metallurgy University of Ballarat ph 03 53279262 fax 03 53279144
Accurel Systems International is looking for SEM Analysts: junior to experienced technicians and engineers
Responsibilities include all aspects of commercial SEM and EDX analysis (instrument operation, sample preparation, customer contact and follow-up, data interpretation etc.).
Knowledge of IC device structure, IC fabrication and packaging a plus.
2+ years hands on experience in the semiconductor industry preferred.
To find out more please check our web site: www.accurel.com
SEM candidates should send resume with references to Regina Campbell: reginac-at-accurel.com or FAX 408-737-3916.
Job description: You will form part of a team of specialists in analytical techniques, providing support to the rapidly growing III-V optoelectronic and microwave device fabrication plant at Caswell Technology. We perform a wide range of activities, including optical microscopy, SEM, TEM, EDX, X-ray techniques and scanning probe microscopies. You will be expected to be able to be competent in several of these techniques and have/develop a degree of expertise in one or more of them. Experience of semiconductor device fabrication techniques would be an advantage.
The post is available immediately.
More details of our company are available at http://www.caswelltechnology.com/
Please contact me if you are interested or would like any further information.
============================================================== Richard Beanland Caswell Technology, Caswell, Towcester, Northants NN12 8EQ
Don't broadcast change of address notices to the Listserver. If you want to change your subscription address, just send a unsubscribe message, followed by a subscribe message...
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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
I run a microscopy unit which is completely digital. I have a problem with printing out images. I am planning to upgrade cameras to 10 or 12 bit megapixel units, but am already restricted by printers. Laser printers seem to deal with greyscales by halftoning at 100-150dpi which is unsatisfactory for small prints. Are there any true greyscale printers available, that manage better than 300dpi and better than 256 grey levels? Is there any advantage in having more than 256 grey levels? Sorry if this is a bit dim, but there are so many self styled experts who give conflicting opinions I end up totally confused. Thank you. Richard Black Nottingham, England richardblack-at-cwcom.net
This is my first message to the list... Caroline, I looked at Scan site and would like to try PDF format... but I can seem to locate the pathway to download? Any thoughts or pointers?
Thanks for the suggestion it looks like a great site. Tim Lyden, Ph.D. Research Scientist
Departments of Internal Medicine/Immunology and Physiology/Cell Biology Ohio State University Davis Medical Research Center 480 W 9th Ave. Columbus, Ohio 43210
I would like to purchase a "Tunneling Electron Microscope".
I have been unable to locate a supplier. If you know of one please let me know. Thanks! Alesia White Darling Secretary, Dr. Amnon Katz University of Alabama Aerospace Engineering and Mechanics Box 870280/205 Hardaway Hall Tuscaloosa, Alabama 35487-0280 ph: 205-348-8525 fax: 205-348-7240 or 205-348-2094
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I have found that the gold does not show up well when you scan the negative itself. So for gold to show up in the scanned image, I usually make one print and scan the print. This works quite well. I guess we will still have to work in darkrooms for quite sometime even though the digital imaging is becoming popular.
Good luck,
Soumitra
} } } I have used my Umax Powerlook III flatbed scanner with transparency } adapter many times to get high quality scans of my TEM negatives but } today it is driving me crazy. I have some great gold labeling of } pretty electron dense granules. On the negative I can clearly see } the gold against the granule matrix but when I scan, the 256 grey } levels compresses all that info into one level of black and therefore } I can't discern the gold. any scan gurus out there who can help? I } have tried playing with the gamma without much luck. thanks in } advance, 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)
***************************************************************** 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
As I recall, the human eye is hard pressed to distinguish more than 6-bit grayscale (64 levels) unless the shades are right next to each other, which they might be in an image. Therefore, printing at 256 gray levels should be more than adequate.
The 10 or 12-bit cameras will still be handy in storing the extra information in your images. You will be able to do some gray scale manipulation to highlight the darker or lighter regions of your image. But when it comes to print time, you will be throwing out all but about 8 bits.
You rightly say that halftoning is a problem on small prints. You need a 16x16 dot cell to render 256 gray levels in halftone. For 1200 dpi printing, that means that you can achieve 75 pixels per inch (not very high). That means a 1024 pixel image would require 13.65 inches to show all that resolution. If the image is printed out smaller, then either grayscale or pixel resolution would be sacrificed. (Printing at 64 gray levels would require a 8x8 cell and could be done at 150 pixels per inch so that the image could fit into 6.8 inches.)
I don't have a dye sub printer here, but their specs would probably meet your needs. You would want to see how well they can render the grayscale. Since they do not halftone, the dpi spec would be the same as the pixel per inch spec. A 300 dpi printer should be able to render full resolution on a 1024 pixel image in 3.4 inches. But check out the grayscale performance to see if it is adequate.
Warren S.
At 07:58 AM 6/29/2000 -0500, you wrote:
} I run a microscopy unit which is completely digital. I have a problem } with printing out images. I am planning to upgrade cameras to 10 or 12 } bit megapixel units, but am already restricted by printers. Laser } printers seem to deal with greyscales by halftoning at 100-150dpi which } is unsatisfactory for small prints. Are there any true greyscale } printers available, that manage better than 300dpi and better than 256 } grey levels? Is there any advantage in having more than 256 grey } levels? Sorry if this is a bit dim, but there are so many self styled } experts who give conflicting opinions I end up totally confused. } Thank you. } Richard Black } Nottingham, England } richardblack-at-cwcom.net
I think this query was posted recently but I haven't been able to dredge up the replies in the list archives - too recent, maybe.
I'm looking for contact info for a colleague, for Balzers or Bal-tec. He inherited a B. carbon evaporator model CED 030 (also: 3U-G03-751/132). Can anyone help out with either contact info for the company, or a copy of the manual?? We will pay copy/ship charges.
Thanks, much.
Ann Hein Lehman Manager, EM Facility Trinity College Hartford CT 06106 v. 860-297-4289 f. 860-297-2538 e. ann.lehman-at-trincoll.edu
Hi Hai Li: Here is some references which claim MgAl2O4 is a good substrate for growing perovskites:
Miura, S., Yoshitake, T., Matsubara, S., Miyasaka, Y., Shohata, N. and Satoh, T., Epitaxial Y-Ba-Cu-O Films on Si with Intermediate Layer by RF Magnetron Sputtering, Appl. Phys. Lett. 53:1967-1969 (1988).
Wu, X.D., Inam, A., Hegde, M.S., Wilkens, B., Chang, C.C., Hwang, D.M., Nazar, L., Venkatesan, T., Miura, S., Matsubara, S., Miyasaka, Y. and Shohata, N., High Critical Currents in Epitaxial YBa2Cu3O7-x Thin Films on Silicon with Buffer Layers, Appl. Phys. Lett. 54:754-756 (1989).
My buddy, Darrell Schlom at Penn State tells me that using perovskite substrates is much better:
Y. Jia, M.A. Zurbuchen, S. Wozniak, A.H. Carim, D.G. Schlom, L-N. Zou, S. Briczinski, and Y. Liu, "Epitaxial Growth of Metastable Ba2RuO4 Films with the K2NiF4 Structure," Applied Physics Letters 74 (1999) 3830-3832.
Best regards, Mike Urbanik www.crystalguru.com
} } Subj: information about MgAl2O4
For a tungsten emitter, my JEOL operators' manual implies to raise the anode closer to the Wehnelt for low keV applications (less than 15keV). The manual does not mention this distance for the LaB6 emitter option. Any suggestions?
TIA and 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/
We also have an Oxford MiniCL detector (attached to our JEOL-8900), however the Research Instruments group of Oxford (CL and Cryo EM stuff) was just sold to Gatan, so you may need to contact Gatan to find out about the product. We are in the middle of some modifications to our unit (extension of the light pipe), and JEOL informed me of the transaction. It doesn't appear on the websites, however it was official as of 6-19-00. Hope the info helps. Sarah
Stafford McKnight 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 } -----------------------------------------------------------------------. } } Stephan, } } We have an Oxford MiniCL detector attached to a JEOL 6300 - } same column as the 6400. This is an inexpensive detector which } suits our needs but there are other suppliers. } No matter who you obtain one from, give some thought to what } ports are free and to your analytical working distance (presume you } have EDS or WDS). On the 6400 this is most likely 15mm - some } ingenuity from Oxford or other supplier will be needed to clear the } pole-piece eg an inclined configuration. It would be worth it - rather } than have to alter the sample height for quant analysis. } } Regards } } Stafford } Stafford McKnight } Geology & Metallurgy } University of Ballarat } ph 03 53279262 } fax 03 53279144
-- Sarah A.W. Lundberg Electron Microanalysis and Imaging Laboratory Department of Geoscience, UNLV 4505 S. Maryland Parkway Box 454010 Las Vegas, NV 89154-4010
Hellow Richard, HP Laserjet 2100TN is used in our laboratory and it has a very high stated resolution of 1200dpi. The printer gives excellent output at its highest setting, and lower. The level of detail I obtain from the printer is sufficient to show all the information from TEM images. However, I believe the final output from a photograph is still slightly better. Also, photographs have very nice glossy or matt finishes which I haven't seen on laser printouts.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ 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 Thu, 29 Jun 2000, richard black 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 run a microscopy unit which is completely digital. I have a problem } with printing out images. I am planning to upgrade cameras to 10 or 12 } bit megapixel units, but am already restricted by printers. Laser } printers seem to deal with greyscales by halftoning at 100-150dpi which } is unsatisfactory for small prints. Are there any true greyscale } printers available, that manage better than 300dpi and better than 256 } grey levels? Is there any advantage in having more than 256 grey } levels? Sorry if this is a bit dim, but there are so many self styled } experts who give conflicting opinions I end up totally confused. } Thank you. } Richard Black } Nottingham, England } richardblack-at-cwcom.net } } } }
Balzers stuff is now handled by a company called Techno Trade. If you can't find them on the 'net or find their address, email me and I'll dig through my files and find their address for you.
Cheers :o)
Paul Grover Chief Microscopist and Bottle Washer Microvista Laboratory Lafayette, IN
My Amray says the same thing. As long as the gun's Whenelt end is the same distance for each emitter type, the spacing should be the same. I used 4mm {=5KV, 6mm {=12KV and 8mm for } 12KV.
gary g.
At 02:47 PM 6/29/00, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I'm selling a perhaps incomplete set of parts for a phase contrast setup for a B&L microscope. I don't have any B&L equipment, and it was part of a lot I got a sale....
ADI - FOUNDRYS OFFER FOR DESIGNERS AND USERS OF CASTINGS
23-24 NOVEMBER 2000 CRACOW – FOUNDRY RESEARCH INSTITUTE
CIRCULAR NO. 1
HOSTED BY: Instytut Odlewnictwa 30-418 Kraków, Zakopiañska 73
CO-ORGANISER: Ministry of Economy Polish Foundrymen’s Technical Association Foundry Economic Chamber SCIENTIFIC COMMITTEE
List of persons invited to participation in the Scientific Committee of the Conference: Prof. Jerzy PIASKOWSKI – Poland Prof. Czes³aw PODRZUCKI – Poland Prof. Edward GUZIK – Poland Prof. Jan R¥CZKA – Poland Prof. Ryszard KOZ£OWSKI - Poland Prof. Mieczys³aw KACZOROWSKI - Poland Prof. Jouko.J. VUORINEN – Finland Prof. Eduard DORAZIL – Czech Republic Prof. Lubomir BECHNY – Slovakia Prof. Jorge A. SIKORA - Argentina dr Franco ZANARDI - Italy dr Matti JOHANSON – Finland dr Jose R. GUIRIDI - Spain dr Konstantin UZ£OW - Ukraine
ORGANISING COMMITTEE Dr Eng. Jerzy TYBULCZUK - Chairman Dr Eng. Adam KOWALSKI – Secretary Dr Eng. Józef TURZYÑSKI M.Sc.Eng. Krystyna £USZCZKIEWICZ Eng. Marta KONIECZNA M.A. Krystyna BANY-KOWALSKA M.Sc.Eng. Andrzej PYTEL Eng. Janusz CUPIA£
PURPOSE AND SUBJECT OF CONFERENCE The purpose of the Conference is to enable the national and foreign research & development centres as well as industrial units to present their latest achievements in developing the heat treatment technology, metallographic examinations, and quality control systems for high-grade ductile iron - ADI in particular.
Therefore you are encouraged to submit your proposals of papers dealing with the following subjects : Foundry New grades of ductile iron, their properties and technologies of production. Methods of spheroidising and inoculation treatments Metals science Electron microscopy. X-ray phase analysis. Quantitative metallography. The techniques of colour etching and their application in metallography. New methods and tools to examine the structure of ductile iron and ADI. Heat treatment Methods of ductile iron heat treatment. Installations. Methods of heat treatment control. Heat treatment parameters. Technologies of ADI fabrication. Quality Quality systems according to ISO and EN Standards used in production of ductile iron and ADI. Control of production process. Statistical quality control.
PRESENTATION OF PAPERS Papers will be presented during the plenary meeting (time for presentation 15-30 minutes). Language of the Conference : English and Polish
INSTRUCTIONS FOR AUTHORS OF THE PAPERS Paper volume : 6-8 pages including tables and figures on A4 paper. Even number of pages. Margins : left - 2.5 cm, right - 2.5 cm, upper - 2.5 cm, lower - 2.5 cm. Write your text in editor Word 6 or Word 97, single line spacing, font Times New Roman 12 pt. The starting line of a new paragraph should be indented by 1 cm.. Figures, photographs and tables should run in the text of the paper. Captions should be typed in bold and centred. Equations should be centred leaving one free spacing above and below the equation. The pages should be numbered consecutively using soft pencil. References : published literature cited in the text should be quoted using a number in square brackets. Abstract : Please start with an abstract of up to 50 words.
SAMPLE OF PAPER FORMAT
On first page (only !) leave free space from the top of 10 single line spacings
TITLE OF PAPER IN BOLD CAPITALS (centred, 14 pt)
space 2 x 1
Name & Surname (bold, centred, 12 pt) Affiliation, e.g. Foundry Research Institute (italics, centred, 12 pt) Place, e.g. Kraków
space 2 x 1 ABSTRACT (bold capitals, centred, 12 pt) space 1 x 1 Text follows - approximately 50 words, indentation of 2.5 cm on the right and left, 12 pt.
space 2 x 1 1. INTRODUCTION (bold capitals, 12 pt) Text justified written in single line spacing. 2. FIRST SUBTITLE (bold capitals, 12 pt) Text justified written in single line spacing. 2.1. Second subtitle (bold, 12 pt) Text justified written in single line spacing. space 3 x 1 REFERENCES (bold capitals, 12 pt) OTHER EVENTS
during the Conference, the universities, industrial plants and companies will have an opportunity to display their products, research methods, manufacturing techniques and computer programmes in the form of short (up to 15 minutes) presentations and/or exhibitions - all those who are willing to take part in the display are kindly requested to agree in advance with the Organising Committee the subject and form of display, the organisers of the Conference also offer the possibility of publishing the ready advertising and information materials in the Conference Proceeding upon previous agreement with the Organising Committee.
Details along with the Conference programme will be circulated early in October 2000 in CIRCULAR NO. 2.
For more information please contact the following persons:
SCHEDULE completed participation forms should be sent by 31 October 2000 technical papers for presentation during the Conference should be sent by 30 September 2000 proposals of promotion during the Conference should be sent by 15 October 2000
REGISTRATION FEE (Note : accomodation is NOT included) full rate: 200 $ reduced rate (for participants presenting papers and posters) 150 $ The cost of promotion is to be agreed.
Please make you cheque payable to : BPH I Oddzia³ w Krakowie Account No. : 10601376-320000033388
Here in Philadelphia we are all looking forward to the upcoming M&M meeting in August. The meeting promises to be the best yet. To make everyone's arrival and traveling as pleasant as possible we recommend you visit the Philadelphia Airport Web site at http://www.phl.org/. Here you will be able to go directly to ground transportation and find the easiest and least expensive ways to get from the airports to the hotels in the city. The ride from the airport to the hotels is approximately 15-20 minutes. Depending on the transportation you choose the prices will range from $8.00 on up. Please feel free to E-mail me with any specific questions you may have and I shall try to assist. We look forward to seeing everyone in August and have a great trip.
I will getting my first SEM/EDS shortly and to obtain the required operating permit from my safety group I need a written procedure for handling LN2. Specifically I need a written procedure for filling the 3 L dewar on the EDS detector from a 50 L dewar mounted on a cart. I plan to make the transfer by using a lab source of N2 to pressurize the 50 L dewar and have obtained all the valves and fittings required to do this from another SEM lab. I would appreciate copies of the procedure. Thanks,
Everett Ramer National Energy Technology Laboratory P.O. Box 10940, Cochrans Mill Road Pittsburgh, PA, USA 15236-0940 Voice: 412-386-4920 FAX: 412-386-4806 ramer-at-netl.doe.gov
The JEOL 840 that I am familiar with (this most likely applies to most JEOL SEM's) sets the tip whenelt distance within a narrow range and as I recall this distance is correct when the auto bias emission current is about 100ua. This makes it possible to fine tune the first cross over with the manual bias control so that at 20kV the emission current would be adjusted with the manual bias control to be about 8-9 and the emission current would be about 10=20uA. The effects of this can be seen viewing the emission pattern. The spot size will be smaller using a high number manual bias number. If the auto bias emission current was say only 50-60uA when the SEM was operated at 1kV accelerating voltage it would be more difficult to fine tune the first cross over spot size because the bias number would be about 0-1 and the emission current would be to low. Increasing the bias number decreases the emission current. If the emission current in auto bias is less than 100uA the whenelt is rotated slightly CW to shorten the distance. Manual bias is the usual way to operate the LaB6 filament while auto bias is usually used with a W filament.
Because the LaB6 source is much brighter than W filament, the first cross over at the source can be fine tuned with the LaB6 cathode which results in smaller spot size on the sample ie. better resolution with sufficient current to have good signal to noise ratio.
If you use a W filament and try to set up manual bias the same way as you would for LaB6 you will see that the signal to noise is worse using high manual bias number (less emission current). The LaB6 filaments are usually operated in manual bias while the W filaments are normally used with auto bias.
On the 840 there are either two accelerating anodes, a high anode for low kV and a low anode for high kV which is the case if you have an elctro static beam blanker installed or a single anode that can be raised for low kV or lowered for high kV operation. The anode in this case snaps into the correct position. Hope this helps.
John Humenansky/Staff Scientist Physical Electronics, Inc. (PHI) 6509 Flying Cloud Drive Eden Prairie, MN 55344 952-828-6387
Hi All, I helped empty on old lab and came across a dozen never-opened bottles of Epon-812 (yes, the real thing). You can well imagine how long they had sat on the shelf. My question to you is....what do I do with it? My gut reaction is to give it to our Life Safety guys to haul away, but if its still good that would be a waste. Short of opening one of the bottles and making a test batch, is there any rule of thumb out there? The bottles have a WPE of 160. They are still in the original plastic wrap and have an EMS label. Maybe Stacie has a thouhgt on this (if she's not overwhelmed with planning for M&M).
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
The Bal-Tec representative is TechnoTrade International. 603-622-5011.
Best,
Angela
At 02:02 PM 06/29/2000 -0400, you wrote:
} I think this query was posted recently but I haven't been able to dredge up } the replies in the list archives - too recent, maybe. } } I'm looking for contact info for a colleague, for Balzers or Bal-tec. He } inherited a B. carbon evaporator model CED 030 (also: 3U-G03-751/132). Can } anyone help out with either contact info for the company, or a copy of the } manual?? We will pay copy/ship charges. } } Thanks, much. } } Ann Hein Lehman } Manager, EM Facility } Trinity College } Hartford CT 06106 } v. 860-297-4289 } f. 860-297-2538 } e. ann.lehman-at-trincoll.edu
--------------------------------------------- Angela V. Klaus
Director, Core Imaging Facility American Museum of Natural History Central Park West at 79th Street New York, NY 10024-5192 USA
We have used Epon-812 that is over 25 years old with no problem. This is valuable stuff so I hope you do not through it out. Somebody out there will surely take it off your hands. I have enough to last me the rest of my career
At 09:04 AM 06/30/2000 -0400, 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
Hi, Everett. I guess I am glad I am not working for a government anymore. Otherwise I would probably be having to fill out similar forms. It would be nice if the folks that review those forms knew what the issues were. I am always afraid that they will know just enough to be dangerous and may actually require some unhealthy procedures. But perhaps they are sensible and only require that you have something officially in writing to document your best practice.
I found the following post in my files from 4 years ago. I hope Larry does not mind me reposting it. His conclusions are quite interesting.
Warren S.
} Wil's recent comment on the safety hazards of distilled water brought to } mind some peculiar safety regulations here in MD. In reference to liquid N ... } officer listening in will recommend new safety procedures requiring } protective booties! } In the end, we can't legislate common sense, nor can we abdicate } responsibility to those above.
Try getting your safety officer to conduct an experiment:
1. Hold out hand, 2. Pour a small volume of liquid N2 over hand 3. Now the interesting bit - put on a glove, and pour the same quantity of liquid N2 into glove. 4. Phone for ambulance.
The point is that a brief contact causes no problems, but if the contact is continued you get a nasty burn.
Gloves, goggles, masks (and shoes) are actually more dangerous when handling liquid N2 than sandals and no protection. And clothes are actually more dangerous than being naked. Get the safety officer to experiment. With a little persuasion you can probably convince the safety officer that when handling liquid N2, everybody should be naked.
More seriously, bureaucrats, administrators and the inexperienced should talk to somebody who has real knowledge.
-------------------------------------------------------------- Dr. Larry Stoter Technesis 17, Rocks Park Road, Uckfield, E. Sussex, TN22 2AT, United Kingdom Larry-at-teknesis.demon.co.uk --------------------------------------------------------------
by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id NAA12520 for dist-Microscopy; Fri, 30 Jun 2000 13:53:29 -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 NAA12512 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Fri, 30 Jun 2000 13:52:59 -0500 (CDT) Received: from navel.ucr.edu (navel.ucr.edu [138.23.225.12]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id NAA12505 for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 30 Jun 2000 13:52:47 -0500 (CDT) Message-Id: {200006301852.NAA12505-at-ultra5.microscopy.com} Received: from batch3122 ([138.23.190.212]) by navel.ucr.edu with SMTP for Microscopy-at-MSA.Microscopy.Com; Fri, 30 Jun 2000 11:48:07 -0700 X-Sender: kalen-at-pop.ucr.edu X-Mailer: QUALCOMM Windows Eudora Pro Version 4.0
Hi All, I'm asking this on behalf of a colleague, so please bear with me if I don't have all of the details. Does anyone out there have experience with Vectashield as an anti-fade agent for viewing immunofluorescence? Specifically, we are looking at plant tissue in which we are using a secondary antibody conjugated to FITC and have found that the controls (treated only with buffer) show "autofluorescence" after mounting with Vectashield, when they were totally non-fluorescent when mounted in buffer. Granted, the Vectashield did expire last month, but this is something that we've seen before with this product. Should I advise my colleague to just take pictures as quickly as humanly possible because we can't seem to rely on this anti-fade agent? I appreciate any input you may have. Have a great Independence Day weekend (for those of you in the US), 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
} Dear List, } Does any one have the dummies guide to electropolishing?? Some years ago } (3), I had a trouble shooting guide for electropolishing. This list had } about 9 or 10 items, problems /solutions on it and it was very } helpful...but it has found one of those very safe filing places (i.e. I } can't find it!) If any one has access to this list or something similar, } I would appreciate it if you could send me a copy. } } Many thanks in advance. } } Dorrance } PS I haven't been keeping up with the weather reports lately but it's 95 } and clear in beautiful Livermore, California. }
} Dear List, } Does any one have the dummies guide to electropolishing?? Some years ago } (3), I had a trouble shooting guide for electropolishing. This list had } about 9 or 10 items, problems /solutions on it and it was very } helpful...but it has found one of those very safe filing places (i.e. I } can't find it!) If any one has access to this list or something similar, } I would appreciate it if you could send me a copy. } } Many thanks in advance. } } Dorrance } PS I haven't been keeping up with the weather reports lately but it's 95 } and clear in beautiful Livermore, California. }
I do not have a "dummies" guide as such, but I do have copies of Bernie Kestel's 66 page report "Polishing Methods for Metallic and Ceramic TEM Specimens". The "polishing" refers to electropolishing. I would be happy to send this out to you if you do not have it already. I also have dozens of other papers dealing with electropolishing and hundreds of papers dealing with specimen preparation. I can email to you a list of the other papers if you'd like to breeze through that. Let me know.
DISCLAIMER: South Bay Technology produces equipment and supplies for specimen prepration 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 "McLean, Dorrance" } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} Dear List, } Does any one have the dummies guide to electropolishing?? Some years ago } (3), I had a trouble shooting guide for electropolishing. This list had } about 9 or 10 items, problems /solutions on it and it was very } helpful...but it has found one of those very safe filing places (i.e. I } can't find it!) If any one has access to this list or something similar, } I would appreciate it if you could send me a copy. } } Many thanks in advance. } } Dorrance } PS I haven't been keeping up with the weather reports lately but it's 95 } and clear in beautiful Livermore, California. }
You can find the information about LaB6 emitter setting and operation in Kimball Physics technical bulletins. I am aware of 7 bulletins being available. Contact Kimball Physics at info-at-kimphys.com . Bulletins are not posted, so you will have to request them. Ask for at least 2: bulletin # LaB6-01B (General guidelines for operating ES-423E LaB6 cathodes) and bulletin # LaB6-03B (Emission drift - LaB6 and gun stability). Kimball Physics guidelines for LaB6 emitter use are comprehensive yet well suited for practical applications.
You are welcome to contact me off line for the fax copy, though I do not have the complete set of bulletins.
Disclaimer: SIA does not have an affiliation with Kimball Physics other than being satisfied customer.
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax -----Original Message----- } From: =shAf= {mshaf-at-darkwing.uoregon.edu} To: Microscopy list {Microscopy-at-sparc5.microscopy.com}
Don't you guys ever check this flow? What if the needle valve or (on the 840A) the solenoid shutoff valve gets blocked? Come on, someone must have done it!
Incidentally, when I pulled apart the solenoid shutoff valve on my 840A, a small amount of corrosion had jammed it permanently "OPEN". This is potentially dangerous, as if the water continues to circulate thru the OL when the instrument is switched off, and if the cooling water is below the dewpoint, condensation on the outside of the OL can wreck it by corrosion.
So if you do decide to check the valve on yours, how about measuring the flow for me?
TIA
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 } -----------------------------------------------------------------------. } } } Hi } } Can anyone advise what the water flows should be in the three } parallel branches (OL; DPs; and power boards) of the 840A) } The manual spec is } 5 l/min total, but I need to know particularly } what the flow should be thru the OL. } } TIA } } rtch } } ps replies welcome from JEOL }
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
I completely agree with the comments by Warren Straszheim and Larry Stoter. A couple more points: 1 Gasoline is rather more dangerous then liquid nitrogen. If anybody would try to impose silly forms and rules and thus curtailing the publics access to gasoline, they be out on their ear fast. Access to gas is available to countless completely untrained people. 2 What evidence is there that serious accidents have occurred by small scale handling of liquid nitrogen. Golden rule of administration: Regulate only when there is a demonstrated need. 3 Its my observation that timid people (intimidated by overstated dangers) are more likely to cause accidents. I instructed first time users of liquid nitrogen that there is a danger when the substance is enclosed, that touching cold metal or pouring it into ones shoes causes burns and that cold tubing can fracture. As important, new users must learn that its quite harmless when a little is splashed and its worth demonstrating that a finger can be stuck into liquid nitrogen for a couple of seconds. This builds handling confidence.
The safety officer's paper wastes time and paper. Users need to learn how to handle the substance; its simple enough. Writing up a procedure for pumping creates a delusion, which has no practical benefits. 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 ABN: 99 724 136 560 www.proscitech.com
On Saturday, July 01, 2000 3:39 AM, Warren E Straszheim [SMTP:wesaia-at-iastate.edu] wrote: } } } Hi, Everett. I guess I am glad I am not working for a government anymore. } Otherwise I would probably be having to fill out similar forms. It would be } nice if the folks that review those forms knew what the issues were. I am } always afraid that they will know just enough to be dangerous and may } actually require some unhealthy procedures. But perhaps they are sensible } and only require that you have something officially in writing to document } your best practice. } } I found the following post in my files from 4 years ago. I hope Larry does } not mind me reposting it. His conclusions are quite interesting. } } Warren S. } } } Date: Sat, 30 Mar 1996 07:57:02 +0000 } } From: "Dr. L. P. Stoter" {LPS-at-teknesis.demon.co.uk} } } } Wil's recent comment on the safety hazards of distilled water brought to } } mind some peculiar safety regulations here in MD. In reference to liquid N } ... } } officer listening in will recommend new safety procedures requiring } } protective booties! } } In the end, we can't legislate common sense, nor can we abdicate } } responsibility to those above. } } Try getting your safety officer to conduct an experiment: } } 1. Hold out hand, } 2. Pour a small volume of liquid N2 over hand } 3. Now the interesting bit - put on a glove, and pour the same quantity of } liquid N2 into glove. } 4. Phone for ambulance. } } The point is that a brief contact causes no problems, but if the contact is } continued you get a nasty burn. } } Gloves, goggles, masks (and shoes) are actually more dangerous when handling } liquid N2 than sandals and no protection. And clothes are actually more } dangerous than being naked. Get the safety officer to experiment. With a } little persuasion you can probably convince the safety officer that when } handling liquid N2, everybody should be naked. } } More seriously, bureaucrats, administrators and the inexperienced should } talk to somebody who has real knowledge. } } -------------------------------------------------------------- } Dr. Larry Stoter } Technesis } 17, Rocks Park Road, Uckfield, E. Sussex, TN22 2AT, United Kingdom } Larry-at-teknesis.demon.co.uk } -------------------------------------------------------------- }
In SEMs the objective lens current required to "bend" the beam is much less than in TEM where the uncooled objective could fry an egg. Many SEMs use no water cooling at all. Cooling would help temperature stability and control drift. I expect that 50ml /minute would do, but such slow flow would soon stop altogether. I suggest that less than 2 liters/ minute indicate blocked tubing. Reverse flushing my help. Otherwise recirculate hot vinegar or 1N HCl for an hour. Cross threading: I never had a safety officer, get him to do it, otherwise you may require a ream of paper applications for pumping acid. Happy now Rich? 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 ABN: 99 724 136 560 www.proscitech.com
-----Original Message----- } From: Ritchie Sims [SMTP:r.sims-at-auckland.ac.nz] Sent: Monday, July 03, 2000 7:14 AM To: Ritchie Sims; Microscopy-at-sparc5.microscopy.com
Dr. Larry Stoter (4 years ago) wrote: (and thanks for sharing it, Jim)
Gloves, goggles, masks (and shoes) are actually more dangerous when handling } liquid N2 than sandals and no protection. And clothes are actually more } dangerous than being naked. Get the safety officer to experiment. With a } little persuasion you can probably convince the safety officer that when } handling liquid N2, everybody should be naked.
I'd like to share my experience with getting naked in a hurry. I was working around midnight in the biochem lab at Okla. State Univ. in '79 and was freezing samples to put in the lyophilizer. My arms were full, carrying numerous flasks plus a dewar of LN2, which I held against my shoulder. As I turned a corner, I slipped on some spilled water (I was wearing those flip flop sandals) and the entire contents of the dewar poured down my back. Fortunately there were no witnesses to my rapid striptease. (I suffered only a minor burn on my upper back.) I heartily agree with Larry that nudity is the way to go if you plan to spill LN2 on yourself!
Cheers :o)
Paul Grover Chief Microscopist and Bottle Washer Microvista Laboratory Lafayette, IN
Hi Ritchie As I remember the relevant cock should be open on 1/4 turnover. It is necessary to check, that the water on exit is not very warm at max current of objective lens. On the other hand, I know the case, when the water was very cold, therefore the condensate accumulated in the lens. I figure, the consequences are clear to you. Regards Victor Sidorenko, ANTRON Co. Ltd., Moscow, Russia
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Hi
Can anyone advise what the water flows should be in the three parallel branches (OL; DPs; and power boards) of the 840A) The manual spec is } 5 l/min total, but I need to know particularly what the flow should be thru the OL.
TIA
rtch
ps replies welcome from JEOL
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
} As important, new users must learn that its quite harmless when a little is } splashed and its worth demonstrating that a finger can be stuck into liquid } nitrogen for a couple of seconds. This builds handling confidence.
I think that this is going a bit far.
Ever had any splashed into your eye? Only needs a drop in the wrong place.
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
I have been reading with interest about the handling of LN2 and am surprised that no one has taken the safety officer's conservative position.
I am all for getting bureaucrats out of our hair but not at the expense of safety. I agree the the extra clothing, goggles and what have you are cumbersome and most of the time un-necessary but I am only reminded of an incident that happened to me about 25 years ago.
I was routinely filling a portable LN2 tank from a larger source. The portable tank had a pressure gauge that would measure tank pressure: when full the gauge read 20 psi; empty 0 psi. The assembly was attached to the tank via a rubber vacuum hose clamped at each end.
The normal procedure was to release the "vent" valve, unclamp the rubber hose, remove the valve assembly, then refill the portable tank from the larger LN2 source. I am sure most of you have seen a similar assembly.
Early one morning (before my coffee), I went through the above procedure. After unclamping the rubber hose, I proceeded to remove the assembly from the tank using both hands to pry it loose. Unfortunately there was a little pressure left and as soon as the hose was released, a stream of LN2 burst from the tank. The noise startled me and in less than a half second I removed my hands from the tank. In less than one second, I realized I was burned and immediately immersed both hands into water from a nearby sink. Too late. In less than one-half second, I received second and third degree LN2 burns on the bottom of my hands. The blisters extended from the bottom of both hands to halfway up my small finger. Worse yet, both hands were bleeding .Apparently, both hands immediately froze and I had cracked the skin by moving them to the sink.
I wish I had gloves at that time.
I worried for the next two weeks about getting gangrene.
Still today, I don't use gloves as they are cumbersome but I would hate to think what would happen if I were sprayed in the eyes from LN2 even for a split second.
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------.
} } Dr. Larry Stoter (4 years ago) wrote: (and thanks for sharing it, Jim) } } Gloves, goggles, masks (and shoes) are actually more dangerous when handling } } liquid N2 than sandals and no protection. And clothes are actually more } } dangerous than being naked. Get the safety officer to experiment. With a } } little persuasion you can probably convince the safety officer that when } } handling liquid N2, everybody should be naked. } } I'd like to share my experience with getting naked in a hurry. I was } working around midnight in the biochem lab at Okla. State Univ. in '79 and } was freezing samples to put in the lyophilizer. My arms were full, carrying } numerous flasks plus a dewar of LN2, which I held against my shoulder. As I } turned a corner, I slipped on some spilled water (I was wearing those flip } flop sandals) and the entire contents of the dewar poured down my back. } Fortunately there were no witnesses to my rapid striptease. (I suffered only } a minor burn on my upper back.) I heartily agree with Larry that nudity is } the way to go if you plan to spill LN2 on yourself! } } Cheers :o) } } Paul Grover } Chief Microscopist and Bottle Washer } Microvista Laboratory } Lafayette, IN
Why? A droplet of liquid N2 would bounce of the eye just like of any other skin and the cushion of gas N2 (Leidenfrost) would help to prevent any damage. I don't suggest to pour liq N2 into an eye and its prudent to wear goggles when working with the stuff overhead, but why would you work with liq N2 high up on your body? Surely when filling an EDS detector on a TEM (higher up than SEM) you would use steps. 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 ABN: 99 724 136 560 www.proscitech.com
On Monday, July 03, 2000 11:46 PM, Ritchie Sims [SMTP:r.sims-at-auckland.ac.nz] wrote: } } } As important, new users must learn that its quite harmless when a little is } } } } splashed and its worth demonstrating that a finger can be stuck into liquid } } } } nitrogen for a couple of seconds. This builds handling confidence. } } } I think that this is going a bit far. } } Ever had any splashed into your eye? Only needs a drop in the wrong } place. } } 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
Normally, it is easier, more efficient and safer to use a small ( {10L) transfer dewar to actually fill an EDS dewar. LN2 itself, as several others have pointed out, is quite safe to handle. The nudity recommendations are only slightly tongue in cheek. Small quantities of LN2 on the skin will 'float' on a vapor phase that forms immediately, avoiding close contact. If the LN2 is able to wedge between skin and gloves or other clothing, a more intimate and hazardous situation can occur.
The real problem with the use of cryogenic hoses is that the exterior metals can also achieve cryogenic temperatures. While they will generally get coated with ice, the underlying metal can burn if touched. When the contents of a dewar are first being drawn, the warm hoses will cause the fluid to evaporate. For the first minute or two, all that will be going into the EDS dewar will be cold vapor. That could cause an accumulation of ice in the EDS dewar that can affect performance.
The use of a small transfer dewar allows you to only put liquid LN2 in the EDS dewar. Also, you have a more immediate control of the flow into the dewar. One other problem using a hose feed system is that you may turn the pressure up too far at first, since there is only vapor passing through. When the fluid starts flowing, the pressure may be high enough to cause excessive 'splashing'.
Allen R. Sampson, Owner Advanced Research Systems, St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
On Friday, June 30, 2000 6:35 AM, Everett Ramer [SMTP:Everett.Ramer-at-netl.doe.gov] 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 will getting my first SEM/EDS shortly and to obtain the required operating permit from my safety group I need a written procedure for handling LN2. Specifically I need a written procedure for filling the 3 L dewar on the EDS detector from a 50 L dewar mounted on a cart. I plan to make the transfer by using a lab source of N2 to pressurize the 50 L dewar and have obtained all the valves and fittings required to do this from another SEM lab. I would appreciate copies of the procedure. } Thanks, } } Everett Ramer } National Energy Technology Laboratory } P.O. Box 10940, Cochrans Mill Road } Pittsburgh, PA, USA 15236-0940 } Voice: 412-386-4920 } FAX: 412-386-4806 } ramer-at-netl.doe.gov } } }
We are looking for an used Oxford cryo-stage, prep-chamber and control panels for our Jeol 6400 SEM. If anybody wants to sell their old one, please let us know.
I am a microscopist who also has the misfortune to be the Departmental Safety Adviser and I have a few comments to make from the point of view of the enemy. While the jokey remarks about the advisability of wearing clothing, protective or otherwise, do have an element of truth, liquid nitrogen has the potential to do a great deal of harm. Last month there was a prosecution in Edinburgh where the Medical Research Council was found guilty of breaches of Health and Safety law after a technician died of asphyxiation while dispensing LN2 in a room with inadequate ventilation and three of his colleagues came close to suffering the same fate while trying to rescue him. The room was fitted with a low oxygen alarm but it was switched off because it went off too frequently and annoyed him. This is an example of familiarity leading to dangerous practices. (His bosses got hammered because they knew about the alarm being switched off.)
To write a procedure you need to think of what could go wrong, as well as the routine safe handling of the LN2, which is basically covered by - would you believe it -common sense. What would happen if the dewar shattered? A full shield should stop LN2 fron getting up your nose or in your mouth and is better than goggles which might trap liquid insude them. Drain holes in the bottom of the outer case of the dewar will reduce the risk of your hand freezing to it. What would happen if the main tank ruptured or fell over? I know that this is very unlikely, but a simple calculation based on the volume of the room and the capacity of the tank will tell you if oxygen depletion to a dangerous level is a significant possibility - and it will look good on your paperwork.
LN2 burns are particularly nasty as you may not be aware of them until the frozen bit thaws out again, by which time it may be too late to prevent serious damage so you might want to consider gloves if you are to handle cold parts of the apparatus. These must be proper cryo-gloves, but it has to be said that they offer limited protection and can in certain circumstances be worse than not wearing gloves at all.
Remember that if it can happen it will,
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
I can't really leave this one alone - it's just begging for a contrary opinion.
The point about gloves holding liquid nitrogen is of course true if they either fill from the wrist or are porous, but you should never wear tight gloves - it should always be possible to shake or flick them off (ideally without leaving fingers inside) and it is possible to get quite impervious gloves. I am extremely concerned about the rejection of need for protection because it could too easily mean that you didn't have any gloves handy when surfaces became cooled. More importantly if you handle super-cooled liquid nitrogen (or countless other cooling mixtures) for freezing samples then you just might forget. I am also interested in the comments about goggles and masks being more dangerous as my understanding was that liquid nitrogen is relatively safe on dry surfaces but presents a greater hazard to the wet surface of the eye - if only by temporary blinding the user or causing them to flinch.
You might also take for granted the nitrogen gas evolving and the consequences could be fatal in an enclosed space - see:
http://www.safetynews.co.uk/archivenews.htm#£25,000 fine for Human Genetics Unit liquid nitrogen fatality
I must admit that apart from the above I have seen little, but I would be interested to hear how a nudist colony would really fare with a major liquid nitrogen leak - remember the horror stories about frying sausages in nudist camps.
I would also like to counter the thought that gasoline is more dangerous than liquid nitrogen. It's different - I carry a 4.5 litre sealed container of gasoline/petrol around in my car quite safely but I wouldn't seal liquid nitrogen like that; open or poorly sealed storage can lead to icing or condensation of oxygen from air (now there's a good one to debate - has anyone heard of that causing an accident?) with possible disastrous consequences; surfaces in prolonged contact with gasoline would present less danger than with liquid nitrogen; and gasoline has a strong odour so I wouldn't expect to be asphyxiated by it so easily (OK it might ignite more easily and be harmful).
Now I must go because I'm on the third page of my triple distilled water risk assessment.
Malcolm Haswell University of Sunderland UK e-mail: malcolm.haswell-at-sunderland.ac.uk
Larry Stoter 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 completely agree with the comments by Warren Straszheim and Larry Stoter. A } } couple more points: } } 1 Gasoline is rather more dangerous then liquid nitrogen. If } } anybody would try } } to impose silly forms and rules and thus curtailing the publics access to } } gasoline, they be out on their ear fast. Access to gas is available to } } countless completely untrained people. } } 2 What evidence is there that serious accidents have occurred } } by small scale } } handling of liquid nitrogen. Golden rule of administration: Regulate only when } } there is a demonstrated need. } } 3 Its my observation that timid people (intimidated by } } overstated dangers) are } } more likely to cause accidents. I instructed first time users of liquid } } nitrogen that there is a danger when the substance is enclosed, that touching } } cold metal or pouring it into ones shoes causes burns and that cold tubing can } } fracture. } } As important, new users must learn that its quite harmless when a little is } } splashed and its worth demonstrating that a finger can be stuck into liquid } } nitrogen for a couple of seconds. This builds handling confidence. } } } } The safety officer's paper wastes time and paper. Users need to learn how to } } handle the substance; its simple enough. Writing up a procedure for pumping } } creates a delusion, which has no practical benefits. } } 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 } } ABN: 99 724 136 560 www.proscitech.com } } } } On Saturday, July 01, 2000 3:39 AM, Warren E Straszheim } } [SMTP:wesaia-at-iastate.edu] wrote: } } } } } } } } } Hi, Everett. I guess I am glad I am not working for a government anymore. } } } Otherwise I would probably be having to fill out similar forms. It would be } } } nice if the folks that review those forms knew what the issues were. I am } } } always afraid that they will know just enough to be dangerous and may } } } actually require some unhealthy procedures. But perhaps they are sensible } } } and only require that you have something officially in writing to document } } } your best practice. } } } } } } I found the following post in my files from 4 years ago. I hope Larry does } } } not mind me reposting it. His conclusions are quite interesting. } } } } } } Warren S. } } } } } } } } } Date: Sat, 30 Mar 1996 07:57:02 +0000 } } } } From: "Dr. L. P. Stoter" {LPS-at-teknesis.demon.co.uk} } } } } } } } Wil's recent comment on the safety hazards of distilled water brought to } } } } mind some peculiar safety regulations here in MD. In reference } } } to liquid N } } } ... } } } } officer listening in will recommend new safety procedures requiring } } } } protective booties! } } } } In the end, we can't legislate common sense, nor can we abdicate } } } } responsibility to those above. } } } } } } Try getting your safety officer to conduct an experiment: } } } } } } 1. Hold out hand, } } } 2. Pour a small volume of liquid N2 over hand } } } 3. Now the interesting bit - put on a glove, and pour the same quantity of } } } liquid N2 into glove. } } } 4. Phone for ambulance. } } } } } } The point is that a brief contact causes no problems, but if the contact is } } } continued you get a nasty burn. } } } } } } Gloves, goggles, masks (and shoes) are actually more dangerous when handling } } } liquid N2 than sandals and no protection. And clothes are actually more } } } dangerous than being naked. Get the safety officer to experiment. With a } } } little persuasion you can probably convince the safety officer that when } } } handling liquid N2, everybody should be naked. } } } } } } More seriously, bureaucrats, administrators and the inexperienced should } } } talk to somebody who has real knowledge. } } } } } } -------------------------------------------------------------- } } } Dr. Larry Stoter } } } Technesis } } } 17, Rocks Park Road, Uckfield, E. Sussex, TN22 2AT, United Kingdom } } } Larry-at-teknesis.demon.co.uk } } } -------------------------------------------------------------- } } } } } Actually, not my comments, although it is a thread to which I } contributed and generally reflects my sentiments. } } Regards, } -- } Larry Stoter } 34, Astwick Road, Stotfold, Hitchin, Herts, SG5 4AU, UK } email: LPS-at-teknesis.demon.co.uk, Home Phone/Fax: +44 (0)1462 733309
as a manufacturer of large chamber SEMs with EDX it would be a pleasure for us to analyze your sample. The max. size of your sample should not exceed 28 inches in diameter and 24 inches in height.
To give you an impression about the technology of large chamber SEMs just click it http://www.visitec-em.de/tokyo.html
----- Original Message ----- } From: Becky Holdford {r-holdford-at-ti.com} To: Microscopy ListServer {microscopy-at-sparc5.microscopy.com} Sent: Saturday, July 01, 2000 2:54 AM
Hi All, there is a serious side to LN2 handling even though many points discussed are valid. I have found most problems associated with LN2 are caused by things other than the LN2 itself. I remember when I first started using LN2 over 25 years ago I was filling a glass vacuum flask when it exploded and filled the room I was in with instant fog, tinsel and plenty of LN2 as a fine spray. I was unharmed apart from fine glass shards and Al foil in my hair. I was wearing no protective gear. The LN2 had also been in my hair as a fine spray thank goodness. I was amazed that I had no effect from the LN2. After that I started to wear thermal type gloves but soon found that these were more dangerous than no gloves at all. There are times when one does have to be careful in handling LN2 and these in my experience have been with 1/ glass containers. 2/ pressurised containers and 3/Accidental contact from surfaces at very cold temperatures.
I am sure that there have also been other experiences that we can learn from so that we can understand how we can avoid them.
Ken Moran.
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"
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Leona, I also have bottles of Epon 812 in my lab, some of which have been opened, and all of which are from the early 1970's. I still use it for certain applications when nothing else works. Epon 812 blocks will pop off some plastic culture wells when nothing else will. Even though the dishes are made of the same material there is some variation from brand to brand and when in doubt I use the old resin. I haven't varied the original recipe and have had no problems. I'll buy it from you if you decide not to keep it.
Mary Gail Engle Electron Microscopy & Imaging Facility University of Kentucky
I was searching through the archives - found numerous helpful suggestions, however no real answer to my recent problem.
Here it is: We were investigating cells in suspension and blood cells in the SEM. Very straight forward - after fixation in suspension the cells were filtered on to polycarbonate filters (0.4 um) with a 1ml syringe.
At this stage I removed the filters from the holder and put them into multiwell plates for additional washes with buffer and dehydration in alcohol. The multiwell plate was placed on a gently shaking rotator. The cells on the filter were dried via liquid substitution, gold coated and observed.
Although the cells survived the procedure structurally quite well I was surprised that we had to hunt a lot to actually find the cells on the filter. The pore size of the filter is too small for human blood cells to escape through - were has 90% of the population gone?
Would it be better to place unfixed cells on the filter?
Is there a way to make them adhere better?
Any suggestion is appreciated - this list is a fantastic source of know-how.
Thank you.
Claudia
Dr. C. Hayward-Costa School of Life Sciences Kingston University Penrhyn Road, Kingston upon Thames Surrey KT1 2EE, UK 44(0)208 547 2000 x 2240 Email: c.hayward-at-kingston.ac.uk
Hi, I use Epofix epoxy. In a very short time after purchase the resin becomes cloudy and a white sediment forms at the bottom of the bottle. I have heard this sediment described as crystals. I have continued to use the resin---carefully avoiding agitation and entrainment of the sediment when I withdraw it from the bottle. Does anyone else have experience with this?
Everett Ramer National Energy Technology Laboratory P.O. Box 10940, Cochrans Mill Road Pittsburgh, PA, USA 15236-0940 Voice: 412-386-4920 FAX: 412-386-4806 ramer-at-netl.doe.gov
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I would require a used / 2nd hand Cambridge S360 IP backplane.Could anybody help me out ?
Thanks, Neville Baker
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
see you at ICEM 15 2002, Durban, South Africa www.icem15.com
From root Mon Jul 3 11:58:24 2000 Return-Path: {Microscopy-request-at-sparc5.microscopy.com} Received: (from daemon-at-localhost) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id LAA16569 for dist-Microscopy; Mon, 3 Jul 2000 11:48:38 -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 LAA16565 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Mon, 3 Jul 2000 11:48:07 -0500 (CDT)
Colleagues,
We are looking into the possibility of mating a Zeiss AttoArc fluorescence lamphouse (for HBO100 bulb) to a Leica DMIRB inverted research microscope. Has anyone had experience doing such a thing?
We can certainly manufacture an adapter to mount the lamphouse to the Leica scope; that's a minor problem. But I am more concerned about the length of the beam path and whether the collector lens in the Atto source will bring the beam to the proper focus point.
I should mention that we have the Leica scope but we haven't yet found an Atto lamphouse and power supply. We have tried to contact Atto but so far we have not gotten a response. Perhaps the Atto rig is sold exclusively for Zeiss scopes? I have done some initial measurements on a Zeiss inverted scope that has an AttoArc lamphouse on it, and I do believe that it would work just fine. But some confirmation from another source would be appreciated!
If anyone has done this before, I would certainly like to hear from you. Please contact me off-list to discuss.
Thanks for your input.
Cheers,
Bob Chiovetti rchiovetti-at-aol.com
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Greetings,
We are currently planning to teach the introductory undergrad SEM course on our JEOL 840A with the digital output. In doing so we are dropping the 4 month/year service contract and most likely we are going to discontinue use of the AMRAY 1200.
I am trying to get a feel of the market for an old (early 1970s) solid-state SEM. The imaging is fantastic - really quite amazing. But there are quite a few limitations - resolution, magnification, only film output (4X5 Polaroid or Sheet film), scan speeds.
Is this a machine that anyone might be able to make use of? One idea I had was this little table top wonder machine would make a great personal SEM, although for me personally why take it home when I can use the 840A when ever I want.
Again, I am not committing this good old 'scope to the open market or the open scrap heap (the other possible avenue for this scope is to call the scrap yard to haul it off) yet, I just need some potential options to through at the Chairman and the Dean.
Any third party folks that are servicing these have a desire for some/all of it?
Geoff
-- Geoff Williams
Electron Microscope Facility Supervisor Biology Department Brooks Hall Central Michigan University Mt. Pleasant, MI 48859
We have a Kreonite automatic photographic paper processor that will go to the scrap heap very soon if no one wants it. You would probably have to arrange for shipping. If anyone would like to have it (and a few spare parts), it can be arranged. Preference will go to a university, but we are interested in getting it out of the storage area. It was working when we stored it. It comes with a cart that was made for it. If you process a lot of paper, this is the unit for you. Let me know this week.
-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 would also like to counter the thought that gasoline is more dangerous } than liquid } nitrogen. It's different - I carry a 4.5 litre sealed container of } gasoline/petrol } around in my car quite safely but I wouldn't seal liquid nitrogen like } that; open or } poorly sealed storage can lead to icing or condensation of oxygen from air (now } there's a good one to debate - has anyone heard of that causing an } accident?) with } possible disastrous consequences; surfaces in prolonged contact with } gasoline would } present less danger than with liquid nitrogen; and gasoline has a strong } odour so I } wouldn't expect to be asphyxiated by it so easily (OK it might ignite more } easily } and be harmful). }
Now there's a thought. To digress a little, we once had an instance where a guy came down to the SEM lab and asked if he could borrow a 5 litre dewar to take some LN2 home and show his kids. When asked for more details his game plan was to sit the dewar on the floor on the passenger side of the car so that he could make sure that it wouldn't fall over. The answer was "No, perhaps you should bring the kids to the LN2...."
The point being that even though LN2 use might seem less of a risk than boiling water when used "properly" and "common sense" is applied, it cannot be assumed without question that everyone's initial mental picture behind these two terms is exactly the same!!
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/
We were having a coffee break in a portacabin room used as a makeshift lab when a delivery lorry about to top up our cryogenic liquid N2 tank reversed into and broke the connecting refill valve . We were observing the resulting stream of liquid N2 spill over the tarmac when one of the guys remembered that his new car was parked in the enclosed area and his new tyres may be damaged . Without a word he pushed through the firedoor , ran the few yards through the increasingly heavy fog and managed to reverse his car in the few feet available out of the way of the liquid nitrogen . But when he went to return he found the fog too heavy to see the few yards to the portacabin , and quickly became disorientated . As panic set in he fell to his knees and ,becoming increasingly short of breath, started crawling- finally to the firedoor which by now was firmly closed as we had all run for safety . Fearing asphyxiation he managed to crawl around the building through the fog and emerged gasping a minute or so later . The contrast to one minute quietly having a coffee and the next to witness this life and death ? struggle and accusations of attempted murder as someone had closed the firedoor caused much merriment at the time . All except for the poor 'victim' ( sorry Leighton if your reading this ) .
I've avoided commenting on LN2 to this point, because one person became quite offended at my comments the 4-5 years ago when this thread was explored in a very similar manner. Ken's comments about shattered thermos is an experience, though, I've shared too many times.
Besides being the glass thermos being a different kind of hazard after breaking, a 5L dewar is very expensive. Just over a year ago I started using a 2 gallon plastic drink dispenser from WalMart. Cost less than $10. It lasted almost a year before the seal around the spigot failed. Of course you can't store LN2 for any length of time but for transport between tank and EDX detector, it does great. BTW, the top vent was permanently removed so no pressure buildup could occur.
Chuck Butterick Engineered Carbons, Inc. Borger, TX
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Hi All, there is a serious side to LN2 handling even though many points discussed are valid. I have found most problems associated with LN2 are caused by things other than the LN2 itself. I remember when I first started using LN2 over 25 years ago I was filling a glass vacuum flask when it exploded and filled the room I was in with instant fog, tinsel and plenty of LN2 as a fine spray. I was unharmed apart from fine glass shards and Al foil in my hair. I was wearing no protective gear. The LN2 had also been in my hair as a fine spray thank goodness. I was amazed that I had no effect from the LN2. After that I started to wear thermal type gloves but soon found that these were more dangerous than no gloves at all. There are times when one does have to be careful in handling LN2 and these in my experience have been with 1/ glass containers. 2/ pressurised containers and 3/Accidental contact from surfaces at very cold temperatures.
I am sure that there have also been other experiences that we can learn from so that we can understand how we can avoid them.
Ken Moran.
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"
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Hello, I've been cleaning and tweaking an older SEM we have in our laboratory. It is an ISI DS-130. I managed to improve the resolution of the lower stage from 2000x mag to 10000x magnification through cleaning the final aperture, cleaning the column liner, replacing the scintillators, and cleaning the anode. However, I would like to improve it up to at least 30000x mag for the lower stage. Could anyone send me some suggestions as how I may improve the performance of this SEM further?
I have an additional problem in using the upper stage of the SEM. It will not focus above 300x in magnification even though it is properly set up according to the manual. Anyone ever come across this problem before? Gordon.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
I have a customer who is considering the purchase of an SEM-EDS system. He has asked me if there are any EDS programs out there that can take the EDS data, and, given some minimal user input, provide a list of possible phases. For example, Fe and S alone and in various proportions may yield - iron sulfide FeS2 or FeS (pyrite or troilite) while the presence of a low level of O might get a result of iron sulfite or maybe iron sulfate, etc.
I know this is out there past the edge of real usable analytical information, but just thought I'd see if anything exists. I'm not talking about anything real fancy like the EBSP techniqes, but basically an "expert" EDS program that will return a list of potential phases or formulas based on a little quantitative and specimen knowledge/input from the real expert.
Like any SEM you need to be sure that you are not undersaturated and are very accurately aligned for maximum signal: use the wave form mode. Performance will be further improved if you run at between 5 and 8mm working distance. Also, as the HT tank needs time to warm up to give stability, do not try for performance until the HT has been on for at least 45 minutes.
The upper stage has two settings, high kV and low kV, where a shorter focal length is usable. In each case the secimen MUST be in exactly the correct position, not to high not too low as there is very little focal length adjustment compared with "normal" out of lens SEM.
Good luck
Steve Chapman Senior Consultant Protrain For professional training and consultancy in EM world wide www.emcourses.com Tel +44 1280 814774 Fax +44 1280 914007
I have also had trouble with exploding glass dewars, I now use the plastic ones from the chem. suppliers for freezing samples.. For filling up our EDX I do that directly out of the 160L liquid nitrogen tanks from our supplier, I use a special liquid nitrogen flexible metal hose with a sintered brass end that lets the gas out the sides and liquid out the end. The 160L tank sits on a special movable dolly, so I unchain it and move it next to the EDX or whatever I need to fill up, it saves spillage and no dewar is needed for EDX fillups. I ordered the equipment from Southland Cryogenics, Inc. but I have seen the same stuff at different cryogenic internet sites.
I had to get 20,000 to 50,000x out of our DS-130 some years ago...I'll see what I remember. Since I don't know the details of your setup, I will just throw some questions at you. What spot size(s) are you using? What size column aperture? I'm pretty sure you'll need to stick with spot HR, or at least STD (Small) to reach 30000x. The TV image will be grainy, but photo scans will be fine. If the image gets TOO noisy, try a larger column aperture. For a given 'Z', a certain combination of spot/aperture will work best. Is your scope on air cushions? If so, are they inflated properly (not OVER inflated)? If not on cushions, do you know what your floor vibration is like? Relocating to ground floor did wonders for ours. You will also need quite high emission current: setup the filament for high-res. (per the manual) and crank up the bias as necessary. It's important to check the mechanical gun alignment frequently...ours drifted quite a bit. Align gun, column aperture, focus, correct astigmatism. In short, pretty standard stuff. I have some good pictures at 50,000, not-so-good pictures at 150,000 and that's on the second floor. Didn't have a functioning 1st stage, so I can't help you there. Wild guess: range of objective current is not switching over properly when you switch to 1st stage? Good luck.
Matt
Matthew J. Lynn, Ph.D. Center for Advanced Microscopy University of Miami (305)284-4736 mlynn-at-miami.edu
On Wednesday, July 05, 2000 4:25 PM, Gordon Vrololjak [SMTP:gvrdolja-at-nature.Berkeley.EDU] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello, } I've been cleaning and tweaking an older SEM we have in our } laboratory. It is an ISI DS-130. I managed to improve the resolution of } the lower stage from 2000x mag to 10000x magnification through cleaning } the final aperture, cleaning the column liner, replacing the } scintillators, and cleaning the anode. However, I would like to improve } it up to at least 30000x mag for the lower stage. Could anyone send me } some suggestions as how I may improve the performance of this SEM further? } } I have an additional problem in using the upper stage of the SEM. It will } not focus above 300x in magnification even though it is properly set up } according to the manual. Anyone ever come across this problem before? } 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
We are having problems with the hardware/software acquiring data from our JEOL JAMP-30. The unit uses an old PDP11. We are wondering if anyone has a similar system and can instruct us how to reformat the hard drive and reload the software on this system. Any help would be appreciated. Thanks. ______________________________________________ Roberto Garcia Senior Analyst, Metallography NC State University / Analytical Instrumentation Facility Campus Box 7531 Room 318 EGRC 1010 Main Campus Dr. Raleigh, NC 27695-7531 (919) 515-8628 (919) 515-6965 Fax rgarcia-at-unity.ncsu.edu http://spm.aif.ncsu.edu/aif http://spm.aif.ncsu.edu/asm/asm_home.htm ____________________________________________
If anyone wants Geoff Williams' Amray 1200, I have some sample holders and tools left over from when we scrapped our Amray 1200 due to water damage. The only cost would be shipping.
Jane L. LaGoy Development Engineer Bodycote IMT, Inc. 155 River Street Andover, MA 01810 978-470-1620 jlagoy-at-bodycote-imt.com
I know this isnt't an EM related question but I'm in search of a couple of LM staining procedures. I'm looking for a good silver stain (s) for looking at neural structures preferably not using chloral hydrate, and a good acid fast procedure for mycobacterium. If anyone can help, I'd appreciate it.
I'm interested in why one would use a glass dewer for transporting LN2 (cost?). Our transportation dewer is metal - don't know how many walls but I assume it's got a vacuum between them. It's been in use for over 20 years and had its share of bumps (I'm sure if it was glass, it'd have been replaced a few times). It's made by Union Carbide, type UC-4.
It works great, has lasted 'forever' and doesn't have the safety issues of glass. Is glass that much cheaper?
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I've avoided commenting on LN2 to this point, because one person became quite offended at my comments the 4-5 years ago when this thread was explored in a very similar manner. Ken's comments about shattered thermos is an experience, though, I've shared too many times.
Besides being the glass thermos being a different kind of hazard after breaking, a 5L dewar is very expensive. Just over a year ago I started using a 2 gallon plastic drink dispenser from WalMart. Cost less than $10.
It lasted almost a year before the seal around the spigot failed. Of course you can't store LN2 for any length of time but for transport between
tank and EDX detector, it does great. BTW, the top vent was permanently removed so no pressure buildup could occur.
Chuck Butterick Engineered Carbons, Inc. Borger, TX
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi All, there is a serious side to LN2 handling even though many points discussed are valid. I have found most problems associated with LN2 are caused by things other than the LN2 itself. I remember when I first started using LN2 over 25 years ago I was filling a glass vacuum flask when it exploded and filled the room I was in with instant fog, tinsel and plenty of LN2 as a fine spray. I was unharmed apart from fine glass shards and Al foil in my hair. I was wearing no protective gear. The LN2 had also been in my hair as a fine spray thank goodness. I was amazed that I had no effect from the LN2. After that I started to wear thermal type gloves but soon found that these were more dangerous than no gloves at all. There are times when one does have to be careful in handling LN2 and these in my experience have been with 1/ glass containers. 2/ pressurised containers and 3/Accidental contact from surfaces at very cold temperatures.
I am sure that there have also been other experiences that we can learn from so that we can understand how we can avoid them.
Ken Moran.
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"
by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id OAA01948 for dist-Microscopy; Thu, 6 Jul 2000 14:16:35 -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 OAA01945 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Thu, 6 Jul 2000 14:16:04 -0500 (CDT) Received: from rwja.umdnj.edu (rwja.UMDNJ.EDU [130.219.4.100]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id OAA01938 for {Microscopy-at-sparc5.microscopy.com} ; Thu, 6 Jul 2000 14:15:52 -0500 (CDT) Received: from umdnj.edu ([130.219.170.159] (may be forged)) by rwja.umdnj.edu (8.8.6 (PHNE_17190)/8.8.6) with ESMTP id PAA20459; Thu, 6 Jul 2000 15:10:09 -0400 (EDT) Message-ID: {3964D997.963C5D7C-at-umdnj.edu}
Phillip Rutledge wrote:
} Microscopists, } } I know this isnt't an EM related question but I'm in search of a couple of LM staining procedures. I'm looking for a good silver stain (s) for looking at neural structures preferably not using chloral hydrate, and a good acid fast procedure for mycobacterium. If anyone can help, I'd appreciate it. } } Thanks, } } Phil Rutledge } USDA/ARS } voice: (410) 778-4136, 2120 } fax: (410) 778-4399 } e-mail: prutledge-at-ars.usda.gov
Dear Phil:
Without knowing more specifics, the Bodain method is a good silver stain for CNS. Details in any good histotechnology text. Can't help with mycobacterium.
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 **********************************************
Dorrance: Have look at ASTM Standard E 1558, Standard Guide for Electropolishing of Metallographic Specimens.
Sam Purdy Tech Center, National Steel Corp. Trenton MI } ---------- } From: McLean, Dorrance } Sent: 30, June 2000, 3:21 PM } To: 'Listserver' } Subject: Mat. Electropolishing. } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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, } } Does any one have the dummies guide to electropolishing?? Some years } ago } } (3), I had a trouble shooting guide for electropolishing. This list had } } about 9 or 10 items, problems /solutions on it and it was very } } helpful...but it has found one of those very safe filing places (i.e. I } } can't find it!) If any one has access to this list or something } similar, } } I would appreciate it if you could send me a copy. } } } } Many thanks in advance. } } } } Dorrance } } PS I haven't been keeping up with the weather reports lately but it's 95 } } and clear in beautiful Livermore, California. } } }
Phil I wonder if Del Rio Hortega's silver stain would be good for your purpose. See Goldblatt & Trump (1965) The application of del Rio Hortega's Silver method to epon-embedded tissue. Stain Technology 40, 105-115.
Add 5ml of 10% AgNO3 to 20ml of 5% aq. sodium carbonate. Horrible precipitate occurs. Add 0.88 ammonia dropwise until precipitate has just cleared. Dilute to 45ml, filter.
Stain free-floating sections at 60oC for 30-120 min (use a solid watch-glass with glass cover). Transfer sections to distilled water to rinse, then dry down to slides from water at 60oC. reduce in 0.5% formaldehyde for 30-60 seconds, wash in distilled water, fix in 5% sodium thiosulphate (hypo), wash again and mount.
Chris
Date sent: Thu, 06 Jul 2000 06:49:00 -0600 } From: Phillip Rutledge {prutledge-at-ars.usda.gov} To: Microscopy-at-sparc5.microscopy.com
Course Announcement
Title: Optical Microscopy and Imaging in the Biomedical Sciences
When: October 11 - October 19, 2000
Where: Marine Biology Laboratory, Woods Hole, MA, USA
Tuition: $2150 (Includes room and board, text, handouts, supplies)
Application Deadline: August 1, 2000
Admission application and information: Carol Hamel, Admissions Coordinator Marine Biological Laboratory 7 MBL Street Woods Hole, MA 02543-1015 (508) 289-7401 Internet: admissions-at-mbl.edu WWW: http://www.mbl.edu (Application forms available via Adobe Acrobat)
Course Director: Colin S. Izzard, State University of New York -at- Albany Phone: [518] 442 - 4367 EMail: csizzard-at-csc.albany.edu
Course Description:
For Whom: Designed primarily for research scientists, physicians, postdoctoral trainees and advanced graduate students in animal, plant, medical and material sciences. Non-biologists seeking a comprehensive introduction to microscopy and video-imaging will benefit greatly from this course as well. There are no specific prerequisites, but an understanding of the basic principles of optics is desirable. Limited to 24 students.
The eight day course consists of lectures, laboratory demonstrations, exercises and discussions that will enable the participant to obtain and interpret microscope images of high quality, to perform quantitative optical measurements, and to produce photographic and video records for documentation and analysis.
Topics to be covered include: principles of microscope design and image formation bright field, dark field, phase contrast, polarized light, differential interference contrast, interference reflection, and fluorescence microscopy confocal scanning microscopy, multiphoton excitation fluorescence microscopy and image deconvolution digital image restoration and 3-D reconstruction video imaging, recording, enhancement, and intensification analog and digital image processing and analysis fluorescent probes and ratiometric-imaging laser tweezers and laser scissors
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.
Students are encouraged to bring their own biological (primary cultures, cell lines, etc.) and material specimens and to discuss individual research problems with the faculty.
This isn't an EM question, but I'm hoping someone can give me an answer. I need postive control slides for PCP fluorescence testing. If anyone knows of any company that sells them, please let me know. Thanks
Email: lpeterso987-at-hotmail.com Name: lauren peterson School: university of michigan
Question: I bought an old student microscope (3 objectives) from a biology lab. It has a mirror and condenser lens (with iris) that can be focused up and down. It doesn't seem to make much difference how the lens is adjusted or how the iris is set -- both just seem to affect the brightness. Is there a procedure for adjusting it to get the best images?
I can help you on both stages. Following the ISI manual can be like many manuals, a little difficult.
The 1st stage is the simplest problem to fix. Select a tall or short specimen holder, mount your sample on the adjustable base of the holder. Turn the base until the top most portion of your sample is at the same height as the top of the holder. Not above!! If you selected the tall holder the 'Mode' button need to be on 2 and for the short holder the 'Mode' button needs to be on 1. Go ahead and put the sample and view. Increase the magnification up to 1 KX, adjust the focus knob. You should be able to find focus.
As for the 2nd stage, it sounds as you are close in your alignments. You should continue to practice. I find that it is difficult to routinely expect to take micrographs above 20 KX. The general rule around here is if you want to image above 20 KX go to the upper stage.
We use a LaB6 filament on our DS-130, I was imaging this week at 30KX on the lower stage and at 100KX on the upper.
Mike ============================================================ Michael Dunlap office (530) 752-0284 University of California lab (530) 752-5489 Chemical Engineering & Material Science Fax (530) 752-9554 110A EU-II mrdunlap-at-ucdavis.edu One Shields Ave. http://www.chms.ucdavis.edu/ Davis CA, 95616 ============================================================
} I have also had trouble with exploding glass dewars, ... } For filling up our EDX I do that directly out of the } 160L liquid nitrogen tanks from our supplier, } I use a special liquid nitrogen flexible metal hose } ...
The 160L dewar I fill from has a "nalgene" hose ... and I've never had a problem with it. However, one plastic hose exploded on me while filling, but it wasn't nalgene, and I can't remember what it was. But, it does underline the use of wearing protective eyewear while working with LN2!
We are planning to acquire a Phillips 300 TEM for our lab located on Long Island, NY. Does anyone have any recommendations for independent service/maintenance contractors who could provide a service contract in this area?
Also, we don't yet have a recirculating water chiller. If anyone has one that they could sell to us, please let us know.
Thanks in advance,
Rick Powell Nanoprobes, Incorporated
********************************************************************** * - PLEASE NOTE OUR NEW ADDRESS - EFFECTIVE FEBRUARY 15, 2000 - * * * * NANOPROBES, Incorporated | US Toll-free: (877) 447-6266 * * 95 Horse Block Road | Tel: (919) 510-0590 * * Yaphank, NY 11980-9710, | Fax: (919) 510-0590 * * USA | rpowell-at-nanoprobes.com * * * * NOW EASY TO FIND ON THE WEB: http://www.nanoprobes.com * **********************************************************************
This is my summary of that thread, reflecting my bias and my logic. Clearly there were divergent opinions. Everett had simply asked for a copy of "handling procedures". Hope that somebody could help him. The question though seems to be "is liquid nitrogen a dangerous substance"? Obviously yes, but so is water - to wit drownings. Danger is a comparative matter. The question "Is a written procedure required?" follows later.
Many common substances are dangerous when mishandled. Petrol/ gas in my opinion is rather more dangerous than liquid nitrogen. Gas is much more common than LN2, but few gas accidents are reported. LN2 accidents become folklore, not because of the greater danger of LN2 but because those accidents demonstrate ingenious, applied stupidity (hi, that would make a fine Uni course). Although gas and its dangers are well known, people inhale it (brain damage and death), start fires with it, store it in crummy plastic containers, siphon it by mouth, use it to re kindle a fire etc. I must agree though, gas is not a good comparison to LN2.
Boiling water is! Arguably its more dangerous than LN2 because it has no insulating layer. So boiling water poured onto skin will burn, whereas LN2 would only do so in extreme cases. Pressurized LN2 must be compared with pressurized boiling water, like in a pressure cooker or autoclave. Boiling water is undeniably dangerous and many, many burns occur every day. Of course its a very commonly used substance, but that means that all, except small children would know of the danger and should use care.
Why then is boiling water not a tightly controlled substance and why is no operator's license required to make a cup of coffee? Ask your safety officer. Our discussion concerns LN2 use in laboratories, by people with more training and understanding of physical properties than that of the general public. Why should these people be required to submit a procedural outline for simple tasks, such as transferring some LN2? Who trained the safety officer to recognize that the procedure makes sense? What is the cost benefit of introducing another bit of paper to the laboratory? How is safety enhanced by a single task written procedure? Most LN2 accidents are caused by lack of knowledge of the material's physical properties, or temporary "insanity"? A six minute briefing of LN2 properties and don'ts would benefits the neophyte and safety rather more than a cabinet full of paper. The required written procedure on decanting LN2 is a fig leaf for the safety officer. Unfortunately safety officers have become part of a bureaucratic system and frequently they are required to perform nonsensical tasks.
Overstated? Well, we now have in Australia legislation in place which makes the importation, sale and trans shipping of Uranyl Acetate virtually impossible. PST has given up on UA and I don't know of anybody who is willing to run that legislative gauntlet for the sale of a few jars of UA! This legislation applies in a country that produces thousands of tons of U Yellowcake (radiological and chem toxicity similar to UA), were jumbo jets carry 370 kg of U metal in the tail fin and were Americium smoke detectors are handled by the public. 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 ABN: 99 724 136 560 www.proscitech.com
} On Friday, June 30, 2000 6:35 AM, Everett Ramer [S} MTP:Everett.Ramer-at-netl.doe.gov] wrote:
} I will getting my first SEM/EDS shortly and to obtain the required operating permit from my safety group I need a written procedure for handling LN2. Specifically I need a written procedure for filling the 3 L dewar on the EDS detector from a 50 L dewar mounted on a cart. I plan to make the transfer by using a lab source of N2 to pressurize the 50 L dewar and have obtained all the valves and fittings required to do this from another SEM lab. I would appreciate copies of the procedure. } Thanks, } } Everett Ramer } National Energy Technology Laboratory } P.O. Box 10940, Cochrans Mill Road } Pittsburgh, PA, USA 15236-0940 } Voice: 412-386-4920 } FAX: 412-386-4806 } ramer-at-netl.doe.gov
Been thinking about your problem and of course suddenly thought that I did not pass on the most important message last time!
If you want better performance from any SEM you must make sure that the emission current is high enough. For DS130 use a current 100uA above the standing current (the current that you have when the filament is off but the selected high voltage is on).
Good luck
Steve Chapman Senior Consultant Protrain For professional training and consultancy in EM world wide www.emcourses.com Tel +44 1280 814774 Fax +44 1280 914007
Try www.kaker.com they have and online demo of electropolishing procedures. You can not search them but it will list all of the ones they have and then you just have to look for your material of interest. ______________________________________________ Roberto Garcia Senior Analyst, Metallography NC State University / Analytical Instrumentation Facility Campus Box 7531 Room 318 EGRC 1010 Main Campus Dr. Raleigh, NC 27695-7531 (919) 515-8628 (919) 515-6965 Fax rgarcia-at-unity.ncsu.edu http://spm.aif.ncsu.edu/aif http://spm.aif.ncsu.edu/asm/asm_home.htm ____________________________________________
Dear List, Here are my 2 cent's worth: 1) Teflon tubing does not become brittle at 77 K, so it is the best inexpensive material for transferring LN2 safely. Using insulating foam tubing around the teflon reduces LN2 boiling to a very acceptible degree. 2) Do not set up an automatic fill for LN2 for an EDS detector; when we tried that, the fill valve froze open, and the resulting LN2-fall broke the seal on the detec- tor dewar, which then became convex on the bottom. (We were able to fix it--I'll be happy to let anyone who is interrested know how.) Naturally, this occurred at the beginning of a long weekend; fortuitous, timely discovery prevented a worse disaster. And another penny's worth: 3) Glass dewars are not used because they're cheap; they're used because the silvered glass is an excel- lent insulator. They are very efficient for containing or transporting LN2, and they hold up very well, unless one drops a screwdriver into one. Stainless steel is tougher, but less efficient and more expensive. Yours, Bill Tivol
The following TEM position has been approved at Michigan State University. The individual selected will supervise a newly funded 200 kV field emission TEM fully equipped with EDS, PEELS, Gatan Image Filter, STEM, digital imaging, etc. Stanley L. Flegler, Acting Director Center for Advanced Microscopy Michigan State University
Full-time transmission electron microscopist. This is a teaching/service/support position at the Center for Advanced Microscopy at Michigan State University. CAM is the central microscopy laboratory for the MSU campus, serving users from a wide variety of disciplines. The appointment will be in the Academic Specialist category, one of the academic support ranks at the University. The appointment will be a 12 month, annual year appointment in the continuing appointment system. Salary will be commensurate with experience. Required: A Ph.D. in the physical sciences or a field of engineering and a minimum of three years experience in TEM instrument operation, maintenance, physical science sample preparation, and teaching. Experience in a multi-user facility and familiarity with biological TEM sample preparation is a plus. The individual must have a demonstrated competence in and a strong commitment to graduate level instruction and be willing to assist others in planning their research programs and/or sample preparation. U.S. citizenship is not required; applicants who are not U.S. citizens or permanent residents must provide documentation evidencing employment authorization in the United States. The position begins effective Spring 2001. Submit a Curriculum Vita, transcripts of academic training, a statement describing your interest in the position, evidence of teaching ability, and arrange for three letters of recommendation to be sent to: Dr. Karen L. Klomparens, Chair, Search Committee, Center for Advanced Microscopy, B4 Center for Integrated Plant Systems, Michigan State University, East Lansing, MI 48824. Applications are due by Sept. 18, 2000. MSU is an Affirmative Action/Equal Opportunity Institution.
I use "dewars" made of foamed polystyrene for transferring small volumes of LN2, and for any work where there is a risk of dropping heavy metal objects into the dewar. They are safe, light, robust, and inexpensive, although they do not have anything approaching the holding time of glass dewars. They should be obtainable through most EM supply houses. If you have difficulty sourcing them, contact me offline.
Just for the record, I have used glass dewars for LN2 for more than 30 years, and have never known one to break under thermal shock.
Chris Jeffree ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 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
Please note the following job posting at Lehigh University:
RESEARCH ENGINEER
The Department of Materials Science and Engineering at Lehigh University is seeking a Research Engineer. This position will be responsible for high spatial resolution x-ray mapping of segregants in alloys using the VGHB603; theoretical spectral simulation using desktop spectrum analyzer; and zeta factor mapping and quantification.
Qualified candidates should have Ph.D. in Materials Science or related field with at least 3 years of postdoctoral experience. Experience with 300KeV Field Emission Gun STEMs is essential.
Anticipated starting date is April 1, 2001. Resumes should be sent to Deanne Hoenscheid, Materials Research Center, Lehigh University, 5 E. Packer Avenue, Bethlehem, PA 18015. Lehigh University is an AA/EOE.
-- ********************************************************* Deanne L. Hoenscheid Phone: (610) 758-3863 Materials Research Center Fax: (610) 758-3526 462 Whitaker Laboratory E-mail: dlh3-at-lehigh.edu 5 E. Packer Avenue Bethlehem, PA 18015 *********************************************************
I believe the problems arise when scratches occur from filling and handling the glass dewars. These can be so fine, as to be difficult to see. (when cutting glass, a fine, chip free scribe does a better job than a heavy one) When the conditions are just right (stress from cooling down, or tipping the dewar up), the vacuum bottle will implode.
We have some old metal outside, plastic inside dewars for transferring LN2 from the storage/supply dewar, to the system dewar. They are not used to store LN2, but are good and rugged for all of the handling they get. They were here when I started, so I do not know where they came from.
We are please to offer the following position at AMD Sunnyvale
REQUIRED EXPERIENCE
Candidate should have a strong Materials Science education and extensive exposure to electron microscopy. At least two years industrial experience is preferred except if candidate did MS or PhD thesis work in electron microscopy. Good team skills are required as well as the capability to handle changes in workflow.
ESSENTIAL JOB FUNCTIONS
Be AMD Sunnyvale's Focussed Ion Beam (FIB) expert. Perform delicate FIB device modifications. Develop sample preparation techniques for Transmission Electron Microscopy using FIB. Represent Materials Technology Development (MTD) department on problem solving teams. Set and drive MTD's FIB roadmap.
Please contact Bryan Tracy
bryan.tracy-at-amd.com 408-749-4819
Bryan Tracy Materials Technology Development Advanced Micro Devices 915 Deguigne Ave, Mailstop 143 Sunnyvale, Ca, 94088
Hello, I am looking for a good review paper concerning immuno EM methods. There was a book suggested recently "Fine Structure Immunocytochemistry", which I am trying to locate through my library here at UNM, but I am really more interested in a few good papers. Thanks in advance, Joyce A. Kotzuk Univ. of New Mexico pathology dept.
} } Dear Lauren, } } While it looks to you like both of these adjustments affect just } brightness, they are actually have a profound influence on the coherence of } the light hitting the sample. To test, move the objectives out of position } and place a piece of white card vertically on the stage so that you can } catch the beam emerging from the condenser. Open and close the condenser } and watch the beam go from a wide angle to a narrow pencil. Open the } condenser then focus it up and down. Notice how the location of the cone } changes. } } The narrower the beam, the more coherent the light impinging on the sample. } The more coherent, the more visible the diffraction effects at edges. For } nearly invisible specimens like cheek cells, the diffraction will enhance } edge information. If you go to far, you will see the bright and dark } fringes which result from the constructive and destructive interference of } diffracted waves hitting the edges. } } 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 } } } } At 03:49 PM 7/6/00 -0500, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Yes, I did show my kids some spectacular show with LN2. You could make beautiful white clouds pouring LN2 in warm water. The most successful demonstration was, when I was using 1 liter calibrated flask with long neck. White cloud coming from the neck forms kind of shape my kids called jinn from "1000 +1 night Shaherezad". We also used LN2 in some shows at traditional New Year Party at my Institute in Russia. One time I was using regular thermos filled with LN2 to transport my samples from Novosibirsk to Moscow (8 hours fly). People at Novosibirsk were short in dry ice, we were using LN2 instead (smile). You could also put a small piece of dry ice into 1.5 ml Eppendorf tube and place it under the door of your boss (a couple of tubes are better). They are blow out with nice sound like a shot from good pistol (but less dangerous). Honestly speaking, I did it one time.
} Now there's a thought. To digress a little, we once had an instance where a } guy came down to the SEM lab and asked if he could borrow a 5 litre dewar } to take some LN2 home and show his kids. When asked for more details his } game plan was to sit the dewar on the floor on the passenger side of the } car so that he could make sure that it wouldn't fall over. The answer was } "No, perhaps you should bring the kids to the LN2...." } } The point being that even though LN2 use might seem less of a risk than } boiling water when used "properly" and "common sense" is applied, it cannot } be assumed without question that everyone's initial mental picture behind } these two terms is exactly the same!!
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
} } * Hello to all. } * I am looking for a supplier of acrylic (Plexiglas) implosion guards for } a 12 } * inch diameter glass bell jar for a vacuum evaporator. I can only seem } to find } * the wire mesh or expanded metal types. I would like to know if anyone } has such } * a thing or knows who fabricates them or supplies them off the shelf. } } * Reply on line or off line direct to me at aberginc-at-cressington.com. } }
Acid alcohol removes tol blue and many other stains. Use 50 to 70% Ethanol with a 1ml of 1N HCl per 20ml, then rinse with water. When used quickly only little destaining occurs and the tol blue differentiates giving nice pinks and blues 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 ABN: 99 724 136 560 www.proscitech.com
On Friday, July 07, 2000 4:48 AM, "Michelle.Taurino-at-aventis.com"-at-sparc5.microscopy.com [SMTP:"Michelle.Taurino-at-aventis.com"-at-sparc5.microscopy.com] wrote: } } } } Hello- } } I am looking for any suggestions on how to remove Tol. blue from Epon } embedded sections? } } I've tried warm water, 100% ethanol, and acetone. } } Any other recommendations? } } Will the Tol. blue interfere with UA/Pb staining if I do not remove the Tol. } Blue?? } } Thank you- } Michelle Taurino } Aventis Pharmaceuticals } Bioimaging and Molecular Histology } Tel-908-231-3357 } Fax-908-231-3962 } e-mail: Michelle.Taurino-at-aventis.com }
Hi, I am looking for 1) possible SEM accessory provider's list (that can supply straining stages) and 2) interaction with somebody with some prior experience with straining stages in SEM; since I want to perform the following experiment: Compression of a coated superally cube with adjustable dimensions (e.g. 10*10*6mm) either by direct compression or by bending. If possible, I would also like to repeat this experiment at various temperatures (say up to 1200C). The above experiment is to be observed under an SEM which is Hitachi S800.
Thanks Rahul Panat ------------------------------------------------------------------------ Rahul Panat Dept of Theoretical & Applied Mechanics Univ of Illinois -at- Urbana-Champaign panat-at-students.uiuc.edu ------------------------------------------------------------------------
I am working on a resolving a bit of a sticky problem. A customer wants to confirm or disprove whether complex Ti/W/Nb carbo-nitrides in a Ni base alloy do indeed contain nitrogen. I have 2 SEMs, both with WDS. The better of the 2 uses an LSM080 crystal for the nitrogen wavelengths, the other has an LOD. My approach so far has been to establish peak ratios on cp titanium and a TiN standard. Some of the numbers look promising, but I suspect an influence on the ratios due to the presence of: W, Nb, and proximity of the matrix. The carbides are small and needle shaped, perhaps 1-2 by 10-15 microns. Any comments or suggestions would be greatly appreciated! Thanks, Chris Holp Reply through: Hm: {mailto:cholp-at-ncweb.com} cholp-at-ncweb.com , Wk: {mailto:holpcr-at-earthlink.net} holpcr-at-earthlink.net , or through the group.
I am looking for low-budget camera w/board: -CCD digital monochrome (630 nm); -12-14 bit dynamic range; -min format 512x512; -shutter; -external triggering; -1 frame/s min rate; -PC-compatible board. Does anybody have any used equipment for sale? Any input on the sources for inexpencive new cameras with such specs will be appreciated.
Best regards
Dmitri Lapotko Luikov Heat and Mass Transfer Institute Minsk, Belarus
We use the Sevier-Munger modification found in Sheehan's Histotechnology text for our neuropath stains. It is more reproducible than the Bodian stain. The protocol is lengthy; will fax if you don't have access to the text.
-----Original Message----- } From: Phillip Rutledge [mailto:prutledge-at-ars.usda.gov] Sent: Thursday, July 06, 2000 8:49 AM To: Microscopy-at-sparc5.microscopy.com
Microscopists,
I know this isnt't an EM related question but I'm in search of a couple of LM staining procedures. I'm looking for a good silver stain (s) for looking at neural structures preferably not using chloral hydrate, and a good acid fast procedure for mycobacterium. If anyone can help, I'd appreciate it.
Alan Berginc 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 to all. } } * I am looking for a supplier of acrylic (Plexiglas) implosion guards for } } a 12 } } * inch diameter glass bell jar for a vacuum evaporator. I can only seem } } to find } } * the wire mesh or expanded metal types. I would like to know if anyone } } has such } } * a thing or knows who fabricates them or supplies them off the shelf. } } } } * Reply on line or off line direct to me at aberginc-at-cressington.com. } } } } } } Regards.........Alan Berginc } Cressington Scientific, INC
Dear Alan,
Please check our web site, http://www.laddresearch.com for catalog # 30110 which I believe is what you are looking for.
Debbie Sicard --
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
EM people/, does anyone know of a supplier who sells the Zoakes Teflon Helix stirrer? I believe that was its name. It provided an efficient way to mix resins without entraining air into the mix. I haven't seen it listed in the any of the recent catalogues.
Hank Adams Manager Integrated Microscopy Core Molecular and Cellular Biology Baylor College of Medicine Houston, Tx 77030
At 01:48 PM 7/6/00 -0500, Michelle.Taurino-at-aventis.com"-at-sparc5.microscopy.com wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Just wash you slids with 100% ETHYL ALCOHOL and after that wash with dd water it should do it
We are seeking a Material Scientist to perform TEM and SEM characterization of materials, particularly nuclear waste forms intended for repository disposal. Measure and evaluate the properties of all types. Participate in planning, developing, and implementing materials research to develop ceramic materials for waste forms and process equipment components.
The successful candidate will: o Conduct and direct materials characterization, with special emphasis on TEM examination of nuclear waste forms. o Analyze, evaluate, document, and communicate results of experiments and programs. o Contribute to and participate in internal and external Laboratory technical reporting and presentations. o Collaborate with other technical staff in the specialty area of materials science. o Ensure that the TEM is properly maintained.
We are seeking candidates with: o Ph.D. or M.S. in Materials Science, Ceramics or related field. o US Citizenship and ability to obtain DOE security clearance required.
Argonne offers a professional environment, excellent working conditions and fringe benefits. Please mail or e-mail your resume, quoting Ref: Material Scientist, ANL-W-1234, to: Argonne National Laboratory-West Human Resources P.O. Box 2528 Idaho Falls, ID 83403-2528. E-mail: hr-at-anlw.anl.gov Equal Opportunity Employer
Deear Chris, You might want to look at my article on N in Ti in the August, 1999 issue of Microscopy Today. By looking carefully at the peak shape of pure Ti vs. TiN , I found the optimum location on the peak to test for the presence of N. The W and Nb should not interfere at this wavelength and you should be able to see if there is N above a detection limit of less than 0.5 weight percent. The other thing that might help is to do an extraction replica of the inclusions, so that they are free of the Ni matrix. Lay this down on a carbon stub to reduce the background and improve your peak-to-background ratio. At 04:34 PM 7/8/00 -0500, you wrote:
} I am working on a resolving a bit of a sticky problem. A customer wants } to confirm or disprove whether complex Ti/W/Nb carbo-nitrides in a Ni base } alloy do indeed contain nitrogen. I have 2 SEMs, both with WDS. The } better of the 2 uses an LSM080 crystal for the nitrogen wavelengths, the } other has an LOD. My approach so far has been to establish peak ratios } on cp titanium and a TiN standard. Some of the numbers look promising, but } I suspect an influence on the ratios due to the presence of: W, Nb, and } proximity of the matrix. The carbides are small and needle shaped, } perhaps 1-2 by 10-15 microns. Any comments or suggestions would be } greatly appreciated! Thanks, Chris Holp Reply through: Hm: } {mailto:cholp-at-ncweb.com} cholp-at-ncweb.com , Wk: } {mailto:holpcr-at-earthlink.net} holpcr-at-earthlink.net , or through the group. Good 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
I recently acquired a unitron MeC3-2313. It is a 30 year old, mono-ocular, metallurgical, inverted, optical microscope. The images are high resolution, bright, and the stage works well. I want to connect it to a digital camera. It came w/ a T-mount and I can get a C-mount from unitron for $625, I suppose I need a digital camera that has a removable lens. I have seen specs for a the SV Micro ($2200) and the Pixera ($1200). Does anybody have any other cameras they would like to suggest or experience w/ these camera? Is the overall image quality good for theses two cameras? I am afraid that the resolution would "not be there" in the color mode for the Pixera. I am not sure I want a consumer camera that has been adapted for a microscope like the Nikon and Kodak, since they include a lens that cannot be re-moved and therefore the minimum magnification work would be "upped" by 3X, if I could go thru the ocular. Plus, I am not sure I can get adaptors for my scope anyway. I would also like to keep my only ocular open, so I can skip the whole digital phase while doing sample inspection. Like to keep the entire endevor under $3K.
We have both of those cameras attached to two different stereomicroscopes. The SV-micro produces a bit better picture but is not as easy to work with especially getting the exposure just right. I like using the Pixera -its quick and easy. We don't do anything critical with these cameras however. But they work well for the price.
For adapters, contact the folks at Diagnostic Instruments. Their web site is http://www.diaginc.com/ccdguide.htm
-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: Smartech [mailto:smartech-at-javanet.com] } Sent: Monday, July 10, 2000 12:44 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Another digital camera question } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } 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 acquired a unitron MeC3-2313. It is a 30 year } old, mono-ocular, } metallurgical, inverted, optical microscope. The images are high } resolution, bright, and the stage works well. I want to } connect it to a } digital camera. It came w/ a T-mount and I can get a C-mount } from unitron } for $625, I suppose I need a digital camera that has a } removable lens. I } have seen specs for a the SV Micro ($2200) and the Pixera } ($1200). Does } anybody have any other cameras they would like to suggest or } experience w/ } these camera? Is the overall image quality good for theses } two cameras? I } am afraid that the resolution would "not be there" in the } color mode for the } Pixera. I am not sure I want a consumer camera that has been } adapted for a } microscope like the Nikon and Kodak, since they include a } lens that cannot } be re-moved and therefore the minimum magnification work } would be "upped" by } 3X, if I could go thru the ocular. Plus, I am not sure I can } get adaptors } for my scope anyway. I would also like to keep my only } ocular open, so I } can skip the whole digital phase while doing sample } inspection. Like to } keep the entire endevor under $3K. } } Thanks } } Ric } }
I am currently searching for a NIST traceable TEM mag. standard. The reason for a NIST traceable standard is that our EM facility is in transition to becoming GMP regulated. I have used the waffle grating replica and catalase crystals in the past and prefer this type of standard but they are not NIST traceable. I am aware that NIST offers polystyrene nanosphere particles for sizing but was hoping to avoid using because of the difficulty in producing a uniform monolayer without aggregate particles. I am also aware that there is a product called MAGICAL and am still waiting from a response from NIST as to whether or not it is traceable. Does anyone know of any 3mm prepared grid for use as a TEM mag std traceable to NIST. I am performing magnification checks from 1,000X through 200,000X. Any info would be much appreciated.
David Cugier Assistant Scientist Microscopy and Microanalysis Abbott Laboratories Abbott Park, Illinois 60064 847-938-6725
I have a Zeiss OPMI-1 binocular microscope. I got it as part of an auction lot. I believe it was used by the ENT department of a medical school.
If it can be restored to working condition, that would be great. If not, I'd be glad to sell it.
I don't know what it's worth, or really what it needs to be restored to working order. If it were pristine and on a stand, that would be another matter, it's clear. On the other hand, I don't know exactly which model it is.
I have several images of it at http://www.threedee.com/sales/zeiss.html . Any assistance would be appreciated!
I do not know what GMP certification, but our lab is QS9000 certified. That is the automotive ISO9001 equivalent. I use the Mag-i-cal sample since all lengths of the sample were traceable to lattice spacings of the Si crystal. John McCaffrey has a letter from NIST, a copy of which that can accompany the Mag-i-cal sample, that states that they do not currently have a standard for calibrating TEMs nor do they have a calibration for the crystalline lattice spacings of Si. They state that the crystalline lattice is an intrinsic property of a material, well characterized, well documented in the literature, and known to 6 decimal places. I have placed this letter in my calibration standards file along with the certification that comes with it. If the GMP is anything like ISO or Qs certification, what is important is it is that you have the paper work and procedures documented and that you follow them.
This should be close enough for government work.
-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: David Cugier [mailto:david.cugier-at-abbott.com] } Sent: Monday, July 10, 2000 3:54 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: NIST traceable TEM magnification standards. } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } 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 currently searching for a NIST traceable TEM mag. } standard. The reason } for a NIST traceable standard is that our EM facility is in } transition to } becoming GMP regulated. I have used the waffle grating } replica and catalase } crystals in the past and prefer this type of standard but } they are not NIST } traceable. I am aware that NIST offers polystyrene } nanosphere particles for } sizing but was hoping to avoid using because of the } difficulty in producing a } uniform monolayer without aggregate particles. I am also } aware that there is } a product called MAGICAL and am still waiting from a response } from NIST as to } whether or not it is traceable. Does anyone know of any 3mm } prepared grid } for use as a TEM mag std traceable to NIST. I am performing } magnification } checks from 1,000X through 200,000X. Any info would be much } appreciated. } } } David Cugier } Assistant Scientist } Microscopy and Microanalysis } Abbott Laboratories } Abbott Park, Illinois 60064 } 847-938-6725 }
On Mon, 10 Jul 2000 RetTek2000-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 } -----------------------------------------------------------------------. } } } }
There is no GMP/GLP certification given by the FDA nor are there FDA certified laboratories. There is only compliance and being subjected to an unannounced FDA audit. Where the various ISO9000 standards are orientated towards improvement of quality and yield, GMP is a different standard orientated towards having the sample used in a legal trial. For example, a company may tolerate 99% yield for economic reasons, however, how many babies on a percentage basis is a doctor allowed to drop a year. Failure to comply with ISO9000 may have some economic consequencies. Caught failing to comply with GMP, the FDA will consider the medical device or drug adulterated until proven otherwise.
J. Roy Nelson, Ph.D. Material Testing Laboratory mtl-at-njcc.com
"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 } -----------------------------------------------------------------------. } } I do not know what GMP certification, but our lab is QS9000 certified. That } is the automotive ISO9001 equivalent. I use the Mag-i-cal sample since all } lengths of the sample were traceable to lattice spacings of the Si crystal. } John McCaffrey has a letter from NIST, a copy of which that can accompany } the Mag-i-cal sample, that states that they do not currently have a standard } for calibrating TEMs nor do they have a calibration for the crystalline } lattice spacings of Si. They state that the crystalline lattice is an } intrinsic property of a material, well characterized, well documented in the } literature, and known to 6 decimal places. I have placed this letter in my } calibration standards file along with the certification that comes with it. } If the GMP is anything like ISO or Qs certification, what is important is it } is that you have the paper work and procedures documented and that you } follow them. } } This should be close enough for government work. } } -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: David Cugier [mailto:david.cugier-at-abbott.com] } } Sent: Monday, July 10, 2000 3:54 PM } } To: Microscopy-at-sparc5.microscopy.com } } Subject: NIST traceable TEM magnification standards. } } } } } } -------------------------------------------------------------- } } ---------- } } The Microscopy ListServer -- Sponsor: The Microscopy Society } } 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 currently searching for a NIST traceable TEM mag. } } standard. The reason } } for a NIST traceable standard is that our EM facility is in } } transition to } } becoming GMP regulated. I have used the waffle grating } } replica and catalase } } crystals in the past and prefer this type of standard but } } they are not NIST } } traceable. I am aware that NIST offers polystyrene } } nanosphere particles for } } sizing but was hoping to avoid using because of the } } difficulty in producing a } } uniform monolayer without aggregate particles. I am also } } aware that there is } } a product called MAGICAL and am still waiting from a response } } from NIST as to } } whether or not it is traceable. Does anyone know of any 3mm } } prepared grid } } for use as a TEM mag std traceable to NIST. I am performing } } magnification } } checks from 1,000X through 200,000X. Any info would be much } } appreciated. } } } } } } David Cugier } } Assistant Scientist } } Microscopy and Microanalysis } } Abbott Laboratories } } Abbott Park, Illinois 60064 } } 847-938-6725 } }
Do any of you have problems getting resin blocks to release from the flat, blue embedding molds? We have used these for years with no trouble but the newer molds don't let the blocks go after just a few embeddings. I have had to cut them out with a razor blade which of course ruins the mold. Has the formula for the molds changed or is it the resin? We use Spurr's and Eponate 12 primarily.
Thank you, Mary Gail Engle Electron Microscopy & Imaging Facility University of Kentucky Medical Center
David Cugier wrote: ============================================================== I am currently searching for a NIST traceable TEM mag. standard. The reason for a NIST traceable standard is that our EM facility is in transition to becoming GMP regulated. I have used the waffle grating replica and catalase crystals in the past and prefer this type of standard but they are not NIST traceable. I am aware that NIST offers polystyrene nanosphere particles for sizing but was hoping to avoid using because of the difficulty in producing a uniform monolayer without aggregate particles. I am also aware that there is a product called MAGICAL and am still waiting from a response from NIST as to whether or not it is traceable. Does anyone know of any 3mm prepared grid for use as a TEM mag std traceable to NIST. I am performing magnification checks from 1,000X through 200,000X. Any info would be much appreciated. =============================================================== This is a really tough question to answer. The Mag*I*Cal TEM calibration standard comes "close" but "close" in this business is not quite enough. But it might be the best there is. You can get information about the Mag*I*Cal at URL http://www.2spi.com/catalog/standards/magical.html
You are correct about the problems associated with using the polystryene calibrated spheres.
It is sold by SPI Supplies as well as some of the other major suppliers of accessories and consumables for EM laboratories.
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 ######################## ============================================
This exactly mirrors the problem I have noticed in the last couple of years. My blue molds formerly lasted for years and now they deteriorate rapidly. We use an epon 812 style embedding medium. I have taken to spraying the molds with teflon spray (708 T.F.E. dry lube - a lot cheaper from the local hardware store than from EM supply houses!) and that seems to help without hurting morphology. but i can't help but wonder what has changed in molds!
} } } Do any of you have problems getting resin blocks to release from the } flat, blue embedding molds? We have used these for years with no } trouble but the newer molds don't let the blocks go after just a few } embeddings. I have had to cut them out with a razor blade which of } course ruins the mold. Has the formula for the molds changed or is } it the resin? We use Spurr's and Eponate 12 primarily. } } Thank you, } Mary Gail Engle } Electron Microscopy & Imaging Facility } University of Kentucky Medical Center
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 might have been a bit more helpful if you had suggested that users should contact NIST and request that they evaluate the MAG*I*CAL, since the industry now requires a traceable standard.
Cheers John
John P. McCaffrey, Ph.D. National Research Council of Canada M-50, Montreal Rd. Ottawa, Ontario K1A 0R6 CANADA
-----Original Message----- } From: Garber, Charles A. [mailto:cgarber-at-2spi.com] Sent: Tuesday, July 11, 2000 2:38 PM To: MICROSCOPY BB
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
David Cugier wrote: ============================================================== I am currently searching for a NIST traceable TEM mag. standard. The reason for a NIST traceable standard is that our EM facility is in transition to becoming GMP regulated. I have used the waffle grating replica and catalase crystals in the past and prefer this type of standard but they are not NIST traceable. I am aware that NIST offers polystyrene nanosphere particles for sizing but was hoping to avoid using because of the difficulty in producing a uniform monolayer without aggregate particles. I am also aware that there is a product called MAGICAL and am still waiting from a response from NIST as to whether or not it is traceable. Does anyone know of any 3mm prepared grid for use as a TEM mag std traceable to NIST. I am performing magnification checks from 1,000X through 200,000X. Any info would be much appreciated. =============================================================== This is a really tough question to answer. The Mag*I*Cal TEM calibration standard comes "close" but "close" in this business is not quite enough. But it might be the best there is. You can get information about the Mag*I*Cal at URL http://www.2spi.com/catalog/standards/magical.html
You are correct about the problems associated with using the polystryene calibrated spheres.
It is sold by SPI Supplies as well as some of the other major suppliers of accessories and consumables for EM laboratories.
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 ######################## ============================================
The Department of Biological Sciences at Northern Illinois University has an immediate opening for a Research Associate in Confocal/Electron Microscopy. Details are in the advertisement below.
Thanks
Michael Parrish, Professor and Chair Biological Sciences Northern Illinois University DeKalb, IL 60115 815-753-1753
Research Associate:
The Department of Biological Sciences at Northern Illinois University has an opening for a Research Associate with expertise in Confocal and Electron Microscopy. The successful candidate will be expected to operate our confocal, transmission and scanning electron microscopes and to supervise both students and faculty in their use. Master's degree in Biology or related field required including at least two years' experience using confocal and electron microscopes in biological or chemical research. Ideally, the successful candidate should be able to calibrate and perform routine maintenance on the instruments. Submit letter of interest, resume and three letters of recommendation to: Dr. Michael Parrish, Chair, Department of Biological Sciences, Northern Illinois University, DeKalb, IL 60115-2861; http:\\www.bios.niu.edu; phone: 815-753-0461; email: biosjobs-at-niu.edu. Review of applications will begin July 28, 2000 and continue until position is filled.
As a relative novice on JEOL 2010 TEM, I was recently advised to use a cold trap in the specimen area. Since I know some people don't use it even though it's mounted on their 'scope, I'd like to hear your opinion on when and how frequently to use it. I understand it improves the vacuum significantly; is there any drawback? Any guidelines or suggestions on the liquid N2 /heat-up cycle? Thank you, Alice.
Alice Dohnalkova Environmental Microbiology Battelle, PNNL Richland, WA (509) 372-0692
HI Alice, Use the cold trap. Use the cold trap. Did I say use the cold trap? Jeol calls it the anti contamination device (ACD). Your vacuum will be better, sample swap time quicker, much quicker. The cold trap will pump (trap) most emissions from your sample. This is a good thing. It keeps them from carbonizing on your apertures or oxidizing the filament. Ever noticed really strange images of your filament that eventually went away? Better vacuum will extend your filament life. I find it hard to believe (I believe you) that users in your facility are not using the ACD. I could rant on a bit longer.... There is one catch, a testament to it's pumping efficiency. When you are finished, you must use the ADC heat cycle. The ACD will have accumulated so many molecules that the ion pump can not handle the gas load if the ACD warms up on it's own. The ACD heat cycle will switch the column over to the diffusion pump for a period of time. In our lab, the last user signed up (weather or not they use the TEM) is responsible for tending to the ACD. In our lab, failure to use the ACD heat cycle is a high crime. It creates a vacuum condition (in our 2010) that causes the gun isolation valve to cycle at ~0.5 Hz. This then causes the bellows to fail, typically in ~10,000 cycles (a long weekend). The effect is cumulative. So now that I said use it. how... fill the dewier, wait ~5min & top it off. Put a foil cap or other loose fitting cap over the fill port. It should stay cold for 7 hours. That is the Spec. on our unit. It varies a bit with # of sample changes. I have seen it cold 11 hours later. Don't ask how I know this :)
Bruce Brinson Rice U.
"Dohnalkova, Alice" 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 } -----------------------------------------------------------------------. } } } As a relative novice on JEOL 2010 TEM, I was recently advised to use a cold trap } in the specimen area. Since I know some people don't use it even though it's } mounted on their 'scope, I'd like to hear your opinion on when and how } frequently to use it. I understand it improves the vacuum significantly; is } there any drawback? Any guidelines or suggestions on the liquid N2 /heat-up } cycle? Thank you, Alice. } } Alice Dohnalkova } Environmental Microbiology } Battelle, PNNL } Richland, WA } (509) 372-0692
I routinely use a combination of LN2 and ion getter pumps on my Zeiss EM 10. The scope is also kept running 24 hours a day, 7 days a week. This combination has afforded me with extremely good vacuum, less downtime, and an incredible life time on tungsten filaments. The filament I am currently using has 430 hours: my record is 520 hours! This of course is also relately to slow saturation and desaturation of the filament whenever I turn off or on the high tension, completely outgassing of film after film exchange and using the scope with liquid nitrogen.
The only disadvantage occurs when the LN2 is depleted and the cold finger around the specimen starts to return to ambient temperature, i.e., there is a substantial rise in vacuum pressure until the cold finger warms up. The advantages I have experienced include long filament life, clean high vacuum, and no contamination on the grid/specimen. I routinely use the scope at magnifications from x1600 to x125,000 and have not notice degradation of image quality, either from hydrocarbon contamination or an aged filament.
I pay about $47.00 for 50 liters of LN2 every 5 to 7 weeks: worth every cent!! One must however get into the habit of using filling the scope dewar about 20 to 30 minutes before using the scope.
Sincerely, Ken ------- Ken Tiekotter, Adjunct Professor The University of Portland Dept. of Biology 5000 N. Willamette Blvd. Portland, OR 97203
Director, MicroImaging/Electron Micrscopy, G50 Legacy Holladay Park Medical Center 1225 NE 2nd Avenue Portland, OR 97232
TEL.: (503) 413-5391
On Tue, 11 Jul 2000, Dohnalkova, Alice 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 } -----------------------------------------------------------------------. } } } } As a relative novice on JEOL 2010 TEM, I was recently advised to use a cold trap } in the specimen area. Since I know some people don't use it even though it's } mounted on their 'scope, I'd like to hear your opinion on when and how } frequently to use it. I understand it improves the vacuum significantly; is } there any drawback? Any guidelines or suggestions on the liquid N2 /heat-up } cycle? Thank you, Alice. } } Alice Dohnalkova } Environmental Microbiology } Battelle, PNNL } Richland, WA } (509) 372-0692 } } }
Hallo Folks! This is a reply to a previous posting of mine regarding a spurious B peak in carbonate spectra plus another matter altogether. Firstly, for Warren S's info, we have a refurbished Kevex detector with a Si (Li) crystal and an ultra thin Moxtex window (3 microns) attached to our JEOL 733. The pulse processing electronics are 1976 vintage Kevex 4050 which give us a pitiful ca. 1200 cps at 25% dead time. C peak occurs in the correct place. To answer Brendan G., the oxygen peak is in the correct place, and everything seems to be in order apart from the spurious peak. Basically, the system is calibrated to give the correct things in the correct place. Now to another problem, and this has to do with the window itself. We have been watching the Cu l:k alpha ratio deteriorate over time since installation 4 months ago from acceptable values over 1 to an alarming 0.44 (intergrated peak ratio specified within WinEDS software). Micro droplets of oil are starting to appear on other wise clean standards (although these are few and dispersed rather than a slick, and I am not alarmed by this considering all other things. I'm actually happy after 1.5 years to be able to analyse things!!), and I suspect that the window is in need of a wash off. The company who outsourced the refurbishment, chose the window, and installed the system have told me that ultra-thin Moxtex windows cannot be cleaned. If this is true, then we have been stitched up. Does anyone out there know anything about these things? Some advice, as usual, very helpful to us L-drivers. 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: m.roberts-at-ru.ac.za SOUTH AFRICA
Bruce has got it right - use the ACD and heat it up overnight. Any cold trap will help remove hydrocarbons and water vapour from your vacuum system.
A few of extra notes. - Fill the trap as soon as you get to the instrument to get it cold. It can be cooling down while you ramp up the HT and load a specimen. - Beware that some dewars throw out a quantity of gas and sometimes liguid N2 shortly after they have been filled from room temperature. Do not sit under the dewar for 5 minutes after first filling. - After you hit the ACD heat button the vacuum system locks up for about 2 hours while the ACD warms up and the trapped gasses are pumped away by the diff pump. Remember to change films and specimen and let them pump down properly before hitting the ACD button.
I am at a materials lab rather than a biological one, our users need the 2010 for small probe work and that shows up any contamination problem. A routine of column bake out and use of the ACD means that I can be sure that contamination problems come from their specimen.
Regards, Ron
On Tue, 11 Jul 2000, Dohnalkova, Alice 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 } -----------------------------------------------------------------------. } } } } As a relative novice on JEOL 2010 TEM, I was recently advised to use a cold trap } in the specimen area. Since I know some people don't use it even though it's } mounted on their 'scope, I'd like to hear your opinion on when and how } frequently to use it. I understand it improves the vacuum significantly; is } there any drawback? Any guidelines or suggestions on the liquid N2 /heat-up } cycle? Thank you, Alice. } } Alice Dohnalkova } Environmental Microbiology } Battelle, PNNL } Richland, WA } (509) 372-0692 } } }
=========================================================================== 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. ============================================================================
While we are on a slight cold trap / ACD thread, I would greatly appreciate your views about the desirability of using cold traps and cold fingers in Field emission SEMs. Some manufacturers have them, others don't and regard them as unnecessary. Some manufacturers exchange specimens by opening the chamber to air, others do it via airlocks. If one is aiming to maximise the performance of a FESEM for low-kV and LTSEM imaging it would seem to me to be desirable not to expose the specimen chamber to atmospheric pressure during specimen changes, and to trap any volatiles to reduce specimen contamination, just as we would normally do in TEM practice. Do users of FESEMs find in practice that there are specimen contamination issues which make the use of cryo-trapping and airlocks desirable, or are these features unnecessary?
Best wishes 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 07710 585 401 email c.jeffree-at-ed.ac.uk SEM / TEM bookings sem-at-ed.ac.uk =====================================================================
I am sending this request for a researcher who is not a memeber of the list so kindly respond to her (Aleksandra Perovic) directly at:
perovic-at-mcmaster.ca
We want to electropolish this alloy which contains the following phases.
1) Q phase: 30 wt% Mg 30 wt% Si 16 wt% Al & 20 wt% Cu.
2) Theta phase: Al2Cu
3) Si:
4) Beta phase: Mg2Si
We have already tried ion milling and also electropolishing with 10% perchloric/methanol, with no success.
If anyone has a recipe for such an alloy we would appreciate hearing from you.
Thanks in advance
Fred
c/o Aleksandra Perovic
******************************************************** Fred Pearson Brockhouse Institute for Materials Research McMaster University 1280 Main St. West Hamilton, Ontario Canada L8S 4M1
We have an Hitachi S4700 FESEM, and we routinely use the LN2 coldfinger. In my experience, it reduces deposition of contamination under the beam, especially during low voltage operation (1-5kV, say).
The disadvantages so far have been vibration introduced by the boiling LN2, although this pretty much disappears after about 20-30 minutes, and the problem of retrieving a sample which has become dislodged inside the chamber. Although the latter occurrence is rare, it obviously requires opening up the entire chamber to atmosphere, which isn't a great thing to do with a chilly cold finger combined with the humidity of mid-Missouri. In this case, you need to wait until the system comes to room temperature, effectively shutting down the scope for several hours.
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: Chris Jeffree [mailto:cjeffree-at-srv0.bio.ed.ac.uk] Sent: Wednesday, July 12, 2000 7:29 AM To: microscopy-at-sparc5.microscopy.com
While we are on a slight cold trap / ACD thread, I would greatly appreciate your views about the desirability of using cold traps and cold fingers in Field emission SEMs. Some manufacturers have them, others don't and regard them as unnecessary. Some manufacturers exchange specimens by opening the chamber to air, others do it via airlocks. If one is aiming to maximise the performance of a FESEM for low-kV and LTSEM imaging it would seem to me to be desirable not to expose the specimen chamber to atmospheric pressure during specimen changes, and to trap any volatiles to reduce specimen contamination, just as we would normally do in TEM practice. Do users of FESEMs find in practice that there are specimen contamination issues which make the use of cryo-trapping and airlocks desirable, or are these features unnecessary?
Best wishes 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 07710 585 401 email c.jeffree-at-ed.ac.uk SEM / TEM bookings sem-at-ed.ac.uk =====================================================================
Dito here. The vendor asked if I was embedding at high temps but we use araldite or 812 at 60 - 65 degree. We sometimes sprayed the molds with high purity silicone which would extend the life after they started sticking. The clear ones also got hard and opaque, along with ambrose silicone molds.
The Australian, and now the various State Governments require permits for the import, storage and transshipping of UA (also a store and radiation safety officer). We will in future drop-ship to the end user direct and therefore not handle UA at all. The crazy rules are laid bare in page, which is linked from our UA item online.
I would like to know from international subscribers (can be back-channel) of any permits required to import or transship UA in other countries. I know that UA can be shipped by normal air freight and through the Postal system anywhere. The USA definition of a radioactive substance is 7x higher than the Australian one. The new legislation will cost Australian labs a good deal of extra time and money. It would be useful to protest to the various Health ministers, State and Federal.
I would also like to know of any significant incident involving UA anywhere. I should point out that radiation damage would not show for years and one would never know the original cause. The radiation though is low, so unless a person would take to sleeping with a jar of UA there is no real danger. The real danger of UA is as a non-cumulative (happily), but severe kidney poison, but this is of course no issue with the Radiation people. 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 ABN: 99 724 136 560 www.proscitech.com
The P2 projector lens of our TEM Siemens Elmiskop 101 is out of order.
We need to replace it as soon as possible.
Could anyone help us ?
Here are the features of our microscope :
Siemens Elmiskop 101 - Fabrication number : 2368 Serial number : M31191-A5 High voltage working between 40 and 100 kV - 60 µA
Thanks in advance for your help
Best regards
Philippe Drouillon Solvay Research and Technology Analytical Technologies Dpt Electron Microscopy and Image Analysis Labs - IT coordinator of the Dpt Rue de Ransbeek, 310 B-1120 Brussels (Belgium) phone : (00 32) 2 264 24 47 mailto:philippe.drouillon-at-solvay.com
} -----Original Message----- } From: Bruce Brinson [SMTP:brinson-at-cnst.rice.edu] } Sent: Wednesday, July 12, 2000 3:00 AM } To: Dohnalkova, Alice } Cc: 'Microscopy-at-MSA.Microscopy.com' } Subject: Re: JEOL 2010 Cold Trap } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 Alice, } Use the cold trap. Use the cold trap. Did I say use the cold trap? } Jeol calls it } the anti contamination device (ACD). Your vacuum will be better, sample } swap time } quicker, much quicker. The cold trap will pump (trap) most emissions from } your } sample. This is a good thing. It keeps them from carbonizing on your } apertures or } oxidizing the filament. Ever noticed really strange images of your } filament that } eventually went away? Better vacuum will extend your filament life. } I find it hard to believe (I believe you) that users in your facility } are not } using the ACD. I could rant on a bit longer.... } There is one catch, a testament to it's pumping efficiency. When you } are } finished, you must use the ADC heat cycle. The ACD will have accumulated } so many } molecules that the ion pump can not handle the gas load if the ACD warms } up on it's } own. The ACD heat cycle will switch the column over to the diffusion pump } for a } period of time. In our lab, the last user signed up (weather or not they } use the } TEM) is responsible for tending to the ACD. } In our lab, failure to use the ACD heat cycle is a high crime. It } creates a } vacuum condition (in our 2010) that causes the gun isolation valve to } cycle at ~0.5 } Hz. This then causes the bellows to fail, typically in ~10,000 cycles (a } long } weekend). The effect is cumulative. } So now that I said use it. how... fill the dewier, wait ~5min & top } it off. Put } a foil cap or other loose fitting cap over the fill port. It should stay } cold for 7 } hours. That is the Spec. on our unit. It varies a bit with # of sample } changes. I } have seen it cold 11 hours later. Don't ask how I know this :) } } Bruce Brinson } Rice U. } } "Dohnalkova, Alice" 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 } } -----------------------------------------------------------------------. } } } } } } As a relative novice on JEOL 2010 TEM, I was recently advised to use a } cold trap } } in the specimen area. Since I know some people don't use it even though } it's } } mounted on their 'scope, I'd like to hear your opinion on when and how } } frequently to use it. I understand it improves the vacuum } significantly; is } } there any drawback? Any guidelines or suggestions on the liquid N2 } /heat-up } } cycle? Thank you, Alice. } } } } Alice Dohnalkova } } Environmental Microbiology } } Battelle, PNNL } } Richland, WA } } (509) 372-0692 }
It seems that it is a good idea to use a cold trap on JEOL 2010 TEM. But I have a concern if anyone has any suggestions. We have a LaB6 JEOL 2010. Once the ACD is hit, the HT is automatically drops down to zero. So for the next day, the HT has to be ramped up again. If we do like this, the HT will have to be ramped up everyday, and I am wondering whether this will do any harm in a long run. Thanks. Regards 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 ========================================
Randy and Listies: I too have an S4700 with a cold finger. But I don't have the time to let the cold finger come to room temperature if I need to retrieve a dropped or stuck sample. Plant A/C keeps the lab humidity below 60% and if I work fast, there is minimal frost buildup. The main chamber is back under working vacuum in less than 1 hour. The scope is equipped with a diffusion pump instead of a TMP. My question: Am I causing any problems by letting the pump system sublime the frost off the cold finger?
"Tindall, Randy D." wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Chris, } } We have an Hitachi S4700 FESEM, and we routinely use the LN2 coldfinger. In } my experience, it reduces deposition of contamination under the beam, } especially during low voltage operation (1-5kV, say). } } The disadvantages so far have been vibration introduced by the boiling LN2, } although this pretty much disappears after about 20-30 minutes, and the } problem of retrieving a sample which has become dislodged inside the } chamber. Although the latter occurrence is rare, it obviously requires } opening up the entire chamber to atmosphere, which isn't a great thing to do } with a chilly cold finger combined with the humidity of mid-Missouri. In } this case, you need to wait until the system comes to room temperature, } effectively shutting down the scope for several hours. } } 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/ }
Hi All, The Anatomy / Cell Biology Department of TEMPLE UNIVERSITY / MED SCHOOL, has three (3) Electron Microscopes it would like to Donate to a good home, for the cost of removing the item you would like. The three are as follows:
1. Philips 300 TEM.
2. JEOL 100B TEM.
3. ETEC CORP. AUTOSCAN SEM.
If you have a need for one or all three microscopes, Please contact Dr. Joanne Orth by E-mail (ASAP) -at- "orthjo-at-astro.temple.edu". The Philips and ETEC were working well a couple of years ago, I have no knowledge of the JEOL.
Microscopy and Microanalysis 2000 is August 13-17 in Philadelphia. Today I received an email reminder to register: } Register at www.peregrine.net/mm2000 When I went to this site, the URL prefix was http:, not https: The latter would indicate a secure server. The text of the web page claimed that it is secure, but Netscape warned that it is not. Since this site is not secure, despite its claim, I did not submit my credit card information online. I advise other registrants to be cautious also.
Don Chernoff
} Questions? Please refer to the MSA web site at } www.msa.microscopy.com } or contact Meeting Management at (708) 361-6045.
Advanced Surface Microscopy, Inc. E-Mail: donc-at-asmicro.com 6009 KNYGHTON RD. Voice: 317-251-1364 INDIANAPOLIS IN 46220 Toll free: 800-374-8557 (in USA) web: http://www.asmicro.com Fax: 317-254-8690 Please use the new web and email addresses shown above. (February 1999)
Here are my observations about cold fingers and cold traps in SEMs. They do a good job of fighting contamination but they do have the vibration and frosting problems noted by others. They require constant refilling to keep them cold. When they warm up the water and oil they collect mostly redistributes to the walls of the chamber and is not pumped away. After using a cold finger cold, the warm cold finger is a major source of contamination.
If a cold finger is frosted during opening a door for dislodging a sample, then pumping the frost should not overburden the vacuum pumps if done occasionally. They are designed to handle the water vapor in the atmosphere. Excessive water vapor pumping may condense water in rotary pump oil. More frequent oil changes are recommended if water vapor is pumped regularly.
I believe that some manufacturers claim cold traps are not needed because of cost and competitive issues. They are selling the merits of their dry pumping systems. However the vacuum system is not the only oil source for contamination. The specimens themselves unless they are freshly cleaned will carry a hydrocarbon scum into the chamber. Cold traps are effective for this problem even on dry pumped systems. Contamination is always observed even in the cleanest, dry-pumped SEM if long scans are used. Cold traps and cold fingers are some of the tools available to control the problem.
Notice: XEI Scientific makes anti-contamination systems for SEMs but not cold traps or cold fingers. See www.SEMCLEAN.com for information.
Ronald Vane XEI Scientific 650-369-0133
-----Original Message----- } From: Chris Jeffree {cjeffree-at-srv0.bio.ed.ac.uk} To: microscopy-at-sparc5.microscopy.com {microscopy-at-sparc5.microscopy.com}
Not a good idea to have the pumps loaded up with so much water. It does not take long to warm the cold finger: A beaker with boiling water on the external finger will rise the internals of the trap within 5 minutes to above room temperature. 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 ABN: 99 724 136 560 www.proscitech.com
On Thursday, July 13, 2000 4:50 AM, Becky Holdford [SMTP:r-holdford-at-ti.com] wrote: } } } Randy and Listies: } I too have an S4700 with a cold finger. But I don't have the time to let the } cold finger come to room temperature if I need to retrieve a dropped or stuck } sample. Plant A/C keeps the lab humidity below 60% and if I work fast, there } is } minimal frost buildup. The main chamber is back under working vacuum in less } than 1 hour. The scope is equipped with a diffusion pump instead of a TMP. } My } question: Am I causing any problems by letting the pump system sublime the } frost off the cold finger? } } "Tindall, Randy D." wrote: } } } } } } Chris, } } } } We have an Hitachi S4700 FESEM, and we routinely use the LN2 coldfinger. } } In } } my experience, it reduces deposition of contamination under the beam, } } especially during low voltage operation (1-5kV, say). } } } } The disadvantages so far have been vibration introduced by the boiling LN2, } } although this pretty much disappears after about 20-30 minutes, and the } } problem of retrieving a sample which has become dislodged inside the } } chamber. Although the latter occurrence is rare, it obviously requires } } opening up the entire chamber to atmosphere, which isn't a great thing to } } do } } with a chilly cold finger combined with the humidity of mid-Missouri. In } } this case, you need to wait until the system comes to room temperature, } } effectively shutting down the scope for several hours. } } } } 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/ } } } } -- } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } Becky Holdford (r-holdford-at-ti.com) } 972-598-1291 (pager) } KFAB Physical Analysis Lab--SEM/FIB/FA } Texas Instruments, Inc. } Dallas, TX } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } }
Gatan’s Inc. is currently looking for an enthusiastic and highly organized person to help maintain and develop our line of state-of-the-art Analytical TEM products.
In this position you will be responsible for maintaining and further improving the overall, system level, performance of our existing TEM instrumentation. You will work extensively with our mechanical, electrical and software engineering groups, as well as our manufacturing and service departments. Experience in these fields, and strong communication skills, are therefore a must.
This is a key position within our organization and will involve some national and international travel. You will be based in Pleasanton, California. Practical TEM experience will be considered as a strong plus.
Gatan offers an exciting, high technology environment within a small, but growing organization.
Interested parties, please submit your resume and salary requirements to Harold A. Brink at hbrink-at-gatan.com.
True. So this could be an argument for using turbomolecular pumps, which are tolerant of water vapour. Is it advisable to subject the cold finger dewar to thermal shock by putting boiling water into it? Chris
} From: jim {jim-at-proscitech.com.au} Send reply to: "jim-at-proscitech.com.au" {jim-at-proscitech.com.au} To: "'Becky Holdford'" {r-holdford-at-ti.com} , Microscopy ListServer {microscopy-at-sparc5.microscopy.com}
Dear all, I have a set of the Journal 'Surface Science' and 'Surface Science reports', 1990-1993 (inclusive). If anyone wants it, please get in touch with me by 20th of this month or it will go in the skip.
Best regards,
Richard Beanland
============================================================== 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."
Could somebody briefly educate me on EBSD (Electron Backscattered ...?) . I am trying to find information for a friend. He would like to know the very basics, capabilities of the technique (for example, resolution) and if there are contract labs that do this. Since he is interested in quantitative texture analysis, crystal orientation and GB analysis I assume that he is talking about getting and analyzing channeling patterns, but I could be wrong. I did not get from him what materials he is interested in. So, any information on EBSD would be appreciated.
That is why I wrote "beaker" taking that little dewar from -180 to +100 could have a shattering result. Would be nice to have a spare for that purpose. Another point to be made: close of the high vacuum system when heating the cold finger, why get the rubbish of the cold finger into your pump, especially if its an ion getter type. Cheers Jim Darley ProSciTech
On Thursday, July 13, 2000 7:48 PM, Chris Jeffree [SMTP:cjeffree-at-srv0.bio.ed.ac.uk] wrote: } True. So this could be an argument for using turbomolecular } pumps, which are tolerant of water vapour. Is it advisable to subject } the cold finger dewar to thermal shock by putting boiling water into } it? } Chris } } From: jim {jim-at-proscitech.com.au} } Send reply to: "jim-at-proscitech.com.au" {jim-at-proscitech.com.au} } To: "'Becky Holdford'" {r-holdford-at-ti.com} , } Microscopy ListServer } {microscopy-at-sparc5.microscopy.com} } Subject: RE: Cold traps and cold fingers in SEM } Date sent: Thu, 13 Jul 2000 11:26:16 +1000 } Organization: ProSciTech } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Not a good idea to have the pumps loaded up with so much water. It does not } } } } take long to warm the cold finger: A beaker with boiling water on the } } external } } finger will rise the internals of the trap within 5 minutes to above room } } temperature. } } 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 } } ABN: 99 724 136 560 www.proscitech.com } } } } On Thursday, July 13, 2000 4:50 AM, Becky Holdford [SMTP:r-holdford-at-ti.com] } } } } wrote: } } } } } } } } } Randy and Listies: } } } I too have an S4700 with a cold finger. But I don't have the time to let } } } the } } } cold finger come to room temperature if I need to retrieve a dropped or } } } stuck } } } sample. Plant A/C keeps the lab humidity below 60% and if I work fast, } } } there } } } is } } } minimal frost buildup. The main chamber is back under working vacuum in } } } less } } } than 1 hour. The scope is equipped with a diffusion pump instead of a } } } TMP. } } } My } } } question: Am I causing any problems by letting the pump system sublime } } } the } } } frost off the cold finger? } } } } } } "Tindall, Randy D." wrote: } } } } } } } } } } } } } } Chris, } } } } } } } } We have an Hitachi S4700 FESEM, and we routinely use the LN2 coldfinger. } } } } } } } } In } } } } my experience, it reduces deposition of contamination under the beam, } } } } especially during low voltage operation (1-5kV, say). } } } } } } } } The disadvantages so far have been vibration introduced by the boiling } } } } LN2, } } } } although this pretty much disappears after about 20-30 minutes, and the } } } } problem of retrieving a sample which has become dislodged inside the } } } } chamber. Although the latter occurrence is rare, it obviously requires } } } } opening up the entire chamber to atmosphere, which isn't a great thing } } } } to } } } } do } } } } with a chilly cold finger combined with the humidity of mid-Missouri. } } } } In } } } } this case, you need to wait until the system comes to room temperature, } } } } effectively shutting down the scope for several hours. } } } } } } } } 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/ } } } } } } } } } } -- } } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } } } Becky Holdford (r-holdford-at-ti.com) } } } 972-598-1291 (pager) } } } KFAB Physical Analysis Lab--SEM/FIB/FA } } } Texas Instruments, Inc. } } } Dallas, TX } } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } } } } } } } } } } } } } ===================================================================== } 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 07710 585 401 } email c.jeffree-at-ed.ac.uk } SEM / TEM bookings sem-at-ed.ac.uk } =====================================================================
Electron BackScatter Diffraction. There's a recent overview and application articles in Journal of Microscopy 195 (3), 170-185 (September, 1999) also quite a bit of info at http://www.hkltechnology.com. These folks make hardware and software for this.
cheers, John
no commercial interest, just a student
"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 } -----------------------------------------------------------------------. } } Could somebody briefly educate me on EBSD (Electron Backscattered ...?) . I } am trying to find information for a friend. He would like to know the very } basics, capabilities of the technique (for example, resolution) and if there } are contract labs that do this. Since he is interested in quantitative } texture analysis, crystal orientation and GB analysis I assume that he is } talking about getting and analyzing channeling patterns, but I could be } wrong. I did not get from him what materials he is interested in. So, any } information on EBSD would be appreciated. } } Thanks } } Jordi Marti } Honeywell
Some years ago I stumbled over an article in the German issue of "Scientific American". I think it was Peter Barham, who wrote about things you can do with physics in the kitchen. So we tried to make ice with LN2 and the result was just wonderful. And you have a nice visual effect and your coworkers will praise you ...
Here is the howto for a small department:
We found strawberry ice the best - take 2 kilos of strawberries, clean them and mash them - take at least 1/2 kilo of sugar - 2 liters of milk - 1/2 liter of sweet cream - add vanilla sugar and skin of a lemon
- precool it in the fridge - fill all the ingredients into a metal bowl, plastic will crack, use gloves - add the LN2, you may need about the same amount as the ingredients - mix and mix and mix ... - important: you have to stir like an idiot to get small ice crystals
- bon appetite
Andreas Loewe
Ps: You'll find more about this in a book by: Herve This-Benckhard, "Les secrets de la casserole", Springer, 1993
=========================================================================== Andreas Loewe e-mail : loewe-at-uni-bonn.de Department of Inorganic Materials Research Phone : (+49-228)-73-4180 University of Bonn Fax : (+49-228)-73-4205 Roemerstr. 164 53117 Bonn ===========================================================================
Does anyone have information regarding performance standards for clinical technicians. ie. How many cases per month per technician? Thanks for any feedback.
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We currently have a position for a scanning electron microscopist in our Mississsauga location.
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Dear Jordi, We have a EBSD system installed here. It is an SEM accessory that picks up the Kikuchi lines that scatter off a flat crystalline surface. It is the same as electron channeling, but picked up from the side by a TV camera. The sample must be strain-free (eg. electropolished) and tilted 70 degrees towards the fluorescent screen that shows the lines. A TV camera behind the screen picks up the lines, enhances and analyses them as the beam is steered across the sample, usually in a square raster at about 0.5 sec. per point. This data can be analysed and displayed a number of ways, such as grain maps, grain boundary maps, histograms of the various parameters, pole figures and many other forms. The HKL web site (www.hkltechnology.com) has a good overview and articles about the use of EBSP, also Oxford, who make a system called Opal, have a promotional CD that covers the technique well. You can request it from Oxford. The resolution depends on the SEM, but you need a high beam current and voltage to generate strong enough lines for the pickup system, so with the high tilt the resolution is about one micron with my W filament SEM. Field emmision SEMs will have higher resolution. The samples on my SEM have to be quite small and thin ( {8 mm.), but we have successfully analysed IF steels, various Al alloys and some minerals. These are well polished with colloidal silica and lightly carbon coated. Please contact me if you need any more information. At 04:38 AM 7/13/00 -0700, you wrote:
} Could somebody briefly educate me on EBSD (Electron Backscattered ...?) . I } am trying to find information for a friend. He would like to know the very } basics, capabilities of the technique (for example, resolution) and if there } are contract labs that do this. Since he is interested in quantitative } texture analysis, crystal orientation and GB analysis I assume that he is } talking about getting and analyzing channeling patterns, but I could be } wrong. I did not get from him what materials he is interested in. So, any } information on EBSD would be appreciated. } } Thanks } } Jordi Marti } Honeywell } 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
EBSD is very much like the production of Kikuchi lines in the TEM. The backscatter electrons while they are still inside the sample act as point sources that are attached to the sample. A fraction of them can diffract off the "front" and "back" side of crystal planes as they come out of the crystal. You get cones of diffraction intensity where the Bragg condition for a given crystallographic plane. The pattern is a fairly week signal on the total backscattered intensity and it carries the crystallographic symmetry and orientation of the illuminated volume. The sample is typically tilted to a high angle ~70 degrees. A piece of film or camera is used to capture the image. The cameras are better, because they can process the image to bring out the details and the analysis automated to determine phase or crystallographic orientation. There are mapping programs that can give you orientational maps of the grain structure of your samples. The resolution is essentially the same as that of a backscattered image. Anything at the surface that distorts or disturbs the quality of the lattice (contamination, oxidation, damage, etc.) will be reflected in the quality of the pattern.
Incidentally, you should take a look at the M&M MM program. Joe Michael has a special session on "Advances in the Instrumentation and Applications of Electron Backscatter Diffraction in the SEM" that should be of interest to you. There are also a number of vendors who have systems. You can see them at the 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
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
} -----Original Message----- } From: Marti, Jordi [mailto:jordi.marti-at-honeywell.com] } Sent: Thursday, July 13, 2000 7:39 AM } To: 'Microscopy' } Subject: EBSD ? } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.ht} ml } } } -------------------------------------------------------------- } ---------. } } } Could somebody briefly educate me on EBSD (Electron } Backscattered ...?) . I } am trying to find information for a friend. He would like to } know the very } basics, capabilities of the technique (for example, } resolution) and if there } are contract labs that do this. Since he is interested in quantitative } texture analysis, crystal orientation and GB analysis I } assume that he is } talking about getting and analyzing channeling patterns, but } I could be } wrong. I did not get from him what materials he is interested } in. So, any } information on EBSD would be appreciated. } } Thanks } } Jordi Marti } Honeywell }
Thanks for the suggestions on vendors and chillers - they were all very helpful, and I will follow up individually. Now, we are faced with the issue of whether we should convert the mercury diffusion pump in our proposed Phillips 300 TEM (it stilll has the mercury pump). If anyone has any opinions or experiences on this that they would like to share, I would be very grateful.
Thanks again,
Rick Powell Nanoprobes, Incorporated
********************************************************************** * - PLEASE NOTE OUR NEW ADDRESS - EFFECTIVE FEBRUARY 15, 2000 - * * * * NANOPROBES, Incorporated | US Toll-free: (877) 447-6266 * * 95 Horse Block Road | Tel: (919) 510-0590 * * Yaphank, NY 11980-9710, | Fax: (919) 510-0590 * * USA | rpowell-at-nanoprobes.com * * * * NOW EASY TO FIND ON THE WEB: http://www.nanoprobes.com * **********************************************************************
Ron Thanks for this - I clearly need some retraining on high vacuum theory! Can you recommend a good review or textbook on this topic that a biologist could understand? Best wishes Chris
} From: "Ronald Vane" {RVaneXEI-at-concentric.net} To: {c.jeffree-at-ed.ac.uk}
Dear All, after receiving my 6th email this afternoon about the surface science journals, I'd like to say they have already found a good home with Rik Brydson in Leeds. Glad to see there is such a thirst for knowledge out there...
Richard
} Dear all, } I have a set of the Journal 'Surface Science' and 'Surface } Science reports', 1990-1993 (inclusive). If anyone wants it, please } get } in touch with me by 20th of this month or it will go in the skip. } } Best regards, } } Richard Beanland
============================================================== 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 have been watching with interest the talk about warming up a cold finger and what comes to mind is WHY?
Having used cold traps and fingers on vacuum systems as well as a wide variety of microscopes for many years, and never force warmed them, why suddenly is this important?
A paper by Hitachi in the 60's related cold finger temperature to vacuum level and contamination rate. The theory went that the vacuum level and contamination rate would fall whilst you were using liquid nitrogen in the traps, but once you stopped using LN2 it would take the same amount of time for the vacuum level and contamination rate to fall back to the original levels. So why heat the cold finger?
I also worked on the design of early SEM cryo systems where once again we do not heat the cold finger; I keep asking why?
Is there a proven scientific reason that helps us out in this area, practical reason I emphasize?
I do find it rather interesting!
Steve Chapman Senior Consultant Protrain For professional training and consultancy in EM world wide www.emcourses.com Tel +44 1280 814774 Fax +44 1280 914007
There have been several messages on these topics lately. The original message was specifically a TEM cold trap question but it has broadened out to SEM cold traps as well.
In answer to Yan's question; JEOL 2010 ACD. We have a JEOL 2010 and as I have previously reported we bake it out overnight. We always shut the HT down at the end of every day and restart it the following one. We have always done this on every instrument we have had. Over the past few years the instruments have become cleaner and higher performance, we leave our 2010 with the lenses set at the 200kV setting and at high (300K - 500K) mag. We do the same with our 4000 HREM and 3000F (but we leave the HT on as it is a FEG). This routine keeps the lenses warm so that they are stable when we want to use them and do not outgas when they increase temperature with higher lens currents.
Operation of cold traps. I would never think of venting a column (or SEM chamber) when the cold trap is cold. Although I am primarily interested in TEM the same principles must apply to a SEM. If you need to vent the chamber to insert a specimen then obviously this may affect your use of cold traps. Cold surfaces trap gasses, they do this by lowering the vapour pressure. If you have ice build up on a surface it will be more difficult to pump than a clean surface, you will have a build up of gasses already on the surface that will not only outgas for a long time but also decrease it's efficiency as a cold trap.
It is recognised that to get a good clean vacuum we must bakeout the vacuum system to increase the vapour pressure of the trapped (absorbed) gasses in the surfaces and pump them away. When the surfaces are then cooled down to room temperature the vapour pressure of any trapped gasses is lower than the vacuum system pressure so the gasses are firmly trapped. If we have gasses already trapped on a cold surface they will be released into the vacuum system (albeit at a reducing rate) all the time that system is pumping. This will give rise to contamination during the operation of the instrument which can only be cured by raising the temperture of the cold trap and cooling it down again.
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. ============================================================================
There are two techniques associated with looking at the angular distribution of emerging radiations from a crystal.
Firstly there is Kossel diffraction, which is demanding experimentally. This requires a material with one dominant atomic element and what is looked at here is the angular distribution of emerging X-rays generated within the excitation volume. Kossel diffraction can be extremely accurate for lattice parameter measurements (I have heard of up to five/six significant figures in accuracy in certain cases).
Then there is EBSD in which inelastic back scattered electrons follow the crystallographic planes a la Kikuchi in the TEM. A scintillator with a high angular solid angle is used to capture the diffraction pattern. EBSD maps requires a scan over an area with a high pixel dwell time, from minutes to hours for a whole map. The technique requires a lot of memory since it records a pattern for point over pre-determined area. The computer usually fits a pre set range of materials of any crystallographic space group i.e. it helps if you know what you have in the first place. The pattern recognition works by an algorithm called a Hough transform, in which the intensity at every point is turned into a pair of angular coordinates. Lines of intensity show up a bright dots in Hough-space and the computer fits the different crystal types to this pattern. Once the whole area is done, it then gives you a scatter plot of all the crystal orientations. The better systems will allow you to do statistical analysis like correlation functions .etc.
A flat sample is quite a stringent condition and the seventy degree surface normal to beam angle means you get BS SEM resolution only parallel to this tilt axis i.e. side to side. Spatial resolution is limited in the other direction to around 1 micron in real terms. If you are trying to map micro precipitates less than a micron, then forget it. So the ideal sample has to be flat and have grains in excess of one micron in size.
As for systems on the market, the ones I have seen are TEXSEM and the Oxford Instruments systems. As far as I am aware, TEXSEM was the first commercial company to do EBSD mapping in the beginning and the others have come after them. TEXSEM was acquired by Edax a couple of years ago, but I am sure Edax can give you information on the EBSD system and maybe their new TEM crystallography mapping system as well. I know the TEXSEM system through Prof David Dingley one the founders on TEXSEM, and also the one who taught me most about SEM techniques when I was at Bristol. I don't know if he is on this listserver, but he's the one person who can tell you anything and everything about EBSD.
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 ********************************************************
I am hoping to upgrade to a better microscope but have a limited budget. I have seen Russian LOMO microscopes advertised on the web. Does anyone have experience with these that they can share - in particular are their objectives up to scratch? Thanks Andrew Logan
1) I presume that just as at atmospheric pressure, materials such as perspex, anodised aluminium, borosilicate glass, stainless steel, gold) differ significantly in their capacity to adsorb water vapour and hydrocarbons under high vacuum conditions? Presumably they would differ not only in their uptake kinetics but also in their desorption kinetics. If one were designing an ultraclean vacuum system, or an ultraclean specimen environment within a specimen chamber, with the objective of minimising deposition to a specimen surface during SEM examination, what materials would be best to use for the chamber surfaces? 2) Presumably the main source of hydrocarbon contamination in an SEM is oil vapour back-streaming from the the rotary and diffusion pump. Turbo pumps, particularly maglev pumps, must reduce this to very low levels while the pumps are running at full speed. However, there must be considerable potential for back streaming from the backing line during rough pumping from air and turbo run- up after specimen changes. This again would seem to me to argue in favour of a specimen air-locked system, so that the specimen chamber is exposed to air and rough-pumping cycles as little as practicable. Does anyone know of any data in the literature comparing the magnitudes of contamination in systems using different pumping and specimen exchange methods? Chris ========================================== Dr. Chris Jeffree University of Edinburgh BIOSEM - Biological Sciences Electron Microscope Facility Institute of Cell and Molecular Biology Daniel Rutherford Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JH, Scotland, UK Tel. #44 (0) 131 650 5345 FAX. #44 (0) 131 650 6563 Mobile 07710 585 401 email c.jeffree-at-ed.ac.uk =========================================
I'm not aware of a standard but at Childrens Hospital in Boston I have processed from beginning to end about 350 EM clinical cases per year for the past fifteen years. That is from embedding through photography and printing with fifteen to thirty photographs per case. The voluum of cases depends on several factors. The type of tissue, the number of blocks per case, possible special techniques (ex. glycogen stains), available equiptment ...tissue processors, knives, microtomes etc. and of course the technologists ability. A number of labs that I have visited require the technologist to limit their involvement to embedding ,sectioning and printing the negatives. The EM lab from a neighboring institution handles about 850 EM clinical cases, far exceding what their technologist could handle and so they use their residents to scope many of the cases. In my own situation I feel our voluum is very high for one individual and the results are reflected in my inability to get the cases out in a timely manner or take a vacation. I have always felt that it was a waste of time for a resident/rotator to come into the lab and scope cases without serious supervision. This has limited our ability to handle more cases but common sense and expierence has told me that someone who doesn't know the ultrastructural pathology, the microscope and is not a technologist and who just comes in the lab to take some photo's is using their time inneficiently. After all, taking an electron micrograph is about communication. If the technologist is capable and has the expierence to understand the pathology, certainly not in making a diagnosis, but in presenting to the pathologist a group of photo's that will stand out and either confirm or alter a suspected diagnosis that certainly is communication and is an efficient cost effective way of running the EM lab. Performance standards? The very words sort of indicate to me that all technologists and labs should be equal . That would take the fun out.
Howard L.Mulhern Childrens Hospital Dept. of Pathology EM Facility mulhern-at-hub.tch.harvard.edu
'cause Hitachi were talking about the cold finger warming under operating conditions. We were discussing opening the chamber when the finger is cold, which would then load up with ice. Also if warm, the roughing pump on immediate re-pumping would have a chance to remove some contaminants from the "warm coldfinger" 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 ABN: 99 724 136 560 www.proscitech.com
On Friday, July 14, 2000 5:50 PM, Steve Chapman [SMTP:PROTRAIN-at-CompuServe.COM] wrote: } } } } Hi } } I have been watching with interest the talk about warming up a cold finger } and what comes to mind is WHY? } } Having used cold traps and fingers on vacuum systems as well as a wide } variety of microscopes for many years, and never force warmed them, why } suddenly is this important? } } A paper by Hitachi in the 60's related cold finger temperature to vacuum } level and contamination rate. The theory went that the vacuum level and } contamination rate would fall whilst you were using liquid nitrogen in the } traps, but once you stopped using LN2 it would take the same amount of time } for the vacuum level and contamination rate to fall back to the original } levels. So why heat the cold finger? } } I also worked on the design of early SEM cryo systems where once again we } do not heat the cold finger; I keep asking why? } } Is there a proven scientific reason that helps us out in this area, } practical reason I emphasize? } } I do find it rather interesting! } } Steve Chapman } Senior Consultant Protrain } For professional training and consultancy in EM world wide } www.emcourses.com } Tel +44 1280 814774 Fax +44 1280 914007
Can anyone on the list help here? We need a protocol for preparation of specimens for a training exercise. We will be doing an assay using Salmonella typhimurium positive control (5x10EE9 cells/ml) and fluorescein-labeled affinity purified antibody to Salmonella CSA-1 (0.5 mg/ml). We have protocols for rehydration/dilution of the reagents and for preparing slides after doing the assay in flasks/vials/plates, but what we would really prefer is something very simple that can be done in-situ on the slide. My first question is if anyone has or can provide a protocol for this and my second question is if anyone can tell us what would be the best dilutions to use for this exercise.
Thank-you in advance,
Erica Valdes US Army Soldier Biological Chemical Command Edgewood CB Center
We offer the Philips EM300 vac. system upgrade, which includes conversion to Hg-less pumping system. Philips Electron Optics (now part of FEI company) was offering the same upgrades, they probably still do.
Benefits one may expect from the upgrade:
1. Vacuum system performance (reliability, pumping speed, etc.) - none.
2. Health hazard reduction for the lab personnel - almost none. The Mercury is not in contact with the outside world, except for a few exotic accident scenarios or when grossly mishandled.
3. Possible future repairs of the vacuum system - upgrade makes for a great advantage, since one will not have to comply with numerous safety rules in order to work with the system containing toxic substance.
4. Speaking of exotic accidents - the probability of such is low, yet is greater than zero nonetheless. Room cleaning is costly and troublesome, shall room become Hg contaminated. An upgrade makes for a great advantage from this standpoint.
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax -----Original Message----- } From: Rick Powell at Nanoprobes {rpowell-at-nanoprobes.com} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
Hello Steve, This all started around the 2010. With our 2010, the ion pump cannot handle the gas load while the ADC warms up on it's own. There is a regeneration cycle that switches over to the DP but should a user fail to start that cycle.... it goes like this... oh the current in the ion pump is too high, goto PD... the DP can out pump the virtual leak (gas load from the ACD regeneration) & the pressure drops. The system crosses over to the ion pump which can't handle the gas load & the cycle repeats at ~0.5 Hz. Along with the switching of pumps the gun isolation valve cycles. It has a bellows seal. Typical cycle life times of a bellows is 10,000. They fail. It is my opinion that this is a silly short coming of the system & could be solved with a simple counting circuit... maybe a ring counter, one shot & an AND GATE. Obviously a little programming could deal with this too. My suggestions to the vender about this issue have not gotten a fix.... so every couple of years we go down for a few days & they buy us a new valve... go figure. BTW I think they now use an o-ring seal in a new valve design. I was told it will not screw into the older machines. For systems that are only pumped by a diffusion or turbo pump, I think this is a non-issue & you can just walk away at the end of the day. This is true of our FESEM.
Bruce Brinson Rice U.
Steve Chapman 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 have been watching with interest the talk about warming up a cold finger } and what comes to mind is WHY? } } Having used cold traps and fingers on vacuum systems as well as a wide } variety of microscopes for many years, and never force warmed them, why } suddenly is this important? } } A paper by Hitachi in the 60's related cold finger temperature to vacuum } level and contamination rate. The theory went that the vacuum level and } contamination rate would fall whilst you were using liquid nitrogen in the } traps, but once you stopped using LN2 it would take the same amount of time } for the vacuum level and contamination rate to fall back to the original } levels. So why heat the cold finger? } } I also worked on the design of early SEM cryo systems where once again we } do not heat the cold finger; I keep asking why? } } Is there a proven scientific reason that helps us out in this area, } practical reason I emphasize? } } I do find it rather interesting! } } Steve Chapman } Senior Consultant Protrain } For professional training and consultancy in EM world wide } www.emcourses.com } Tel +44 1280 814774 Fax +44 1280 914007
} Hello, I'm a grad student attending the M&M2K meeting in Philadelphia next } } month, and I'm looking for a roommate. I'm a non-smoking, non-snoring female } } arriving Aug. 12. PLEASE REPLY TO gcelio-at-arches.uga.edu . } } Thanks, } --Gail Celio
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
Could I get opinions based on experience? I have a nice scanner with a SCSI interface. The vender says my 20 MHz SCSI card exceeds the scanner through put. The scanner would be the bottle neck on the SCSI buss no mater how fast the SCSI bus or drive is. So my question : once you have the image on your hard drive, is in terms of disk I/O for processing, by the time you go from the SCSI bus to the mother board & back, is there a still a speed advantage to SCSI over current IDE technology. Or put another way, can I justify buying a super duty SCSI HD & interface in the name of processing speed & what would be the relative improvement?
During my LM analysis of serial, toluidine blue-stained plastic sections of decalcified in tact mouse inner ears, I have encountered what appear to be dark-staining, elongated "crystals" in the supranuclear region of some Deiter's (support) cells in the organ of Corti. This region of the cells typically has numerous round lysosomes that stain dark blue; the vast majority of support cells have the usual round lysosomes present. Deiter's cells with the long crystals show crystals also in serial sections of the same cell. The appearance of the spikey "crystals" in serial sections of the same cell, the specificity of the inclusions for the support cells only, and the limited supranuclear location within these highly polarized cells suggest that these inclusions are not artifacts of staining. I have observed them in wildtype and mutant animals, but not in every ear of any genotype. I have done relatively little EM analysis of the ears so far, and no samples with these inclusions have been checked yet by EM. Are there lysosomal abnormalities that may result in their contents becoming crystalline? Why do relatively few Deiter's cells have the crystals? Apparently the genetic background of the mouse gene knock-out models I am analyzing includes the development of hearing deficits with aging. Is it possible that these "deviant Deiter's cells" may be related to impending changes in the inner ear due to aging (the oldest animals I've evaluated are 21 weeks old)? Have such inclusions been reported in Deiter's cells (or other cell types)? I hope someone has an answer! Thanks for your help or suggestions,
Emma Lou Cardell, Research Associate Department of Cell Biology, Neurobiology, and Anatomy University of Cincinnati Medical College 231 Albert Sabin Way Cincinnati, OH 45267-0667
Vacuum Methods in Electron Microscopy Wilbur C. Bigelow Portland Press, London, 1994
Ron Vane
-----Original Message----- } From: Chris Jeffree {cjeffree-at-srv0.bio.ed.ac.uk} To: Ronald Vane {RVaneXEI-at-concentric.net} Cc: microscopy-at-sparc5.microscopy.com {microscopy-at-sparc5.microscopy.com}
An exciting and challenging opportunity is available with Gatan.
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Gatan Inc. has an immediate opening for a person to handle GIF and PEELS Product management. Individual should have either GIF or PEELS experience, a strong TEM background in Biological or Materials science, good computer skills, a willingness to travel and the vision and drive to help determine the future of these products. The position is based in Pleasanton, CA and carries a salary comensurate with experience as well as a bonus plan and Corporate success share plan. Please contact Ian Cotton, Director of Marketing at 925-224-7343 or email your resume to icotton-at-gatan.com.
********************************* Ian Cotton Marketing Director, Gatan, Inc. 5933 Coronado Lane Pleasanton, CA 94588 USA Phone 925-224-7343 Fax 925-463-0204 E-mail icotton-at-gatan.com Surf the Web to www.gatan.com *********************************
We've had the same experience but no answers. We have a bottle right now that is about one fourth filled with 'crystals'. Once it gets this full we dispose of it and move on to a fresh bottle. I always wonder about the quality of our mounts once the crystals start forming but it seems to work OK.
We store our bottles at room temperature and in the solvents cabinet.
Anybody else have any ideas?
Diane Ciaburri General Dynamics Pittsfield MA 01210 diane.a.ciaburri-at-gdds.com (413)494-2847
Hi, I use Epofix epoxy. In a very short time after purchase the resin becomes cloudy and a white sediment forms at the bottom of the bottle. I have heard this sediment described as crystals. I have continued to use the resin---carefully avoiding agitation and entrainment of the sediment when I withdraw it from the bottle. Does anyone else have experience with this?
Everett Ramer National Energy Technology Laboratory P.O. Box 10940, Cochrans Mill Road Pittsburgh, PA, USA 15236-0940 Voice: 412-386-4920 FAX: 412-386-4806 ramer-at-netl.doe.gov
by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id MAA17995 for dist-Microscopy; Fri, 14 Jul 2000 12:22:26 -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 MAA17992 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Fri, 14 Jul 2000 12:21:55 -0500 (CDT) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id MAA17985 for {Microscopy-at-sparc5.microscopy.com} ; Fri, 14 Jul 2000 12:21:44 -0500 (CDT) Received: from [166.16.19.7] (helo=campfire.gdds.com) by net2.gendyn.com with esmtp (Exim 2.12 #1) id 13D943-000Dl6-00 for Microscopy-at-sparc5.microscopy.com; Fri, 14 Jul 2000 13:15:51 -0400 Received: from ccmail.gdds.com (ccmail.gdds.com [166.16.19.25]) by campfire.gdds.com (8.8.8/8.8.8) with SMTP id NAA24556 for {Microscopy-at-MSA.Microscopy.com} ; Fri, 14 Jul 2000 13:20:28 -0400 (EDT) Received: from ccMail by ccmail.gdds.com (ccMail Link to SMTP R8.31.00.5) id AA963594544; Fri, 14 Jul 2000 13:09:05 -0400 Message-Id: {0007149635.AA963594544-at-ccmail.gdds.com} X-Mailer: ccMail Link to SMTP R8.31.00.5
Thank you to all who sent ideas and procedures for de-encapsulating plastic encapsulated IC's. I haven't received any samples yet so haven't been able to try out any ideas. --The old hurry up and wait routine.
I'm always astounded by how helpful and knowledgeable everyone is. Thank you! and thanks for managing this listserver, Nestor!
Diane Ciaburri General Dynamics Pittsfield MA 01210 diane.a.ciaburri-at-gdds.com (413)494-2847
Does any one know the current phone number and or web site of Denton Vacuum? The ones I am using : 888-336-8661, www.dentonvacuum.com are not responding. I need some advice on one of their old evaporators and possibly puchase a couple of thermocouples (vacuum gage). Why do they call it a thermocouple? The TEM service person that was here didn't think that was a proper name. Thanks.
Some turbo pumped vacuum systems stop the turbo and rough pump through it when evacuating. I feel you are correct in your comments about such systems.
Other designs implement a valved vacuum bypass so that the chamber is directly roughed to about 100 microns. When, at this point, the flow is more random, the valves switch the turbo in series between the chamber and the roughing pump and continue to high vacuum. The turbo is allowed to run continuously, minimizing backstreaming.
The only disadvantage I know to this scheme is the added expense of controling circuitry and the (major) expense of a large hi-vac gate valve.
My old ETEC was plumbed like this and I replaced the dif pump with a Leybold mag-lev turbo (too much vibration from the ball bearing pumps). "Cooked" very little carbon on to the specimen surfaces and the vacuum (excluding outgassing specimens) was typically 1.0 E-6 Torr or better.
Hi All, I have a colleague who needs to do real-time confocal or multi-photon. Her minimum wavelength requirement is 488, although additional wavelengths would be useful. We are located in New York City, but she is open to anywhere in the northeast. If anyone can help her, please contact her directly:
Dr. Geri Kreitzer Dyson Vision Institute Weill Medical College of cornell University 1300 York Ave New York, NY 10021
(212)746-2277
e-Mail: ggurlan-at-med.cornell.edu
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
I used Denton's site yesterday and today. No problem. Gregory W. Erdos, Ph.D. Assistant Director, Biotechnology Program PO Box 110580 University of Florida Gainesville, FL 32611
Hi - I am looking for a good/successful protocol (from fixation to polymerization) for LR white immunocytochemistry for cell culture. Can anyone help?. Thanks very much in advance.
G. Ning EM Facility Medical College of Wisconsin 414-456-8344 Fax 414-456-6535
Hi All We desperately need some help with our SEM. Our filament "appeared" to have blown during operation (I was out of the room for 5 mins and came back to a pretty green screen). I initially assumed that the electronics were playing up, which sometimes occurs, and all the settings are thrown out. While tweaking the Contrast/brightness controls, some kind of warning flashed on the screen. However, it dissappeared too quickly for me to figure it out, and it has not reappeared. We changed our filament after no emission current reading was found. No luck. The technicians then advised us to change and recheck the filament settings in the cap. Alas, still nothing. We have, however, found that the LC reading shoots up to about 190ua, during the 4th filament check but still no image on the screen. Between changing , cleaning & checking filaments, there has been an eerie inconsistency with the LC. Between three of us, we have twiddled here and twiddled there without success. Help!!!!! Any info would be greatly appreciated. Nazlia
The newer IDE technologies(Ultra DMA 66) approaches the "burst speed" read/write capabilities of the newer SCSI technologies. However, the newest emerging Ultra DMA 100 technology provides data transfer rates beyond SCSI. These new drives are going to hit the market any day now. Of course, you will need to buy a DMA 100 controller card to go along with the DMA 100 hard drive, unless you opt to buy an new computer with the capabilities built into the motherboard. If memory serves me right, Quantum Corp. patented the technology, but I think Maxtor is going to be first to the market with the drives. Look to Promise Technologies(www.promise.com) for the appropiate controller card. Good luck.
Gary M. Easton Scanners Corporation Third Party SEM Service 410-857-7633 x102 ----- Original Message ----- } From: "Bruce Brinson" {brinson-at-cnst.rice.edu} To: "MSA Listserver" {microscopy-at-sparc5.microscopy.com} Sent: Friday, July 14, 2000 11:53 AM
Bruce writes ...
} I have a nice scanner with a SCSI interface. The vender } says my 20 MHz SCSI card exceeds the scanner throughput. } The scanner would be the bottle neck on the SCSI buss } no matter how fast the SCSI bus or drive is. } ...
I believe your "vendor" sells bridges too :o)
A slower SCSI device will not bottleneck the SCSI bus. The exception to this is putting a 68pin non-ultrawide (i.e. non-U2W) device on a U2W bus when you have U2W devices on it. This will slow "U2W" down to "UW". However, note I am talking about the wide 68pin bus (40Mbs), not the 50pin bus common to scanners. Even as slow as scanners are, the modern ones are fast-SCSI compatible ... meaning, even if they're not fast, they know how to communicate with a fast-SCSI controller and not slow it down. Of course ... this is the way it is supposed to work. If your scanner's fireware is not communicating with the controller properly, then who knows ... BUT generally speaking, your vendor is incorrect.
1. The numbers that you want are readily available in a number of Vacuum science books. Probably the best one with the most data about different materials is the one by Roth. Sticking coefficients, desorption, adsorption, and chemisorption of the different gases depend on the gas, temperature, material, and the surface roughness. That is why the relative partial pressures of different gases are so different at different vacuum regimes and vacuum histories (e.g. baked, not baked). If you want a great material to coat the inside of your vacuum system with that has very good properties, try gold. I once had a UHV system that was gold plated on the inside. It was also very pretty.
2. You will only get hydrocarbon (HC) contamination from the mechanical pump if you pump the system into the molecular range with this pump. The pump speed for HCs is extremely high. If you pump the system through the turbo pump, while the T-pump is starting, the mechanical pump is still in the viscous range. It doesn't take the T-pump to get up to enough speed before the pump speed will be sufficient to prevent the backstreaming of the mechanical pump oil. When a T-pump system vents, what is important is that there is a valve between the mechanical pump and T-pump to prevent oil from being sucked out of the pump and back into the chamber. A properly designed T-pump system should also have a gas inlet to help prevent this in case of a valve failure when a power failure occurs. On T-pumped systems, this is what you need to examine to see whether the system can be contaminated with pump oil. Essentially, turn off the power (hypothetically, of course) and see what happens. If it does not have several systems in place which includes valves and automatic venting in the correct places, don't buy the system.
With respect to pump down times, the pressure follows exponential curves. These curves have dominant regimes. Initially, the volume gas is pumped, but that is the very fast evacuation stage. Then it transitions over to virtual leaks (i.e. trapped gas), then desorption off walls, then outgassing of materials, then rate of gas diffusion through materials (permeation). If you plot the log of the pressure with time, you will see these changeover points. The longer the system is under vacuum, the better it gets, assuming that there are no leaks that set up a steady state condition. Your ultimate pressure is determined by a simple equation, SP=Q. S is the pump speed (liters/sec), P(Torr) is the pressure, and Q(Torr-liters/sec) is the throughput. The throughput is the sum of all leaks and outgassing at your ultimate pressure. These are all temperature dependent and will increase at higher temperatures -that's why you bake out.
If you pre-pump with an exchange system that is not designed to minimize backstreaming, it can lead to contamination of the system when you finally open the chamber valve.
If you have a chamber that pumps to the 10^-6 Torr region, and you can operate in the low 10^-5 Torr range while it gets into the 10^-6 range with time, then the only thing that the exchange system really buys you is time.
-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: Chris Jeffree [mailto:cjeffree-at-srv0.bio.ed.ac.uk] } Sent: Friday, July 14, 2000 8:12 AM } To: microscopy-at-sparc5.microscopy.com } Subject: Specimen and chamber contamination } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } 1) I presume that just as at atmospheric pressure, materials such } as perspex, anodised aluminium, borosilicate glass, stainless } steel, gold) differ significantly in their capacity to adsorb water } vapour and hydrocarbons under high vacuum conditions? } Presumably they would differ not only in their uptake kinetics but } also in their desorption kinetics. If one were designing an } ultraclean } vacuum system, or an ultraclean specimen environment within a } specimen chamber, with the objective of minimising deposition to a } specimen surface during SEM examination, what materials would } be best to use for the chamber surfaces? } 2) Presumably the main source of hydrocarbon contamination in an } SEM is oil vapour back-streaming from the the rotary and diffusion } pump. Turbo pumps, particularly maglev pumps, must reduce this } to very low levels while the pumps are running at full speed. } However, there must be considerable potential for back streaming } from the backing line during rough pumping from air and turbo run- } up after specimen changes. This again would seem to me to argue } in favour of a specimen air-locked system, so that the specimen } chamber is exposed to air and rough-pumping cycles as little as } practicable. Does anyone know of any data in the literature } comparing the magnitudes of contamination in systems using } different pumping and specimen exchange methods? } Chris } ========================================== } Dr. Chris Jeffree } University of Edinburgh } BIOSEM - Biological Sciences Electron Microscope Facility } Institute of Cell and Molecular Biology } Daniel Rutherford Building } King's Buildings, Mayfield Road } EDINBURGH, EH9 3JH, Scotland, UK } Tel. #44 (0) 131 650 5345 } FAX. #44 (0) 131 650 6563 } Mobile 07710 585 401 } email c.jeffree-at-ed.ac.uk } ========================================= }
I am currently looking into a confocal microscope system. Based on my Internet search, I have a feeling that most of the confocal microscope systems are primarily designed for "biology". My application will be investigation of (i)microstructure of ceramic and metallic coatings (mostly porous), and (ii) surface (including fracture surface) topology. I do not have any experience in confocal microscopy. I would like to get some of your opinions about confocal microscope systems for materials science applications.
I am working on Mycobacterium marinum infection of frogs (Rana pipiens) which gives them a chronic tuberculosis-like disease. The frogs develop granulomas. We have done EMs of these frog granulomas and find certain cell types that we cannot identify (macrophages vs heterophils?). If any of you have expertise with amphibian tissues, could you get in touch with me? We could send you pictures or come down and see you if you are close by. We would greatly appreciate any help we can get. Thanks in advance.
You probably have a high voltage problem as indicated by the high LC current. Possibly a capacitor in the oil tank or defective high voltage cable.
The repairs are not for a novice or user as the voltages are potentially lethal. Call a service engineer.
Regards,
Earl Weltmer
Nazlia Samodien 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 } We desperately need some help with our SEM. } Our filament "appeared" to have blown during operation (I was out of the } room for 5 mins and came back to a pretty green screen). I initially } assumed that the electronics were playing up, which sometimes occurs, and } all the settings are thrown out. While tweaking the Contrast/brightness } controls, some kind of warning flashed on the screen. However, it } dissappeared too quickly for me to figure it out, and it has not } reappeared. We changed our filament after no emission current reading was } found. No luck. The technicians then advised us to change and recheck the } filament settings in the cap. Alas, still nothing. We have, however, found } that the LC reading shoots up to about 190ua, during the 4th filament check } but still no image on the screen. Between changing , cleaning & checking } filaments, there has been an eerie inconsistency with the LC. Between three } of us, we have twiddled here and twiddled there without success. Help!!!!! } Any info would be greatly appreciated. } Nazlia } } _____________________________________________________ } } Nazlia Samodien } } Cardiovascular Research Unit } Dept. of Cardiothoracic Surgery } University of Cape Town Medical School } Anzio Rd, Observatory, 7925, South Africa } } Tel. +27 21 406 6398 (office) } +27 21 406 6476 (Secretary) } Fax + 27 21 448 5935 } } Email: ctssamodien-at-samiot.uct.ac.za
Email: jyr15102-at-glaxowellcome.com Name: Jason Remillard School: University of Alberta
Question: Good Day;
My name is Jason Remillard, and I am a pharmacy student from the University of Alberta in Edmonton, Alberta, Canada. I am working as a summer student at GlaxoWellcome in Toronto, Ontario and one of my projects requires the use of the light microscope. Since I am not well trained in this area, I have a couple of questions that you may be able to help me with.
The purpose of my project is to help develop a faster method of determining particulate count for Flonase(TM) Nasal Spray (fluticisone propionate) - a corticosteroid nasal spray suspension for allergic rhinitis and sinusitis. The current counting method is labour intensive, and time consuming. It involves using a regular light microscope, and counting the particles in the field (and there are a lot). Based on this data, the particulate count for the entire bottle is determined. This is currently being done by the Quality Control Labs, who want to speed it up.
I have the following questions to ask:
Have you ever done any work with fluticisone propionate before? I thought that by the use of oil immersion microscopy, I would be able to get better magnification of the area, and have to count less particles. I would still be able to calculate particle count, but how would I go about using oil to increase the refractive index?
I have developed a computer program to process electron diffraction ring patterns. The program can be downloaded FREE from
http://www.mfa.kfki.hu/~labar/
I hope you will enjoy it. Future versions with enhanced functionality are coming. In case of any problem or suggestion or remark, do not hesitate to contact me.
Best regards:
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
} } The phone number I have for Denton is 856-439-9100. I recently used the number so it should be good. } } } } At 12:48 PM 07/14/2000 -0500, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
wouldn't this be a good candidate for some type of optical particle counting method (hiac type)?? or maybe a coulter type method?
just my 2c...
b- ******************************************
Email: jyr15102-at-glaxowellcome.com Name: Jason Remillard School: University of Alberta
Question: Good Day;
My name is Jason Remillard, and I am a pharmacy student from the University of Alberta in Edmonton, Alberta, Canada. I am working as a summer student at GlaxoWellcome in Toronto, Ontario and one of my projects requires the use of the light microscope. Since I am not well trained in this area, I have a couple of questions that you may be able to help me with.
The purpose of my project is to help develop a faster method of determining particulate count for Flonase(TM) Nasal Spray (fluticisone propionate) - a corticosteroid nasal spray suspension for allergic rhinitis and sinusitis. The current counting method is labour intensive, and time consuming. It involves using a regular light microscope, and counting the particles in the field (and there are a lot). Based on this data, the particulate count for the entire bottle is determined. This is currently being done by the Quality Control Labs, who want to speed it up.
I have the following questions to ask:
Have you ever done any work with fluticisone propionate before? I thought that by the use of oil immersion microscopy, I would be able to get better magnification of the area, and have to count less particles. I would still be able to calculate particle count, but how would I go about using oil to increase the refractive index?
At 12:48 PM -0500 7/14/0, "rlvaughn-at-unmc.edu"-at-sparc5.microscopy.com wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Rick, I have some recent literature from Denton, and the telephone # listed is: (856)439-9100 The web site you have looks OK. I have and e-mail for them:info-at-dentonvacuum.com YOu can also get in touch with Dick Daniel, he is the sale/sevice fellow here in the NY Metro area. His number is" (201)847-8845. He is very helpful.
I have nofinfncial interests in Denton Vacuum.
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
3rd Euroconference on NANOSCIENCE FOR NANOTECHNOLOGY
16 - 19 September 2000 Somerville College, Oxford
See: http://nano.org.uk/euro.htm
Special funds are available to support attendance by younger scientists (under the age of 35) from EU countries.
NOTE: Deadline for submission of abstracts: 1 August 2000.
Under the contract providing EU sponsorship, the organisers are requested to make special efforts to attract participants from the following target groups:
- Researchers aged 35 years or under at the time of the conference. - Researchers whose place of work is in a less-favoured region. - Women researchers. - Researchers who work in industry.
The sponsorship includes funding to facilitate participation of researchers under the age of 35 who are citizens of EU countries. In order to take advantage of the low cost air fares which are available for journeys which include a Saturday night stay, the meeting is being organised over a "long weekend" (Saturday to Tuesday). Only the lowest cost air fares can be reimbursed, within the available funds.
INTERNATIONAL STEERING COMMITEE Professor K. Schaumburg, University of Copenhagen Professor J.T. Devreese, University of Antwerp Professor G.D.W. Smith, University of Oxford
PARTIAL LIST OF INVITED SPEAKERS Professor G.A.D. Briggs, Oxford University Professor J. Chapman, University of Glasgow Professor J.M. Cooper, University of Glasgow Professor P.J. Dobson, Oxford University Dr. J. Gimzewski, IBM Zurich Dr. J.A. Liddle, Lucent Technologies Professor R. Palmer, University of Birmingham Professor A. Persoons, University of Leuven Dr. A.K. Petford-Long, Oxford University Professor J.B. Pethica, Oxford University Dr. Rasmita Raval, University of Liverpool Dr. T. Spiller, Hewlett Packard Professor M. Welland, University of Cambridge
SPECIAL EVENTS Meeting of the ORCHYD Network for Organic-Inorganic Hybrid Devices. Meeting of the U.K. Foresight Panel Task Force on Nanotechnology Meeting of Summer School working group 'Self Organisation of Nanoparticles' Workshop session on "The interaction of Organic Molecules with Surfaces". 'Consulting the Community' workshop session: "The impact of nanotechnology on society". Visits to nanoscience laboratories and nanotechnology spin-out companies in the Oxford area. Displays of books and product literature relating to nanoscience and nanotechnology. Nanotechnology Networks: It is hoped to announce the successful applicants for EPSRC networks in nanotechnology, the application deadline for which is June 2000. These will be interdisciplinary networks including the full range of scientific and engineering disciplines. It is expected that successful networks will be actively looking to extend their membership at this meeting.
CONFERENCE VENUE Somerville College is one of the constituent colleges of Oxford University. With a superb location a few minutes walk from the city centre, and close to the University's main science and engineering laboratories, it provides an excellent, self-contained location for the conference.
PUBLICATION OF PROCEEDINGS Selected papers from the conference will, after peer review, be published as a special issue of Materials Science and Technology. This issue will be available for purchase by delegates at the special discounted price of £20 per copy.
CALL FOR PAPERS Contributed papers are sought, both for oral and poster presentation, in all areas of nanoscience and nanotechnology. Deadline for submission: 1st August 2000. Please state whether you wish to submit an oral presentation or a poster. Specified Format:
* Abstracts to fit on a single A4 page (21cm wide x 29.7 cm high), with 2cm margins to left and right, 3cm margins top and bottom. * Use Times font, 12 point size. * Start title on a line 3.5 cm from the top of the page. * Put title in bold, with upper/lower case print (i.e. only initial letters of title word capitalised). * Put names of authors on next line, with the name of the presenting author underlined. * Put addresses of authors on subsequent lines. * Leave gap of two lines before commencing text. * Do not indent at start of paragraphs, leave gap of one line between paragraphs. * Put any references at end, in numerical order. * Send abstract either in hard copy form, or by email as a Word attachment (specify Word 97).
Send abstracts to: Julie Hutcheon, Events and Membership Manager, The Institute of Nanotechnology, 9, The Alpha Centre, Stirling University Innovation Park, Stirling FK9 4NF, Scotland, UK Email: julie-at-nano.org.uk
CONFERENCE SCHEDULE The conference starts officially at 6 p.m. on Saturday 16th September, and ends at 4.30 p.m. on Tuesday 19th. The first meal provided is the evening meal on the 16th, and the last meal is lunch on the 19th. Therefore the 'nightly' accommodation and meals charge refers to a 24-hour period, beginning with the evening meal on one day, and ending with lunch on the following day.
GRANT INFORMATION Grants are available to assist attendance by younger scientists from the European Union. Reimbursement can be provided for a limited number of researchers aged 35 years or younger, who are nationals of a member state of the European Union or of an associated state (Iceland, Liechtenstein, Norway and Israel). EARLY REGISTRATION IS ESSENTIAL IN ORDER TO ENSURE GRANT ASSISTANCE.
Please indicate on your registration form if you wish to apply for grant support, and include an estimate of your expected travel costs. Note that only the LOWEST COST return fares to Oxford can be reimbursed.
The organisers will notify you as soon as possible if your grant application has been accepted. Funds are limited, so early application is essential. Reimbursement can only be made retrospectively, AFTER the conference has taken place. You must provide the conference organisers with evidence of your citizenship, your age at the start of the conference, and the actual expenditure incurred. Please bring with you to the conference your passport or other original evidence of citizenship and date of birth, plus original receipts for all expenditure incurred.
Registration and payment may be made online - see http://www.nano.org.uk/euro.htm
REGISTRATION FEE Industry / Government participants: £300 University participants: £200 Registered students: £100
RESIDENTS AT SOMERVILLE COLLEGE An inclusive charge of £65 per night is payable for residential accommodation (in single rooms) at Somerville College, together with all main meals, morning coffee and afternoon tea.
I disagree that uranyl acetate is fairly innocuous. Using the appropriate dosimeters, we have measured increases of alpha, beta, and gamma radiation over background levels with the lid of the reagent jar open. Increased counts were still measured with the closed jar behind an alpha shield at some distance. We ask our scientific and support staff to take some necessary precautions for weighing uranyl acetate and other uranium salts (wear dust mask and latex/vinyl gloves, use a shield in front of the balance, etc.).
Of course, while the dilute stain solution poses no radiation hazard, it should be treated as heavy metal waste for disposal purposes. These are my own suggestions and not necessarily that of USDA.
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-commserver.srrc.usda.gov
-----Original Message----- } From: jim [mailto:jim-at-proscitech.com.au] Sent: Wednesday, July 12, 2000 8:21 AM To: microscopy-at-sparc5.microscopy.com
Hello Microscopist & Lab Managers, well it's old news to a lot of you but the time has come for us to try to support service contracts with user fees. I guess we should be grateful to have deferred this expense to our users for this long. If we pick a rate based historical usage, all is well? I don't think so. The old cost vs demand has to come into play. My questions to those of you that have lived this experience:
1. What was the impact of significant increases in user fees with respect to hours logged? 2. Did use come back to historical levels after the sticker shock wore off? 3. Has anyone on the university level actually been able to support their contracts & related expenses with user fees.
Your experiences & insights would be greatly appreciated. Also I recall that there was an meeting being organized on this & similar subjects at M&M 2000. Would the the organizers please contact me.
just to say thanks for all your formaldehyde recipes, they solved my polymerisation problem just in time so we had plenty of formaldehyde to take on our field-collection trip.
cheers Elizabeth McKenzie -------------------------------------------------- Geomicrobiology and Electron Microscopy Laboratory Room S9 Cramer Hall 1721 SW Broadway Portland State University Portland OR97201
I have a professor here that wants to determine the subsurface crack shape in a ceramic by placing a drop of fluorescent dye-penetrant onto indentations. The cracks are forced open after the dye is allowed to penetrate and the pre-existing crack area will be covered with dye penetrant. Our reference says to use external ultraviolet illumination in an optical microscope to distinguish the pre-cracked area.
It is my understanding that we will need a mercury vapor light and a fluorescent dye penetrant that is made for weld crack detection. Why do I need a mercury vapor light? Where would I find out about the basics of this technique? Will I also need some sort of filter for the microscope?
Has anyone out there done something similar? What supplies and microscope attachments did they use?
Post-Doctoral Research Position Department of Cell Biology and Anatomy University of Calgary Calgary, Alberta, Canada
A post-doctoral fellowship position is available to investigate structure-function relationships of the mammalian cell nucleus. Specifically, the goal of the research is to determine at high resolution the spatial relationships of transcriptionally active genes with sub-nuclear domains, and how particular gene families are recruited to these domains. The research will involve dynamic fluorescence microscopy, digital confocal microscopy and energy filtered transmission electron microscopy, together with traditional biochemical and molecular biological approaches. Interested applicants should refer to the following recent publications: Kruhlak et al (2000) J. Cell Biol. 150, 41-51; Boisvert et al (2000) J. Cell Biol. 148, 283-292; Hendzel et al (1998) Mol. Biol. Cell 9, 2491-2507. The position includes salary, benefits and the opportunity to work in a well-equipped and dynamic environment in the Cancer Biology Research Group. Send C.V. and the names of three references to: Dr. David P. Bazett-Jones Department of Cell Biology and Anatomy 3330 Hospital Dr Calgary, AB T2N 4N1 Canada TEL: (403) 220-3025, FAX (403) 270-0737 email bazett-at-ucalgary.ca
Hello Lou, Slow down... Trying to look good? Obviously something has been lost in the translation & someone has given you the shaft in the past. I am looking for a solution to things beyond my control. I Personally have no control over the rates unless I can produce a solution (like yourself?). I view my self as a (under paid) scientist & I am very pissed off at our university's attitude toward supporting research facilities. I live eat & breath research. We drag in mega bucks for tools & nothing for staff or support. I am hoping to confirm by the voices of experience that this is not a solution. Ya know a defense without data doesn't go far. FYI our past (93-2000) fees were $5/hour on state of the art TEM, SEM, AFM ... come on whine about that! Mean while if our locals will get their bowl in a up roar like you seem prone to do, we may get a solution. I appreciate the sincerity of your support even though you missed the target.
Bruce
Lou Solebello wrote:
} Do you imply keeping fees the same as in past contracts? Forget it } guy.....that mentality is putting scientists out of work. Nothing goes down } in prices, and the COL has increased. USA scientists are already under paid } and over worked because of people like you who want to look good at the } expense of others. } ----- Original Message ----- } From: Bruce Brinson {brinson-at-cnst.rice.edu} } To: MSA Listserver {microscopy-at-sparc5.microscopy.com} } Sent: Monday, July 17, 2000 7:59 AM } Subject: CvsD,Fees } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 Microscopist & Lab Managers, } } well it's old news to a lot of you but the time has come for us to try } } to support service contracts with user fees. I guess we should be } } grateful to have deferred this expense to our users for this long. } } If we pick a rate based historical usage, all is well? I don't think } } so. The old cost vs demand has to come into play. My questions to those } } of you that have lived this experience: } } } } 1. What was the impact of significant increases in user fees with } } respect to hours logged? } } 2. Did use come back to historical levels after the sticker shock wore } } off? } } 3. Has anyone on the university level actually been able to support } } their contracts & related expenses with user fees. } } } } Your experiences & insights would be greatly appreciated. } } Also I recall that there was an meeting being organized on this & } } similar subjects at M&M 2000. Would the the organizers please contact } } me. } } } } thanks, } } Bruce Brinson } } Rice U. } } } } } }
High pressure mercury vapour lamps are used in most fluorescence microscopes because they are very intense sources producing strong emissions only at UV, blue and green wavelengths. A great many fluorescent dyes are excited by wavelengths in the visible spectrum, and various lamps can be used to excite them if the intensity is great enough. Fluorescein, for example can be excited by blue light, and rhodamine by green. For many applications, e.g. using rhodamine isothiocyanate, all you need for illumination is a powerful source of green light, such as a tungsten halogen lamp, and a filter which transmits green efficiently but cuts all other wavelengths. An interference filter would be the best choice. For viewing or photography you will need a filter which efficiently blocks green light, but efficiently transmits orange/red. Again an interference filter is best, but you will get a result with a red or orange photographic filter.
For fluorescence applications at low magnification, using a macro setup or a low power binocular microscope, and where the dye concentrations are fairly high, black UV tubes, such as those battery-powered hand lamps sold to read security marks can provide enough intensity to be useful for excitation of a wide range of fluorescent dyes. If you use these to excite rhodamine, keep the incident light off the orange barrier filter - red, orange and yellow dyed glass photographic filters are usually fluorescent themselves.
Chris
} From: "rgriffin-at-eng.uab.edu"-at-sparc5.microscopy.com To: microscopy-at-sparc5.microscopy.com
Denton's web site is up and running at www.dentonvacuum.com. You can call or fax at: Phone: (856) 439-9100 Fax: (856) 439-9111 Thermocouple gauges are called that because they measure the conductive heat loss from a heated filament with a thermocouple attached to the filament. Your TEM tech is probably more familiar with Pirani gauge, which measures the conductive heat loss by measuring the resistance of a heated filament. The Pirani gauge is commonly used on European and Asia instruments, where American instruments are more likely to use thermocouples.
Allen R. Sampson, Owner Advanced Research Systems, St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
On Friday, July 14, 2000 12:48 PM, "rlvaughn-at-unmc.edu"-at-sparc5.microscopy.com [SMTP:"rlvaughn-at-unmc.edu"-at-sparc5.microscopy.com] wrote: } Does any one know the current phone number and or web site of Denton } Vacuum? } The ones I am using : 888-336-8661, www.dentonvacuum.com are not } responding. } I need some advice on one of their old evaporators and possibly puchase a } couple of thermocouples (vacuum gage). Why do they call it a thermocouple? } The TEM service person that was here didn't think that was a proper name. } Thanks. } } Rick Vaughn } rlvaughn-at-unmc.edu } } }
From root Mon Jul 17 14:16:29 2000 Return-Path: {Microscopy-request-at-sparc5.microscopy.com} Received: (from daemon-at-localhost) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id NAA23550 for dist-Microscopy; Mon, 17 Jul 2000 13:35:14 -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 NAA23547 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Mon, 17 Jul 2000 13:34:44 -0500 (CDT) 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 NAA23540 for {microscopy-at-sparc5.microscopy.com} ; Mon, 17 Jul 2000 13:34:32 -0500 (CDT) Received: from inveresk (dialup-108.publab.ed.ac.uk [129.215.38.108]) by renko.ucs.ed.ac.uk (8.8.7/8.8.7) with SMTP id TAA27752; Mon, 17 Jul 2000 19:28:52 +0100 (BST) Message-Id: {200007171828.TAA27752-at-renko.ucs.ed.ac.uk}
Our experience was that in 1986, when charges were first imposed on our users, numbers of users crashed and stayed low, even when rates of charge were insufficient to cover the bills. When charges were raised towards realistic rates usership fell even further. Even in today's environment, where people know that charges are a fact of life, and that they should be raising them in grant funding, users are rarely prepared to include funding for em in their proposals. They want em free at the point of use, or they leave em out of the equation, or get some of it done as a favour by someone in another institution who doesn't apply charges.
Date sent: Mon, 17 Jul 2000 09:59:08 -0500 } From: Bruce Brinson {brinson-at-cnst.rice.edu} Send reply to: brinson-at-rice.edu Organization: Rice University To: MSA Listserver {microscopy-at-sparc5.microscopy.com}
Hi! I don't know much about the specificsof your test but you will need a source of UV light, such as mercury lamp, to excite the fluorescing material. Go to the Bio Dep't and talk to them about fluorescence microscopy. I would be very cautious about interpreting the results. Crack penetrating fluid have very low viscosities and surface tensions in order to penetrate the crack. Once you have broken open the specimen, I would expect the fluid to creep over the newly generated surfaces. A better way, I would think, would be to obtain a low viscosity epoxy resin ( back to the Bio Dep't), add some fluorescent dye, and vacuum impregnate the specimen. After curing the epoxy, break open the specimen. The cracked surfaces will be covered with a solid fluorescing compound that will not creep. A complication will be that the epoxy may cement together the cracked surfaces and fracture will occur elsewhere. Another approach woud be to use image analysis techniques. See Jon Russ' book "Image Processing Handbook" or "The Image Processing Tool Kit" from Reindeer Games, PO Box 37188, Raleigh, NC 27627.
I have no financial intersest in the company.
Sam Purdy Technical Center, National Steel Corp 1745 Fritz Dr, Trenton,MI 48183 spurdy-at-nationalsteel.com
Several years ago, we had some precision X-sections done using a microtome. This was done to create TEM samples of gold metalization in a low-k dielectric. Since that time, we've been unable to find the service offered in an outside lab. Does anyone know a lab offering this type of X-sectioning? Any info would be appreciated.
Steve Brockett Customer Returns and Failure Analysis Manager TriQuint Semiconductor 2300 NE Brookwood Parkway Hillsboro, OR 97124 503-615-9303 sbrockett-at-tqs.com
} Leica S 360 SEM, $35,000 } Purchased 1989, LaB6 filament. } Cathode luminescence, Robinson BSE, Solid state BSE, } Oxford 3 position EDS detector (BE, thin window, windowless), 4PI system for EDS & digital image acquisition. } Buyer pays for the move. All systems are operational and available for demo. } } Hitachi H-600 TEM, $20,000 (without CCD camera)- $50,000 (with CCD camera) } Purchased 1988, } 100 keV, W filament, } Gatan Model 791 digital camera YAG with Digital Micrograph ( 2 years old). } Buyer pays for move. All systems are operational and available for demo. } } Topcon DS 130 F, $40,000 } Purchased 1989 } Schottky field emission gun SEM, } Dual stage system } Resolution (top stage- 10 Angstroms at 30keV, 40 Angstroms at 1keV) (bottom stage-20 Angstroms at 30keV, 100 Angstroms at 1keV), Robinson BSE on lower stage, Oxford Be window EDS detector on lower stage, 4PI system for EDS and digital image acquisition, IP30 frame over imaging system. } Buyer pays for move. All systems are operational and available for demo. } } } contact: } } John Blackson } The Dow Chemical Company } Midland, MI 48667 } 517-636-6316 office } 517-638-6443 fax } jhblackson-at-dow.com } or
} Steve Rozeveld } The Dow Chemical Company } Analytical Sciences Laboratory } 1897 Bldg., Door E43 } Midland, MI 48667 } 517-636-5167 office } 517-638-6443 fax } sjrozeveld-at-dow.com }
Anyone interested in the Yearbooks of the New York Microscopical Society for 1955, 1956, 1957, 1958,1961, and 1968? All or nothing. I'll even pay the postage for domestic mailings.
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.)
Wondering if you can recommend a good penetrant and epoxy to use and where to get it. I am in the middle to source for the penetrant to perform solder joint crack (BGA etc), molded parts crack and plating crack. Perhpas you can advice if this method will be the simplest method to identify the crack/craze line.
My understand from our local supplier is they have an external accessory that can attach to the scope. The cost of the external features is about USD5000 (Olympus). The bulb is mercury lamp that can last only 200 hours.
Regards, YH Koh (MQE Engineer) Motorola CGISS Penang, Bayan Lepas FIZ, Phase 3, 11900 Penang, Malaysia. Tel : 60-4-8504089 Fax: 60-4-6124903
-----Original Message----- } From: Purdy, Sam [mailto:SPurdy-at-nationalsteel.com] Sent: Tuesday, July 18, 2000 7:27 AM To: Microscopy-at-sparc5.microscopy.com
Hi! I don't know much about the specificsof your test but you will need a source of UV light, such as mercury lamp, to excite the fluorescing material. Go to the Bio Dep't and talk to them about fluorescence microscopy. I would be very cautious about interpreting the results. Crack penetrating fluid have very low viscosities and surface tensions in order to penetrate the crack. Once you have broken open the specimen, I would expect the fluid to creep over the newly generated surfaces. A better way, I would think, would be to obtain a low viscosity epoxy resin ( back to the Bio Dep't), add some fluorescent dye, and vacuum impregnate the specimen. After curing the epoxy, break open the specimen. The cracked surfaces will be covered with a solid fluorescing compound that will not creep. A complication will be that the epoxy may cement together the cracked surfaces and fracture will occur elsewhere. Another approach woud be to use image analysis techniques. See Jon Russ' book "Image Processing Handbook" or "The Image Processing Tool Kit" from Reindeer Games, PO Box 37188, Raleigh, NC 27627.
I have no financial intersest in the company.
Sam Purdy Technical Center, National Steel Corp 1745 Fritz Dr, Trenton,MI 48183 spurdy-at-nationalsteel.com
In the past, EM/microscopy facilities I've been part of and used in Australia have not charged fees, and most still don't charge users within their institutions, or have a very minimal charge, e.g. $5/hour for confocal just to put something into laser replacement, and $500-$1000 per year for use of all EM facilities, including costs to prepare tissue, etc. To external users, government facilities are supposed to charge commercial rates, so they don't compete unfairly with fully commercial facilities. I believe it's similar in the USA.
But "full cost recovery" is coming, even for users of facilities within their own institutions. It has come to a few facilities, and the result is a dramatic fall-off in users. Even when fairly minimal fees are imposed, this breaks the bank for some people, especially for undergrad student projects.
To be more specific: 1. What was the impact of significant increases in user fees with respect to hours logged? Hours logged for TEM dropped to 30% and continued to decline. Hours on SEM dropped 50% and stayed low.
2. Did use come back to historical levels after the sticker shock wore off? No, though there was a little recovery.
3. Has anyone on the university level actually been able to support their contracts & related expenses with user fees. In my experience in 5 institutions, no, even when high fees were charged.
I don't know how to fight this, you need strong support from a large user base. Even then, you can be beaten by the bean counters if they are in ascendency.
Rosemary White Microscopy Centre CSIRO Plant Industry GPO Box 1600 Canberra, ACT 2601 Australia
Ed, You were the first to reply and they are yours for the museum. Should go out today.
Don Marshall
} From COURYHOUSE-at-aol.com Tue Jul 18 01:10:29 2000 } } From: COURYHOUSE-at-aol.com } Received: from COURYHOUSE-at-aol.com } } Date: Tue, 18 Jul 2000 01:06:41 EDT } Subject: Re: new york microscopical society } To: dmrelion-at-WORLD.STD.COM } } } we would like to have these for the museums collection. } many will get use of them. } thanks Ed Sharpe archivist for SMECC } } Coury house / SMECC } 5802 w Palmaire Ave } Glendale AZ 85301 } } { { Subj: new York microscopical society } Date: 7/17/00 9:16:22 PM US Mountain Standard Time } From: dmrelion-at-world.std.com (donald j marshall) } 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 } -----------------------------------------------------------------------. } } } Anyone interested in the Yearbooks of the New York Microscopical Society } 1955, 1956, 1957, 1958,1961, and 1968? All or nothing. I'll even pay the } postage for domestic mailings. } } Don Marsha } } }
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.)
One of our researchers is having trouble with cryo-thin sections jumping back onto the face of the block. Apparently this is a static problem. Does anyone use an anti-static device for this? If so, what is available. Also any thoughts for this cure would be appreciated. Thanks.
I am wondering if someone could help me out with trying to use John Russ's Image toolkit for putting on scale bars for digital plates. I have light micrographs that were taken at 20 and 40x magnification, and also 20 and 40x magnification of calibration grid slides of 25 and 250 micrometers from MicrobrightField Inc. These calibration images I tried to set up a txt file using calibrate by marking out the points from one grid bar to the next, and specifying it as either 25 or 250 micrometers depending on which calibration grid I used. I saved this file and imported it on a light micrograph of similar magnification. When I do this importing, and then use the magnification bar option, the bar does not reflect the calibration scale. Also, if I just use the magnification bar on the original calibration grid, it comes out to be 7micrometers for each division rather than the 25 or 250 that I have specified. If someone could suggest where I am going wrong, I would very much appreciate it. I hope I haven't completely confused people!
Thanks, Susan
Susan Carbyn Electron Microscopy Technician Atlantic Food and Horticulture Research Centre Agriculture and Agri-Food Canada 32 Main Street, Kentville, Nova Scotia B4N 1J5 Canada
i've been in a "cover costs or die" situation for the past 6 years. at first usage stayed about the same...then dropped off as my user base found cheaper and better alternatives on the equipment side of the equation (a happier and more accomodating microscopist they'll never find!), at other less "cost recovery" oriented universities. i was then funded by the NSF to update some of the lab (a new FESEM in particular). with this came a new influx of users. i'm now able to cover nearly all of the lab expenses (contracts, supplies, upgrades, and salary). i still have a slush fund to cover unsponsored research and classroom activities, but have imposed a user fee for specialized training courses.
hope this helps... b- **************************
} Date: Mon, 17 Jul 2000 12:19:25 -0500 } From: Bruce Brinson {brinson-at-cnst.rice.edu} } Reply-To: brinson-at-rice.edu } Organization: Rice University } X-Mailer: Mozilla 4.06 [en] (Win95; U) } To: Lou Solebello {microls1297-at-mindspring.com} , } MSA Listserver {microscopy-at-sparc5.microscopy.com} } Subject: Re: CvsD,Fees } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Are you sure that the scale is wrong? There could be two different kinds of problems here. One is that the tool kit draws a line whose length is determined by the size of the image, and labels it with the length in the calibrated measurement units. This might not (would not, in general) be the same length as the distance that you originally marked for establishing the calibration. The other problem might be that the calibration is in fact wrong, possibly something wrong with the file, etc., but the first explanation is more likely. If you want to create markers that are exactly a certain length then the best way to do it is to draw the line in a blank image, label it as you wish, crop to just the region around the label and line, and save it as a file. Then copy and paste this onto your images. The calibration routine in the tool kit is primarily aimed at making measurements come out in the right units, and the scale marker was more-or-less an afterthought. Hope this helps and doesn't add to the confusion.
I agree, its certainly not innocuous, but neither are many common household goods. The questions really are: is it more dangerous than other substances, which are not subject to permits and would regulations and permits improve safety in any significant way? Regulations should be relative to risks. I think its absurd that we as a reseller are required to have permits and many other regulations under the new Australian system. A reseller is not opening the jar, or even the outer metal container. A 25g glass jar of UA is enclosed in heavy Al foil (to produce low energy Al X-rays) and then enclosed in a tin can, which shields most of the remaining radiation. What about the end-user? Should they need a permit? To see that in perspective, just imagine running a lab that requires a couple dozen permits for things like OsO4, the aldehydes, cacodylic acid and many more substances including liquid nitrogen and I suggest boiling water. You'd better hire a secretary and obtain more funds to pay for those permits. I have a full discussion of the topic -at-
www.proscitech.com/uranyl.htm
The other point you are making about it being a heavy metal was rebutted recently and nicely in this forum. Gold, platinum and iridium are very heavy metals. In fact you cannot get heavier than Iridium! You can eat these heavy elements in great quantity if you can afford it and suffer no ill effect. "Heavy metal" has become a slur, but its meaningless. Uranyl acetate is fully water soluble and is not cumulative in organisms. So, however toxic as a chemical poison (and less so radiologically), its not a problem in the environment, unless you are emitting very high doses. Some locations will have laws dis- allowing the discharge of tiny quantities of a 1% UA solution; so people accumulate the material and create a problem. Much smarter to discharge UA down the sink with plenty of water. Please read up on our site if you are interested in the topic. Regarding your safety precautions: I think that the people handling 200 litre drums of uranium oxide (yellow-cake) or underground uranium miners would find your precautions highly entertaining. Yet its these people who need to be concerned, not laboratory staff who weigh out 0.25grams of UA once a month. 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 ABN: 99 724 136 560 www.proscitech.com
On Tuesday, July 18, 2000 12:41 AM, Ingber, Bruce F. [SMTP:bingber-at-commserver.srrc.usda.gov] wrote: } } } I disagree that uranyl acetate is fairly innocuous. Using the appropriate } dosimeters, we have measured increases of alpha, beta, and gamma radiation } over background levels with the lid of the reagent jar open. Increased } counts were still measured with the closed jar behind an alpha shield at } some distance. We ask our scientific and support staff to take some } necessary precautions for weighing uranyl acetate and other uranium salts } (wear dust mask and latex/vinyl gloves, use a shield in front of the } balance, etc.). } } Of course, while the dilute stain solution poses no radiation hazard, it } should be treated as heavy metal waste for disposal purposes. These are my } own suggestions and not necessarily that of USDA. } } 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-commserver.srrc.usda.gov } } -----Original Message----- } } From: jim [mailto:jim-at-proscitech.com.au] } Sent: Wednesday, July 12, 2000 8:21 AM } To: microscopy-at-sparc5.microscopy.com } Subject: want to know national permit requirements for Uranyl } Acetate } } ------------------------------------------------------------------------ } The Microscopy ListServer-Sponsor: The Microscopy Society 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 also like to know of any significant incident involving UA anywhere. } I should point out that radiation damage would not show for years and one } would never know the original cause. The radiation though is low, so unless } a person would take to sleeping with a jar of UA there is no real danger. } The real danger of UA is as a non-cumulative (happily), but severe kidney } poison, but this is of course no issue with the Radiation people. } 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 } ABN: 99 724 136 560 www.proscitech.com }
} 1. . . . If you want a great } material to coat the inside of your vacuum system with that has very good } properties, try gold. I once had a UHV system that was gold plated on the } inside. It was also very pretty.
Dear Scott, Maybe it's my high-voltage upbringing, but I'd be careful about using high-Z materials (except for apertures). The scattering and brehmsstrahlung production cross-sections are much higher for high-Z, so stray electrons and x-rays will be higher. This leads to high background for microanalysis and (for sufficiently high beam energies) x-rays emerging from the column. Yours, Bill Tivol
I have a good experience and can recommend DiATOME Static line (this particular one is Static line II), the ionizer with variable voltage. If you're sectioning with a diamond cryo knife, it should really help. Let me know if you need more info. Alice.
Alice Dohnalkova Environmental Microbiology Battelle, PNNL MS P7-50 Richland, WA 99352 tel. (509) 372-0692 fax (509) 376-1321
-----Original Message----- } From: Chris Edwards [SMTP:fishon-at-umich.edu] Sent: Tuesday, July 18, 2000 5:55 AM To: Microscopy-at-sparc5.microscopy.com
One of our researchers is having trouble with cryo-thin sections jumping back onto the face of the block. Apparently this is a static problem. Does anyone use an anti-static device for this? If so, what is available. Also any thoughts for this cure would be appreciated. Thanks.
Seems I am stucked with another sample for XTEM. :) I was preparing a sample, metal thin film on MgO (100) substrate. I am having a hard time to get it thin, monitoring the thickness with a Si piece on top. The problem is that the MgO has a bad habit of chipping even before the Si get thin enough to be light transparent. I observed a piece of the substrate after ion milling in TEM and there was a network of dislocations. Probably introduced by the polishing on diamond films. Does someone else tried to do tripod polishing with MgO to make a wedge shaped sample? Thanks.
Carlos
o-------------------------------------------------------o | Carlos Kazuo Inoki | | Department of Physics - University at Albany | | 1400 Washington Ave.- Albany - NY - 12222 | o-------------------------------------------------------o
A trick which I have found useful during tripod polishing is to use a 1/4" thick mirror instead of a clear glass on the polishing wheel. The mirror allows you to easily see if there are any contamination particles on the surface before placing the polishing disk on the glass. I got mine made at the local glass company for $6.50. I specifically asked them to insure that there weren't any scratches on the surface.
Please forgive if this is common knowledge...
************************************************************** Kim W. Pierson, Ph.D. Dept of Physics & Astronmoy University of Wisconsin-Eau Claire (715) 836-5009 FAX 836-3955
Our small educational TV production company is working on a half hour video project on VIRUSES. It is intended for classroom and library use and is for middle school age kids. We are looking for electron microscope photographs of various viruses including TMV, HIV, Herpes, Hepatitis B, and Rabies. We are also interested in images showing the invasion of a bacterium by a bacteriophage. More generic images of viruses could also be used in certain portions of the program. Our preferred format for delivery is 35mm slides, but we are flexible on that point. Reasonable image clarity is important, but images can be either B&W or colorized. We can cover all expenses, and offer a modest fee for image use. We will need a signed non-exclusive release from the owner (broad in rights, but limited to this series), and we can credit the appropriate person in the credits of the program. Images can be scanned and e-mailed to us for review, or we can log on to your site to view them. Please e-mail me at Stnehouse-at-aol.com if you would like to discuss this further.
I believe that my book "Vacuum Methods in Electron Microscopy" (Portland Press, London, 1994. ISBN I85578-052-6) provides a discussion of viscous (p. 29) and molecular (p.32) flow, and of the significance of these two modes of flow in the overall evacuation process (Ch. 2) that can be understood by anyone, who has had basic training in any field of science. Dr. Audrey M. Glauert was the Editor who monitored the writing and publication of this book, and since she is a biologist you can be sure it can be understood by biologists.
The use of specimen anticontamination devices is also discussed (pp. 12), as well as procedures for warming them up (p.400).
Reviewers' comments and a general description of this bok can be found at http://pup.princeton.edu, and http://www.2spi.com/catalog/books/book48.html
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
Maddeningly, my e-mail program destroyed my microscopy mailbox earlier today, so I lost all correspondence from several people who were interested in this microscope.
I started the bidding at $1,500, which I thought was a reasonable mid-point given the range of offers ($20 .. $600 .. $1,200 .. $2,000) I'd received on it in e-mail.
thanks much! the collection grows ed sharpe archivist for smecc
{ { Subj: Re: new york microscopical society Date: 7/18/00 5:56:08 AM US Mountain Standard Time From: dmrelion-at-world.std.com (donald j marshall) To: COURYHOUSE-at-aol.com CC: Microscopy-at-sparc5.microscopy.com
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Ed, You were the first to reply and they are yours for the museum. Should go out today.
Date sent: Mon, 17 Jul 2000 17:50:02 -0400 To: Microscopy Listserver {microscopy-at-sparc5.microscopy.com} } From: Wil Bigelow {bigelow-at-engin.umich.edu}
Hi,
I have been asked to section undecalcified bone that has been embedded in histo-technik resin at a section thickness up to 20 um. Has any one any experience or know of any references, in cutting with this medium that they would like to share eg type of knife/microtome, optimum thickness, mounting sections onto slides and staining. any info would be greatly appreciated thanks in advance
Phil Mr Philip Mutch, School of Biomedical Science, E Floor Medical School, University of Nottingham, Nottingham, NG7 2UH. UK. E-mail Philip.Mutch-at-nottingham.ac.uk
Hi, does anybody knows where I could purchase a specific cytokinin antibody? thanks in advanced Nuria
------------------------------------------------------------------------------ Nuria Cortadellas Unitat de Microscopia de Transmissio Serveis Cientifico-Tecnics Universitat de Barcelona C/ Sole i Sabaris, 1-3 08028 Barcelona Tel: +34 3 402 13 52 Fax: +34 3 402 13 98 E-mail: nuriac-at-giga.sct.ub.es
Title: Optical Microscopy and Imaging in the Biomedical Sciences
When: October 11 - October 19, 2000
Where: Marine Biology Laboratory, Woods Hole, MA, USA
Tuition: $2150 (Includes room and board, text, handouts, supplies)
Application Deadline: August 1, 2000
Admission application and information: Carol Hamel, Admissions Coordinator Marine Biological Laboratory 7 MBL Street Woods Hole, MA 02543-1015 (508) 289-7401 Internet: admissions-at-mbl.edu WWW: http://www.mbl.edu (Application forms available via Adobe Acrobat)
Course Director: Colin S. Izzard, State University of New York -at- Albany Phone: [518] 442 - 4367 EMail: csizzard-at-csc.albany.edu
Course Description:
For Whom: Designed primarily for research scientists, physicians, postdoctoral trainees and advanced graduate students in animal, plant, medical and material sciences. Non-biologists seeking a comprehensive introduction to microscopy and video-imaging will benefit greatly from this course as well. There are no specific prerequisites, but an understanding of the basic principles of optics is desirable. Limited to 24 students.
The eight day course consists of lectures, laboratory demonstrations, exercises and discussions that will enable the participant to obtain and interpret microscope images of high quality, to perform quantitative optical
measurements, and to produce photographic and video records for documentation and analysis.
Topics to be covered include: principles of microscope design and image formation bright field, dark field, phase contrast, polarized light, differential interference contrast, interference reflection, and fluorescence microscopy confocal scanning microscopy, multiphoton excitation fluorescence microscopy and image deconvolution digital image restoration and 3-D reconstruction video imaging, recording, enhancement, and intensification analog and digital image processing and analysis fluorescent probes and ratiometric-imaging laser tweezers and laser scissors
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.
Students are encouraged to bring their own biological (primary cultures, cell lines, etc.) and material specimens and to discuss individual
} 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.
Yes, 1,2 dichloroethane is a carcinogen; it's also a hepatotoxin. However, so is chloroform, and I don't know that there is a significant difference in the toxicities of these two compounds. Neither is as toxic as carbon tetrachloride.
} } 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. }
I have used formvar in chloroform and in 1:1 chloroform-acetone; either will work. Yours, Bill Tivol
} 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????) }
Dear Christine, The magnetic properties of Co should not affect EDS. The incoming electrons have sufficient energies that it takes fields on the order of kilogauss to deflect them (typical EM lens fields). If you can get an undistorted image, the fields in your sample will not affect the electrons' path. Because electrons scatter strongly, the volume of the specimen where x-rays are generated may be much larger than you would predict from the size of the beam. Also, scattering within the specimen area may result in parts of the specimen being illuminated be stray electrons and producing x-rays. You do not mention such relevant parameters as the beam and grain sizes, which lines you're looking at, or the incident electron energy. I might be able to say more if I knew these things. Yours, Bill Tivol
} This doesn't seem to cover it. Gold and platinum are heavy metals. } So are we to treat them as we treat osmium tetroxide? } Where does light end and heavy begin anyway? There is a real } need here for a proper definition of the risk. } } } } } The problem is that any form of osmium is a heavy metal, and therefore } } should be treated as one would cadmium or arsenic. It is not innocuous. } }
Dear Chris, Don, etc., It's not the atomic mass, but the chemistry that's important. Many heavy elements mimic the effects of lighter--and essential--elements, so Cd and Hg can mimic Zn, Ra can mimic Ca, and Tl can mimic K, etc. In all these cases, the heavier element has sufficiently different properties that it interferes with the function of something that the lighter element is involved with. As was pointed out, OsO4 is very toxic, but Os metal is not. In fact, Os metal has properties very much like Pt. (Again to ill- ustrate the importance of chemistry, Pt metal is harmless, but cis-platin is a toxic chemotherapy agent). Yours, Bill Tivol
I have a customer from the chemistry department that wants to obtain some SEM images of zeolite crystals in a flyash material (fine grain powder). My question is how to prep this material to get it into the probe? Any ideas or suggestions would be appreciated.
Thanks
Roy Beavers Southern Methodist University Dept. of Geological Sciences Electron Microprobe Lab P.O. Box 750395 Dallas, Tx 75275 voice: 214-768-2756 fax: 214-768-2701 E-mail: rbeavers-at-mail.smu.edu
I am in need of an AFM for the purpose of measuring the pit structure of compact disc and DVD stampers and replicas. I am considering a Q-Scope 350 from Quesant. I am wondering if anyone has any experience, good or bad, with this system or anything else from this company. Also, I am open to other suggestions if anyone has any.
When working with fly ash in the past, I would usually use a double-sticky carbon tab on an SEM stub and press it down into the ash, then gently blow off the loose particles with compressed air. Double-sided carbon tape would probably work, although the tabs seem less likely to engulf very small particles with excess adhesive.
Another alternative might be a product called "Mikrostik", a clear liquid you paint onto a stub and which dries to a tacky finish. It's good for fine particulates. It's available from Ted Pella (cat.no. 16033) and possibly other places, as well. (No financial interest in Ted Pella here.)
A problem with fly ash is that it is difficult to avoid charging, since it's basically a pile of spheres and irregularly shaped chunks in not-very-continuous contact with each other. A low vacuum SEM makes this much easier, but at the expense of high resolution. Try to blow off as many particles as possible, so the ones left are in tight contact with the adhesive. If that's not an option, you will have to coat your stub from different angles, preferably on a tilting, rotating stage in the sputter coater/vacuum evaporator.
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: Beavers, Roy [mailto:rbeavers-at-post.cis.smu.edu] Sent: Wednesday, July 19, 2000 10:49 AM To: 'Microscopy Listserver'; Microprobe List (E-mail)
List,
I have a customer from the chemistry department that wants to obtain some SEM images of zeolite crystals in a flyash material (fine grain powder). My question is how to prep this material to get it into the probe? Any ideas or suggestions would be appreciated.
Thanks
Roy Beavers Southern Methodist University Dept. of Geological Sciences Electron Microprobe Lab P.O. Box 750395 Dallas, Tx 75275 voice: 214-768-2756 fax: 214-768-2701 E-mail: rbeavers-at-mail.smu.edu
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When working with fly ash in the past, I would usually use a double-sticky carbon tab on an SEM stub and press it down into the ash, then gently blow off the loose particles with compressed air. Double-sided carbon tape would probably work, although the tabs seem less likely to engulf very small particles with excess adhesive.
Another alternative might be a product called "Mikrostik", a clear liquid you paint onto a stub and which dries to a tacky finish. It's good for fine particulates. It's available from Ted Pella (cat.no. 16033) and possibly other places, as well. (No financial interest in Ted Pella here.)
A problem with fly ash is that it is difficult to avoid charging, since it's basically a pile of spheres and irregularly shaped chunks in not-very-continuous contact with each other. A low vacuum SEM makes this much easier, but at the expense of high resolution. Try to blow off as many particles as possible, so the ones left are in tight contact with the adhesive. If that's not an option, you will have to coat your stub from different angles, preferably on a tilting, rotating stage in the sputter coater/vacuum evaporator.
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: Beavers, Roy [mailto:rbeavers-at-post.cis.smu.edu] Sent: Wednesday, July 19, 2000 10:49 AM To: 'Microscopy Listserver'; Microprobe List (E-mail)
List,
I have a customer from the chemistry department that wants to obtain some SEM images of zeolite crystals in a flyash material (fine grain powder). My question is how to prep this material to get it into the probe? Any ideas or suggestions would be appreciated.
Thanks
Roy Beavers Southern Methodist University Dept. of Geological Sciences Electron Microprobe Lab P.O. Box 750395 Dallas, Tx 75275 voice: 214-768-2756 fax: 214-768-2701 E-mail: rbeavers-at-mail.smu.edu
Does anyone have a positive experience getting a TEM viewing screen re-coated? I've got a small focusing screen for a Philips 410 , viewed with binoculars, that needs re-coating with minimal grain size. If you have used a vendor successfully, would you please forward their contact information to me?
Thank you 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
I used to buy boats from LKB for 6 mm knives. Now I can't locate LKB and the other companies seem to sell only for larger knives. Does anyone know where I can get boats for 6 mm knives? I really don't want to go back to the tape method. Thanks. Joyce Craig Chicago State University
Here's the summary (long) for all those interested in deencapsulating plastic encapsulated ICs. I have no prefferences as I haven't tried any yet, but thought the fuming sulfuric acid might be 'fun'. Thanks again!
The way this is generally done is to mill the plastic down on a grinding wheel to the point where only a fairly thin layer of plastic remains. Then, using a plasma etcher, and a mixture of oxygen to CF4 (for example, 30% oxygen/70% CF4), whereby the oxygen etches away the plastic and the CF4 etches away the glass frit that is usually found in the plastic, you can remove the remaining plastic (package) without damaging the device itself. Different people like to use different gas ratios, of course, and that is probably a function, at least to some degree, of the concentration of glass frit in their particular plastic.
The SPI Plasma Prep II unit, as shown on URL http://www.2spi.com/catalog/instruments/etchers1.html in the world, is probably the most widely used unit for doing this type of operation. It is inexpensive and highly reliable, and requires virtually no maintenance.
Chuck ____________________________________________________________________________ ******************************************************************************** *************************** 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. ___________________________________________________________________________ ******************************************************************************** *************************
Diane: attached is a text document outlining the procedure my FA lab uses. Yellow fuming nitric is usually the acid of choice. If you can get a few extra parts to practice on, that would be best. And you're right, plasma takes FOREVER. If I can be of any more help, please let me know.
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-598-1291 (pager) KFAB Physical Analysis Labs--SEM/FIB/FA Kilby Center West Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
TOOLS, EQUIPMENT, & SUPPLIES 1. Milling machine and appropriate end mills 2. Stabilo SuperFine OH pen or equivalent 3. Fume hood properly equipped for exhausting acid fumes and solvent vapors (see reference 3.4) 4. Explosion proof hot plate (see reference 3.4) 5. Ultrasonic cleaning apparatus 6. An optical microscope capable of 100X to 500X magnification, equipped with a lighting system. 7. Chemical resistant latex gloves 8. Chemical resistant laboratory coat 9. Chemical resistant safety glasses (see reference 3.5) 10. Hand tools (tweezers, scalpels and etc.) 11. Plastic micro-pipette 12. Fuming red nitric acid 13. Yellow nitric acid 14. Methyl alcohol 15. Acetone
1. RECORDING OF PACKAGE MARKINGS Record all of the device markings that are on the top and bottom sides of the devices prior to starting any of the decap operations. 2. CAVITY MILLING Determine the exact location of the chip within the package and mark the top of the device package showing the chip perimeter, using a Stabilo SuperFine OH pen and a straight edge. A SAM plot or X-ray image may be used to help determine the exact location of the chip and also to determine the thickness of the mold compound covering the chip. Note: This should be done on devices having large chips. Devices with small chips (less than 0.125 inches in their longest dimension) do not require this step. Mill a cavity out of the plastic package that is centered over the chip. The size of the milled cavity should typically be .050 to .100 inches larger than the length and width dimensions of the chip. The depth of the milled cavity depends on the thickness of the mold compound and the location and loop height of the bond wires. During the milling operation use a vacuum line to pick up the loose plastic particles generated. Caution: do not mill into the bond wires or the chip. Mill counter bores on devices with chip dimensions greater than 0.400 inches on a side. These counter bores should be made on one or more levels within the bond pad perimeter and at the outermost corners of the cavity (this is necessary to facilitate etching of the mold compound at the corners of the chip before the sides are exposed and subsequent damage to the leadframe). Care must be taken during the milling operation to avoid excessive pressure on the mill resulting in filler induced damage to the chip P.O. The end mill should not bind, bend, or "smoke" during the milling operation. 3. PACKAGE ETCHING All etching must be performed in a chemical hood that meets the requirements defined in references #.3 and 4.Heating of acid or device prior to application of acid must be done using an explosion proof hot plate that meets the requirements of reference #4. Obtain the appropriate acid for use on the mold compound being removed. Following are the acids that have been identified for the removal of the various mold compounds. Mold Compound Acid/Temperature Shinetsu Red fuming nitric acid at 140-150 degrees Celsius Plascon & Sumitomo Red fuming or yellow nitric acid 140-150 degrees Celsius Note: Fuming sulfuric acid reacts with exposed aluminum bond pad metallization and may result in ball bond discontinuity thus hampering further analysis.
· When using red fuming nitric acid it may be helpful to start the etching process using a mixture of red and yellow nitric acids in order to slow down the etch process until a "residue crust" is formed over the cavity and then switch to the red nitric acid. · Apply the acid in drops using a plastic micro-pipette. The drops should be placed in the center and at the corners of the cavity in approximately a 1:1 ratio. · Allow the acid to react with the mold compound and form a crust of dissolved compound. Caution: Do not allow the crust to dry out completely before adding additional drops of acid. · Remove the dissolved material using cotton swabs or by rinsing with acetone when the dissolved materials threaten to spill over the cavity. Caution: Rinse the device immediately with acetone if acid spills onto the package pins. · Soak the device in acetone for a minimum of 10 minutes, followed by a spray of methanol to remove loose residue and to clean the residue from the cavity rim. · Perform a thorough microscopic inspection to determine whether all necessary areas of the chip are exposed. · If dried mold compound residue persists on the chip surface, use the following in the order shown to attempt removal: · Solvent bath (such as methyl alcohol) in ultrasonic cleaner · Several drops of room temperature fuming sulfuric acid applied to the chip (with the chip at room temperature) for several seconds then rinse the device in DI water. · Several drops of fuming sulfuric acid applied to the chip with the chip on a 100 degree Celsius hot plate. Note: Fuming sulfuric acid will attack aluminum bond pads and is therefore the method of last resort.
_____________________________________________________________________ ********************************************************************* 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.
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 used to do failure analysis on semi conductor memories which were starting to be made of plastic/epoxy with glass rods about 20 years ago.
I have some technics and possible help but its too much to write. Basically you drill a small hole about 0.1" deep then heat the IC on a hot plate and then you drop your acid to remove the plastic. I dont know chemistry, I'm and Electronics Technician. I did this work with a meterial sicentist, my mentor. We used fumming sulfuric acid and fuimg nitric acid, also some type of organic pink and blue solutions to stop some of the acids etching effect.
The company back then was Burroughs Corp. today is Unysis.
I presently work for the U S Department of Energy in New York City. My phone number is 212 6203650, I'll be happy to walk through some ideas and things I learned.
Regards Peter Roiz ______________________________________________________________________ **********************************************************************
Perhaps it's time to comment on this thread.
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)
I have been asked to pass the word on a possible upcoming EM Tech. position at UC Santa Cruz.
The position has not yet been formalized but will involve doing biological TEM in the lab of Yishi Jin, a recently funded Howard Hughs investigator. Jin does serial sectioning of C. elegans and needs someone who is familiar with the trials and tribulations of biological TEM and would be willing to see this position as a longer term commitment.
The exact job description and salary etc. have not been set. It will probably be a Hughs title with some EM experience required, probably not more than a BA needed, experience and trainability might count more than degrees.
Be aware that the cost of living in Santa Cruz is significantly higher than most places in the US.
If you would like more information regarding this position when it finally materializes, please send me your contact information and I will see that you get the announcement.
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
I have imaged replicas from compact discs an CD-R without any problems using Multimode and D3000 (both in contact mode) from di, for Measuring the dimensions of the pits (height, width...) this technique is also very useful. Other AFM should work in the same manner.
Those of you who are still waiting for accommodation in Philadelphia and who are appalled at the room rates might like to know about the Windsor Hotel (A Big Ten Discounted Hotel, which is how Iknow about it). http://www.windsorhotel.com/ Rooms are $119 per night and they have availability for the week we are at the M&M conference. Just a little info. to save a little money!
It is 5 blocks from the convention center. Check out the map at: http://maps.yahoo.com/py/maps.py?BFCat=&Pyt=Tmap&newFL=Use+Address+Below&addr=1700+Benjamin+Franklin+Parkway&csz=Philadelphia%2C+PA%2C+19103&country=us&Get%07Map=Get+Map This is no worse that the Wyndham which is about 6 blocks away.
Hope this is useful to someone.
Cheers 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"
In looking for a substitute for dichlorethane, there is another hazard. Chloroform-acetone mixtures can be explosive, especially where even traces of base (in the glass of the container, even) might generate dichlorocarbene, leading to a runaway reaction leading to an explosion. Storage in particular is out of the question, and we always make sure that these two do not share the same waste bottle.
I asked our Head of Chemical Safety, and he tells me that the use of this mixture was specifically discontinued round about 1975, when he was doing his exams. I also remember reading about this in "Chemistry in Britain" long ago, but this was certainly before 1981, which is as far back as our literature database goes.
+------------------------------------------------------------------------+ | 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 | +------------------------------------------------------------------------+
For our quality system, I need to get a process going whereby I can monitor the performance of our SEM/EDS system over a period of time. Things that I am doing are e.g. resolution tests, checking the EDS calibration and so on.
I would really appreciate input from you and methods/checks you have found that work.
If this has been discussed before, maybe give me a pointer to when, so I can search the archives more effectively.
Date sent: Wed, 19 Jul 2000 14:59:39 -0500 } From: Joyce Craig {j-craig-at-csu.edu} Send reply to: j-craig-at-csu.edu Organization: Chicago State University To: microscopy listserver {microscopy-at-sparc5.microscopy.com}
Another option for hotels in Philly is the Hilton Garden Center located at the convention center. I was able to get rates of $104/night through travelocity.
The Microscopy Society of America
Those of you who are still waiting for accommodation in Philadelphia and who are appalled at the room rates might like to know about the Windsor Hotel (A Big Ten Discounted Hotel, which is how Iknow about it). http://www.windsorhotel.com/ Rooms are $119 per night and they have availability for the week we are at the M&M conference. Just a little info. to save a little money!
It is 5 blocks from the convention center. Check out the map at: http://maps.yahoo.com/py/maps.py?BFCat=&Pyt=Tmap&newFL=Use+Address+Below &addr=1700+Benjamin+Franklin+Parkway&csz=Philadelphia%2C+PA%2C+19103&cou ntry=us&Get%07Map=Get+Map This is no worse that the Wyndham which is about 6 blocks away.
Hope this is useful to someone.
Cheers 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"
Dear All, I am a first year Ph.D student in Entomology at FPRC, Buckinghamshire college, UK. My project is got to compare the gut structure of several Cerambycid woodborers and their relevance in taxonomy. I have given some of the wood borers which I am gonna work with. Can any one tell me of which fixative( with its chemical constituents) will be the best for me to use on for each beetles in order to get best results?????
Hylecoetus dermestoides Lymexylon navale (The above two species belongs to Lymexylidae but are closely related to the family cleridae). Nacerdes melanura (oedemeridae), Rhagonycha fulva (oedemeridae?), Phymatodes testaceus Hylotrupes bajulus , Stromatium barbatum ( both species belongs to cerambycidae-subfamily: cerambycinae).
We urgently require a carbon evaporator capable of shadowing. If anyone in this area has surplus equipment they would like to sell/unload, please contact me off-line.
Thanks,
Karen Rethoret Biology Department Microscopy Laboratory York University, Toronto 416-736-2100 x33289
I've done this before and it works very well. It is important to make sure that the dye is completely dry before opening the crack. Otherwise, the dye will creep across your surface of interest (yes, I've done that).
The materials we've used are from Magnaflux's Zyglo line: www.magnaflux.com
They can probably help with selection of materials and drying requirements.
These chemicals are typically used for liquid penetrant testing of materials as a nondestructive method of finding cracks and surface discontinuities. The general procedure includes 1)surface cleaning, 2)application of the dye for a prescribed period of time, 3)cleaning the dye from the surface (but not out of cracks), 4)applying a 'developer' to the surface to draw the dye out of the cracks, and 5)examination under a black light.
Since you want to look at the opened cracks, the developer step probably won't be appropriate.
The dye has a low viscosity and low surface tension so it will creep into cracks well, but to improve on this, you might want to pressurize a submersed specimen to about 2 atm. Or possibly drawing a vacuum on a submersed specimen would pull out bubbles trapped in the cracks.
The light used for fluorescent liquid penetrant testing is typically a black light consisting of a current regulating transformer, a mercury arc bulb and a deep red-purple filter (for safety and to pass appropriate wavelengths). This is a fairly common type of industrial black light. The light is supposed to produce an intensity of at least 800 microwatts per square cm at the test surface. I use one of these black lights with my stereomicroscope (not attached to the microscope - just next to it).
Hope this helps.
Diane Ciaburri General Dynamics Pittsfield MA 01210 diane.a.ciaburri-at-gdds.com (413)494-2847
I have a professor here that wants to determine the subsurface crack shape in a ceramic by placing a drop of fluorescent dye-penetrant onto indentations. The cracks are forced open after the dye is allowed to penetrate and the pre-existing crack area will be covered with dye penetrant. Our reference says to use external ultraviolet illumination in an optical microscope to distinguish the pre-cracked area.
It is my understanding that we will need a mercury vapor light and a fluorescent dye penetrant that is made for weld crack detection. Why do I need a mercury vapor light? Where would I find out about the basics of this technique? Will I also need some sort of filter for the microscope?
Has anyone out there done something similar? What supplies and microscope attachments did they use?
by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id KAA07389 for dist-Microscopy; Thu, 20 Jul 2000 10:21:21 -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 KAA07384 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Thu, 20 Jul 2000 10:20:50 -0500 (CDT) 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 KAA07376 for {microscopy-at-sparc5.microscopy.com} ; Thu, 20 Jul 2000 10:20:39 -0500 (CDT) Received: from wadsworth.org (ithaca [172.16.1.89]) by newton.wadsworth.org (8.8.8/8.8.8) with ESMTP id LAA26387 for {microscopy-at-msa.microscopy.com} ; Thu, 20 Jul 2000 11:15:02 -0400 (EDT) Sender: tivol-at-wadsworth.org Message-ID: {39771609.BC8579A5-at-wadsworth.org}
Warren E Straszheim wrote:
} On the side, I had a chemist tell me some years back that the reason that } chloroform got a bad rap was because of the minute amounts of carbon } tetrachloride that were almost inevitably present. He said chloroform was } not so bad in its purest form. Any comments? }
Dear Warren, I looked up the three compounds--CCl4, CHCl3, and (CH2Cl)2-- in the Merck Index (11th Edition). CCl4 is the worst: "Poisoning by inhalation, ingestion or skin absorption", followed by a list of nasty symptoms; CHCl3 is not nearly as bad; however, "Inhalation of large doses may cause hypotension, respiratory and myocardial depression and death. Banned by FDA from use in drug, cosmetic and food pack- aging products in 1976.", although it was in use in cough syrups and throat lozenges before that; (CH2Cl)2 does not have a section on Human Toxicity, but under Caution, "Vapors may produce irritation of respira- tory tract and conjunctiva, corneal clouding, CNS depression, liver and kidney impairment." All three "may reasonably be anticipated to be carcinogenic." The 12th edition lists fewer symptoms for (CH2Cl)2 and CHCl3 and does not have a Human Toxicity section for CCl4, although the same quotation and nasty symptoms are listed in the Caution section. I'm sure from the above that CCl4 impurities in CHCl3 would make it much more toxic, but CHCl3 is not innocuous. Of course, all these compounds should be used in a hood. Yours, Bill Tivol
} For our quality system, I need to get a process going whereby I can monitor the performance of our SEM/EDS system over a period of time. Things that I am doing are e.g. resolution tests, checking the EDS calibration and so on. } } I would really appreciate input from you and methods/checks you have found that work. }
Dear Loukie, Resolution tests for Si(Li) detectors--the usual ~145 eV resolution EDS detector--are done with the Mn K-alpha line, so evaporating Mn on a formvar grid, placing it in the scope, and measuring the FWHM of the peak will give you the resolution of your system. Using a 55Fe source with the detector not mounted in the scope will test the resolution of the crystal apart from noise introduced by the scope, e-beam, etc. If these two mea- surements differ, and the resolution from the latter measurement is OK, then the detector is OK, but something in the probe system is lowering the resolution. I use Al evaporated onto a formvar-coated Cu grid for calibra- tion. I set up the conditions such that I get roughly equal count rates from Al and Cu K-alpha lines by putting the beam near a grid bar and tweaking the appropriate adjustments on the analyser according to the manual. The software for all these measurements was included in the analyser we bought nearly 20 years ago, and, I'm sure, is included in every system now on the market. These checks take ~10 min each at most. Yours, Bill Tivol
A post-doc in our Chemical Engineering department came to me with this need - to quote:
"Is there a way to distinguish between white polymer "buds" and microorganisms in a black-and-white SEM picture at a magnification of 8000X ? Both the cultured "bugs" and "buds" have the same size, shape, and appearance in/on spherical carbon/polymer beads that are sliced in half, fixed, mounted, and then viewed under the SEM".
As you can see, it would appear that the polymer beads have surface textures/features ("buds") that are of comparable "shape" and "size" to the microorganisms cultured on the surface of the beads. I have yet to see the photos, so I can not comment on just what he is, or is not, seeing. The sample prep, as I understand it (never having done it!):
1. fixation in 1% glutaraldehyde with 0.1 molar cacodylate (excuse any misspellings) 2. post-fix of 1% osmium tetroxide in 0.1 molar cacodylate 3. dehydration in ethanol 4. critical point drying 5. sputter coating
(whew!.....a lot more complicated than powder on a carbon sticky tab!!)
So, based on this information, can any of you suggest a/other (pre-SEM) processing technique(s) (stains?) that he and our microscopist (in the Biology department) might utilize to help "differentiate" these "buds" from the cultured microorganisms?
Winton
PS: you can direct your feedback to me or the listserv - in either case I will compile the responses and have them available should any of you out there have need for them
PS #2: I have an SEM too, with a Be-windowed EDX detector (= no light-element detection)
--
Winton C. Cornell, Ph.D. Department of Geosciences The University of Tulsa Tulsa, OK 74104 phone: 918-631-3248 fax: 918-631-2091
Does anyone have a positive experience getting a TEM viewing screen re-coated? I've got a small focusing screen for a Philips 410 , viewed with binoculars, that needs re-coating with minimal grain size. If you have used a vendor successfully, would you please forward their contact information to me?
Dear Doug,
Some years ago we had a very good experience with Grant
Scientific. We talked to Dana Dunkelberger; his phone # then was
(803) 892-2841--I'd make it better than even that the area code
has been changed. We sent him blanks for both standard high-
brightness and high-resolution focussing screens and have been
using them ever since. The quality is excellent and the prices were
OK at several hundred dollars each screen. I also have a procedure
we have used to coat screens ourselves. Other than as a satisfied
customer, I have no relationship with Grant Scientific.
} From: "Tindall, Randy D." {TindallR-at-missouri.edu} To: "'Beavers, Roy'" {rbeavers-at-post.cis.smu.edu} Copies to: "'microscopy-at-sparc5.microscopy.com'" {microscopy-at-sparc5.microscopy.com}
Please reply to: kimwin-at-eecs.umich.edu
A colleague of mine (Kim Winnick) has a ion miller that needs service and I was wondering if anyone knows of any service organization that could help him out.
Ion Mill System Description: Manufactured by CVC (Commonwealth Scientific Corp): Millatron IV G Ion Beam Source with a Millatron 400 Power Supply, an ID-3500 Beam Controller and a Single Wafer Load Lock Assembly.
ADVThanksANCE
Cheers 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"
LKB is now part of Leica and the 6mm (actually, they're 6.4mm) boats or "Trufs" are available from Leica distributors like ourselves in cases of 500, part #16840042.
Best regards, Steven Slap
****************************** Energy Beam Sciences, Inc. The Laboratory Microwave Company http://www.ebsciences.com Adding Brilliance to Your Vision ******************************
-----Original Message----- } From: Joyce Craig [SMTP:j-craig-at-csu.edu] Sent: Wednesday, July 19, 2000 4:00 PM To: microscopy listserver
I used to buy boats from LKB for 6 mm knives. Now I can't locate LKB and the other companies seem to sell only for larger knives. Does anyone know where I can get boats for 6 mm knives? I really don't want to go back to the tape method. Thanks. Joyce Craig Chicago State University
I am a student with a great deal of SEM-EDX mapping data that was obtained on an Amray 1830 SEM with a Noran Pioneer Series II x-ray analyzer with micro-Z detector. I am trying to use this data to look at the differences between uranium precipitates obtained in a batch study with apatite (Ca5(PO4)3(F,OH)) and potassium. I'm getting interesting trends based on the area% association of U-Ca-K-P and pH. However, there is a huge difference between the sum of all of the area%s reported for a given sample (which includes a "no element" area%). The total summed area % for these sample/scans ranges from 20% to 87%. The elemental analyses of these precipitates indicated that most (97 to 99.9 wt%) of any given sample was made up of U, K, Ca, and P. Can anyone explain to me what it means when the total area% of all of the possible associations mapped sums to much less than 100%? What are the possible differences between samples or scans that sum to near 100% and those that sum to 20%? Tami Thomas - tthomas-at-ch2m.com -
Please reply directly;Somayyah is not on this listserver.
} Fabian-Baber is a small production and communication company in Primos, } Pennsylvania that creates educational materials. We are currently producing } a ten-part video series for middle school children on the human body, which } includes segments such as The Brain & Nervous System, Cells, Genes & } Heredity, The Immune System, etc. We have just begun an arduous search for } a broad variety of interesting, creative, and dynamic images and animations } of the inside of the human body. } One of the things we are looking for are images using a variety of } microscopy techniques, both still and moving, of any aspect of the human } body. As we are a small company which produces only educational material, } our budget is fairly modest, however we still strive to produce high quality } material. Thus we are looking for images donated by researchers and } research institutions. Donors of course will be acknowledged in the } credits. If necessary, we do have some funds allotted for obtaining images } and footage for this series. } If you are interested in helping this project or learning more about it, } please get in touch with me, Somayyah, at the contact listed below. I would } be most appreciative of any assistance you could offer. Thank you in } advance for your time and attention. } } Somayyah Siddiqi } Fabian-Baber Communications, Inc. } 525 Mildred Ave. } Primos, PA 19018 } e-mail: sam-at-fabian-baber.com } phone: 610 623 7812 } fax: 610 623 8970
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
Hello, Can anyone out there help me. I am not sure if this is the right forum for this question. I am still looking for a film scanner for 4X5 film. The work is mainly technical macro work in the magnification range 1X to 50X, the subject matter being mainly aero engine parts. We have decided that we do not need more than about 1000dpi as most of our scans will be at about 300dpi as the prints are for reports and are rarely enlarged. So far I have seen good reports on the Microtek and the Agfa Duoscan. Any comments and also any comments on others such as the Umax Powerlook 3 or the Epson would be welcome. Thanks in advance for any help. Cheers Richard Black
Email: cresponc-at-moffitt.usf.edu Name: Nichole Crespo School: University of South Florida
Question: I have a little experience with fluorescence microscopy and am looking for a good course (a week or so) on fluorescent microscopy techniques (fixation, confocal microscopy, etc.) Know of any?
Hi Knowledgeable Ones, A few weeks ago I sent in a question regarding problems a colleague of mine had been having with Vectashield and autofluorescence on tissue that had been labeled with a secondary antibody conjugated to FITC, and received many emails of support and suggestions. For anyone curious, the theme was two part. The first thing to try was a new mounting medium. Secondly a different fluorophore (especially favoured was Alexa 488). Thanks to all of you who responded. Here's the question. We decided to try a different mounting medium, specifically Citifluor. Moments before I sat down to write this, there was a small scream of exasperation from the microscope area as my colleague noticed that what she had just used was Citifluor non-fluorescent immersion oil. We quickly looked up its specifications in the catalog, and its usefulness in this application is still nebulous. Does anyone have any thoughts/advice? Thanks again, 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
There is a complex interaction between the beam and the specimen. Usually standardisation is performed in spot mode or by simply selecting the highest magnification buttons. If you repeat the standardisation on a homogeneous standard using a lower magnification your results will be different to a spot analyses. There are many reasons including: the spatial resolution for your atomic number specimen (using a spot) will be several micrometers in area, so the area analysed is not the actual. Furthermore, the beam deflection system does not necessarily deliver the same number of electrons at various magnifications, there will for instances be different losses at the apertures. To do area analyses you would at least need to also standardise using the same area. 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 ABN: 99 724 136 560 www.proscitech.com
On Friday, July 21, 2000 4:33 AM, Thomas, Tami/SEA [SMTP:tthomas-at-ch2m.com] wrote: } } } I am a student with a great deal of SEM-EDX mapping data that was obtained } on an Amray 1830 SEM with a Noran Pioneer Series II x-ray analyzer with } micro-Z detector. I am trying to use this data to look at the differences } between uranium precipitates obtained in a batch study with apatite } (Ca5(PO4)3(F,OH)) and potassium. I'm getting interesting trends based on } the area% association of U-Ca-K-P and pH. However, there is a huge } difference between the sum of all of the area%s reported for a given sample } (which includes a "no element" area%). The total summed area % for these } sample/scans ranges from 20% to 87%. The elemental analyses of these } precipitates indicated that most (97 to 99.9 wt%) of any given sample was } made up of U, K, Ca, and P. } Can anyone explain to me what it means when the total area% of all of the } possible associations mapped sums to much less than 100%? What are the } possible differences between samples or scans that sum to near 100% and } those that sum to 20%? } Tami Thomas - tthomas-at-ch2m.com -
This equipment in almost new condition. If some one needs this it is in vary good shape.
I was helping Bob Taig sort out a Raman Spectrometer. It uses a 4 or 5 watt laser(tunable I think but I am not sure) and a frequency doubler through a 1 meter long 4 reflector monocromater with a cooled detector and cooled photomultliplier. It uses an Olympus BH2 microscope with transparent and opaque lighting and 100 X.95, 50x.80 and 5X.13 Plan objectives. The microscope has 4 stages and a flip top 1.3 condenser.
It was removed from a working lab when the entire lab was dismantled. It is in almost new condition. The plastic covering on the microscope stage is still attached and almost perfect. This is normally discarded at installation. It was billed in February of 1988 for a little over a quarter million dollars.
It is marked Yobin Yvon and Mole systems.
It appears to work by sending the laser into the monocromater and into the microscope and the light is reflected off the sample back out the same path as it came in and is split off in the monocromater to the detector.
It a appears all MSDOS computers, software and documentation is intact. The only thing we could find missing was the video camera that mounted on the microscope and the table that held the monocromater and microscope above the laser and the mirrors and lenses that manipulated the light from what came out of the laser to what the moncromater needed.. The Microscope is set up to feed directly into a video camera. A binocular head could be used if some one wanted to set it manually insted of remotely.
There is a lot of very nice equipment here if someone has a use for it. It has some very nice hardware for a optical bench.
If you are interested contact Bob Taig : 580-765-9727 ustrade-at-fullnet.net The equipment is located at Ponca City Oklahoma.
I have some pictures I will have up tomorrow at http://www.couger.com/gcouger/raman/
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
Randy and Kent I also use the double sticky (conductive) tape procedure to mount zeolites and fly ash specimens. The only additional caution I would add, is to avoid blowing the specimen across the stub. Because of the significant difference in shape/size of the zeolite crystals and the fly ash agglomerates, you may change the distribution of the particles in the specimen that you examine. For example, if bigger less dense flocs of fly ash exist, they may readily blow away. Rather than blowing onto the stub, I try to get several representative micro samplings and disperse those across the tape surface, and then blow away the excess to achieve a "mono-layer" of particles. Good Luck, Brad
} ---------- } From: Kent Hampton[SMTP:khampton-at-oz.oznet.ksu.edu] } Sent: Thursday, July 20, 2000 11:44 AM } To: Tindall, Randy D. } Cc: 'microscopy-at-sparc5.microscopy.com' } Subject: RE: Images of Zeolites } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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: "Tindall, Randy D." {TindallR-at-missouri.edu} } To: "'Beavers, Roy'" {rbeavers-at-post.cis.smu.edu} } Copies to: "'microscopy-at-sparc5.microscopy.com'" } {microscopy-at-sparc5.microscopy.com} } Subject: RE: Images of Zeolites } Date sent: Wed, 19 Jul 2000 12:45:58 -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 } } -----------------------------------------------------------------------. } Here, we routinely work with starches and do so in a similar } manner. Regular double sided tapes works fine for us. To help get } some area with a "mono-layer" of particles we use a spatula to } place a small pile of the powder on one edge on the stub and then } blow across the stub spreading the powder. } } } } When working with fly ash in the past, I would usually use a } double-sticky } } carbon tab on an SEM stub and press it down into the ash, then gently } blow } } off the loose particles with compressed air. Double-sided carbon tape } would } } probably work, although the tabs seem less likely to engulf very small } } particles with excess adhesive. } } } } Another alternative might be a product called "Mikrostik", a clear } liquid } } you paint onto a stub and which dries to a tacky finish. It's good for } fine } } particulates. It's available from Ted Pella (cat.no. 16033) and } possibly } } other places, as well. (No financial interest in Ted Pella here.) } } } } A problem with fly ash is that it is difficult to avoid charging, since } it's } } basically a pile of spheres and irregularly shaped chunks in } } not-very-continuous contact with each other. A low vacuum SEM makes } this } } much easier, but at the expense of high resolution. Try to blow off as } many } } particles as possible, so the ones left are in tight contact with the } } adhesive. If that's not an option, you will have to coat your stub from } } different angles, preferably on a tilting, rotating stage in the sputter } } coater/vacuum evaporator. } } } } 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: Beavers, Roy [mailto:rbeavers-at-post.cis.smu.edu] } } Sent: Wednesday, July 19, 2000 10:49 AM } } To: 'Microscopy Listserver'; Microprobe List (E-mail) } } Subject: Images of Zeolites } } } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } List, } } } } I have a customer from the chemistry department that wants to obtain } some } } SEM images of zeolite crystals in a flyash material (fine grain powder). } My } } question is how to prep this material to get it into the probe? Any } ideas or } } suggestions would be appreciated. } } } } Thanks } } } } Roy Beavers } } Southern Methodist University } } Dept. of Geological Sciences } } Electron Microprobe Lab } } P.O. Box 750395 } } Dallas, Tx 75275 } } voice: 214-768-2756 } } fax: 214-768-2701 } } E-mail: rbeavers-at-mail.smu.edu } } } } } } } } } Kent Hampton } } } Department of Entomology } khampton-at-oz.oznet.ksu.edu } Phone: 785 532-4746 }
We purchased a new 15cm diam. screen a year ago from the Electron Optics division of Phillips S.A for about R12000. They had quoted an amount of around R13500 for having our phospherous screen resurfaced. This amount probably included the shipping of the screen to and from the Phillips agents. Obviously, from South Africa everything appears to escalate, the further across the ocean we go, due to our exchange rates. However, since then, an American based company (Sorry I don't have the name) has taken over Phillips and Zeiss has been given the contract for the service of the equipment. Just something to consider before you make your decision. Nazlia
I have a March Plasmod (Plasma Etcher) which no longer produces the rf needed to generate the plasma. Does anyone have a surplus/broken/obsolete unit that could possibly still have operable electronics that they want to get rid of? I'd like to try to repair mine before declaring it junk. (if it can't be repaired economically, perhaps someone will need the parts on it that are still good.)
Email: ay813-at-chebucto.ns.ca Name: Robert Harnum
Question: 1. Which books do you recommend that a person new to light microscopes read regarding the use and care of light microscopes? I cannot find very much about the care of light microscopes. I want to look at bacteria.
We purchased a new 15cm diam. screen a year ago from the Electron Optics division of Phillips S.A for about R12000. They had quoted an amount of around R13500 for having our phospherous screen resurfaced. This amount probably included the shipping of the screen to and from the Phillips agents. Obviously, from South Africa everything appears to escalate, the further across the ocean we go, due to our exchange rates. However, since then, an American based company (Sorry I don't have the name) has taken over Phillips and Zeiss has been given the contract for the service of the equipment. Just something to consider before you make your decision. Nazlia
Date sent: Fri, 21 Jul 2000 08:14:13 -0500 To: Microscopy-at-sparc5.microscopy.com } From: ay813-at-chebucto.ns.ca ()
Tami writes ...
} ... } ... I am trying to use this data to look at } the differences between uranium precipitates } ... } ... However, there is a huge difference between the sum } of all of the area%s reported for a given sample } (which includes a "no element" area%). The total summed } area % for these sample/scans ranges from 20% to 87%. } ... } Can anyone explain to me what it means when the total area% } of all of the possible associations mapped sums to } much less than 100%? } What are the }
The results of this type of spacial elemental analysis should have resulted in two other calculated percentages ... (1) undefined pixels and (2) multiply defined pixels. Depending on the software, the first is generally included in the total you report, but not the second. It is even possible neither have been included in the total. Do you have access to these numbers, or the pixel maps???
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/
The following tip was passed to me by a user in the Scottish Agricultural College for spreading a monolayer of starch granules:
Dip a polished acrylic plastic rod into the powder and roll it gently on the surface of a double-sided carbon tab. The method has two advantages - grains are electrostatically attracted to the surface of the rod forming a monolayer, and the rod applies enough pressure to ensure that the particles are stuck down across a broad base, improving continuity of the metal coating on the grain with that of the tab surface and stub, minimizing charging.
It works for other particulates too, but can only be recommended for hard particulates that will withstand light compressive forces without damage.
Chris
} From: "Huggins, Bradley J" {HUGGINBJ-at-bp.com} To: "Tindall, Randy D." {TindallR-at-missouri.edu} , "'Kent Hampton'" {khampton-at-oz.oznet.ksu.edu} Copies to: "'microscopy-at-sparc5.microscopy.com'" {microscopy-at-sparc5.microscopy.com}
richard writes ...
} ... } I am still looking for a film scanner for 4X5 film. } ...
Flatbed scanners will not adapt very well to the optical density of films. At best, they may not achieve 3.0. If yours are TEM films then you probably want a scanner which is capable of near OD=4. The only scanner I'm aware of, rated at OD=3.8, also reasonably priced is the Polaroid 4x5 Ultra (~$4500). I realize this is much more expensive than the scanners you are asking about, although they should be adequate for SEM films.
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/
Does anyone have any recommendations for a conductive epoxy (other the Ag paint) that can be used to mount specimens to a stud that will be used for both polishing and subsequent analysis? (i.e., mechanically durable, water resistant and stable under ion bombardment)
Any info would be appreciated.
Brenda I. Prenitzer, Ph.D. Member of Technical Staff Cirent Semiconductor (Lucent Technologies) 9333 S. John Young Parkway 6D-Lab Orlando, FL 32819-8612
For Sale: Noran Oz Confocal Laser Scanning Microscope System with a new Krypton-Argon laser, red HeNe, SGI O2 (acquisition), SGI Indy R5000 (analysis) and an Olympus IX 50 inverted microscope with 20x, 60x and 100x Plan Apo lens'. This system is only 4 years old and fully operational. Some highlighted features are:
AOD laser scanning, allows for very fast acquisition rates (up to 480 frames/sec), for live localization, and slow scan for fixed material.
Triple label capability (488, 568 and 633nm excitation) with broad and narrow band emission filter sets.
Two and Three dimensional analysis software by Intervision, Tiff and RGB formats (amoung others).
Deconvolution software allows for thinnest possible z-sectioning for light microscopy.
Primary dichroic filter wheel upgrade for precise fluorophore registration.
We are offering this system for $180,000, buyer to pay shipping. I will personally give free on-site setup and training. Please contact Michael Delannoy for further information (410) 955-1365.
Michael Delannoy Assistant Director JHMI Microscopy Facility Johns Hopkins School of Medicine Baltimore, Maryland
Dear Winton, With my experience of looking at organic materials, there is usually a sulfur peak from the protein. If your specimen prep doesn't wash it away and your sputter coat is thin, you might use the EDX to distinguish bacteria from polymer. BTW, you should be able to look at both the polymer and bacteria with no fixing or dehydrating, ad this should simplify identification. At 11:20 AM 7/20/00 -0500, you wrote:
} Microscopists: } } A post-doc in our Chemical Engineering department came to me with } this need - to quote: } } "Is there a way to distinguish between white polymer "buds" and } microorganisms in a black-and-white SEM picture at a magnification of } 8000X ? Both the cultured "bugs" and "buds" have the same size, } shape, and appearance in/on spherical carbon/polymer beads that are } sliced in half, fixed, mounted, and then viewed under the SEM". } } As you can see, it would appear that the polymer beads have surface } textures/features ("buds") that are of comparable "shape" and "size" } to the microorganisms cultured on the surface of the beads. I have } yet to see the photos, so I can not comment on just what he is, or is } not, seeing. The sample prep, as I understand it (never having done } it!): } } 1. fixation in 1% glutaraldehyde with 0.1 molar cacodylate (excuse } any misspellings) } 2. post-fix of 1% osmium tetroxide in 0.1 molar cacodylate } 3. dehydration in ethanol } 4. critical point drying } 5. sputter coating } } (whew!.....a lot more complicated than powder on a carbon sticky tab!!) } } So, based on this information, can any of you suggest a/other } (pre-SEM) processing technique(s) (stains?) that he and our } microscopist (in the Biology department) might utilize to help } "differentiate" these "buds" from the cultured microorganisms? } } Winton } } PS: you can direct your feedback to me or the listserv - in either } case I will compile the responses and have them available should any } of you out there have need for them } } PS #2: I have an SEM too, with a Be-windowed EDX detector (= no } light-element detection) } } } -- } } Winton C. Cornell, Ph.D.
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 are also pricing scanners now. If anyone out there has experiences with the Umax Powerlook 3000, particularly used to scan TEM negatives, I'd be grateful for your insights.
Thanks, Wharton Sinkler UOP LLC Des Plaines, IL
} -----Original Message----- } From: michael shaffer [SMTP:mshaf-at-darkwing.uoregon.edu] } Sent: Friday, July 21, 2000 10:06 AM } To: richard black; Microscopy news group } Subject: RE: Film Scanners } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } richard writes ... } } } ... } } I am still looking for a film scanner for 4X5 film. } } ... } } Flatbed scanners will not adapt very well to the optical density of } films. At best, they may not achieve 3.0. If yours are TEM films } then you probably want a scanner which is capable of near OD=4. The } only scanner I'm aware of, rated at OD=3.8, also reasonably priced is } the Polaroid 4x5 Ultra (~$4500). I realize this is much more } expensive than the scanners you are asking about, although they should } be adequate for SEM films. } } 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/ } }
Dear Brenda, We, Electron Mictroscopy Sciences, did offering this TWO PART CONDUCTIVE SILVER PAINT, product number 12642-14. Please contact us at 1-800-523-5874. Meatime, we believe some other EM supply houses also carrying this product. Regards, Bang Nguyen
I would strongly guess that the answer will lie with how the area fractions are determined as Michael Sshaffer (shAf) has suggested. I am not familiar with the Noran system so I can only offer general comments.
Depending on the collection conditions, it is possible that you have a lot of pixels with no x-ray counts, or counts that are indistinguishable from background. What are you conditions and counts per pixel?
I would certainly think that with a proper map that you would approach 100% coverage. Perhaps you can put some images up on a web site so those of us interested might take a look at them. Or you might e-mail a copy _just to those of us interested_. Do not send a copy to the whole list!
Warren S.
At 12:33 PM 7/20/2000 -0600, you wrote: } I am a student with a great deal of SEM-EDX mapping data that was obtained } on an Amray 1830 SEM with a Noran Pioneer Series II x-ray analyzer with } micro-Z detector. I am trying to use this data to look at the differences } between uranium precipitates obtained in a batch study with apatite } (Ca5(PO4)3(F,OH)) and potassium. I'm getting interesting trends based on } the area% association of U-Ca-K-P and pH. However, there is a huge } difference between the sum of all of the area%s reported for a given sample } (which includes a "no element" area%). The total summed area % for these } sample/scans ranges from 20% to 87%. The elemental analyses of these } precipitates indicated that most (97 to 99.9 wt%) of any given sample was } made up of U, K, Ca, and P. } Can anyone explain to me what it means when the total area% of all of the } possible associations mapped sums to much less than 100%? What are the } possible differences between samples or scans that sum to near 100% and } those that sum to 20%? } Tami Thomas - tthomas-at-ch2m.com -
---------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
I am tasked with setting up an electronic library of SEM images, and I'm looking for good database software applications for cataloging and viewing images. Multi-user access and strong software technical support are primary requirements. What are my options?
} Greetings! } } Does anyone have any recommendations for a conductive epoxy (other the Ag } paint) that can be used to mount specimens to a stud that will be used for } both polishing and subsequent analysis? (i.e., mechanically durable, water } resistant and stable under ion bombardment) } } Any info would be appreciated. } } 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
You might contact Ablestick Adhesive in Gardena, CA. I worked for them on summer many (and I do mean many) years ago, they made silver epoxy and copper epoxy that might fill the bill.
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 **********************************************
Sorry to be fielding another question so soon after my "bugs and polymer beads" question; this time, however, I am on a very different track.
In a paper on nanowire arrays (nano-scale nickel wires) I note a technique entitled "nuclear track etching" by which porous templates were constructed in single-crystal muscovites. Apparently a collimated alpha-particle beam was used to generate the porous templates. (in which the Ni was later electrochemically deposited)
Do any of you know what sort of instrumentation this requires? We would like to produce sub-micron-scale holes in polymers for specialized battery development, and right now we're searching for a technique to do this. (nuclear etching?)
Winton
PS: the paper on nanowires does not mention the instrumentation
.respond directly to me or to the listserv
--
Winton C. Cornell, Ph.D. Department of Geosciences The University of Tulsa Tulsa, OK 74104 phone: 918-631-3248 fax: 918-631-2091
At 08:45 AM 7/21/00, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Try the Powerlook III. It does a nice job with chromes and negs. Since the TEM neg does not have a color cast, best scanning is by setting the scanner to transparent positive and reverse the image in Photoshop.
Depending on what you intend to do with the image file, this scanner should do it all. They run about $1100 with SCSI interface or a bit more with USB.
gary g.
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Modern surfers use PC boards. You can too at http://photoweb.net ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
The silver epoxy that we use for the small angle cleavage technique and for mounting samples on SEM stubs is H-22 epoxy from Epoxy Technology in Billerica, Mass. This epoxy is viscous, has a long working time, and needs to be heat cured.
-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: Friday, July 21, 2000 11:21 AM } To: 'List Server' } Subject: Conductive Epoxy? } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: 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 } } } -------------------------------------------------------------- } ---------. } } } } Greetings! } } Does anyone have any recommendations for a conductive epoxy } (other the Ag } paint) that can be used to mount specimens to a stud that } will be used for } both polishing and subsequent analysis? (i.e., mechanically } durable, water } resistant and stable under ion bombardment) } } Any info would be appreciated. } } 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 } }
Try Thumbsplus at www.cerious.com This is a great program. It is shareware and relatively inexpensive.
-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: default [mailto:byron.johnson-at-medtronic.com] } Sent: Friday, July 21, 2000 1:46 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: image database 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/MicroscopyListse} rver/FAQ.html } } } -------------------------------------------------------------- } ---------. } } } I am tasked with setting up an electronic library of SEM } images, and I'm } looking for good database software applications for cataloging and } viewing images. Multi-user access and strong software } technical support } are primary requirements. What are my options? } }
I have a open post-doc position and I am looking for a "hands-on" microscopist who can do excellent science. The position is in the materials science department, university of illinois, urbana-Champaign. I'd appreciate your help in passing this along to potential applicants.
Here is the job ad: ****************************************************************************
Postdoctoral Research Associate in In-Situ Thin Film Growth and Electron Microscopy
Applications are invited for a postdoctoral associate position at the Dept. of Materials Science and Engineering of University of Illinois at Urbana and Champaign to begin as soon as possible. The position will involve in-situ thin film growth in a UHV transmission electron microscope and study of interface structures using advanced electron microscopy at the Frederick-Seitz Materials Research Laboratory. A vita and three letters of recommendation should be sent to Prof. J.M. Zuo, Dept. of Materials Science and Engineering, University of Illinois at Urbana and Champaign, 1304 W Green Street, Urbana, IL 61801; 217-333-2736 (fax); (217) 244-6504 (phone); jianzuo-at-uiuc.edu (email). The successful candidate should have experience with UHV instruments and thin-film deposition. Experience with RHEED and transmission electron microscopy is strongly desired. A Ph. D. in Physics or Materials Science is required. The appointment is initially for one year and may be renewed for second year.
To be advertised in Physics today ****************************************************************************
Sorry to be fielding another question so soon after my "bugs and polymer beads" question; this time, however, I am on a very different track.
In a paper on nanowire arrays (nano-scale nickel wires) I note a technique entitled "nuclear track etching" by which porous templates were constructed in single-crystal muscovites. Apparently a collimated alpha-particle beam was used to generate the porous templates. (in which the Ni was later electrochemically deposited)
Do any of you know what sort of instrumentation this requires? We would like to produce sub-micron-scale holes in polymers for specialized battery development, and right now we're searching for a technique to do this. (nuclear etching?)
Winton
PS: the paper on nanowires does not mention the instrumentation
.respond directly to me or to the listserv Dear Winton, In order to etch tracks with alpha particles, you need to give them kinetic energies of at least a few MeV--nuclides which undergo alpha decay typically produce ~5 MeV alphas. Thus, either a suitable radionuclide source or a cyclotron would be required. However, even with this kind of source there may be problems. The text for my radiation sciences course, Nuclear and Radiochemistry, by G. Friedlander, et al., has a section on nuclear track detectors in which it is stated, "For each material there is a critical value of specific ionization, (dE/dx)c, below which tracks are not registered. For example, (dE/dx)c is ~ 13 MeV mg^-1 cm^2 for mica and ~4 MeV mg^-1 cm^2 for Lexan, two of the most widely used detectors. As a result mica does not register ions with A {~30, Lexan is insensitive to ions with A {~12." Alphas have A = 4. Now this text is old (1981) and not too good, so the technique for producing appropriate tracks may have been improved or Friedlander may be incorrect for the kind of tracks you need. Also, the tracks are produced when the incident particles ionize and/or displace atoms in the target, so the tracks may not be as small as you would like. Bear in mind that electrons incident on a specimen generate x-rays throughout a teardrop-shaped volume about 1 micron across. Although alphas do not have as great a spread as electrons, much of the ionization produced is from secondary particles, and these will be electrons. If the paper you refer to tells you how to generate suitable tracks given a source of alphas, then you are probably OK. Good luck. Yours, Bill Tivol
At 10:46 AM 7/21/00, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Consider ImageAXS. It is set up to manage your image database and also to allow you to set up CDs with thumbnails and full resolution files.
The other option is ThumbsPlus--but Cerious Software is rather dysfunctional based on my experience with them.
Other options are Canto Version 5 Cumulus, Kudo Image Browser {http://www.imspace.com} , Extensis/CreativePro Portfolio {http://creativepro.com} . See {http://riecks.com/redbarns.html} or {http://riecks.com/crash/} . I believe the Kudo software is still $49 for the web download version. You can download a demo version for free and give it a spin. Then there is Gallerymaker (www.rjhsoftware.com) which produces thumbnails and webpages, all of which are customizable. Cheap as I remember too. I just use it to batch process thumbs from whole directories.
*snip* Winton A brand of commercially-available polycarbonate filters is made using this method. Nuclepore have pores mostly with perfectly circular cross sections sizes from about 50nm to 12µm. I am writing from home so unable to check, but I think they are marketed by Whatman. Your usual supplier of filter media should be able to source.
Chris
} We } would like to produce sub-micron-scale holes in polymers for } specialized battery development, and right now we're searching for a } technique to do this. (nuclear etching?) } } Winton } } PS: the paper on nanowires does not mention the instrumentation } } .respond directly to me or to the listserv } Dear Winton, } In order to etch tracks with alpha particles, you need to give them } kinetic energies of at least a few MeV--nuclides which undergo alpha decay } typically produce ~5 MeV alphas. Thus, either a suitable radionuclide } source } or a cyclotron would be required. However, even with this kind of source } there may be problems. The text for my radiation sciences course, Nuclear } and Radiochemistry, by G. Friedlander, et al., has a section on nuclear } track } detectors in which it is stated, "For each material there is a critical } value of } specific ionization, (dE/dx)c, below which tracks are not registered. For } example, (dE/dx)c is ~ 13 MeV mg^-1 cm^2 for mica and ~4 MeV mg^-1 } cm^2 for Lexan, two of the most widely used detectors. As a result mica } does not register ions with A {~30, Lexan is insensitive to ions with } A {~12." } Alphas have A = 4. Now this text is old (1981) and not too good, so the } technique for producing appropriate tracks may have been improved or } Friedlander may be incorrect for the kind of tracks you need. Also, the } tracks are produced when the incident particles ionize and/or displace } atoms } in the target, so the tracks may not be as small as you would like. Bear } in } mind that electrons incident on a specimen generate x-rays throughout a } teardrop-shaped volume about 1 micron across. Although alphas do not } have as great a spread as electrons, much of the ionization produced is } from secondary particles, and these will be electrons. If the paper you } refer } to tells you how to generate suitable tracks given a source of alphas, then } you are probably OK. Good luck. } Yours, } Bill Tivol } } }
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 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
Brenda I. Prenitzer wrote: ================================================================ Does anyone have any recommendations for a conductive epoxy (other the Ag paint) that can be used to mount specimens to a stud that will be used for both polishing and subsequent analysis? (i.e., mechanically durable, water resistant and stable under ion bombardment) ================================================================= The softness of silver generally makes it less than desired as a conductive filler in this kind of application because of silver smearing, although silver filled epoxies are available from SPI and others such as described on URL http://www.2spi.com/catalog/spec_prep/cond_adhes2.html
Can you consider using a conductive copper-filled diallyl phthalate such as is available from both SPI and others. This is a thermoset resin system (it requires a mounting press) and you can get an excellent metallographically polished surface. See URL http://www.2spi.com/catalog/spec_prep/metall2a.html#12 It is certainly mechanically durable, water resistant and so far as it is possible for any polymer, it is also stable under ion bombardment.
You would then literally glue the polished mount to the SEM mount of your choice, which you would then prepare using your standard procedures.
This "solution" permits you to have the best of both worlds, first, a mounting medium that was specifically designed for this kind of application plus a final SEM mount ready to put into your SEM.
Disclaimer: SPI offers both silver filled epoxies as well as conductive metallographic mounting media. But similar materials are available from some of the other major firms supplying the microscopy and microanalysis market as well.
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
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William H. Roberts wrote: ====================================================== I have a March Plasmod (Plasma Etcher) which no longer produces the rf needed to generate the plasma. Does anyone have a surplus/broken/obsolete unit that could possibly still have operable electronics that they want to get rid of? I'd like to try to repair mine before declaring it junk. (if it can't be repaired economically, perhaps someone will need the parts on it that are still good.) ======================================================
If what you are asking about is the pair of power tubes, then new ones can be purchased from SPI Supplies, our part number 11022-AC for the pair of power tubes. Details can be found on the SPI website given below. This was a well built unit and there is not reason why it can't continue to give you good service for years into the future.
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 tasked with setting up an electronic library of SEM images, and I'm } looking for good database software applications for cataloging and } viewing images. Multi-user access and strong software technical support } are primary requirements. What are my options?
Another very powerful database management option is Ugather®. UGather® is a free multimedia database manager that can catalog images, QuickTime movies, and audio files and store information related to each file. The databases can be imported easliy into UPresent®. UPresent® is a free multimedia presentation application designed for flexible, interactive control of presentations. It is available for Windows and Mac OS. You can download it from http://upresent.umn.edu/. I have no commercial interests, just a satisfied user. 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
Allied High Tech has just introduced an image acquisition, measurement, DATABASE, archiving and report generation software package (Micro-Vision and Micro-Vision Explorer). The software is modular so the database portion (Micro-Vision Explorer) can be purchased separately.
Micro-Vision Explorer has thumbnails for each image and search features for the data you enter into the customizable fields. All fields are set up by an administrator to meet your needs. Unique fields can be created for text input or numeric input. The fields can have drop down menus if you choose. Each field can have specific attributes such as: flagged for required input, read only, archive, clear last input plus more. The search features include: file name, date, specific field search, text contained any where in the properties, fuzzy searches plus much more. It supports an unlimited number of images. It has built in archiving capability with volume control. Micro-Vision Explorer has an easy to use Windows interface and will run on Windows 95, 98, NT and 2000 computers. All of the popular image formats are supported. The software can be networked and password protection can be implemented.
In addition to images, Micro-Vision Explorer is also capable of indexing, searching and achieving plain text, Lotus AMI Pro, HTML, Microsoft Word, Excel, Works, RTF, PowerPoint 97, Adobe Acrobat PDF, Word Perfect versions 5.0 thru 7, and Word Star versions 4 to 6 and 2000.
If you require more information please let me know off-line.
I work for Allied High Tech and have a financial interest in this and all products Allied High Tech sells.
Sincerely,
Ed Hirsch
************************************************* 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: default [mailto:byron.johnson-at-medtronic.com] Sent: Friday, July 21, 2000 12:46 PM To: Microscopy-at-sparc5.microscopy.com
I am tasked with setting up an electronic library of SEM images, and I'm looking for good database software applications for cataloging and viewing images. Multi-user access and strong software technical support are primary requirements. What are my options?
Job description for analytical engineer, Defect and Thin Film Characterization Laboratory (DTCL), Applied Materials, Inc. Santa Clara, CA
Job Summary Active participation in product division contamination control and defect reduction programs. Prioritized by value to the product divisions, conduct internal and external defect characterization focused on defect root cause analysis. Interact with external experts and DTCL members to leverage value to product divisions. Document and communicate root cause solutions and strategic information to internal customers and strategic personnel in product divisions and functions.
Minimum Requirements: MS in Material Science/ Physics with 3 years hands-on experience in analytical instrumentation applied for defect/ thin film analysis. Must demonstrate: (1) Good hands-on capabilities as well as sound theoretical understanding of SEM, EDX, WDX, and FIB applications for particle defect/ thin film analysis. In addition, practical experience with Auger is a plus. (2) Experience in wafer surface defect detection using light scattering technique, e.g. Applied Materials WF736, Excite, Tencor 62XX and SP1-TBI. (3) Experience in TEM sample preparation and bulk samples cross-sectioning. (4) Ability to work independently with minimum of supervision. Must have good technical documentation/computer skills and is a team player.
Please send resume via e-mail to: richard_savoy-at-amat.com
I am seeking a Transmission Electron Micoscopy Position in the Chicago, IL or Los Angeles, CA area. I have 34 years of experience as a Biological Transmission Electron Microscopist in research and clinical areas, have held MSA certification for 21 years, and have a Bachelors Degree. If you know of positions in the above areas or know of resources for job postings in Electron Microscopy, please contact me via the following:
Elaine R. Sundin 6033 N. Sheridan Rd. #38L Chicago, IL 60660 Phone 773-728-8083 e-mail djemge-at-aol.com
TNT (Trends In Nanotechnology) 2000 will take place this year in the UNESCO World heritage city of Toledo. The aim of this conference will be to focus on the applications of Nanotechnology and to bring together in a Forum many of the major European groups working in this field. www.cientifica.com {http://www.cientifica.com} TNT 2000 will be held in the Monastery 'San Pedro Mártir' in the historic city of Toledo, Spain from the 16th - 20th October 2000. The aim of this conference will be to focus on the applications of Nanotechnology and to bring together in a Forum various groups working in this field. 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". The first announcement is available here as a MS Word or Acrobat pdf document 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". Currently 2 workgroups, "Nanofabrication: printing" and "Nanofabrication: Self-Assembly" from the 5th MEL/ARI-NID program (European Commission) have decided to actively participate at this conference. They will hold their NID (Nanotechnology Information Devices) meeting during TNT2000. Scientific Program The scientific program will cover a wide range spectrum of Nanotechnology research and related areas including keynote lectures, oral presentations, posters and a product/instrument exhibition. 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 TNT'2000, following each session, large breaks will be set-up in order to allow these discussions. This allows a high degree of interactivity both between research groups, and between researchers and exhibitors. The TNT conference will allow either Academic institutions or Industrials (such as IBM, Lucent, Philips, Thomson, Sony, etc.) to present their research and foster cooperation between both. Key topics * Nanotubes * Local Electromagnetic Phenomena * Quantum + DNA Computing * Devices and Machines * Nanoscale Integration * Nanobiology * Nanotechnology for space Applications * Molecular Nanotechnology * Nanomedicine * Nanostructure Behaviour Modelling * Nanomaterials * Nanofabrication Tools and Techniques * Ultimate Limits of Measurement
More information about the Workshop (Keynote Speakers, Accommodation, Committees, schedule, social events, etc.) is available at: http://www.cientifica.com/ {http://www.cientifica.com/}
If you need more information, please do not hesitate to contact me. Regards Tim E. Harper
***************************************************************** 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/}
Dear Byron, You will find that the Quartz Imaging Corp. (www.quartzimaging.com) image databasing software is very complete, easy to use and access and fully supported. It has several options for intranet or web-based access and is the fastest I've ever seen. At 12:46 PM 7/21/00 -0500, you wrote: } } I am tasked with setting up an electronic library of SEM images, and I'm } looking for good database software applications for cataloging and } viewing images. Multi-user access and strong software technical support } are primary requirements. What are my options? } 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 Brenda, I use the Cu-filled Kulzer Technovit 5000 cold-curing resin to mount and polish metal samples that I do not want to carbon-coat. I believe it is made in Germany, but imported by Energy Beam Sciences.} } Greetings! } } Does anyone have any recommendations for a conductive epoxy (other the Ag } paint) that can be used to mount specimens to a stud that will be used for } both polishing and subsequent analysis? (i.e., mechanically durable, water } resistant and stable under ion bombardment) } } Any info would be appreciated. } } Brenda I. Prenitzer, Ph.D.
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
} A colleaque needs to know the value of an ISI Super IIIA that was } dontated to their institution (probably for tax credit purposes to } the donor). } } If someone is able to give a reasonable figure to this individual, } please send the figure to the following address: } } DWARD-at-flemingc.on.ca } } Thank you on his behalf. -- #################################################################### 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 ####################################################################
Gary, Let me know where I can get an O2 for only $750, also where I can get a confocal system (not pcm) for under 200K. The point is we are not dumping the system, it is heavily used, check out www.med.jhu.edu/micfac/reserve.cgi, for availability, we simply need a Uv laser and rather than retro-fitting we decided to buy a new one. I would put any of our deconvoluted confocal images up agaisnt any system out there and feel confident that they are as good if not better.
Mike D.
On Fri, 21 Jul 2000, 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 10:46 AM 7/21/00, 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 tasked with setting up an electronic library of SEM images, and I'm } } looking for good database software applications for cataloging and } } viewing images. Multi-user access and strong software technical support } } are primary requirements. What are my options? } } } Consider ImageAXS. It is set up to manage your image database } and also to allow you to set up CDs with thumbnails and full } resolution files. } } The other option is ThumbsPlus--but Cerious Software is rather } dysfunctional based on my experience with them. } } Other options are Canto Version 5 Cumulus, Kudo Image Browser } {http://www.imspace.com} , } Extensis/CreativePro Portfolio {http://creativepro.com} . See } {http://riecks.com/redbarns.html} or {http://riecks.com/crash/} . I believe } the Kudo software is still $49 for the web } download version. You can download a demo version for free and give it a spin. } Then there is Gallerymaker (www.rjhsoftware.com) which produces thumbnails } and webpages, all of which are customizable. Cheap as I remember too. I just } use it to batch process thumbs from whole directories. } } There are other options too. } } gg } } }
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I'm passing this along if anyone is interested. Reply to CMI..
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If you are interested in pursuing a career with us please email your resume to rklein-at-cmiinternational.com or fax your resume to R. Klein -at- (847) 439-4425. If you would like to discuss potential opportunities at the Denver X-ray Conference please leave a note at the front desk of the Denver Marriot Tech Center Hotel. Look us up at our web site which is: cmiinternational.com.
} Date: Mon, 24 Jul 2000 17:04:44 -0700 } To: MICHAEL DELANNOY {delannoy-at-mail.jhmi.edu} } From: Gary Gaugler {gary-at-gaugler.com} } Subject: Re: Confocal sale } } I just sold an Indigo 2 Extreme complete system for $900. There } are numerous O2 boxes around for between $600-$1200, depending } on whether it is a 5K or 10K CPU and how much memory it has. } The O2 is a nice toaster for sure. But it is rather dated. } } Any inverted scope is not going to have the image quality and } resolution of a vertical unit. My worst experience was with an } Olympus IMT-2. I dumped it for $6K and was glad to be rid of it. } } If your system is working OK for you, all the better. I was just } trying to point out the impact of technology advancement on } after-the-purchase value. } } gary g } } } At 12:15 PM 7/24/00, you wrote: } } } Gary, } } I don't know where you estimates come from, but let me know where } } I can get an O2 for $750. Anyway we aren't dumping this system, } } this system is still making money for our facility (check out our reservation } } scheduler www.med.jhu.edu/micfac/reserve.cgi), the Noran Oz is always booked. } } The reason for the sale is to get a confocal system with a Uv laser, we don't } } want to retro fit this one, so it seemed simplier to sell it. I would put } } any of our deconvoluted confocal images up against any other system } } available, } } and feel confident that the imaging is as good if not better. } } } } } } On Fri, 21 Jul 2000, Gary Gaugler wrote: } } } } } How many Mercedes come with this? The O2 is worth about } } } $750. The Indigo 5000 is not worth more than $750. And the } } } inverted Olympus is barely worth a mention. } } } } } } No wonder you want to dump it. } } } } } } gg } } } } } } } } } } } } At 08:19 AM 7/21/00, 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 } } } } -----------------------------------------------------------------------. } } } } } } } } } } } } } } } } For Sale: } } } } Noran Oz Confocal Laser Scanning Microscope System with a new } } } } Krypton-Argon laser, red HeNe, SGI O2 (acquisition), SGI Indy R5000 } } } } (analysis) and an Olympus IX 50 inverted microscope with 20x, 60x and } } 100x } } } } Plan Apo lens'. This system is only 4 years old and fully operational. } } } } Some highlighted features are: } } } } } } } } AOD laser scanning, allows for very fast acquisition } } } } rates (up to 480 frames/sec), for live localization, and } } } } slow scan for fixed material. } } } } } } } } Triple label capability (488, 568 and 633nm excitation) } } } } with broad and narrow band emission filter sets. } } } } } } } } Two and Three dimensional analysis software by Intervision, } } } } Tiff and RGB formats (amoung others). } } } } } } } } Deconvolution software allows for thinnest possible z-sectioning } } } } for light microscopy. } } } } } } } } Primary dichroic filter wheel upgrade for precise fluorophore } } } } registration. } } } } } } } } We are offering this system for $180,000, buyer to pay shipping. } } } } I will personally give free on-site setup and training. Please contact } } } } Michael Delannoy for further information (410) 955-1365. } } } } } } } } } } } } Michael Delannoy } } } } Assistant Director } } } } JHMI Microscopy Facility } } } } Johns Hopkins School of Medicine } } } } Baltimore, Maryland } } }
} Date: Mon, 24 Jul 2000 17:07:20 -0700 } To: MICHAEL DELANNOY {delannoy-at-mail.jhmi.edu} } From: Gary Gaugler {gary-at-gaugler.com} } Subject: Re: Confocal sale } } Check out this link for an almost complete O2/10K system. } It is in the range I quoted. } } http://cgi.ebay.com/aw-cgi/eBayISAPI.dll?ViewItem&item=388534054 } } gg } } } At 12:15 PM 7/24/00, you wrote: } } } Gary, } } I don't know where you estimates come from, but let me know where } } I can get an O2 for $750. Anyway we aren't dumping this system, } } this system is still making money for our facility (check out our reservation } } scheduler www.med.jhu.edu/micfac/reserve.cgi), the Noran Oz is always booked. } } The reason for the sale is to get a confocal system with a Uv laser, we don't } } want to retro fit this one, so it seemed simplier to sell it. I would put } } any of our deconvoluted confocal images up against any other system } } available, } } and feel confident that the imaging is as good if not better. } } } } } } On Fri, 21 Jul 2000, Gary Gaugler wrote: } } } } } How many Mercedes come with this? The O2 is worth about } } } $750. The Indigo 5000 is not worth more than $750. And the } } } inverted Olympus is barely worth a mention. } } } } } } No wonder you want to dump it. } } } } } } gg } } } } } } } } } } } } At 08:19 AM 7/21/00, 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 } } } } -----------------------------------------------------------------------. } } } } } } } } } } } } } } } } For Sale: } } } } Noran Oz Confocal Laser Scanning Microscope System with a new } } } } Krypton-Argon laser, red HeNe, SGI O2 (acquisition), SGI Indy R5000 } } } } (analysis) and an Olympus IX 50 inverted microscope with 20x, 60x and } } 100x } } } } Plan Apo lens'. This system is only 4 years old and fully operational. } } } } Some highlighted features are: } } } } } } } } AOD laser scanning, allows for very fast acquisition } } } } rates (up to 480 frames/sec), for live localization, and } } } } slow scan for fixed material. } } } } } } } } Triple label capability (488, 568 and 633nm excitation) } } } } with broad and narrow band emission filter sets. } } } } } } } } Two and Three dimensional analysis software by Intervision, } } } } Tiff and RGB formats (amoung others). } } } } } } } } Deconvolution software allows for thinnest possible z-sectioning } } } } for light microscopy. } } } } } } } } Primary dichroic filter wheel upgrade for precise fluorophore } } } } registration. } } } } } } } } We are offering this system for $180,000, buyer to pay shipping. } } } } I will personally give free on-site setup and training. Please contact } } } } Michael Delannoy for further information (410) 955-1365. } } } } } } } } } } } } Michael Delannoy } } } } Assistant Director } } } } JHMI Microscopy Facility } } } } Johns Hopkins School of Medicine } } } } Baltimore, Maryland } } }
Dear All, Just wondering if anyone out there uses a nitrogen bubble system to agitate their film processing solutions in tanks. We are considering such a system but I would like a clearer idea about the set-up before going ahead and asking our workshop to make the system. At present we use Kodak 4489 film and ordinary tanks with Kodak D19 developer and Ilford Hypam, this works fine except occasionally when people forget to agitate the film.
If anyone has such a system which is being used for sheet negs I would appreciate a rough description how it works. ie
1) what the layout of tubes on the bottom of the tanks is like
2) what kind of control system is used to set the bubbling intervals (presumably something like a solenoid on a timer connected through a regulator on to a N2-filled cylinder?)
Are there any problems or drawbacks to the systems which are in use? Are they worth the trouble of setting up and maintaining?
Best regards,
Richard
Richard Easingwood South Campus Electron Microscope Unit School of Medical Sciences, University of Otago PO Box 913, Dunedin NEW ZEALAND
Proscitech offers very pure, very fine silver powder in 20 g tubes. This can be mixed with any suitable compound, which has several advantages. I believe that using silver powder for mixing in with epoxy and similar applications has several advantages. Obviously ProSciTech has an interest in the use of this product. 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 ABN: 99 724 136 560 www.proscitech.com
-----Original Message----- } From: Mary Mager [SMTP:mager-at-interchange.ubc.ca] Sent: Tuesday, July 25, 2000 1:46 AM To: Microscopy-at-sparc5.microscopy.com
On Tue, 25 Jul 2000, Richard Easingwood wrote:
Hi Richard,
We have been using N2 burst in our developing tanks since long before I came here over 20 years ago and I think that it is a great facility. We are a large multi user facility that can be processing between 200 and 2000 sheets a week in two different processing lines. The only problem we have is that occasionally users do not check that there is N2 in the gas bottle before processing and hence it does not work. The importance of correct agitation is obvious and lack of agitation is easily spotted when a users has got it wrong (very rare).
We use systems bought from Kodak 30(ish) years ago. There are 6 tubes with small holes in the bottom of our deep tanks, a solenoid valve controlled by a timer and a N2 bottle with regulator. The timer can be set for off, continous and timed (with variable burst times and off intervals). We generally use 1 second burst every 10 seconds. The gas pressure is adjusted to ensure we have the minimum flow with a good agitation (around 10 psi).
An added advantage is that I can reload the films during the developing time instead of hanging around, occasionally shaking the films. Maybe it does not save that much time but it seems like it late at night.
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 } -----------------------------------------------------------------------. } } } Dear All, } Just wondering if anyone out there uses a nitrogen bubble system to agitate } their film processing solutions in tanks. We are considering such a system } but I would like a clearer idea about the set-up before going ahead and } asking our workshop to make the system. At present we use Kodak 4489 film } and ordinary tanks with Kodak D19 developer and Ilford Hypam, this works } fine except occasionally when people forget to agitate the film. } } If anyone has such a system which is being used for sheet negs I would } appreciate a rough description how it works. ie } } 1) what the layout of tubes on the bottom of the tanks is like } } 2) what kind of control system is used to set the bubbling intervals } (presumably something like a solenoid on a timer connected through a } regulator on to a N2-filled cylinder?) } } Are there any problems or drawbacks to the systems which are in use? Are } they worth the trouble of setting up and maintaining? } } } Best regards, } } Richard } } } Richard Easingwood } South Campus Electron Microscope Unit } School of Medical Sciences, University of Otago } PO Box 913, Dunedin } NEW ZEALAND } } Telephone: 64-03-479 7301 Facsimile: 64-03-479 7254 } mailto:richard.easingwood-at-stonebow.otago.ac.nz } http://anatomy.otago.ac.nz:800/Department/EMUnit.html } http://www.otago.ac.nz/anatomy/emunit/ } } } } } }
=========================================================================== 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. ============================================================================
Just an additional comment on this. Late '99 we evaluated several database packages: ImageCentral from Advanced Imaging Concepts (Princeton, NJ), ElectroImage (ElectroImage, Great Neck, NY) and Archive4Images (Long Island Instruments, East Meadow, NY). Each system has its pros and cons. The ElectroImage database package has since been upgraded with Java and Web enabling; ImageCentral is a network-enabled package; Archive4Images includes some processing tools. ElectroImage is an outstanding product, but has a limited number of fields for searching, whereas ImageCentral has a much larger number of fields--the single feature that the Experimental Pathology group found to be most important. None of these packages is inexpensive--expect $1500 to $4K for cost of the database alone. If cost is not a factor, check out these systems. Of course, you can always develop your own using Paradox or Access. On the non-scientific side, check out the current version of ULead's ImagePals--I have been using this at home since v.2 came out a very long time ago.
Roger Moretz, Ph.D. Dept of Toxicology BI Pharmaceuticals, Inc.
I have no interest in any of the products, vendors or suppliers other than to have evaluated or used the product(s) somewhere in my life. On Fri, 21 Jul 2000 12:46:21 -0500, default 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 tasked with setting up an electronic library of SEM images, and I'm } looking for good database software applications for cataloging and } viewing images. Multi-user access and strong software technical support } are primary requirements. What are my options? } }
Roger C Moretz, Ph.D.
_______________________________________________________ Say Bye to Slow Internet! http://www.home.com/xinbox/signup.html
Anyone out there know of any antibody that would be specific for mouse endothelial cells?? We are trying to identify endothelial cells in tumors as well as cultured endothelial cells. I know a lot of them work for LM after various treatments. I seem to have had no luck so far staining at EM level. I would appreciate all the input.Thanks in advance..... Neelima Shah.............. Biomedical Imaging Core Facility Uni of Pennsylvania Philadelphia, Pa.
There has been a creeping effect in postings over time where individuals are listing items "for sale" on the listserver.
Let me once again remind you all of our rules (from the FAQ sheet you ALL received) that this is not permitted and it applies to everyone (including Universities) not just Commerical organizations.
The general public does not know but every posting that I see (I do miss some) that breaks the Listserver rules generates a PRIVATE warning from me. If the transgressions continue it will culminate with exclusion (ie. filtering/banning) of an individual's privilege of posting to the Listserver. For the record, nearly a dozen individuals have been barred from posting (but they may still receive mail) for extended periods.
Returning to the subject of providing a forum for listing/offering used equipment it is obvious that the community needs an electronic forum for this function.
To this end, I will create a WWW site which will allow this and will announce it as soon as it is operational.
It will be seperate from the Listserver and will be purely WWW based. The model for this is the Vendor news site which I created over a year ago and continues to operate successfully.
Exerpt from the Microscopy Listserver Rules ---------------------------------------------------- Can I post an Advertisement / Commerical / Marketing Survey Message(s)? ---------------------------------------------------
No, that does not fit within the bounds of this discussion forum.
This listserver is not intended to be a Sales/Marketing/Survey mechanism for organizations, but rather it is an open discussion area about microscopy and microanalysis problems and solutions.
If you are an organization and have equipment you wish to donate, or sell, for nominal cost (i.e. no profit) then this is generally an acceptable posting. If you are not sure then send a copy of the announcement in question to Zaluzec-at-MSA.Microscopy.Com and I will give you my opinion. An example of this type would be an old decommissioned instrument which someone is trying to give away for removal/shipping costs, that would fit within the bounds of the purposes of this list.
If you are interested in using the Internet for Commerical Advertising of Microscopy/Microanalysis Related Products/Services, you may wish to contact MSA at it's WWW site (http://www.msa.microscopy.com) or the MSA Business Office (MSABusinessOffice-at-MSA.Microscopy.Com). These alternative Internet services, are provided independently of the Listserver Operation, which MSA provides as a FREE service to the WorldWide Microscopy and Microanalysis Community. Any funds derived from the above are used to defray the costs of running MSA's Internet site.
If you wish to purchase a copy of the MSA Membership directory/Mailing list please contact the MSA Business Office .
There is a mechanism to post Commerical NEWS to the Microscopy Community. If you have something of a purely Commerical Nature and wish to make it known to the community you may FREELY use the following WWW site (just follow the on-line instructions)
We have a nitrogen burst system that we use for the developing tank. It consists of an open metal rack with five metal tubes at the bottom, each with a series of tiny holes spaced about 1-1.5 cm apart and facing the bottom of the tank. Our plexiglass negative rack fits inside the gas rack, and both fit in our 9x5.5 inch developing tank. The rack is connected to a nitrogen tank via a burst regulator (Arkay Corp.) with variable burst frequency and duration.
One caution: about a year ago we noticed that our developed negatives were coming out uneven, even when they were exposed to an absolutely uniform electron beam. On examination, some of the holes were found to be encrusted with metal salts. Once cleaned out, the negatives became uniform again. This was only after many (15+) years of operation, so it isn't a big problem if you watch out for it.
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
Since we (and probably others) use this film, I'm curious what is the hazardous/toxic part. Is it the Na sulfite solution, the goop on the film, or the coater?
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 think both. I know on the pieces of film, there was some kind of warning about getting the goop (Na sulfite) on your skin. Before we went to the coaterless film (type 53) way back when, we had a special hazmat can for the coaters.
Would anyone have a wavelength sensitivity curve for photoconversion of diamonobenzidine?
M. Malecki, M.D., Ph.D. Director and Principal Investigator
Molecular Imaging Laboratories University of California at San Diego address: 1500 Suite Bonner Hall, 9500 Gilman Drive, La Jolla, CA 92093-0368 phone: 8588223373 fax: 8588223715 pager: 8586161420 e.mail: mmm-at-ucsd.edu e.pager: 1620024619-at-alphapage.airtouch.com www site: http://mil.ucsd.edu/ ftp site: mil1.ucsd.edu
Harry Ekstrom wrote: ============================================================ I think both. I know on the pieces of film, there was some kind of warning about getting the goop (Na sulfite) on your skin. Before we went to the coaterless film (type 53) way back when, we had a special hazmat can for the coaters. ============================================================ Over the thirty plus years we have used the Polaroid P/N film , the type that took the sodium sulfite clearing solution (but we hardly use any today) , it was my recollection that we had cases of what seemed like contact dermatitis from more than several employees, with some being sensitive to the chemicals in the coaters. The sodium sulfite solution seemed to be a mild irritant to others. I don't recall anyone being affected by the chemicals in the composite film pack, but then again, the strong warning might have given everyone a special "heads up". But I would avoid having any of these kinds of chemicals come in contact with your skin.
Another thing I found out is that one can develop a sensitivity to at least the coater chemicals, that is one might not be initially reactive to them, but with time, one could develop a sensitivity to them.
Disclaimer: Digital microscopy has put an almost complete stop to the sales of these kinds of film products.
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 ######################## ============================================
Did you try PECAM-1/CD31 (rat) from Pharmingen? It must work well at least using a pre-embedding protocol without permeabilization.
Greetings, Michael Reiner
University of Cologne, Germany Department of Anatomy I
} Anyone out there know of any antibody that would be specific for mouse } endothelial cells?? We are trying to identify endothelial cells in tumors } as well as cultured endothelial cells. I know a lot of them work for LM } after various treatments. I seem to have had no luck so far staining at EM } level. I would appreciate all the input.Thanks in advance..... } Neelima Shah.............. } Biomedical Imaging Core Facility } Uni of Pennsylvania } Philadelphia, Pa. } } } } } http://www.MED.upenn.edu/morphlab/ } } è
Hello Microscopists I have a client who wants to image (SEM) thermophile bacteria in situ in a bioleaching environment. Any one have any experience in this regard? We have tried the conventional prep procedure: .2u filter GA-----CPD, but with no luck. Any pointers will be appreciated
How large are your samples? Are you going to do UV cure or heat? I have procedures for embedding cells on coverslips as well as "normal" samples that need flat-embedding....let me know which samples you have!
Tamara Howard CSHL
On Tue, 25 Jul 2000, Sarah Peters 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 appreciate any hints, tips, or procedures available on flat embedding using LR White. } Thanks! } S. Peters } } Sarah Peters NRC04 } Oregon Hearing Research Center } 3181 SW Sam Jackson Pk Rd } Portland, Or. 97202 } Ph:(503) 494-2942 } Fax: (503) 494-5656 } } } }
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
To all you struggling managers of multi-user or service facilities:
This is a reminder that there will be a special "EXPERTS" session to discuss topics of interest in this area. The session will be help from 9:00 to 12:00 on Wednesday, August 16 in Room 106.
Format of the session will be as follows: 1) 3 topics recommended by you will be initially presented with each introduced by an expert on that topic. 2) The order of the discussion will be announced in advance so those of you with conflicts will have an idea of when to attend this session. 3) The initial introduction of a topic will be short (5-10min) so as to leave maximum time for questions and discussion from attendees. 4) We will try to limit open discussion on any one topic to no more than 30min so that all 3 topics can be discussed in approximately a 2 hr. time period. The remaining time can be used to revisit one of the topics or introduce new topics from the floor for discussion.
In an earlier E-mail, I requested suggestions for topics be sent to me. Of the ones received, those listed below seemed to be the most popular.
1. Maintenance of equipment: service contracts ; service contracts vs. third-party vs insurance company or self-insure; timely service
2. Justification of costs/ cost recovery….electronic bookkeeping and billing
3. login systems for instruments/lab security/training
You will notice that they are quite broad and may have to be narrowed down by the "expert" introducing them in order to have a meaningful discussion. I am now looking for the "experts" to introduce topics 1 and 2. Keith Darlington from Elf ATOCHEM North America, Inc has gratiously consented to introduce topic #3. Please E-mail me if you would like to introduce topics 1 or 2 or can recommend someone to do so.
Also, if any of you have information relating to the above topics that can be made available to those attending, please bring handouts or send the info to me and I will have it copied for distribution.
There has been requests to record the session so that those not in attendance can benefit from the discussions. I will try to arrange this if appropriate recording equipment is available.
One final note...I will not arrive at the meetings until Monday so would appreciate a volunteer to put up notices about the session throughout the convention hall on Sunday. Please contact me if you are willing to do this and I will get the notices to you.
Many thanks and PLEASE send me names of individuals I can ask to introduce topics. Also, if you have suggestions about the format of the session, please contact me...there is still plenty of time to modify it.
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
by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id JAA08852 for dist-Microscopy; Wed, 26 Jul 2000 09:32:57 -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 JAA08848 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Wed, 26 Jul 2000 09:32:27 -0500 (CDT) Received: from denexc2.timet.com ([63.102.13.36]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id JAA08841 for {Microscopy-at-sparc5.microscopy.com} ; Wed, 26 Jul 2000 09:32:16 -0500 (CDT) Received: by denexc2.timet.com with Internet Mail Service (5.5.2650.21) id {NJ3XTDDN} ; Wed, 26 Jul 2000 08:31:53 -0600 Message-ID: {90F2CBD1ADD9D3119A3A00A0C9A94BA302E770-at-henexc1.timet.com} {Microscopy-at-sparc5.microscopy.com}
We must obtain and pay for a permit for any item put into our process waste stream. Any item not permitted, is considered a "hazardous" waste, and must be disposed of at great cost.
Also, all waste water must either be ponded or reclaimed, this is now becoming a large expense for our facility.
Our enviormental department examined each departments waste stream and in cooperation with the individual departments (thank goodness) made some determinations as to where we could save not only permit costs but water usuage.
The processing of the p/n film uses a rather significant amount of water for my department, not to mention the occassional print production.
Also, you can check Polaroid's site for it's MSDS sheets for all it's products.http://www.polaroid.com/service/msds/index.html
One positive outcome of this, my request for a new metallograph with a high end digital camera has been approved!
Anyway, that's the basic scenario. No Federales came in and spanked us, rather it was a cost-saving(??) measure implemented by our enviormental group.
Back to my original question: What type of Polaroid do you all use?
William T. Giles Sr. Electron Microscopist Met. Lab. Coordinator Henderson Technical Laboratory TIMET PO Box 2128 Henderson NV 89009 Ph: (702)566-4436 Fax: (702)564-9038 E-mail: Bill.Giles-at-timet.com
} -----Original Message----- } From: Marie E. Cantino [SMTP:cantino-at-uconnvm.uconn.edu] } Sent: Tuesday, July 25, 2000 8:53 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: Polaroid Film Choices } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Since we (and probably others) use this film, I'm curious what is the } hazardous/toxic part. Is it the Na sulfite solution, the goop on the } film, } or the coater? } } Marie }
We have been using an old model Leedal Sink with nitrogen burst system for about 12-14 years. Other than having the temperature gauge (which regulates the temperature of the developer) replaced, the system has been great! It has metal tanks that have metal rods with holes on the bottom (where the nitrogen comes through). You regulate the duration of the burst and the interval between the bursts with a timer built into the sink. Hoses connect the tanks to the nitrogen tank. The developing and fixing tanks float in water that is temperature-regulated. This system gives good, even distribution of the chemicals over the negatives.
(The best part is I got this refurbished sink, racks, darkroom lights, etc. for $750.00! Can't complain.)
Peggy Sherwood MGH-Wellman Labs of Photomedicine (W224) 55 Fruit Street Boston, MA 02114
Dear listers, We have been using AGFA scientia 23D56 (3.25x4 ") film for years now until AGFA discontinued that product. I would greatly appreciate any help in finding a suitable replacement for this film
I need to measure the dislocation density in an Al-30Si alloy.
I've been told there is a condition that will give the optimum dislocation contrast which will give some proportion of the total dislocations.
Does anyone know what these conditions are and the proportion that this gives (and a reference if possible?)
Also any comments on other methods that don't require one to measure the thickness of the sample e.g. simply counting the intersections of dislocations with the surface and dividing by the area?
Thank-you,
Simon Hogg
University of Sheffield Department of Engineering Materials Sir Robert Hadfield Building Mappin Street Sheffield S1 3JD Tel. 0114 222 5934 Fax. 0114 222 5943
Where can I find prepared microscope slides that use flourescence? Or, where can I find information about how to prepare my own slides with flourescent stains? I would like such samples for research on 3D microscopy.
} Back to my original question: What type of Polaroid do you all use?
Oh yea ... the question.
Type 52 is what we use when we aren't grabbing digital ... but even then, printing four 1024x800s to the dye-sub proves more cost effective ... needless-to-say, my type 52 is all past its use-by date.
Type 52 is also ASA400, so you will have to re-calibrate your exposure. It also doesn't require the smelly "goop".
I've studied Goldstein's book to try and accurately compute the true spot size of a SEM beam based on the myriad of factors which influence it. At issue is how to determine the actual spot size of a beam, in microns, at the specimen.
SEMs from various makers use relative numbers for spot size. These indicate the relative size of the beam from "really small" to "largest." "Really small" means low specimen current but high resolution and the converse for the large spot size. If one is doing x-ray analysis, the spot size may or may not be an issue.
If the area being analyzed is large, the spot size is most likely not all that important. But for the analysis of small feature size and sub-micron semiconductor devices, true spot size is an issue. If the beam covers more than the area of interest, the x-ray data will not accurately reflect the composition of the intended material.
Therefore, is there some basic limitation of the applicability of quantitative SEM x-ray analysis based on the size of the specimen? If so, what is this limit? Also, is there some way to accurately define the physical size of a beam at the specimen? If so, what is this method?
For those of you planning on attending the Microscopy Society of America in Philadelphia August 13-17 (coming up in a few weeks), I wanted to alert you to the following program setup.
***** Please note: due to a scheduling snafu, David DeRosier's talk will be in the morning whereas the other two talks are scheduled for the afternoon session *****
There are 3 biological tutorials being presented:
David DeRosier: DeRosier's Guide to Three-Dimensional Reconstruction of Helical Structures Tuesday 11:00 AM Room 103
Liz Wilson-Kubalek: Structural Analysis of Proteins on Lipid Substrates Tuesday 3:00 PM Room 103
Steve Ludtke: Single Particle Analysis of Macromolecules and Complexes: How to Get Started Tuesday 4:00 PM Room 103
Sincerely, Gina Sosinsky Session Chair for Biological Sciences Tutorial
P.S. For those of you who yearn to go beyond electrons, there is also a joint biological/physical sciences tutorial being given by Dave Piston from Vanderbilt University on Multi-Photon Excitation Microscopy: An Old Idea in Quantum Theory Applied to Modern Scientific Problems. Monday 3:00 PM Room 103 ***************************************** * Gina Sosinsky, PhD. * * University of California at San Diego * * 365 San Diego Supercomputer Center * * 9500 Gilman Drive * * La Jolla, CA 92093-0505 * * * * Note new area codes: * * 858-534-6264 (office phone & * * voice mail) * * 858-534-4583 (lab phone) * * 858-822-0861 (fax) * * gsosinsky-at-ucsd.edu (email) * *****************************************
You can replace the Agfa Scientia Film with Kodak´s SO-163 EM Film without any Problems. Greetings, Michael
Michael Reiner University of Cologne, Germany Dept. of Anatomy I _______________________________________________________________________ 1.000.000 DM gewinnen - kostenlos tippen - http://millionenklick.web.de IhrName-at-web.de, 8MB Speicher, Verschluesselung - http://freemail.web.de
} From: "Gary Gaugler" {gary-at-gaugler.com} } } I've studied Goldstein's book to try and accurately compute } the true spot size of a SEM beam based on the myriad of } factors which influence it. At issue is how to determine the } actual spot size of a beam, in microns, at the specimen. } } SEMs from various makers use relative numbers for spot } size. These indicate the relative size of the beam from } "really small" to "largest." "Really small" means low } specimen current but high resolution and the converse } for the large spot size. If one is doing x-ray analysis, } the spot size may or may not be an issue. } } If the area being analyzed is large, the spot size is } most likely not all that important. But for the analysis } of small feature size and sub-micron semiconductor devices, } true spot size is an issue. If the beam covers more than } the area of interest, the x-ray data will not accurately } reflect the composition of the intended material. } } Therefore, is there some basic limitation of the applicability } of quantitative SEM x-ray analysis based on the size of } the specimen? If so, what is this limit? Also, is there } some way to accurately define the physical size of a } beam at the specimen? If so, what is this method? }
Gary,
It was a good deal different set up but the idea should be the same. We were working with a scanning gamma ray device and the resolution was measured in hundredths of an inch but we built test samples that we knew the size of the metal slug and we knew how far it was between them. We would do a scan and see if we could resolve them.
It would be a lot tougher to build the model for what you want to do than for us. We used drill bits in Plexiglas. But if you coat various spacing grids with something that shows up in your X-ray scans and then polish the peaks off you should be able to get islands of various sizes and spacing to do your calibration.
We were primarily using gamma rays for image reconstruction using a Raydon
transform but we did some work with fluoresced x-rays. The gamma beam was about 3/32 of an inch. The customer did some direct imaging of Plutonium intrusion into various rocks with it as well. Better him than me.
The device was a gamma source and a detector with the subject on a rotary table that would move in the X and Z axis.
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
*From: "S.C.Hogg" {S.C.Hogg-at-sheffield.ac.uk} *To: Microscopy-at-sparc5.microscopy.com *Date sent: Wed, 26 Jul 2000 18:58:25 +0100 *Subject: TEM dislocation density measurment in Al *Priority: normal
*------------------------------------------------------------------------ *The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello,
It seems to me that the proportion of the total dislocation density which are people talking about is very rough estimate. But for sure two beam condition is the proper one to visualise dislocations. So, if you want to be acurate try to obtain an image of dislocation structure in the same area using two perpendicular g vectors, for example in the [001] zone axies. Your accuracy in dislocation density estimate depends also on deformation mode like slip systems involved, cross slip etc. so using more g vectors you'll be obtaing higher accuracy.
Good luck,
Witold Z.
* *Hello, * *I need to measure the dislocation density in an Al-30Si alloy. * *I've been told there is a condition that will give the optimum *dislocation contrast which will give some proportion of the total *dislocations. * *Does anyone know what these conditions are and the proportion *that this gives (and a reference if possible?) * *Also any comments on other methods that don't require one to *measure the thickness of the sample e.g. simply counting the *intersections of dislocations with the surface and dividing by the *area? * *Thank-you, * *Simon Hogg * * *University of Sheffield *Department of Engineering Materials *Sir Robert Hadfield Building *Mappin Street *Sheffield *S1 3JD *Tel. 0114 222 5934 *Fax. 0114 222 5943 * * %%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%% Witold Zielinski, Ph.D. Warsaw University of Technology Department of Materials Science and Engineering 02-507 Warszawa, Woloska 141 POLAND
I was asked before I leave to post this job opening. It's been great reading about all our trials and tribulations. I have probably ran into some of you at the meetings which were always fun. Dr. Rodriguez-Sierra is attempting to go to the meetings to place an advertisement at the Forums' booth. Thanks for all the help in the past.
Rick Vaughn
The Electron Microscopy Core Research Facility at the University of Nebraska Medical Center (UNMC) in Omaha, NE has an opening for a Biomedical EM Technologist to direct the laboratory and support research projects that involve transmission 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 electron microscopy. Additional courses or experience in immunocytochemistry, Cell Biology, and digital imaging is desirable. Operating knowledge of transmission 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 goals with respect to preparation for a variety of biomedical samples for TEM, operation of TEM, darkroom developing and printing, digital image capture and reporting. The technologist will also assist the faculty in the laboratory section of EM graduate courses.
UNMC offers a competitive salary and benefits package. UNMC is a equal opportunity employer.
If interested, please submit a cover letter and resume to: Dr. Jorge F. Rodriguez-Sierra Department of Cell Biology and Anatomy University of Nebraska Medical Center, Omaha, NE 68198-6395 Fax: 402-559-7328 Phone 402-559-6259 e-mail: jrodrigu-at-unmc.edu
We have been recommended Kodak SO-163 3 1/4" x 4" Film to replace, it has a similar speed, transfer to this type should be accomplished with minimal changes to exposure and development. Alternatively the Kodak 4489 which is slower film, which has been widely used for biological applications where high contrast is desirable. we can supply if required Boxes of 100, please contact off line.
Kind Regards,
Christine Fitzgerald
In message {Pine.HPP.3.93.1000726100957.15489F- 100000-at-merle.acns.nwu.edu} , Arun Kumar Subramanian {ksu973-at-merle.acns.nwu.edu} writes } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Simon Hogg wrote: } I need to measure the dislocation density in an Al-30Si alloy. } } I've been told there is a condition that will give the optimum } dislocation contrast which will give some proportion of the total } dislocations. } } Does anyone know what these conditions are and the proportion } that this gives (and a reference if possible?) } } Also any comments on other methods that don't require one to } measure the thickness of the sample e.g. simply counting the } intersections of dislocations with the surface and dividing by the } area?
Hi Simon,
There are a number of problems to consider if you want to measure dislocation density by TEM. (i) The dislocations may rearrange during preparation of the thin foil because of image forces. (ii) Some of the dislocations may be out of contrast under the diffraction conditions used. (iii) There may be a preponderance of certain Burgers vectors. (iv) The line vectors may not be evenly distributed in space.
I think most dislocation density measurements have been made under some simplifying assumptions that make them - as Witold Zielinski wrote - very rough estimates. Images are taken with a given two-beam condition. If, for instance, the material is fcc and the reflection is 200, it is then assumed that 1/3 of the dislocations will be out of contrast for this reflection. It is further assumed that the stereological equation (which assumes a random line vector distribution) is valid. A result obtained under such assumptions is not necessarily very exact.
As Witold wrote, you can try to image all dislocations by taking several images under different diffraction conditions. (You can also do this in a single image with a hollow-cone technique). This still leaves you with points (i) and (iv).
With respect to (i) you will e.g. in pure Al find that the dislocation density varies with foil thickness because relatively more dislocations are lost from the thinner parts of the foil. In other words use as thick a foil as possible.
With respect to (iv): the method that requires measurement of the foil thickness gives you the advantage that you can count intersections with lines of various orientations (or with a circle). If you count intersections with the surface, you have to assume that the line vectors are evenly distributed in space (consider the extreme case that all dislocation lines are parallel to the surface).
Instead of counting individual dislocations in a TEM it is also possible to measure line broadening in selected area channelling patterns in a scanning microscope or in X-ray diffractograms.
Best regards, Jorgen
***************************** J. B. Bilde-Soerensen Senior Research Scientist, Ph. D. Materials Research Department Risoe National Laboratory DK-4000 Roskilde Denmark
The beam size plays an important role in determining the size of a particle or feature one can analyze. Obviously the beam diameter must be smaller than the location to be analyzed and typically larger beam diameters indicate hagher current and count rates. The real determinant of what size can be analyzed without influence from surounding material is what "analysis volume" size is generated. The diameter of the analysis volume will never be smaller than the beam. It is usually much larger due to scattering within the specimen. Parameters which influence the analysis volume size and shape include the composition of the material being analyzed, the surface condition of the specimen, the angle of the incident beam, and the potential of the beam (kV). Hope I didn't forget one... If I recall correctly, Goldstein, et al, does cover this effect in the book.
Don Chernoff has available (http://www.small-world.net/efs.htm) a program which models the analysis volume. A similar but less flexable program came with my (former) Kevex EDS in the mid 80s. I have not tried the EFS software, nor have I any ties to small-world.
Arkay makes an affordable gas burst controller as well as water jacket/ tank systems for sheet film developing. Leedal also produces excellent jacket/tank systems. Both are available with gas burst plenums as required. Neither Leedal or Regal/Arkay has an active website that I could find. I will forward information to anyone who wishes to contact me directly.
George Laing National Graphic Supply scisales-at-ngscorp.com
} Dear All, } Just wondering if anyone out there uses a nitrogen bubble system to agitate } their film processing solutions in tanks. We are considering such a system } but I would like a clearer idea about the set-up before going ahead and } asking our workshop to make the system. } } If anyone has such a system which is being used for sheet negs I would } appreciate a rough description how it works. ie } } 1) what the layout of tubes on the bottom of the tanks is like } } 2) what kind of control system is used to set the bubbling intervals } (presumably something like a solenoid on a timer connected through a } regulator on to a N2-filled cylinder?) } } Are there any problems or drawbacks to the systems which are in use? Are } they worth the trouble of setting up and maintaining? } } } Best regards, } } Richard } } } Richard Easingwood } South Campus Electron Microscope Unit } School of Medical Sciences, University of Otago } PO Box 913, Dunedin } NEW ZEALAND } } Telephone: 64-03-479 7301 Facsimile: 64-03-479 7254 } mailto:richard.easingwood-at-stonebow.otago.ac.nz } http://anatomy.otago.ac.nz:800/Department/EMUnit.html } http://www.otago.ac.nz/anatomy/emunit/ } } } } } }
=========================================================================== 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. ============================================================================
Simon, My advice would be to do a series of two-beam dark field images with the Bragg reflection (g) chosen to optimise the g.b condition (b = Burgers vector). First find out the dominant dislocation types in Al & Si alone (or any other similar alloy) as a guide for the Burgers vectors you could possibly expect. If you can, try a reflection which makes g.b non zero for all dislocation types (giving contrast for all dislocations), and then subsequently select reflections that give g.b=0 for each type of dislocation (or g.b non-zero for one dislocation type). This procedure will allow you to deduce population densities for each dislocation type. If the dislocation density is too high i.e. you cannot resolve individual dislocations, then try a weak beam dark field imaging instead e.g g-3g (3g set to Bragg, g imaged). Contrast in these images come from the more highly strained crystal regions close to the dislocation core and one can narrow the contrast down to the order of nanometers.
A note on your definition of dislocation densities. You mention dividing by an area. Does this mean you are calculating the areal density or a volume density? The former is usually attributed to growing thin films, and most of the dislocation emanate from the initial interface. In that case you if you are looking at a cross-section, you will need to know the thickness, whereas a plan-view sample doesn't. For volume densities again you will need the thickness too. Areal densities are pretty meaningless in 3D.
Dislocation analysis is a fantastic area with a huge history, and I would advise you take a look at two books. Firstly Hirsch, Howie, Nicholson, Whelan & Pashley (Electron Microscopy of Thin Crystals), described as the Bible of transmission electron microscopy, has several chapters related to imperfect crystals, and the black and white photos are quite beautiful, even just to look at. Any time invested with this book pays off big. A more modern book is Williams & Carter (Transmission Electron Microscopy) and is extremely helpful. Again a superb selection of pictures and diagrams will help you with the theory and practice. ******************************************************** 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 ********************************************************
Whatever for? What would the environmental folks disagree about with the 55 processing? The sulphite? That's sprayed on fruit at the grocery store. The coater? I hate that myself, but it's no worse than regular darkroom chemicals.
T 52 won't replace T 55, as it has no negative. I don't think T 53 or T 54 do either, but I'm not sure. The T 55 negative is an *excellent* negative (in spite of some folks' comments otherwise) with about 3X the resolution of the T55 positive and 2 more zones of contrast. No positive-only film or digital camera can equal it. If you have to get rid of T 55, you're better off changing camera backs and using 4X5 negative sheet film. Good photo stores should have this, or the big mail-order companies like Helix or Shutan (in the back of photography mags).
Phil
} Howdy List, } } I need to stop using Polaroid T-55 p/n sheet film in my lab(good bye old } friend). The enviormental guys got to us. } } The question is, what to replace it with? T-52, T-53 or T-54. } } We will be using it primarily in our old Cambridge 250, inverted } metallograph, and some macro stand usage. } } Its use will be limited as our new JEOL SEM and inverted metallograph are } going digital. } } Any suggestions? } } } William T. Giles } Sr. Electron Microscopist } Met. Lab. Coordinator } Henderson Technical Laboratory } TIMET } PO Box 2128 Henderson NV 89009 } Ph: (702)566-4436 } Fax: (702)564-9038 } E-mail: Bill.Giles-at-timet.com
-- }}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{ 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)
} } } Where can I find prepared microscope slides that use flourescence? Or, where } can I find information about how to prepare my own slides with flourescent } stains? I would like such samples for research on 3D microscopy. } } Sincerely, } } Jennifer Delille } } jen-at-sl3d.com ********************** Jennifer, Its unclear what you wnat to do....Are you planning to work with biological samples that have been labelled with fluorescent markers, or is this a materials-related project? For biological samples, "normal" glass slides are fine. You usually do need to treat the slides in some way to get your samples (sections of tissue) to stick well enough to survive the multiple steps involved in the labelling process. The traditional method is to sub (coat) the slides with a gelatin-chrom alum solution. You can also use slides that have been treated by the manufacturer (silanized or the mysterious but useful "plus" slides).
The fluorescent labelling protocols depend on what you wih to look at. some are quite simple, such as the use of propidium iodide to label nuclei, others are multi-step, antibody protocls. You can find descriptions of these in the many good texts about immuncytochemistry.
If you have some materials-related project in mind, someone else will have to address that, I'm atrictly a biologicals person.
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
Its some time since I've looked at Goldstein's book, but I am sure you have missed a chapter. First, its important to note that in microanalyses the area irradiated is always smaller than the area you are collecting X-rays from. Its all related to the penetrating envelope, Monte Carlo patterns, kV used and average atomic number of the area under the beam. Typically, using a small spot, you may at 15kV collect X-rays from a 20 um radius and depths if your specimen is biological and only from 3um if its moderately heavy in atomic numbers. A good analyst would standardize all parameters, but probe current is more important than spot size. A dedicated microprobe is equipped with a lightmicroscope which has two main functions: Adjusting the specimen height to the correct position (taking advantage of the lightmicroscope's poor depths of view (or field) and visualising the beam spot so this can be centered, so the spot is were we think it is. The beam is visualised through the lightmicroscope by using a fluorescent mineral (Willemite). Centering using deflection coils and adjusting the beam (or spot) size is easy with that scope. Without an attached lightmicroscope, a small intense beam could be measured later by the burnmark left on a sensitive specimen. As an aside, geologists new to microanalysis are generally appalled by the poor quality of the lighmicroscope. They are rarely fond of learning to use atomic number contrast and to do their lightmicroscopy before a probe session. However, I wonder about the usefulness of the proposed exercise, because the relative beam size is usually less important because X-ray resolution is much less than beam size anyway. All of the above applies of course to SEMs/Probes since in a TEM (thin sections) the penetrating envelope is chopped off and the beam diameter plus a bit becomes the area analysed. Don't rush off and buy a TEM with analytical facilities, they have other disadvantages in microanalyses. Hope this clarified something . . I should also disclaim: ProSciTech supplies WDS/EDS standards. 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 ABN: 99 724 136 560 www.proscitech.com
On Thursday, July 27, 2000 10:23 AM, Gary Gaugler [SMTP:gary-at-gaugler.com] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I've studied Goldstein's book to try and accurately compute } the true spot size of a SEM beam based on the myriad of } factors which influence it. At issue is how to determine the } actual spot size of a beam, in microns, at the specimen. } } SEMs from various makers use relative numbers for spot } size. These indicate the relative size of the beam from } "really small" to "largest." "Really small" means low } specimen current but high resolution and the converse } for the large spot size. If one is doing x-ray analysis, } the spot size may or may not be an issue. } } If the area being analyzed is large, the spot size is } most likely not all that important. But for the analysis } of small feature size and sub-micron semiconductor devices, } true spot size is an issue. If the beam covers more than } the area of interest, the x-ray data will not accurately } reflect the composition of the intended material. } } Therefore, is there some basic limitation of the applicability } of quantitative SEM x-ray analysis based on the size of } the specimen? If so, what is this limit? Also, is there } some way to accurately define the physical size of a } beam at the specimen? If so, what is this method? } } Any thoughts on this? } } gary g. }
} Therefore, is there some basic limitation of the applicability } of quantitative SEM x-ray analysis based on the size of } the specimen? If so, what is this limit? Also, is there } some way to accurately define the physical size of a } beam at the specimen? If so, what is this method?
The applicability for the spot size regarding x-ray analysis would have as much to do with the energy of the beam, i.e., the interaction volume for x-ray generation (not to mention the possibility of x-ray fluorescence within the surrounding material.
The best method for determining the spot size for any beam parameter, is to examine the y-modulated intensity for a "knife edge". For example, a perfect orthoganal slope would indicate infinitely small .. for anything else you would determine the 'x' distance at 80% and 20% of the 'y' intensity (although these numbers could be debated).
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've studied Goldstein's book to try and accurately compute } the true spot size of a SEM beam based on the myriad of } factors which influence it. At issue is how to determine the } actual spot size of a beam, in microns, at the specimen. }
As others have already said, the relevant size is that of the volume from which x-rays are generated, and that is larger than the beam due to scattering within the specimen.
} } SEMs from various makers use relative numbers for spot } size. These indicate the relative size of the beam from } "really small" to "largest." "Really small" means low } specimen current but high resolution and the converse } for the large spot size. If one is doing x-ray analysis, } the spot size may or may not be an issue. }
If the spot size is small compared to the distances that the electrons will typically be scattered, then the x-ray emission volume will be nearly independent of spot size, but if the relevant scattering distance is small, then the spot size will affect the spatial resolution of the analysis. Fur- thermore, the emission volume will be smaller if the over- voltage for the line of interest is low. This is because scat- tering through large angles--which leads to large trans- verse momentum and large effective spot size--results in large transfer of kinetic energy, thus lower (or zero) cross section for the production of the line of interest. So if you use the k-lines and a relatively low voltage, the x-rays will be emitted from a smaller volume than if you use l- lines with the same voltage or k-lines with a higher vol- tage.
} If the area being analyzed is large, the spot size is } most likely not all that important. But for the analysis } of small feature size and sub-micron semiconductor devices, } true spot size is an issue. If the beam covers more than } the area of interest, the x-ray data will not accurately } reflect the composition of the intended material. }
Yes, although knowing the characteristics of the emission volume could allow you to deconvolute that effect and get a better idea of the compositions of the small areas of interest. That is, if you know the compo- sition of one area and the apparent composition of an adjacent area, you could use a Monte Carlo simulation to compute the part of the apparent composition due to x-rays generated within the first area. There are many such simu- lation programs out there, and you want one which will calculate not only the paths of the electrons, but also the x-ray production. I do not know which of the simulation programs will do this, but perhaps their authors will tell you.
} } Therefore, is there some basic limitation of the applicability } of quantitative SEM x-ray analysis based on the size of } the specimen? If so, what is this limit? Also, is there } some way to accurately define the physical size of a } beam at the specimen? If so, what is this method? }
Yes. It depends on specimen composition and what is of interest. Yes. Either simulation or doing an experiment with a specimen that has well characterized features of known size and composition. This specimen should be as similar to what you're trying to measure as possible to assure that both features of interest and matrix effects are well modelled. In the case of semiconductor devices, perhaps the Ge stripes in Si matrix of a MAGICAL test specimen could be used. (I have no connection with MAGICAL except as a user.) Yours, Bill Tivol
Dear Gary, Yes, there certainly is a size limit to a specimen or phase or particle in the specimen to get accurate SEM x-ray analysis. I use the Small World's Electron Flight Simulator to get a good estimate of the size of the electron interaction volume in a given material. These are all Monte Carlo mathematical simulations, since the SEM spot is not a spot, but a fuzzy disk and the volume is a randomly generated approximation based on the factors influencing the interaction of the electrons and material of the solid. What is seldom mentioned is that the the x-rays generated by the primary electrons can also spread laterally in the solid and these x-rays can spread much further than the initial excitation volume and excite secondary x-rays several microns away. These secondary x-ray effects vaary widely, dependant on which elements are being excited and fluoresced. There are two ways to test these effects. The first is to carefully scan the SEM beam across a sharp interface between two materials, such as Ni plated on polished Cu, and record the shape of the Ni and Cu line scans. The length of the curve dropping between 100% Cu and 0% Cu will tell you the size of the beam interaction volume in the solid (which is not the spot size, but the size of the x-ray generation volume). The second is to drill a small hole in a copper planchet and stick a tungsten aperture over the hole. Slowly scan the beam over the Cu, W and Faraday cup and see where you can see Cu and W x-rays. This gives you some idea of the beam spread and x-ray scatter. At 05:22 PM 7/26/00 -0700, you wrote:
} I've studied Goldstein's book to try and accurately compute } the true spot size of a SEM beam based on the myriad of } factors which influence it. At issue is how to determine the } actual spot size of a beam, in microns, at the specimen. } } SEMs from various makers use relative numbers for spot } size. These indicate the relative size of the beam from } "really small" to "largest." "Really small" means low } specimen current but high resolution and the converse } for the large spot size. If one is doing x-ray analysis, } the spot size may or may not be an issue. } } If the area being analyzed is large, the spot size is } most likely not all that important. But for the analysis } of small feature size and sub-micron semiconductor devices, } true spot size is an issue. If the beam covers more than } the area of interest, the x-ray data will not accurately } reflect the composition of the intended material. } } Therefore, is there some basic limitation of the applicability } of quantitative SEM x-ray analysis based on the size of } the specimen? If so, what is this limit? Also, is there } some way to accurately define the physical size of a } beam at the specimen? If so, what is this method? } } Any thoughts on this? } } gary g.
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 everyone, I have a TEM Philips CM12 and one problem of this type of microscope is to know all the rotation angles between the SAED pattern and the image in every magnification. Surely it is possible to use a test specimen to determine this angles, but may be there is someone had done this and he have this list and he can help me.
Thank you everyone, Elena Belluso
---------------------------------------------------- Elena BELLUSO Dipartimento di Scienze Mineralogiche e Petrologiche Via Valperga Caluso, 35 I-10125 TORINO - ITALIA tel:(39) 011 670 7135 - fax: (39) 011 670 7128 e-mail: belluso-at-dsmp.unito.it http://www.dsmp.unito.it ----------------------------------------------------
Our roughing pump which service our JSM6300 has began making a rumbling noise which I might believe is bearings ... altho it could be the pump's bearings or the motor's. Has anyone serviced this problem?
tia and 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/
Would someone direct me to websites and/or texts about in situ hybridization techniques? I'll be working with gene expression in shoot meristems from plant material (in vivo and in vitro). For regular LM I use glutaraldehyde+paraformaldehyde as a fixative and Historesin (Leica) for embedding, but I don't think this method would be appropriate. I'd appreciate some input. Thank you very much in advance
Are you sure the endothelial cells you're after express PECAM1/CD31 in appreciable amounts. I have found a great deal of disparity in CD31 staining in mouse tissue. I would suggest vWF or factor VIII or AP2 as potential substitutes.
Ramin Rahbari Pfizer Global Research & Development Drug Safety Evaluation 2800 Plymouth Road Ann Arbor, MI 48105 Voice (734) 622-3383 Fax (734) 622-5001 Ramin.Rahbari-at-WL.COM
-----Original Message----- } From: Michael Reiner [mailto:Elektronenmikroskopie-at-web.de] Sent: Wednesday, July 26, 2000 5:32 AM To: Neelima Shah; Microscopy-at-sparc5.microscopy.com
Hello Neelima Shah!
Did you try PECAM-1/CD31 (rat) from Pharmingen? It must work well at least using a pre-embedding protocol without permeabilization.
Greetings, Michael Reiner
University of Cologne, Germany Department of Anatomy I
} Anyone out there know of any antibody that would be specific for mouse } endothelial cells?? We are trying to identify endothelial cells in tumors } as well as cultured endothelial cells. I know a lot of them work for LM } after various treatments. I seem to have had no luck so far staining at EM
} level. I would appreciate all the input.Thanks in advance..... } Neelima Shah.............. } Biomedical Imaging Core Facility } Uni of Pennsylvania } Philadelphia, Pa. } } } } } http://www.MED.upenn.edu/morphlab/ } } è
Jackson, D.P. 1991. In situ hybridization in plants. In: Molecular Plant Pathology - A Practical Approach. (sorry, I don't have the page numbers)
McFadden, G.I. 1995. In situ hybridization. Meth. Cell Biol. 49:165-183
Good luck!
Tamara Howard CSHL
On Thu, 27 Jul 2000, Adriana Pinheiro Martinelli Rodriguez 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 member: } } Would someone direct me to websites and/or texts about in situ hybridization techniques? I'll be working with gene expression in shoot meristems from plant material (in vivo and in vitro). } For regular LM I use glutaraldehyde+paraformaldehyde as a fixative and Historesin (Leica) for embedding, but I don't think this method would be appropriate. } I'd appreciate some input. } Thank you very much in advance } } Adriana } }
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I have recently inherited a great deal of equipment (stage warmers, micro-incubators, perfusion chambers, etc) distributed by Medical Systems Corp. of Greenvale, NY. I've been told that they're still productive, yet I've been unable to locate any current contact information. Does anyone know whether they still exist and how I might be able to contact them?
Warren Davis Cell Imaging Facility School of Medicine University of Utah 40 N. 2030 E. Bldg. 585, Rm. 55 Salt Lake City, UT 84112 Ph: (801) 587-7964 email: warren.davis-at-path.utah.edu
by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id OAA12567 for dist-Microscopy; Thu, 27 Jul 2000 14:44:55 -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 OAA12563 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Thu, 27 Jul 2000 14:44:25 -0500 (CDT) Received: from risc4.numis.nwu.edu (risc4.numis.nwu.edu [129.105.122.70]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id OAA12556 for {Microscopy-at-sparc5.microscopy.com} ; Thu, 27 Jul 2000 14:44:14 -0500 (CDT) Received: from localhost (ldm2-at-localhost) by risc4.numis.nwu.edu (8.9.3 (PHNE_18979)/8.9.3) with ESMTP id OAA19622 for {Microscopy-at-sparc5.microscopy.com} ; Thu, 27 Jul 2000 14:53:25 -0500 (CDT)
A postdoctoral position is open immediatesly for work on Environmental Catalysts using UHV-TEM. The position will involve looking at the the surface structure of a number of different oxides as a function of gas treatments using our UHV instrument (see http://www.numis.nwu.edu .) Experience in catalysis, UHV systems and a strong background in TEM (not just a few courses) preferred. Send a CV and letters of recommendation (or referees names) to
Laurence Marks Department of Materials Science and Engineering Northwestern University ldm2-at-risc4.numis.nwu.edu
What is frequently measured in TEM is scalar dislocation density (the sign of dislocations is not taken into account), even though this is not the only thing that it is possible to measure (e.g. excessive dislocation density).
Concerning the scalar dislocation density (which is simply called dislocation density), it is better to use such diffraction conditions when there are several strong reflections present, then most of the dislocations should be visible. I would recommend to use this condition. However, as Witold Zielinski wrote, you can use two-beam conditions but then you should take several images at different conditions and analyze what is visible and what is not. Alternatively, using two beam conditions a correction factor can be introduced (see ref. 1 below).
It is also very important to know the thickness of the analyzed area, you will need it to calculate dislocation density in your material.
Another important consideration is rearrangement of dislocations due to the stress relaxation after removal of the load and during thinning due to the image force. Therefore, it is better to analyze thicker area of the sample. You can calculate this force (ref 3) and estimate the thickness of the sample where you should perform your measurement for a particular material.
Total error of your measurements, which you can estimate (e.g. ref 10), will be around 20%.
I would recommend to look through this references. If you won't find everything you need, you can contact me directly and I will fax you some additional information.
1. Hirsch et al. 1965 Electron Microscopy of thin crystals
2. D.G. Brandon, Y. Komem Metallography, 1970, v.3, 111
3. A. Seeger, Handbuch der Physik, 1955, v. VII/1, 560.
4. J.W. Steeds, Roc. Roy. Soc., 1966, A292, p.343
5. J.E. Bailey, Phil. Mag. 1963, v. 8, p.223.
6. J.R. Hancock, Phil. Mag., 1968, 18, p. 1235
7. Baker T.N., ed. 1983 Yield, flow and fracture of polycrystals London, Applied Science, 1983
8. Ecob R.C., J. Microscopy, 137, 1985, p. 3
9. Ivanov A.I.Y., Mezhennyi O., Ostrov A.E., Fomicheva E.I. Zavodskaya laboratoriya, 53 (2), 1987, p.43 (this is in Russian, but there is a copy of this paper in English).
10. Staker M.R. and Holt D.L Acta metall. 20, 1972, p. 569
11. Finally, there is an excellent book in Russian, may be you can find somebody to translate to you several pages from there. Utevskii L.M. "Diffraction elecron microscopy in metallurgy", Moscow, Metallrgiya, 1973.
Good luck,
Evgenia
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****************************************** Evgenia Pekarskaya Materials Science Mail Stop 138-78 California Institute of Technology Pasadena, CA, 91125, USA tel: 1-626-3953571 fax: 1-626-7956132
For those of you attending Microscopy & Microanalysis 2000 in Philadelphia next month-
Once again, MSA's Education Committee is organizing mini-seminars or tutorial demonstrations known as "Exhibitor Demonstrations". These sessions are conducted by Exhibitors in their booths on Tuesday evening from 5:00 to 6:00 pm (or some fraction thereof). To attend one of these demonstrations you must sign up at MSA's Education Booth, now part of the MSA "Mega-Booth", before Tuesday noon. You will be issued a ticket that will allow you back into the Exhibit Hall after it is closed to regular attendees that evening. A list of titles and a short description of each demonstration will be available at the Education Booth. While you are there, see what else the Education Committee has to offer! Below is a current list of participating Exhibitors:
Advanced Microscopy Techniques Digital Instruments/Veeco Metrology Group E.A. Fischione Instruments EDAX Electron Microscopy Sciences Energy Beam Sciences Evex Analytical FEI Company Illumea Corp. KS Electron Technologies LEO Electron Microscopes Microbiology International/Syncroscopy Micro Photonics Microscopy/Marketing & Education Motic NORAN Instruments Photon Technology International SELA USA South Bay Technology Ted Pella TSL
See you in Philly-
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* ****************************************************************************
At 05:59 AM 7/27/00, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Yes, it is covered but does not seem to lead one to a finite conclusion. The big variable is electron optics. My current FESEM has a flat pole piece. My LaB6 SEM has a 60 degree conical lens. Other units have different angles. The lens design must play some major portion of the whole operation of getting from the emitter to the specimen. It is a tight system.
} Don Chernoff has available (http://www.small-world.net/efs.htm) a program } which } models the analysis volume. A similar but less flexable program came with } my } (former) Kevex EDS in the mid 80s. I have not tried the EFS software, nor } have } I any ties to small-world. } } Woody White } McDermott Technology, Inc.
Since so many variables come into play, it is not surprising that details are a bit vague in the book. Haven't referred to it in quite a while and I forget the exact wording... Taming the variables is where the statistical software is handy.
I can't imagine differing lens construction making a difference in the analysis volume other than a conical lens permitting higher tilt angles for certain specimens which will cause the volume to skew along the surface (the incident beam angle variable). Have I missed something?
Since the beam diameter is typically MUCH smaller than the analysis volume, variations in its size will not significantly affect the x-ray analysis resolution. One exception to this is WDS analysis. Sometimes HUGE beam currents are desired. This can lead to a(fat)beam that will limit x-ray resolution. Generally speaking, if you can see a crisp SE image at the desired magnification, the lens configuration and beam diameter are not part of the analysis volume equation. At this point you may or may not have enough beam current to achieve the desired count rate. Rule of thumb: Only if you must increase the beam current/spot size (to raise the count rate) until the SE image is no longer well resolved is it time consider beam size affecting the outcome.
Woody ------------------------------------------------------------- At 05:59 AM 7/27/00, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Yes, it is covered but does not seem to lead one to a finite conclusion. The big variable is electron optics. My current FESEM has a flat pole piece. My LaB6 SEM has a 60 degree conical lens. Other units have different angles. The lens design must play some major portion of the whole operation of getting from the emitter to the specimen. It is a tight system.
} Don Chernoff has available (http://www.small-world.net/efs.htm) a program } which } models the analysis volume. A similar but less flexable program came with } my } (former) Kevex EDS in the mid 80s. I have not tried the EFS software, nor } have } I any ties to small-world. } } Woody White } McDermott Technology, Inc.
Hello I am observing a double image from my TEM filament. The microscope has been aligned over and over. If anyone has seen this please help Thanks Wilbur
A few days ago I reminded everyone about the abuses that were starting to creep into the Listserver Re: Selling of Items.
At that time I promised to create something which would address that issue. Well I've bitten the bullet and got the prototype completed and running.
There now exists a WWW site where any individual can post an electronic advertisement to dispose of SURPLUS equipment (ie not new instrument sales). This site is open to all individuals, organizations and vendors. The URL is:
All postings to the Surplus Equipment Forum are reviewed (by guess who) to make sure they are not junk/spam/bogus postings but not reviewed for accuracy.
There are no charges to use this Forum, however, I am suggesting an honor system. If you use the Forum to dispose of excess equipment and you receive more than shipping costs, then you should consider voluntarily donating an appropriate amount to the Society to help defray operating costs of this site. I will not monitor this aspect as I have neither the time nor the inclination to do so.
Remember, if you have items that you are giving away for at most nominal shipping costs, then by all means continue to use the Microscopy Listserver. It was created with that spirit in mind and I wish that tradition to continue.
This is clearly an experiment. There will of course be some problems and growing pains and if it doesn't work then I've only lost a few hours of time. However, I expect that it could be reasonably successful and fill a need in our community. So have at it.
Cheers....
Nestor Your Friendly Neighborhood SysOp..
.. as he heads off to the sun set to get dinner and a beer....
Does some one know of a source for: Soft Touch II grey tips Catalog Number #00593, manufactured by the Boehringer Mannheim Company of Indianapolis, IN (for use with Soft Touch II Lancet Device) in or near the Washington, D. C. -- Northern Virginia -- Maryland area?
They are still being manufactured, as per our conversation with the manufacturer (last month - June) even though it is an older item.. However the manufacturer only ships to wholesalers. Many suppliers no longer carries them. . .
OR perhaps someone in the DC area has switched to a newer Lancet device and might still have some grey tips in stock..
Also need a local source for frosted slides 25x75 mm x 1mm thick.
OR would there be a source of help near Atlanta, GA? I'll be in Atlanta in conference (July 30-August 4). The need is immediate -- I am traveling with an Olympus CH30 phase contrast microscope, performing live blood cell demonstrations and these are no longer available from our own supplier. Any information would be greatly appreciated. Thank you in advance...
Gratefully, Ann Soleman, BA, BS, ThM. Certified Nutritional Microscopist
Wellness-4-All Post Office Box 250 Oakton, VA 22124 USA www.wellness4all-at-eyionline.com + Visit My Web Site + passcode: wellness4all E-mail: wellness4all-at-juno.com E-mail: RASoleman-at-aol.com E-mail: anns-at-eyionline.com Voice Mail: 1-877-881-2593 Electronic Fax: 1-877-881-2593 Phone 1-703-591-1232 ________________________________________________________________ YOU'RE PAYING TOO MUCH FOR THE INTERNET! Juno now offers FREE Internet Access! Try it today - there's no risk! For your FREE software, visit: http://dl.www.juno.com/get/tagj.
we are having serious problems with the resolution of our EDS detector - the peak height-width ratio is terrible, around 450 counts. Consequently, our peaks look more like rolling hills and one peak blurs into another. I have callibrated the machine using an Al-Cu standard, checked that the detector is kept cool, sample is tilted at 30 degrees, etc etc. I also got another microscopist to come and have a look to check my methods and he also concluded there was a serious problem.
Is there anything else I could try before I phone the Noran technicians and pay them a lot of $$$ to fix/replace the detector?
regards Liz McKenzie -------------------------------------------------- Geomicrobiology and Electron Microscopy Laboratory Room S9 Cramer Hall 1721 SW Broadway Portland State University Portland OR97201
At 03:50 PM 7/27/00, you wrote: } Since so many variables come into play, it is not surprising that details } are a bit vague in the book. Haven't referred to it in quite a while and } I forget the exact wording... } Taming the variables is where the statistical software is handy. } } I can't imagine differing lens construction making a difference in the } analysis volume other than a conical lens permitting higher tilt angles } for certain specimens which will cause the volume to skew along the } surface (the incident beam angle variable). Have I missed something? } } Since the beam diameter is typically MUCH smaller than the analysis } volume, variations in its size will not significantly affect the } x-ray analysis resolution. One exception to this is WDS analysis. } Sometimes HUGE beam currents are desired. This can lead to a(fat)beam } that will limit x-ray resolution. Generally speaking, if you can see a } crisp SE image at the desired magnification, the lens configuration } and beam diameter are not part of the analysis volume equation. } At this point you may or may not have enough beam current to achieve } the desired count rate. } Rule of thumb: Only if you must increase the beam current/spot size } (to raise the count rate) until the SE image is no longer well resolved } is it time consider beam size affecting the outcome. } } Woody
The link below shows what I am trying to analyze.
http://photoweb.net/ICxsect.jpg
I know that PSG, Al and poly si are correctly marked. The poly gate is what is confusing me. Its contrast is like the surrounding oxide while its interior is like the poly layer....or it could be metal over poly.
Based on these rather small feature sizes, will quantitative x-ray analysis produce viable results? If I probe the small central area of the gate, will I get the results of that area or the results of it and the surrounding material?
when the pumps start to go bad, it seems to be preceded by leaking at the motor-pump interface. The unit goes downhill from there.
If you don't have any leaking oil, does the pump sound like knocking? I've heard this and found that it was due to an air leak. If the exhaust close off control on the pump is closed, and the knocking persists, it means there is an air leak somewhere. You should see this in poorer vacuum readings.
Have you ever changed the oil?
gary g.
At 09:39 AM 7/27/00, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We recommend nitrogen-burst agitation for one second out of every three seconds. This timing is built into our Autoprocessor. We will have a small model system in our booth at M & M in Philadelphia.
Best regards, Steven Slap
****************************** Energy Beam Sciences, Inc. The Laboratory Microwave Company http://www.ebsciences.com Adding Brilliance to Your Vision ******************************
Thank you very much for mineralogical samples I have received from Martin J. Roe.
Regards, Elena
---------------------------------------------------- Elena BELLUSO Dipartimento di Scienze Mineralogiche e Petrologiche Via Valperga Caluso, 35 I-10125 TORINO - ITALIA tel:(39) 011 670 7135 - fax: (39) 011 670 7128 e-mail: belluso-at-dsmp.unito.it http://www.dsmp.unito.it ----------------------------------------------------
Part-time Laboratory Assistant Position - 20 hours/week
The American Museum of Natural History is seeking a Part-Time Laboratory Assistant for its state-of-the-art core imaging and image processing facility. The Interdepartmental Laboratory houses an Hitachi Cold Field Emission Scanning Electron Microscope/PGT EDS, a Zeiss Laser Scanning Confocal Microscope, and image processing/high quality output resources.
Major responsibilities include assisting scientific staff with imaging and x-ray microanalysis projects, and training users in independent operation of microscopes and computers. Other laboratory duties include minor equipment maintenance, occasional specimen preparation, and other general lab-related tasks.
A Bachelors degree in biology is preferred, along with some knowledge of or interest in other scientific disciplines such as geology or materials science. Candidates who are currently enrolled in a scientific undergraduate or graduate program will also be considered. The position requires some knowledge of SEM and strong computer skills. Other qualifications include willingness to learn, flexibility, and excellent organizational and interpersonal skills.
Hourly rate is commensurate with experience. Interested candidates can contact me directly.
--------------------------------------------- Angela V. Klaus
Director - Interdepartmental Laboratories American Museum of Natural History Central Park West at 79th Street New York, NY 10024-5192 USA
I'm seeking recommendations for 20-21 color monitors to use for histological image analysis. Important parameters are high resolution and color reproduction, refresh rates of at least 85Hz and , of course, reliability. I've heard some good things about Viewsonics and the Sony GDM-500PS. Are there any I should avoid?
Thanks, Brett
Brett M. Connolly, Ph.D. Merck Research Laboratories Department of Pharmacology WP26A-3000 PO Box 4 West Point, PA 19486 Ph. 215-652-2501 FAX 215-652-2075 e-mail: brett_connolly-at-merck.com
Microsope Folks, I am going to have the fun of doing a minor rebuild on an old ISI 100B SEM. Alas, the manual seems to not have arrived with this old beast and I am in need of pointers to where I can obtain a copy. Thanks in advance!
I just got this Oxford EDS Link QX2000 attached to JEOL JSM 5300. I wonder if someone can enlighten me on way the Gross Integral and Net Integral are calculated from the EDS Link QX2000 and also what is that STRB?
Hi, The rotation calibration is described in Edington, using MoO3 samples. If you have any kind of sample for which a crystallographic direction is unambiguously known with respect to some linear feature of the sample then you can use that too. It is not hard to do this calibration. Just make double exposures (SAED and BF image) at each magnification for each camera length you are interested in. I think you can use a short cut: once the entire rotation calibration is done for a single camera length, then you need only do one magnification at any other camera length to get the whole thing - on the Philips microscopes I've used in the past, diffraction patterns do not rotate with change in camera length.
Milo
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Hi, It's likely caused by EMI, but could also be caused by a ground loop, or a noisy power supply.
Regards, Dave Harrison
On 27 Jul 00, at 17:21, waltew82-at-eng.uab.edu wrote:
} Hello I am observing a double image from my TEM filament. The microscope has } been aligned over and over. If anyone has seen this please help Thanks Wilbur }
By chance, has anyone run any calibration coatings with the Hummer VII using an Au/Pd target? The coating thickness is dependent on the thickness setting but also on the nm/minute rate. While the system is not meant to be absolute, it does provide a general idea of thickness. The question is whether there is one or more rate settings which have corresponding tables or relationships to end-point thickness settings? I am running at 80mTorr.
Hello Brett, I personally have a PS 790 & it's predecessor (both 19" View Sonic). I am quite happy with them. Max. refresh rates are 90hz at 1280x1024 & 76Hz at 1600x1200.... a no $ interest disclaimer applies here. The price of large monitors has dropped significantly which puts good monitors in the price range of not so good units. When looking at large monitors one thing to be weary of is a spatial non linearity across the screen (distortion). Unless your trying to make measurements on the screen with a hand ruler this is generally not noticeable. The larger the monitor, the more difficult this is to control. It is not uncommon to find small magnets glued to the CRT. This is obviously not an adjustable parameter. An easy way to look for/qualify the problem is to look to see if a round object in the middle is round or oval by the edge. This can vary with individual monitors. I have an expensive 19" (not VS) that has this problem to a greater extent than my 2 VS monitors in which it exist just a bit. Actually I hadn't even checked them 'til just now. The other thing I have run it to (and not recently) is color or brightness non-uniformity across the screen. Again this is often not noticed in normal computer applications. Try putting up a white screen & working with the brightness & contrast to check for this problem.
good luck, Bruce Brinson Rice U.
"Connolly, Brett" 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 seeking recommendations for 20-21 color monitors to use for histological } image analysis. Important parameters are high resolution and color } reproduction, refresh rates of at least 85Hz and , of course, reliability. } I've heard some good things about Viewsonics and the Sony GDM-500PS. Are } there any I should avoid? } } Thanks, } Brett } } Brett M. Connolly, Ph.D. } Merck Research Laboratories } Department of Pharmacology } WP26A-3000 } PO Box 4 } West Point, PA 19486 } Ph. 215-652-2501 } FAX 215-652-2075 } e-mail: brett_connolly-at-merck.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 I am observing a double image from my TEM filament. The microscope has } been aligned over and over. If anyone has seen this please help } Thanks } Wilbur
} } There now exists a WWW site where any individual can post } an electronic advertisement to dispose of SURPLUS equipment (ie not } new instrument sales). This site is open to all individuals, } organizations and vendors. The URL is: }
} } This is clearly an experiment. There will of course be some problems } and growing pains and if it doesn't work then I've only lost a few hours } of time. However, I expect that it could be reasonably successful } and fill a need in our community. So have at it. }
This is a wonderful thing, I'm sure it will work and will be much appreciated.
Will it be appropriate to post queries from those seeking items also, such as me in my ceaseless quest for 840 accessories?
cheers (at least three)
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 are having serious problems with the resolution of our EDS detector - the peak height-width ratio is terrible, around 450 counts. Consequently, our peaks look more like rolling hills and one peak blurs into another. I have callibrated the machine using an Al-Cu standard,
You do not say what your total count rate is or whether it is the same as it used to be for the standard you are using. I assume that both count rate and dead time are what they should be. I also assume that the FWHM of the Mn k-alpha peak is } 150 eV (regardless of height).
checked that the detector is kept cool, sample is tilted at 30 degrees, etc etc. I also got another microscopist to come and have a look to check my methods and he also concluded there was a serious problem.
When this happened to us, it turned out that the vacuum had deteriorated. After having sent the detector back for reconditioning, we designed and installed a valve which could be attached to the EM column pumping system. This enabled us to warm up and pump out the detector ourselves. After about 15 years, however, this no longer worked, and we had to send the detector for a pro- fessional cleaning.
Is there anything else I could try before I phone the Noran technicians and pay them a lot of $$$ to fix/replace the detector?
Get estimates for the various repair options. We decided to have Doug Connors of TNAS clean and overhaul our unit, since his price for that service was the cheapest. After the bakeout and pumpdown he performed, the resolution had returned to 147 eV (from ~200 eV when we sent it in). His charges for other services--which we hoped would not be needed--were comparable to Noran's and others'. We have no affiliation with Doug or TNAS except as customers. Good luck. Yours, Bill Tivol
} } } Our roughing pump which service our JSM6300 has began making a } rumbling noise which I might believe is bearings ... altho it could be } the pump's bearings or the motor's. Has anyone serviced this problem? }
I guess that this isn't your problem, but a couple of times I've had alarming sharp knocking noises from my belt-drive JEOL pumps, which have been completely cured with a few squirts of drive-belt dressing compound from an aerosol can. Seems to be some sort of sticky tacky lubricant, works well for me.
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
From looking at the image, I'd estimate that the features you need to analyse are about 1/2 micrometer for the body of the gate, and 100 nm for the surrounding material. For sufficiently low voltage and a sufficiently small beam you might have an emission volume small enough to obtain an analysis from the body. It's less likely that a meaningful analysis can be obtained just from the surrounding layer. However, you might be able to measure apparent elemental concentrations as a function of po- sition and calculate the concentrations in the layer with the assumption that both body and layer have uniform compositions--this is not a trivial assumption.
I know that PSG, Al and poly si are correctly marked. The poly gate is what is confusing me. Its contrast is like the surrounding oxide while its interior is like the poly layer....or it could be metal over poly.
Here may be a problem. In order to reduce the size of the emission volume, you need to lower the voltage; however, if the voltage is too small, there might not be a enough overvoltage to obtain sufficient yield of a suitable x-ray line from the elements in the layer.
Based on these rather small feature sizes, will quantitative x-ray analysis produce viable results? If I probe the small central area of the gate, will I get the results of that area or the results of it and the surrounding material?
Could be. Probably just from the body with the right exsperimental parameters. Good luck. Yours, Bill Tivol
You can do in situ hybridisation on plant material embedded in wax, LRWhite, LRGold, BMM (butyl methyl methacrylate) and probably historesin. I haven't looked for references on in situs in non-embedded material, but there is no reason why you couldn't do this in the same way people do antibody labelling of plant material for immunofluorescence. I've seen bacteria processed for in situs without resin embedding.
Good review article: McFadden (1989) Cell Biol Int Rep 13:3-21 covers use of wax and resin for in situs.
Quite a few people still use FAA as a fixative, which is OK for tissue work but makes my cell biologist hair stand on end! However, you usually need to do protease digestion if you fix with aldehydes as the probes have a hard time getting in through a highly cross-linked protein matrix.
If you fix in 4% paraformaldehyde and embed in BMM, you can do both antibody labelling and in situs on the same material. See Kronenberger et al. (1993) Cell Biology International 17:1013-1021 for BMM embedding protocol for in situs.
cheers, Rosemary
} } Dear list member: } } Would someone direct me to websites and/or texts about in situ } hybridization techniques? I'll be working with gene expression in shoot } meristems from plant material (in vivo and in vitro). } For regular LM I use glutaraldehyde+paraformaldehyde as a fixative and } Historesin (Leica) for embedding, but I don't think this method would be } appropriate. } I'd appreciate some input. } Thank you very much in advance } } Adriana
Rosemary White Microscopy Centre CSIRO Plant Industry GPO Box 1600 Canberra, ACT 2601 Australia
The Gross Integral and NetIntegral values on Oxford Instruments (ex Link) EDS systems refer to the total number of counts in a set energy window (Gross Integrals) and the number of counts in the energy window after subtracting the background (Net Integrals). The background is calculated by drawing a line between the top of the first window channel and the top of the last window channel, all the area under that line is assumed to be the background.
STRB is the zero energy strobe.
Have fun with your QX2000,
Ron
On Sat, 29 Jul 2000, Fauzi Mohd Som wrote:
} I just got this Oxford EDS Link QX2000 attached to JEOL JSM 5300. I wonder } if someone can enlighten me on way the Gross Integral and Net Integral are } calculated from the EDS Link QX2000 and also what is that STRB? } } TQ. } } } }
=========================================================================== 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. ============================================================================
We are looking for TEM capability to run a few thin film cross section samples. The films are ~100nm thick, & we need full crystallographic analysis, as well as imaging of the top surface, interface with substrate etc.
If anyone has these capabilities, preferably in Europe, can you contact me off list?
Thanks
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/
Bill - you are an optimist. Very much doubt that anything close to 2 um diameter or less can be truly quantitative (in SEM type instrument; within +/- 1% of reality). If you lower the kV you run out of oomph, if the specimen is light (not needing much oomph, then the volume is even greater. I like optistist because they persevere. 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 ABN: 99 724 136 560 www.proscitech.com
On Monday, July 31, 2000 7:53 AM, "wft03-at-health.state.ny.us"-at-sparc5.microscopy.com [SMTP:"wft03-at-health.state.ny.us"-at-sparc5.microscopy.com] wrote: } } } Dear Gary, } } The link below shows what I am trying to analyze. } } http://photoweb.net/ICxsect.jpg } } From looking at the image, I'd estimate that the features you need to } analyse } are about 1/2 micrometer for the body of the gate, and 100 nm for the } surrounding } material. For sufficiently low voltage and a sufficiently small beam you } might have } an emission volume small enough to obtain an analysis from the body. It's } less likely } that a meaningful analysis can be obtained just from the surrounding layer. } However, } you might be able to measure apparent elemental concentrations as a } function of po- } sition and calculate the concentrations in the layer with the assumption } that both body } and layer have uniform compositions--this is not a trivial assumption. } } I know that PSG, Al and poly si are correctly marked. The poly } gate is what is confusing me. Its contrast is like the surrounding } oxide while its interior is like the poly layer....or it could be metal } over poly. } } Here may be a problem. In order to reduce the size of the emission } volume, you } need to lower the voltage; however, if the voltage is too small, there } might not be a enough } overvoltage to obtain sufficient yield of a suitable x-ray line from the } elements in the layer. } } Based on these rather small feature sizes, will quantitative x-ray } analysis produce viable results? If I probe the small central } area of the gate, will I get the results of that area or the results } of it and the surrounding material? } } Could be. Probably just from the body with the right exsperimental } parameters. } Good luck. } Yours, } Bill Tivol } }
Broadened peaks can be caused by electromagnetic interference. We experienced this problem several years ago when we decided to "tidy-up" the maze of cables lying around on the floor. Without thinking, we neatly coiled up the detector signal cable and hung it on a clip attached to the back of the EDS monitor. Soon after, we began realizing that our peak width had increased significantly. It was only after much pain and grief that we realized what we had done. The intense electromagnetic field generated by the flyback coil in the monitor was the culprit. As soon as we un-coiled the cable the peak width returned to normal.
Look for sources of fields such as monitors, fluorescent lights, etc. Moving them as little as a foot or so can make a big difference.
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: E. J. McKenzie [SMTP:elizm-at-pdx.edu] } Sent: Friday, July 28, 2000 12:31 PM } To: microscopy-at-sparc5.microscopy.com } Subject: EDS resolution } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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, } } we are having serious problems with the resolution of our EDS detector - } the } peak height-width ratio is terrible, around 450 counts. Consequently, our } peaks look more like rolling hills and one peak blurs into another. I have } callibrated the machine using an Al-Cu standard, checked that the detector } is kept cool, sample is tilted at 30 degrees, etc etc. I also got another } microscopist to come and have a look to check my methods and he also } concluded there was a serious problem. } } Is there anything else I could try before I phone the Noran technicians } and } pay them a lot of $$$ to fix/replace the detector? } } regards } Liz McKenzie } -------------------------------------------------- } Geomicrobiology and Electron Microscopy Laboratory } Room S9 Cramer Hall } 1721 SW Broadway } Portland State University } Portland } OR97201 } } ph:503 725 3362 } fax:503 725 3025 }
Shut down the instrument and remove the drive belt from the mechanical pump/Motor. Restart the instrument to power the vacuum pump motor to isolate which is faulty. Chances are that it is the vacuum pump and you will hear the motor running quietly. If it is the motor, replacement of the motor is required. If the noise is coming from the vacuum pump you could change the oil, and that is worth a try. If one or more of the bearings are defective they are easily replaced but you will need a gasket kit.
If you are under a service contract that would be the best place to start. Hope this helps
John Humenansky/Staff Scientist Physical Electronics, Inc. (PHI) 6509 Flying Cloud Drive Eden Prairie, MN 55344 952-828-6387
I have been invited to present an overview talk on elemental microanalysis in botany during the 7th International Conference on Nuclear Microprobe Technology and Applications (ICNMTA'2000), Bordeaux 10-15 September 2000. I would like to cover recent applications in research areas such as plant physiology, agriculture and environmental pollution. Special emphasis will be made on an update of nuclear microprobe applications, following the research summarised during the Santa Fe conference in 1996 and published in Nucl. Instr. Meth. B130 (1997) 335. I also intend to report on problems solved using other microanalytical techniques such as EDX, SIMS, LMMS, SXRFM and EELS. I will greatly appreciate your cooperation in that matter as I do not want to miss any related work in the overview. Please send me any information of relevance, a reprint, fax copy or just an e-mail with the list of publications. Due to time restrictions I would be grateful for your fast reply.
Best regards
Jolanta Mesjasz-Przybylowicz
Dr Jolanta Mesjasz-Przybylowicz Materials Research Group National Accelerator Centre PO Box 72 Faure 7131 South Africa E-mail: mesjasz-at-srvnac3.nac.ac.za Fax: +27-21-8433543 Phone: +27-21-8433820
We recently purchased a Nikon Coolpix 990 for documentation use, but I am absolutely loving the resolution and capability of this camera. Does anybody know where I can find an adapter to use it on my various scopes throughout the lab? I know that they make some lenses for this camera, but does anybody know of any third party adapters or methods besides some duct tape and a steady hand holding to an eyepiece (just kidding...)?
Thanks for your help, ~Jonathan Dunlap
Jonathan Dunlap Electronic Components and Materials Analytical Laboratory Manager Osram Sylvania Inc. 816 Lexington Avenue Warren, PA 16365 Ph: 814-726-6991 Fax: 814-726-6956
} Bill - you are an optimist. Very much doubt that anything close to 2 um } diameter or less can be truly quantitative (in SEM type instrument; within +/- } 1% of reality). If you lower the kV you run out of oomph, if the specimen is } light (not needing much oomph, then the volume is even greater. } I like optistist because they persevere. }
Guilty. On the HVEM--much different, but with relatively large beam size (~0.5 mu-m)--I was able to distinguish a feature ~0.25 mu-m across. You're correct about quantitation.
} } I know that PSG, Al and poly si are correctly marked. The poly } } gate is what is confusing me. Its contrast is like the surrounding } } oxide while its interior is like the poly layer....or it could be metal } } over poly.
To distinguish between oxide and a metal surrounding the poly, all you would need is a qualitative analysis. That's what I am optimistic about. You're right about the greater volume for less dense materials. Since the range of electrons is roughly the same in units of mg/cm^2 for all materials with the same number of protons as neutrons in the nucleus, scattered electrons will travel further. The hope is that there is enough oomph and small enough volume at a particular energy. This could happen if there is a good low-energy (m or n) line for a heavier element which is not either on top of a k or l line for a lighter element and which is not buried in noise or too low a cross section. Yours, Bill Tivol
} I have a TEM Philips CM12 and one problem of this type of microscope is to } know all the rotation angles between the SAED pattern and the image in every } magnification. Surely it is possible to use a test specimen to determine } this angles, but may be there is someone had done this and he have this list } and he can help me. } Dear Elena, The Mag-i-cal specimen has instructions for doing this. I am not affiliated with this specimen or its vendor(s) except as a user. Yours, Bill Tivol
To the wealth of knowledge on the Microscopy Listerver,
We are looking into purchasing a digital camera for our TEM... We have a Philips 208S and do mostly diagnostic work here in the EM lab related to kidney biopsies...
I have looked at two cameras.. The AMT 1K and the Gatan Dual view 1K.. They both deliver acceptable images.. I was just wondering what kind of problems have occurred with both cameras after that have been set up and in operation? i.e. customer service? problems? breakdowns?
As was mentioned, the Mag-i-cal sample is really good for determining the roation calibration. You can either buy or make a sample of MoO3 by heating wire or foil in air until you get white smoke and waving a coated grid around in the smoke.
You do have a 180 degree ambuguity in the diffraction pattern. To eliminate that, go to a convergent beam mode and underfocus the pattern (decrease the condenser lens strength) until you see the BF image in the disk. That will correlate with both the image and the diffraction pattern.
-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: Belluso elena [mailto:belluso-at-dsmp.unito.it] } Sent: Thursday, July 27, 2000 12:33 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: rotation angle between SAED and image in TEM } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: 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 everyone, } I have a TEM Philips CM12 and one problem of this type of } microscope is to } know all the rotation angles between the SAED pattern and the } image in every } magnification. Surely it is possible to use a test specimen } to determine } this angles, but may be there is someone had done this and he } have this list } and he can help me. } } Thank you everyone, } Elena Belluso } } } } ---------------------------------------------------- } Elena BELLUSO } Dipartimento di Scienze Mineralogiche e Petrologiche } Via Valperga Caluso, 35 } I-10125 TORINO - ITALIA } tel:(39) 011 670 7135 - fax: (39) 011 670 7128 } e-mail: belluso-at-dsmp.unito.it } http://www.dsmp.unito.it } ---------------------------------------------------- } }
Dear Gary, I had a look at your image and I recently had a similar problem with the analysis of fine layers in a GaAs/Al device. I was able to get good analyses by lowering the acc. voltage to 3 to 5 kV. In that case I used the L lines of Ga and As and the K line of Al to probe each layer in turn. My Monte Carlo program tells me that the x-ray generation volume for carbon, aluminum and silicon is 0.2 microns in depth and diameter at 3 kV, giving you the resolution you need to analyse the edge and centre of the gate, assuming that C, O, Al and Si are all the elements present. The 3 kV is almost a doubling of the highest energy line analysed (1.74 keV Si Ka line) and should excite all the lines you need. At 08:29 PM 7/27/00 -0700, you wrote:
} At 03:50 PM 7/27/00, you wrote: } } Since so many variables come into play, it is not surprising that details } } are a bit vague in the book. Haven't referred to it in quite a while and } } I forget the exact wording... } } Taming the variables is where the statistical software is handy. } } } } I can't imagine differing lens construction making a difference in the } } analysis volume other than a conical lens permitting higher tilt angles } } for certain specimens which will cause the volume to skew along the } } surface (the incident beam angle variable). Have I missed something? } } } } Since the beam diameter is typically MUCH smaller than the analysis } } volume, variations in its size will not significantly affect the } } x-ray analysis resolution. One exception to this is WDS analysis. } } Sometimes HUGE beam currents are desired. This can lead to a(fat)beam } } that will limit x-ray resolution. Generally speaking, if you can see a } } crisp SE image at the desired magnification, the lens configuration } } and beam diameter are not part of the analysis volume equation. } } At this point you may or may not have enough beam current to achieve } } the desired count rate. } } Rule of thumb: Only if you must increase the beam current/spot size } } (to raise the count rate) until the SE image is no longer well resolved } } is it time consider beam size affecting the outcome. } } } } Woody } } The link below shows what I am trying to analyze. } } http://photoweb.net/ICxsect.jpg } } I know that PSG, Al and poly si are correctly marked. The poly } gate is what is confusing me. Its contrast is like the surrounding } oxide while its interior is like the poly layer....or it could be metal } over poly. } } Based on these rather small feature sizes, will quantitative x-ray } analysis produce viable results? If I probe the small central } area of the gate, will I get the results of that area or the results } of it and the surrounding material? } } gary g. } 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'm seeking recommendations for 20-21 color monitors to use for histological } image analysis. Important parameters are high resolution and color } reproduction, refresh rates of at least 85Hz and , of course, reliability. } I've heard some good things about Viewsonics and the Sony GDM-500PS. Are } there any I should avoid? }
I use Viewsonic P185 monitors. I have 7 of them. I understand there is an updated model (perhaps the P817?). I much prefer them to Sony. Sony uses an aperture grill supported by wires that create two very fine lines visible on the screen. We use the monitors at our graphics workstations, on our Hitachi S3500n SEM and on our Gatan CCD camera on the Philips TEM.
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
Just wanted to add my 2 cents worth...I agree with Jonathan Dunlap - the Nikon CoolPix 990 is a great digital camera. Ours was purchased for departmental use so it's used out in the field, on the copy stand, etc. Everyone has been very pleased with the images. best regards, Beth
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
I just saw that Bruce mentioned measurements with a ruler on the computer screen:
I don't think, that is a viable method of measuring something. The glass on the monitors is quite thick to withstand the enormous forces at the center of the screen (at 1kg/cm2 the screen glass has to be thick!!). Since the image is created on the inside, while the ruler is on the outside, the angle at which you look at the ruler plays a significant role. Just place a ruler on the screen and move your head. The measurements will change by much more than the distortion of the screen (unless it's really bad).
For accurate measurements, one should always (in my opinion) use software to do the measurements. That makes the measurement completely independent of display devices.
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: Bruce Brinson [mailto:brinson-at-cnst.rice.edu] Sent: Saturday, July 29, 2000 9:35 PM To: Connolly, Brett Cc: 'MSAMicroscopy'
Hello Brett, I personally have a PS 790 & it's predecessor (both 19" View Sonic). I am quite happy with them. Max. refresh rates are 90hz at 1280x1024 & 76Hz at 1600x1200.... a no $ interest disclaimer applies here. The price of large monitors has dropped significantly which puts good monitors in the price range of not so good units. When looking at large monitors one thing to be weary of is a spatial non linearity across the screen (distortion). Unless your trying to make measurements on the screen with a hand ruler this is generally not noticeable. The larger the monitor, the more difficult this is to control. It is not uncommon to find small magnets glued to the CRT. This is obviously not an adjustable parameter. An easy way to look for/qualify the problem is to look to see if a round object in the middle is round or oval by the edge. This can vary with individual monitors. I have an expensive 19" (not VS) that has this problem to a greater extent than my 2 VS monitors in which it exist just a bit. Actually I hadn't even checked them 'til just now. The other thing I have run it to (and not recently) is color or brightness non-uniformity across the screen. Again this is often not noticed in normal computer applications. Try putting up a white screen & working with the brightness & contrast to check for this problem.
good luck, Bruce Brinson Rice U.
"Connolly, Brett" 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 seeking recommendations for 20-21 color monitors to use for histological } image analysis. Important parameters are high resolution and color } reproduction, refresh rates of at least 85Hz and , of course, reliability. } I've heard some good things about Viewsonics and the Sony GDM-500PS. Are } there any I should avoid? } } Thanks, } Brett } } Brett M. Connolly, Ph.D. } Merck Research Laboratories } Department of Pharmacology } WP26A-3000 } PO Box 4 } West Point, PA 19486 } Ph. 215-652-2501 } FAX 215-652-2075 } e-mail: brett_connolly-at-merck.com }
we have several CoolPix here with adapters for different scopes - the folks at Diagnostic Instruments are very helpful in discussing and selecting components. You can then purchase through one of their distributors. Be aware that depending on your microscope you'll need some relatively expensive components to couple to the photo port because the CP has a non removeable lens. For others it is as simple as an adapter tube. Make sure you also get the Nikon shutter release bracket (unless the 990 has a remote shutter release - the 800/950 doesn't)
regards, Wolfgang Ziegler
At 10:34 AM 7/31/00 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi. I am a studant from Brazil and I´m trying to measure the grain size of my samples (They are Niobium 9%Tantalun). I know there are a lot of softwares available on the internet, but I don´t know where and if there is a specific program to analise the grain size of metals. Please, remember that I work at Brazil, so I don´t have any money buy a software. We allways try to do the best we can.
Thank you very much..... João Paulo Barros Machado.....
Please answer this email for: jpbm-at-easygold.com.br
A few weeks ago, I asked if there was interest in having an Emispec Users Group meeting at M&M. Emispec is willing to have one if there is interest. So far, I have not received very many replies. I will have to let Emispec know what the interest level is and if there isn't much, it will not fly.
If you have an Emispec system and you have an interest in having this meeting at M&M, please let me know.
Please pass this message on to Emispec users and have them respond to me.
If you already responded to me, it wouldn't hurt to do so again.
Thanks,
-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
FALL 2000 COURSE ANNOUNCEMENT - Transmission Electron Microscopy (BIO. 221-E2)
NASSAU COMMUNITY COLLEGE (Long Island, NY)
A fourteen week, Fall 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 ONE EVENING PER WEEK, Thursdays, starting at 5:30pm. Classes will begin on Sept. 7 and end on Dec. 14, 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.
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} .
For those without www access, the catalog description is specified below. If you have further questions, you should e-mail me directly at the address below.
Interested individuals should register early (by Aug. 10) since the course is limited to a total enrollment of ten (10) students.
Questions regarding the actual registration process can be directed to our registrar at (516) 572-7355. ________________________________________________________________________________
CATALOG DESCRIPTION BIO 221: Transmission Electron Microscopy -- 4 credits Prerequisites: BIO 109-110 (Intro. Bio.) or equivalent, CHE 151-152 (Inorganic Chem.) An introduction to the basic principles of transmission electron microscopy including tissue preparation, microscope (TEM) operation, black & white photography, and micrograph interpretation. The entire laboratory is devoted to the development of skills and preparative techniques involved with the operation of an actual transmission electron microscope. (3 lecture, 3 laboratory hours). Laboratory fee applies. ________________________________________________________________________________
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}
Thanks very much to all who have responded to this initial posting. It seems to me that the data I have received confuses me for than before. But the ideas presented are great to consider. Proving them out is quite another matter.
At 09:14 AM 7/30/00, you wrote: } Gary, } } It is common for IC manufacturers to use metals in direct contact with poly } gates thus reducing the sheet resistance of the poly especially for } interconnects such as in your image. It could be that the similar contrast } means a titanium-silicide or something like that. Therefore, to excite the } higher x-ray lines will require higher beam voltage and you are correct in } wondering about the volume which is generating the detected x-rays. } } I have had much better success analyzing semiconductor cross sections with } Auger than EDS. Auger's built-in ion gun sputtering provides a good means } of cleaning the face of the cross section, the resolution (for imaging and } analyzing) is certainly adequate for this size structure and once you have } identified the elements present over an area, a line scan nails the elements } present in the gate. } } I have some question about the labels on the photo. Are you really sure } about those?
I offer a second image at
http://photoweb.net/icxsect2.jpg
which is a different and augmenting image of the IC structure.
I am not sure of the identification of each layer but I am pretty sure of them. However, I can be dissuaded by alternate opinions. The main point here is I think to keep in mind that this is a fifteen year plus construction...it is not modern. Therefore, what would be vogue or state-of-the-art back then?
The ability of x-ray analysis to quantify the various layers seems to be easy or impossible, based on the feedback I have received. On first blush, it does seem difficult based on the small geometric area available for probing. I do not yet have an x-ray capability but will be receiving one soon. It is a 133eV light element dewar system. It will be interesting to see how it performs in this application.
I forgot to add the URL to a second image. Here it is:
http://www.photoweb.net/icxsect2.jpg
This and the previous image are BSE. The Z contrast in these images is telling, I think. The problem is that they are not conclusive. The particular problem is the area at the substrate surface. I think that this is a gate area, surrounded by field oxide. But I am not sure. What is really troubling is the central portion of the structure in the center of the image. It has the same contrast as the poly Si....but how could such a structure be made?
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BOC Gases, has a unique opportunity for an Electron Microscopist/Materials Engineer located at it's state of the art Technical Center in Murray Hill, NJ.
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Electron Microscopist/Materials Engineer
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BOC Gases is an equal opportunity employer.
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When I had resolution problems recently (which turned out to be due to a "leaky" detector) one of the first things my service engineer asked me to check was to make sure that the detector wasn't touching the pole piece (which can cause a ground loop?) Try backing the detector out a little to see if the peak width changes. I doubt it will help, but it is easy to try. Matt
"E. J. McKenzie" {elizm-at-pdx.edu} on 07/28/2000 12:30:59 PM
To: microscopy-at-sparc5.microscopy.com cc:
Hi everyone,
we are having serious problems with the resolution of our EDS detector - the peak height-width ratio is terrible, around 450 counts. Consequently, our peaks look more like rolling hills and one peak blurs into another. I have callibrated the machine using an Al-Cu standard, checked that the detector is kept cool, sample is tilted at 30 degrees, etc etc. I also got another microscopist to come and have a look to check my methods and he also concluded there was a serious problem.
Is there anything else I could try before I phone the Noran technicians and pay them a lot of $$$ to fix/replace the detector?
regards Liz McKenzie -------------------------------------------------- Geomicrobiology and Electron Microscopy Laboratory Room S9 Cramer Hall 1721 SW Broadway Portland State University Portland OR97201
Folks does anyone know where I could obtain a service manual (not a user manual) for a Microm 505 cryostat Tx Simon
---------------------------------------------- Simon C. Watkins Ph.D. MRC Path Associate Professor Cell Biology and Physiology Director Center for Biologic Imaging University of Pittsburgh BSTS 225 3500 Terrace ST Pittsburgh PA 15261 Tel:412-648-3051 Fax:412-648-8330 URL: http://sbic6.sbic.pitt.edu --------------------------------------------
We manufacture semiconductors. I work in the Analytical Services cross-sectioning these daily. These images you sent are of a bulk silicon process. To best describe the layers, I will start at the silicon and work my way up. On the second picture the field oxide comes to a point in two locations allowing the metal above to make contact to the silicon. The first picture shows this layer as the full thickness along the silicon. The next layer is the 1st metal(not poly silicon) that is making a contact in the second image and just a first metal line on the first image. The metal has a undoped oxide over it that gives the different z contrast. Then there is an oxide over that, could be PSG. Then there is a second metal layer(labeled Poly Si on the first picture). On top of that is a layer of oxide(which is labeled Al on the first picture). Then on top of everything is either a nitride or polyimide capping the chip. The metals are easy to identify because of the grain structure. There are not any gates shown on these two photos. Gary if you would like to give me a call I can explain the layers in more detail. I can also give you information on how they are processed and what to look for.
Mark Windland Honeywell Solid State Electronics Center Plymouth, Minnesota 612-954-2845 ------------------- } Thanks very much to all who have responded to this initial } posting. It seems to me that the data I have received confuses } me for than before. But the ideas presented are great to } consider. Proving them out is quite another matter. } } At 09:14 AM 7/30/00, you wrote: } } Gary, } } } } It is common for IC manufacturers to use metals in direct contact with } poly } } gates thus reducing the sheet resistance of the poly especially for } } interconnects such as in your image. It could be that the similar } contrast } } means a titanium-silicide or something like that. Therefore, to excite } the } } higher x-ray lines will require higher beam voltage and you are correct } in } } wondering about the volume which is generating the detected x-rays. } } } } I have had much better success analyzing semiconductor cross sections } with } } Auger than EDS. Auger's built-in ion gun sputtering provides a good } means } } of cleaning the face of the cross section, the resolution (for imaging } and } } analyzing) is certainly adequate for this size structure and once you } have } } identified the elements present over an area, a line scan nails the } elements } } present in the gate. } } } } I have some question about the labels on the photo. Are you really sure } } about those? } } } I offer a second image at } } http://photoweb.net/icxsect2.jpg } } which is a different and augmenting image of the IC structure. } } I am not sure of the identification of each layer but I am } pretty sure of them. However, I can be dissuaded by alternate } opinions. The main point here is I think to keep in mind } that this is a fifteen year plus construction...it is not modern. } Therefore, what would be vogue or state-of-the-art back then? } } The ability of x-ray analysis to quantify the various layers } seems to be easy or impossible, based on the feedback I } have received. On first blush, it does seem difficult based } on the small geometric area available for probing. I do not } yet have an x-ray capability but will be receiving one soon. } It is a 133eV light element dewar system. It will be interesting } to see how it performs in this application. } } gary } }
Does anyone out there have a Kevex-ray EDX detector, 2003 model? We don't have a manual (or any literature, for that matter) and I just wondered if I could obtain a photocopy or acrobat file from someone. I've also contacted Noran.
Thanks for all the hints on fixing the peak problems - it seems to be working fine now (210eV FWHM for Mn peak), I just have to figure out what went right!
regards Elizabeth McKenzie -------------------------------------------------- Geomicrobiology and Electron Microscopy Laboratory Room S9 Cramer Hall 1721 SW Broadway Portland State University Portland OR97201
I did not comment on your last image, as I needed more info. before making a statement. The second image is easy to comment on. The gate structure is the line which is the same contrast as the substrate. It starts towards the left of the image, over "field oxide", drops down over the "birds beak" and continues across the gate oxide. Then it rises over the other "birds beak", and terminates over the "field oxide". The darker thin line at the surface of the substrate (connecting the two "bird beaks") is the gate oxide. The gate appears to be polysilicon, but I would need more information to say for sure.
I am having problems removing a white, spotty, crystalline- like residue from my XTEM sample. Has anyone out there experienced similar problems? Any suggestions would be appreciated. I summarize my prep procedure below, as well as the methods I used to try to remove the residue. (It should be noted that my sample is a sandwich sample prepared using Gatan G-1 epoxy.)
1. Mount sample on pyrex polishing stub with superglue. 2. Polish with alumina lapping films. 3. Final polish with polycrystalline diamond suspension on a final polishing cloth. 4. Soak in acetone to dissolve superglue and release sample from stub.
After the above procedure there was the aforementioned residue. I attempted removing it by the following methods: 1. Rinsed/soaked in ethanol. 2. Rinsed/soaked in water. 3. Rinsed/soaked in acetone overnight. 4. Rinsed/soaked in hexane. 5. Sonicate in acetone. 6. Sonicate in water.
} From time to time we have recommended cyanoacrylate super glues for use with tripod polishing and related methods. The trouble with using consumer products is that they sometime change without warning. We have experienced poor results with glues we have recommended in the past. No point in naming names. After performing an evaluation on replacement glues, we now recommend:
1). RP30 CYANOACRYLATE ADHESIVE - $6.75/1 oz. bottle
by AMERICAN CHEMICAL CO. 2630 South Irving Avenue Minneapolis, MN 55408-1047 (612)-374-1767 Contact: Bill Reker FAX: (612)-377-3590 Web: www.glueit.com E-mail: info-at-glueit.com
2). 310035 INSTANT SUPERGLUE ETHYL CYANOACRYLATE - $5.98/1 oz. bottle
by ND INDUSTRIES 750 Wylly Avenue, Suite 3 Sanford, Fl. 32773 (800)-521-2663 Contact: Laurie Russell
Sold to us via: Moreau Marketing & Sales 501 Shepherd Street, Suite #4 Winston Salem, NC 27103 Contact: Robert Moreau (336)-659-6674 FAX: (336)-659-6683
For further details, please contact Mark Hudson at (845)-892-2441 (currently using RP30) or Stan Klepeis at (845)-892-2192 (currently using 310035).
Ron Anderson, IBM, Hopewell Jct., New York, USA. anderron-at-us.ibm.com
IBM Analytical Services; http://www.chips.ibm.com/services/asg
It seems as if you may be contaminating your sample with the superglue after using the acetone to clean the sample. However, what you describe sounds much like colloidal silica particles.
Try using IPA or methanol after acetone. Acetone leaves residue on samples and does not come clean easily.
If for some reason you neglected to mention the use of colloidal silica, I am 98 percent sure that is what you see. If that is the case, MicroOrganic Soap works well to remove colloidal silica from samples. You may also wish to switch the type of colloidal silica you are using.
I hope this works for you.
Sincerely,
Gary Liechty Manager, Technical Products
Allied High Tech Products, Inc. 2376 E. Pacifica Place Rancho Dominguez, CA 90220
310-635-2466 310-762-6808 Fax 800-675-1118 Toll Free, US and Canada
www.alliedhightech.com
Equipment and Consumables for Metallurgical Sample Preparation ----- Original Message ----- } From: "john david whitaker" {jwhitake-at-u.washington.edu} To: "MSA listserver" {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, August 01, 2000 10:39 AM
Dear Microscopy Listserv,
I have a colleague that is doing a light microscopic comet assay (single cell gel electrophoresis) on glass slides using respiratory cells from sputum. She has been using silver staining (w/ a kit from Trevigen) and is having difficulty getting clean preps (ie without precipitates or extraneous staining). Does anyone have any tips or suggestions? She's tried doing the assay with fluorescent preps on our confocal microscope, but the specimen fades too quickly to acquire useful data.
Yours, Doug .................................................................... : 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://swehsc.pharmacy.arizona.edu/exppath/ Home of: "Microscopy and Imaging Resources on the WWW"
Arizona State University Center for High Resolution Electron Microscopy Research Specialist
The Center for High Resolution Electron Microscopy at Arizona State University has an open position for a Research Specialist. The Center provides electron microscopy resources for the university and external community, including a range of transmission and scanning electron microscopes, as well as specimen preparation facilities. The successful applicant will be expected to assist with the day-to-day operation and maintenance of microscopes within the Center. Primary instrument responsibility will include but not be limited to the JEOL 2000FX transmission electron microscope (TEM), the JEOL 840 Scanning EM, and the soon-to-be acquired JEOL 6300 SEM with liquid-helium CL system. Other tasks will include training and supervision of researchers in proper use of the instruments, including observance of safety procedures, and authorization of users. Some limited opportunity to assist and advise researchers in designing and carrying out experiments, including active participation and collaboration in experiments with data collection and analysis as required.
Minimum qualifications: Bachelors or Masters degree or equivalent training in Physical or Materials' Sciences or closely related discipline, with at least three but preferably five years' additional experience with operation and maintenance of scanning and/or transmission electron microscopes. Experience with design, construction and testing of electronic circuitry, and knowledge of vacuum systems would be desirable.
Application deadline: August 15, 2000, or until position filled.
Arizona State University is an equal opportunity employer. Women and minorities are encouraged to apply.
Send resume and the names and addresses of three references to John Wheatley. Please contact John via one of the following ways--preferably e-mail:
John C. Wheatley, Lab Manager Arizona State University Center for Solid State Science Tempe, AZ 85287-1704
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
| ------------------------------------------------------------------------ | The Microscopy ListServer -- Sponsor: The Microscopy Society 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. 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 | | ****************************************************************** |
Nelson Conti 164 Ferne Court Palo Alto, CA 94306
Gen Pei- I've unsubscribed myself one time. I believe that you need to send an e-mail to the MSA Listserver (address = ? ), and just indicate "unsubscribe" in the subject heading. To find the address, go to the MSA Listserver Web address, and it should be possible to find the e-mail address for the MSA Listserver. Sorry, I don't recall it off-hand. Good luck with unsubscribing. Nelson Conti P.S. When in doubt - you can also re-send your message, but direct it to Nestor Zaluzec. I'm sure that he can tell you precisely how to unsubscribe.
_______________________________________________________ Say Bye to Slow Internet! http://www.home.com/xinbox/signup.html
Have any of you had any experience with the installation of a wireless lan affecting the operation of your microscopes in any way? I would very much appreciate your advice in this regard.
Sincerely,
Mick Thomas ------------------------------------------------- Mick Thomas UHV-STEM Laboratory E-1 Clark Hall Cornell University Ithaca, NY 14853
I have not yet seen any effects, with the Apple Airport system installed for my computers as well as a GigaHertz wireless broadcasting systems which are carrying video signals from one room to another.
The Computer monitors however are a different story.
Nestor
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Sorry for the previous blank message. Nestor thinks it was just a fluke.
We have an Astromed cooled CCD camera, which we would like to use. However, we have no manuals for it. If you have manuals or other information that would allow us to put this camera into operation please let me know "off line".
Thank you, Alwyn Eades .
The camera has two labels: Model number TE2W (serial 117) Model number P86121 (serial 7451/8/14)
There are two boxes of electronics labelled: Box 1 Model TE2/PSU (serial 102) Box 2 Model DDE/2200 (serial 116) 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
Hello everyone; I have just started looking at a GFP-transformed fungus growing on plant surfaces, using regular fluorescence microscopy. I can clearly see the fungus using two filter sets: a UV set (ex/em 365/} 420) and a blue filter set (ex/em 470/} 515). While the fluorescence in blue light is much brighter, stuff that is blue/blue-white with UV illumination is green in the blue light. is the "extra" green in the blue light likely to be where the GFP is not as concentrated, and hence doesn't show up in the UV, or is it more likely to be some other compound that has similar excitation/emission characteristics? I have done a fair bit of reading on the subject, but am still feeling a bit vague, and would really appreciate hearing from those of you who have some first hand experience with GFP.
thanks in advance shea
Dr. S.Shea Miller Agriculture & AgriFood Canada Eastern Cereal and Oilseed Research Centre Rm. 2068 Neatby Building Central Experimental Farm Ottawa, Ontario Canada K1A 0C6 Phone: (613) 759-1760 Fax: (613) 759-1701 E-mail: millers-at-em.agr.ca
With regard to the posting I made on this subject, Chuck Garber sent me the following note wherein he raises a number of important points. Chuck is correct in all regards. The product we were using was not DURO. We have no experience with Duro. We have made specific, brand name, recommendations in the past and my note was to share our recent experiences regarding these products. The products we were using were not "garage level" products.
So, IF, and only IF, you are using a product solely because of a recommendation from our lab, we now make new recommendations.
Ron
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Hi Ron,
I guess I should give some kind of disclaimer statement first, that in no way am I suggesting that what you are saying it not correct. If this was your perceived experience, then I am sure it is correct and I am not questioning it.
And it is a fact that there are some garage level operations where people take from 5 gallon pales and "fill" the 1 oz plastic bottles and put on a label and put it into distribution, without any real testing of the integrity of their packaging or any of the other things you and I might think one should be doing to insure the integrity of their product.
But I can assure you, without hesitation, the cyanoacrylate that is made by Duro, which is indeed a consumer products company, but it is also "top in class" when it comes to this type of product, has been offered by us (e.g. SPI) for years and we have never ever had anyone every voice even the slight complaint about it. We also decided a long time ago that once opened, the shelf life of these products can be very short, and consequently, we decided to offer the product in the smaller 3g package.
I go through the trouble to point this out to you because I see things from a slightly different perspective possibly than you do:
1] For one thing, someone even at IBM, having to place an order with either of the two mentioned companies, given the cost of placing an order period of any size, what is the overhead going to be? But if someone was ordering the Duro product from someone like SPI, then it just gets added in with a bunch of other things and the real cost is almost nothing.
2] For someone in Mexico, the situation is far worse. You have a lot of influence on people because of your reputation. Your posting could make someone in Mexico City think that in order to get acceptable results, they have to place a direct order in the USA for one of these "special" cyanoacrylates and in situations with limited research budgets, it just seems to me like they have better ways to be spending their money.
So while the Duro product might not be any better than the two mentioned, the point is that because of the distribution and transportation costs, if people are led to believe that they really do need something that can not be procured from their traditional sources of purchasing, they end up wasting a lot of money unnecessarily.
Of course if you had any hard evidence that the Duro product was in some way inferior, that would surely be another story. But chemically, these are all the same identical materials, they all have the same identical MSDS information and the all perform identically. When ever I have heard of someone having problems it has been because they were using product from a previously opened package.
Feel free to use this information however you feel might be approriate, either in whole or in part. But I did want to share my views with you on your posting, which I hope you won't mind my having done.......
Chuck
Ron Anderson, IBM, Hopewell Jct., New York, USA. anderron-at-us.ibm.com
IBM Analytical Services; http://www.chips.ibm.com/services/asg
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Hi all, We would like to add an individualized computer password system to our FEI/Philips CM-10 TEM. We would like a user to be able to type in their password which would then trigger a switch tied into the high tension and turn the HT on. The system would ideally then have a timer to record "on" time and a function to record this time with the user ID. Connecting a timer to the high tension is not a problem...the computer password part is where we need help. Please contact me if you have a system such as this on a FEI/Philips CM-10 or even on another make TEM which might work on a CM-10. Thanks, 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
by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id PAA15187 for dist-Microscopy; Wed, 2 Aug 2000 15:52:00 -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 PAA15181 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Wed, 2 Aug 2000 15:51:29 -0500 (CDT) Received: from guru.palm.cri.nz (guru.palm.cri.nz [161.66.1.22]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id PAA15174 for {microscopy-at-sparc5.microscopy.com} ; Wed, 2 Aug 2000 15:51:02 -0500 (CDT) Received: from mailscan.hort.cri.nz (mailscan.hort.cri.nz [161.66.1.30]) by guru.palm.cri.nz (8.10.0/8.10.0) with ESMTP id e72Kidl26285 for {microscopy-at-sparc5.microscopy.com} ; Thu, 3 Aug 2000 08:44:39 +1200 (NZST) Received: from hra1.marc.cri.nz (unverified) by mailscan.hort.cri.nz (Content Technologies SMTPRS 4.1.5) with ESMTP id {Ta142011e834dcc4abc46-at-mailscan.hort.cri.nz} ; Thu, 3 Aug 2000 08:38:51 +1200 Received: from HRA1/SpoolDir by hra1.marc.cri.nz (Mercury 1.47); 3 Aug 00 08:44:46 GMT+1300 Received: from SpoolDir by HRA1 (Mercury 1.47); 3 Aug 00 08:44:31 GMT+1300
Date sent: Wed, 02 Aug 2000 11:20:54 -0400 } From: "Shea Miller" {millers-at-em.agr.ca} To: confocal-at-listserv.acsu.buffalo.edu, microscopy-at-sparc5.microscopy.com
Well, I guess Mark and I would have to see a higher magnification image, detailing the polysilicon, the "birds beak" area, and the substrate. This way we could tell whether this is a gate, with gate oxide, or a diffusion contact. I am assuming that your cross section is well into the middle area of the polysilicon line, and not just at the edge.
OK...I think that I have this figured out. I got some good input from several folks who were right about some portions and some who were perhaps wrong. So was I. But I think that I have it sorted out now.
I really can't say that this is a double poly structure quite yet until my x-ray system is available (Rontec 133eV SHV). Even then, it may not be able to discern the constituents in the small gate area. We'll see.
In the meantime, I will put an annotated pix on my site which shows what I think is the real identification of each layer. It still may not be 100% right....but I think that it will be quite close.
I am looking for a programme (or a collaboration) to compute powder XRD or SAED taking in account the size effect due to the platelet shape of small crystals in a polycristalline (non textured?) powder (hexagonal Bravais lattice).
In a first, simple, approach we can consider only the beam broadening of the bulk reflections (‰+/size in that particular direction). A more accurate computation would take in account that the smallest crystals are no longer a sum of complete unit cells but a supercell.
Simulation of powder patterns with CrystalMaker or Carine show that it exist a strong overlapping between reflections for isotropic crystal some nanometers in size.
Do somebody know if such a programme is available or where somebody could perform a few simulations for me?
Best regards
Philippe Buffat
__________________________________________________________________ Philippe Buffat Ecole Polytechnique Federale de Lausanne (EPFL) Centre Interdepartemental de Microscopie Electronique Address: EPFL-CIME, Batiment MX-C, CH-1015 Lausanne, Switzerland Phone: +41(21)693 29 83 Fax: +41(21)693 44 01 (Central European Time) E-mail: philippe.buffat-at-epfl.ch, WWW URL http://cimewww.epfl.ch/ ______________________________ Eudora F2.1 ___________________________
As we approach the upcoming meeting I would like to take this opportunity as the LAC Chair to let you know that this meeting promises to be one of the best yet. With a sell out of Exhibit space and pre registration growing Philadelphia is the place to be this August. To all of our surprise, we have sold out on most social events. We do have left just 75 Spirit of 76 Wednesday Evening Dinner Cruise tickets left which we would like to open up to those of you whom have not purchased any. Joining this trip will afford you the relaxing experience of floating down the Delaware River on an elegant ship filled with wonderful food, and lively music. Sail by Penn's Landing, Old Christ's Church, the Ben Franklin Bridge, the Flagship Olympia, and much much more. The cruise includes a five entry buffet with all of the condiments, salad, desserts and coffee and tea. There are 2 dance bands as well as entertainment on board. Transportation will be from all of the local hotels and convention center. All of this on a usually boring Wednesday night for only $55 for everything. Please E-mail me directly or call if you are interested so we may make all of the arrangements. I look forward to hearing from you.
Our university is looking to do some imaging of DNA and DNA/protein interactions. We do not have a AFM and are looking to contact anyone in the NJ, NY, CT, or PA area so that we could visit a lab and possibly contract out some work or work in collaboration. If you are aware of any company/university that has AFM, could you please contact myself or Jack Gaynor at these email addresses: mganger-at-aol.com or jack.gaynor-at-montclair.edu.
Thanks in advance,
Mike Ganger Montclair State University mganger-at-aol.com
Dear Sir/Madam: Do you know of any vendors who sell older or discontinued Nikon parts? I'm looking for a dic nosepiece for a diaphot 200? Any suggestions. TIA
This message is in reply to Philip Oshel at U. of Wisconsin:
I've always been told that the brown goop that is easy to get on your hands while pulling apart the photo & negative of the Polaroid T55 film is a known carcinogen and to wash your hands immediately upon contact.
Jane L. LaGoy Development Engineer Bodycote IMT, Inc. 155 River Street Andover, MA 01810 978-470-1620 jlagoy-at-bodycote-imt.com
I would like to ask you how to stain non-collagenous fibres in paraffin-embedded human tissues (i. e. liver), by using Fast Green stain. Have someone the method?
I read some paper about this stain, but these reported a pre-incubation with 0.01% Fast green in saturated picric acid solution for 15 min, and then after some washes a new incubation with Fast green at the concentration of 0.04% in saturated picric acid solution for 30 min.
Why is important the pre-incubation?
Many thanks, Fabio Grizzi
Dr. Fabio Grizzi Direzione Scientifica, Istituto Clinico Humanitas Via Manzoni 56 20089 Rozzano, Milan, Italy
Curious. I've never heard this, but I won't disagree. It's photodeveloper chemicals, same basic sort of stuff used in photography darkrooms. I haven't seen any warnings about these chemicals in the packaging materials/information sheets. It is common sense to keep it off your hands, and to wash your hands if you get it on them, but I haven't had any problems not getting the goop on my hands. And I'd have several trophies for clumsiness if I hadn't dropped and broken them all ...
Phil
} This message is in reply to Philip Oshel at U. of Wisconsin: } } I've always been told that the brown goop that is easy to get on your hands } while pulling apart the photo & negative of the Polaroid T55 film is a known } carcinogen and to wash your hands immediately upon contact. } } Jane L. LaGoy } Development Engineer } Bodycote IMT, Inc. } 155 River Street } Andover, MA 01810 } 978-470-1620 } jlagoy-at-bodycote-imt.com
-- }}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{ 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)
The MSDS on the 55 developer states that the developer does not contain any listed carcinogens by OSHA, IARC or NTP. The major danger seems to be the corrosiveness of the alkali from Potassium and Lithium hydroxide. Ingestion obviously is a danger due to these and other components. Polaroid has it's MSDS's up on it's website at:
http://www.Polaroid.com/service/msds/index.html
Gloves and/or handwashing are very much in order.
} I've always been told that the brown goop that is easy to get on your hands } while pulling apart the photo & negative of the Polaroid T55 film is a known } carcinogen and to wash your hands immediately upon contact.
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Date sent: Wed, 02 Aug 2000 11:20:54 -0400 } From: "Shea Miller" {millers-at-em.agr.ca} To: confocal-at-listserv.acsu.buffalo.edu, microscopy-at-sparc5.microscopy.com
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Fellow microscopists,
Have any of you had any experience with the installation of a wireless lan affecting the operation of your microscopes in any way? I would very much appreciate your advice in this regard.
Sincerely,
Mick Thomas ------------------------------------------------- Mick Thomas UHV-STEM Laboratory E-1 Clark Hall Cornell University Ithaca, NY 14853
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I have not yet seen any effects, with the Apple Airport system installed for my computers as well as a GigaHertz wireless broadcasting systems which are carrying video signals from one room to another.
The Computer monitors however are a different story.
Nestor
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Fellow microscopists,
Have any of you had any experience with the installation of a wireless lan affecting the operation of your microscopes in any way? I would very much appreciate your advice in this regard.
Sincerely,
Mick Thomas ------------------------------------------------- Mick Thomas UHV-STEM Laboratory E-1 Clark Hall Cornell University Ithaca, NY 14853
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Well, I guess Mark and I would have to see a higher magnification image, detailing the polysilicon, the "birds beak" area, and the substrate. This way we could tell whether this is a gate, with gate oxide, or a diffusion contact. I am assuming that your cross section is well into the middle area of the polysilicon line, and not just at the edge.
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On Tue, 1 Aug 2000 19:09:28 -0400, Gen Pei 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. 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 | | ****************************************************************** |
Nelson Conti 164 Ferne Court Palo Alto, CA 94306
Gen Pei- I've unsubscribed myself one time. I believe that you need to send an e-mail to the MSA Listserver (address = ? ), and just indicate "unsubscribe" in the subject heading. To find the address, go to the MSA Listserver Web address, and it should be possible to find the e-mail address for the MSA Listserver. Sorry, I don't recall it off-hand. Good luck with unsubscribing. Nelson Conti P.S. When in doubt - you can also re-send your message, but direct it to Nestor Zaluzec. I'm sure that he can tell you precisely how to unsubscribe.
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Hello everyone; I have just started looking at a GFP-transformed fungus growing on plant surfaces, using regular fluorescence microscopy. I can clearly see the fungus using two filter sets: a UV set (ex/em 365/} 420) and a blue filter set (ex/em 470/} 515). While the fluorescence in blue light is much brighter, stuff that is blue/blue-white with UV illumination is green in the blue light. is the "extra" green in the blue light likely to be where the GFP is not as concentrated, and hence doesn't show up in the UV, or is it more likely to be some other compound that has similar excitation/emission characteristics? I have done a fair bit of reading on the subject, but am still feeling a bit vague, and would really appreciate hearing from those of you who have some first hand experience with GFP.
thanks in advance shea
Dr. S.Shea Miller Agriculture & AgriFood Canada Eastern Cereal and Oilseed Research Centre Rm. 2068 Neatby Building Central Experimental Farm Ottawa, Ontario Canada K1A 0C6 Phone: (613) 759-1760 Fax: (613) 759-1701 E-mail: millers-at-em.agr.ca
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Hello everyone; I have just started looking at a GFP-transformed fungus growing on plant surfaces, using regular fluorescence microscopy. I can clearly see the fungus using two filter sets: a UV set (ex/em 365/} 420) and a blue filter set (ex/em 470/} 515). While the fluorescence in blue light is much brighter, stuff that is blue/blue-white with UV illumination is green in the blue light. is the "extra" green in the blue light likely to be where the GFP is not as concentrated, and hence doesn't show up in the UV, or is it more likely to be some other compound that has similar excitation/emission characteristics? I have done a fair bit of reading on the subject, but am still feeling a bit vague, and would really appreciate hearing from those of you who have some first hand experience with GFP.
thanks in advance shea
Dr. S.Shea Miller Agriculture & AgriFood Canada Eastern Cereal and Oilseed Research Centre Rm. 2068 Neatby Building Central Experimental Farm Ottawa, Ontario Canada K1A 0C6 Phone: (613) 759-1760 Fax: (613) 759-1701 E-mail: millers-at-em.agr.ca
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Hi all, We would like to add an individualized computer password system to our FEI/Philips CM-10 TEM. We would like a user to be able to type in their password which would then trigger a switch tied into the high tension and turn the HT on. The system would ideally then have a timer to record "on" time and a function to record this time with the user ID. Connecting a timer to the high tension is not a problem...the computer password part is where we need help. Please contact me if you have a system such as this on a FEI/Philips CM-10 or even on another make TEM which might work on a CM-10. Thanks, 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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On Tue, 1 Aug 2000 19:09:28 -0400, Gen Pei 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. 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 | | ****************************************************************** |
Nelson Conti 164 Ferne Court Palo Alto, CA 94306
Gen Pei- I've unsubscribed myself one time. I believe that you need to send an e-mail to the MSA Listserver (address = ? ), and just indicate "unsubscribe" in the subject heading. To find the address, go to the MSA Listserver Web address, and it should be possible to find the e-mail address for the MSA Listserver. Sorry, I don't recall it off-hand. Good luck with unsubscribing. Nelson Conti P.S. When in doubt - you can also re-send your message, but direct it to Nestor Zaluzec. I'm sure that he can tell you precisely how to unsubscribe.
_______________________________________________________ Say Bye to Slow Internet! http://www.home.com/xinbox/signup.html
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I have not yet seen any effects, with the Apple Airport system installed for my computers as well as a GigaHertz wireless broadcasting systems which are carrying video signals from one room to another.
The Computer monitors however are a different story.
Nestor
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BOC Gases, has a unique opportunity for an Electron Microscopist/Materials Engineer located at it's state of the art Technical Center in Murray Hill, NJ.
BOC Gases, a division of The BOC Group Inc., was founded in 1888 and is a global leader in the manufacturing and distribution of industrial, medical, and specialty gases and related equipment. Poised for new growth opportunities, BOC currently operates in more than 50 countries, providing customers with gases and technology which are used to support life, freeze food, forge steel, shape glass, refine petroleum, create computer chips, and clean wastewater and smokestack emissions, in addition to hundreds of other applications.
Electron Microscopist/Materials Engineer
Responsibilities
The successful candidate will support the materials characterization needs of the Gases Technology R&D organization using and maintaining analytical equipment, such as, Scanning Electron Microscopy, X-ray Diffraction. Metallography. Characterization includes metallic surfaces, adsorption/catalysis materials as well as other materials related to gases delivery equipment and vessels. This position is within the analytical services part of the Materials Technology Group. More specifically, the person will have the responsibility to:
Provide scanning electron microscope (SEM), environmental scanning electron microscope (ESEM), and metallography services.
Maintain the SEM and ESEM equipment in good operating condition.
Develop new techniques to support researchers needs.
Perform failure analysis and provide technical assistance for material selection projects.
Experience/Education
A B.S. or M.S. in the field of materials science and engineering with practical working experience with scanning electronic microscope techniques. Experience in x-ray diffraction a plus.
A knowledge of a wide variety of materials. Experience with mechanical testing, failure mechanisms, corrosion, and materials, especially metals and polymers. 5-10 years work experience in a materials laboratory.
Work experience in the chemical processing industry a plus.
Ability to perform failure analysis.
Ability to multitask is essential.
Send application to: BOC Gases, Attn: Recruiting - SEM-NM, 575 Mountain Avenue, Murray Hill, NJ 07974. E-mail: jobs-at-us.gases.boc.com
fax: 908-771-1702. Along with opportunities for professional growth we offer competitive compensation and comprehensive benefits.
BOC Gases is an equal opportunity employer.
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Sorry for the previous blank message. Nestor thinks it was just a fluke.
We have an Astromed cooled CCD camera, which we would like to use. However, we have no manuals for it. If you have manuals or other information that would allow us to put this camera into operation please let me know "off line".
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Hi all, We would like to add an individualized computer password system to our FEI/Philips CM-10 TEM. We would like a user to be able to type in their password which would then trigger a switch tied into the high tension and turn the HT on. The system would ideally then have a timer to record "on" time and a function to record this time with the user ID. Connecting a timer to the high tension is not a problem...the computer password part is where we need help. Please contact me if you have a system such as this on a FEI/Philips CM-10 or even on another make TEM which might work on a CM-10. Thanks, 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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Sorry for the previous blank message. Nestor thinks it was just a fluke.
We have an Astromed cooled CCD camera, which we would like to use. However, we have no manuals for it. If you have manuals or other information that would allow us to put this camera into operation please let me know "off line".
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Well, I guess Mark and I would have to see a higher magnification image, detailing the polysilicon, the "birds beak" area, and the substrate. This way we could tell whether this is a gate, with gate oxide, or a diffusion contact. I am assuming that your cross section is well into the middle area of the polysilicon line, and not just at the edge.
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When I had resolution problems recently (which turned out to be due to a "leaky" detector) one of the first things my service engineer asked me to check was to make sure that the detector wasn't touching the pole piece (which can cause a ground loop?) Try backing the detector out a little to see if the peak width changes. I doubt it will help, but it is easy to try. Matt
"E. J. McKenzie" {elizm-at-pdx.edu} on 07/28/2000 12:30:59 PM
To: microscopy-at-sparc5.microscopy.com cc:
Hi everyone,
we are having serious problems with the resolution of our EDS detector - the peak height-width ratio is terrible, around 450 counts. Consequently, our peaks look more like rolling hills and one peak blurs into another. I have callibrated the machine using an Al-Cu standard, checked that the detector is kept cool, sample is tilted at 30 degrees, etc etc. I also got another microscopist to come and have a look to check my methods and he also concluded there was a serious problem.
Is there anything else I could try before I phone the Noran technicians and pay them a lot of $$$ to fix/replace the detector?
regards Liz McKenzie -------------------------------------------------- Geomicrobiology and Electron Microscopy Laboratory Room S9 Cramer Hall 1721 SW Broadway Portland State University Portland OR97201
ph:503 725 3362 fax:503 725 3025
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I'm trying to find references for vital staining with fluorobora stains (or fluorabora). There don't seem to be any, so I don't know if this is because they don't work, or because they just didn't catch on. Has anyone any experience of using them, and could anyone give me any references? Thanks.
Lesley Weston.
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Hi , i wanted to know if anyone knew of any SEM/EDS courses in Mexico or Texas. Thanks.
Rafael Peña.
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Well, I guess Mark and I would have to see a higher magnification image, detailing the polysilicon, the "birds beak" area, and the substrate. This way we could tell whether this is a gate, with gate oxide, or a diffusion contact. I am assuming that your cross section is well into the middle area of the polysilicon line, and not just at the edge.
Darrell
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Fellow microscopists,
Have any of you had any experience with the installation of a wireless lan affecting the operation of your microscopes in any way? I would very much appreciate your advice in this regard.
Sincerely,
Mick Thomas ------------------------------------------------- Mick Thomas UHV-STEM Laboratory E-1 Clark Hall Cornell University Ithaca, NY 14853
I am currently being tasked to bring my Amray 3200C SEM into compliance with ISO9000. A major issue with my supervision is the accuracy of the micron bar given with every picture - and what proceedure do I use for callibration. If you have any insight or suggestions, I would appreciate it. Thanks Gary Weaver Gary.Weaver-at-MW.Boeing.com
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Hi all, We would like to add an individualized computer password system to our FEI/Philips CM-10 TEM. We would like a user to be able to type in their password which would then trigger a switch tied into the high tension and turn the HT on. The system would ideally then have a timer to record "on" time and a function to record this time with the user ID. Connecting a timer to the high tension is not a problem...the computer password part is where we need help. Please contact me if you have a system such as this on a FEI/Philips CM-10 or even on another make TEM which might work on a CM-10. Thanks, 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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Hello everyone; I have just started looking at a GFP-transformed fungus growing on plant surfaces, using regular fluorescence microscopy. I can clearly see the fungus using two filter sets: a UV set (ex/em 365/} 420) and a blue filter set (ex/em 470/} 515). While the fluorescence in blue light is much brighter, stuff that is blue/blue-white with UV illumination is green in the blue light. is the "extra" green in the blue light likely to be where the GFP is not as concentrated, and hence doesn't show up in the UV, or is it more likely to be some other compound that has similar excitation/emission characteristics? I have done a fair bit of reading on the subject, but am still feeling a bit vague, and would really appreciate hearing from those of you who have some first hand experience with GFP.
thanks in advance shea
Dr. S.Shea Miller Agriculture & AgriFood Canada Eastern Cereal and Oilseed Research Centre Rm. 2068 Neatby Building Central Experimental Farm Ottawa, Ontario Canada K1A 0C6 Phone: (613) 759-1760 Fax: (613) 759-1701 E-mail: millers-at-em.agr.ca
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Date sent: Wed, 02 Aug 2000 11:20:54 -0400 } From: "Shea Miller" {millers-at-em.agr.ca} To: confocal-at-listserv.acsu.buffalo.edu, microscopy-at-sparc5.microscopy.com
I'm trying to find references for vital staining with fluorobora stains (or fluorabora). There don't seem to be any, so I don't know if this is because they don't work, or because they just didn't catch on. Has anyone any experience of using them, and could anyone give me any references? Thanks.
You will still need Nikon Part# 82014 (Optical Coupler 28mm to C-Mount) available only from your local Nikon Microscope Dealer.
Regards,
Lawrence Kordon Nikon, Inc. nikon-at-jagunet.com
"Purdy, Sam" 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 } -----------------------------------------------------------------------. } } Jonathon: } } Contact Diagnostic Instruments, www.diaginc.com or info-at-diaginc.com. } They manufacture couplings to connect cameras and microscopes. } } Sam Purdy } Staff Specialist } National Steel Corp.
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With regard to the posting I made on this subject, Chuck Garber sent me the following note wherein he raises a number of important points. Chuck is correct in all regards. The product we were using was not DURO. We have no experience with Duro. We have made specific, brand name, recommendations in the past and my note was to share our recent experiences regarding these products. The products we were using were not "garage level" products.
So, IF, and only IF, you are using a product solely because of a recommendation from our lab, we now make new recommendations.
Ron
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Hi Ron,
I guess I should give some kind of disclaimer statement first, that in no way am I suggesting that what you are saying it not correct. If this was your perceived experience, then I am sure it is correct and I am not questioning it.
And it is a fact that there are some garage level operations where people take from 5 gallon pales and "fill" the 1 oz plastic bottles and put on a label and put it into distribution, without any real testing of the integrity of their packaging or any of the other things you and I might think one should be doing to insure the integrity of their product.
But I can assure you, without hesitation, the cyanoacrylate that is made by Duro, which is indeed a consumer products company, but it is also "top in class" when it comes to this type of product, has been offered by us (e.g. SPI) for years and we have never ever had anyone every voice even the slight complaint about it. We also decided a long time ago that once opened, the shelf life of these products can be very short, and consequently, we decided to offer the product in the smaller 3g package.
I go through the trouble to point this out to you because I see things from a slightly different perspective possibly than you do:
1] For one thing, someone even at IBM, having to place an order with either of the two mentioned companies, given the cost of placing an order period of any size, what is the overhead going to be? But if someone was ordering the Duro product from someone like SPI, then it just gets added in with a bunch of other things and the real cost is almost nothing.
2] For someone in Mexico, the situation is far worse. You have a lot of influence on people because of your reputation. Your posting could make someone in Mexico City think that in order to get acceptable results, they have to place a direct order in the USA for one of these "special" cyanoacrylates and in situations with limited research budgets, it just seems to me like they have better ways to be spending their money.
So while the Duro product might not be any better than the two mentioned, the point is that because of the distribution and transportation costs, if people are led to believe that they really do need something that can not be procured from their traditional sources of purchasing, they end up wasting a lot of money unnecessarily.
Of course if you had any hard evidence that the Duro product was in some way inferior, that would surely be another story. But chemically, these are all the same identical materials, they all have the same identical MSDS information and the all perform identically. When ever I have heard of someone having problems it has been because they were using product from a previously opened package.
Feel free to use this information however you feel might be approriate, either in whole or in part. But I did want to share my views with you on your posting, which I hope you won't mind my having done.......
Chuck
Ron Anderson, IBM, Hopewell Jct., New York, USA. anderron-at-us.ibm.com
IBM Analytical Services; http://www.chips.ibm.co
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With regard to the posting I made on this subject, Chuck Garber sent me the following note wherein he raises a number of important points. Chuck is correct in all regards. The product we were using was not DURO. We have no experience with Duro. We have made specific, brand name, recommendations in the past and my note was to share our recent experiences regarding these products. The products we were using were not "garage level" products.
So, IF, and only IF, you are using a product solely because of a recommendation from our lab, we now make new recommendations.
Ron
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Hi Ron,
I guess I should give some kind of disclaimer statement first, that in no way am I suggesting that what you are saying it not correct. If this was your perceived experience, then I am sure it is correct and I am not questioning it.
And it is a fact that there are some garage level operations where people take from 5 gallon pales and "fill" the 1 oz plastic bottles and put on a label and put it into distribution, without any real testing of the integrity of their packaging or any of the other things you and I might think one should be doing to insure the integrity of their product.
But I can assure you, without hesitation, the cyanoacrylate that is made by Duro, which is indeed a consumer products company, but it is also "top in class" when it comes to this type of product, has been offered by us (e.g. SPI) for years and we have never ever had anyone every voice even the slight complaint about it. We also decided a long time ago that once opened, the shelf life of these products can be very short, and consequently, we decided to offer the product in the smaller 3g package.
I go through the trouble to point this out to you because I see things from a slightly different perspective possibly than you do:
1] For one thing, someone even at IBM, having to place an order with either of the two mentioned companies, given the cost of placing an order period of any size, what is the overhead going to be? But if someone was ordering the Duro product from someone like SPI, then it just gets added in with a bunch of other things and the real cost is almost nothing.
2] For someone in Mexico, the situation is far worse. You have a lot of influence on people because of your reputation. Your posting could make someone in Mexico City think that in order to get acceptable results, they have to place a direct order in the USA for one of these "special" cyanoacrylates and in situations with limited research budgets, it just seems to me like they have better ways to be spending their money.
So while the Duro product might not be any better than the two mentioned, the point is that because of the distribution and transportation costs, if people are led to believe that they really do need something that can not be procured from their traditional sources of purchasing, they end up wasting a lot of money unnecessarily.
Of course if you had any hard evidence that the Duro product was in some way inferior, that would surely be another story. But chemically, these are all the same identical materials, they all have the same identical MSDS information and the all perform identically. When ever I have heard of someone having problems it has been because they were using product from a previously opened package.
Feel free to use this information however you feel might be approriate, either in whole or in part. But I did want to share my views with you on your posting, which I hope you won't mind my having done.......
Chuck
Ron Anderson, IBM, Hopewell Jct., New York, USA. anderron-at-us.ibm.com
I have been trying to study the ultra-structure (TEM) of strach granules (in wheat and maize seeds) that contains amorphus and crystaline layers. Unfortunately, I am unable to look at these layers under TEM even after keeping the tissues longer time in fixative (over night in 3%gluteraldehyde) and embedding medium (LR white/Spurs for 2 days).
I also tried the the above procedure with isolated starch granules in low melted agar but the result is the same - poor penetration of embedding resin as a result I am getting broken starch granules without any ultrastructural details.
I would appreciate it if anybody could suggest me how to obtaine a reasonably good ultra-structural (TEM) details of starch granules.
With thanks,
Siddique
Get Your Private, Free E-mail from MSN Hotmail at {http://www.hotmail.com/} http://www.hotmail.com
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Below is the program for the Facility Management session at M&M 2000. I am trying to record the sessions so we can summarize as much of the discussion as possible and make it available through the list for those of you who cannot attend. For those of you that can attend, please bring any informationin the form of handouts that you feel helpful in discussion of the topics. I would also appreciate this information sent to me as an attachment so that it can be added to the final summation of the session. Hope to see many of you in Philadelphia. Debby Sherman
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
--------
Experts Session on Facility Management
Wednesday, Aug. 16 9:00AM to Noon Room 106
The following topics will each be briefly introduced by a presenter followed by an open discussion period.
*9:00am: Multi-user Facilities…Managing Users Keith Darling, Ph.D. Manager, Analytical Research Dept, Atofina Chemicals, Inc.
*9:45am: Justification of costs/cost recovery….Electronic bookkeeping and billing. Gregory W. Erdos, Ph.D. Ass’t Director, U. of Florida Biotechnology Program Scientific Director, Electron Microscopy Core Lab
Break
*10:40am: Equipment Maintenance: Vender service contracts vs. Independent providers vs. Insurance company/self-insure options Paul J. Wirtz President, Expense Reduction Analysts International
*Times approximate…Additional discussion starting at 11:30am.
by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id KAA03087 for dist-Microscopy; Fri, 4 Aug 2000 10:58:32 -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 KAA03084 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Fri, 4 Aug 2000 10:58:02 -0500 (CDT) Received: from mail.psych.uic.edu (mail.psych.uic.edu [128.248.41.50]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id KAA03076 for {microscopy-at-msa.microscopy.com} ; Fri, 4 Aug 2000 10:57:20 -0500 (CDT) Received: by mail.psych.uic.edu with Internet Mail Service (5.5.2650.21) id {QGD8M3N5} ; Fri, 4 Aug 2000 10:53:54 -0500 Message-ID: {B98B03BD7E44D411ACE300D0B773E8CA442456-at-mail.psych.uic.edu} "'Dick Burry'" {burry.1-at-osu.edu} , "'Don Theyken'" {dtheyken-at-mindspring.com} , "'Dr. A. J. Tousimis'" {trc-at-tousimis.com} , "'Dr. Ed Daniels'" {ewdaniels-at-earthlink.net} , "'Dr. Hector Caruncho'" {cmhector-at-usc.es} , "'Dr. Hsi-Yuan Yang'" {hyhy-at-ccms.ntu.edu.tw} , "'Dr. Jaci Sagen'" {jsagen-at-miamiproject.med.miami.edu} , "'Dr. Marietta Issidorides'" {mrissid-at-compulink.gr} , "'Dr. P. Chien'" {p-chien-at-nwu.edu} , "'Dr. Sookja Chung'" {skchung-at-hkucc.hku.hk} , "'John Kriho'" {jk-at-meadowsinfo.com} , "'John Robinson'" {robinson.21-at-osu.edu} , "'Kathy Wolken'" {wolken.1-at-osu.edu} , "'Laura Kriho'" {lkriho-at-ucar.edu} , "'Lily Yang'" {lsyang77-at-cm1.hinet.net} , "'Lisa Adler'" {horizonco-at-mindspring.com} , "'List Server'" {microscopy-at-sparc5.microscopy.com} , "'Murat Elcin'" {elcin-at-science.ankara.edu.tr} , "'Nicholas Kriho'" {idklaus1-at-uic.edu} , "'Ralph J. Kriho'" {rjkriho-at-hotmail.com}
} ---------- } From: Hasemeier, Cynthia } Sent: Friday, August 04, 2000 10:39 AM } To: beth bourland; Fran Layne; Ginny; Harry; JDB; Judy Sobiesk; Kathleen } McMullan; marksr; mikedeb; Pauline; Ron; Ted; Fitzgibbon, Genevieve; } Madison, Sybil; Manning, George; Montgomery, John; Kriho, Virginia; Mason, } Sally; Peterson, Jim; Rodriguez, Walter } Subject: Birdcage } } Subject: THE BIRDCAGE } } There once was a man named George Thomas, a pastor in a small New England } town. One Easter Sunday morning he came to the Church carrying a rusty, } bent, old birdcage, and set it by the pulpit. Several eyebrows were raised } and, as if in response, Pastor Thomas began to speak. "I was walking } through town yesterday when I saw a young boy coming toward me swinging } this bird cage. On the bottom of the cage were three little wild birds, } shivering with cold and fright. } } I stopped the lad and asked, 'What you got there son?' } } "Just some old birds," came the reply. } } "What are you gonna do with them?" I asked. } } "Take 'em home and have fun with 'em," he answered. I'm gonna tease 'em } and pull out their feathers to make 'em fight. I'm gonna have a real good } time." } } "But you'll get tired of those birds sooner or later. What will you do?" } } "Oh,! I got some cats," said the little boy. "They like birds. I'll take } 'em to them." } } The pastor was silent for a moment. "How much do you want for those birds, } son?" } } "Huh?!!! Why, you don't want them birds, mister. They're just plain old } field birds. They don't sing - they ain't even pretty!" } } "How much?" the pastor asked again. } } The boy sized up the pastor as if he were crazy and said, "$10?" The } pastor reached in his pocket and took out a ten-dollar bill. He placed it } in the boy's hand. In a flash, the boy was gone. } } The pastor picked up the cage and gently carried it to the end of the } alley where there was a tree and a grassy spot. Setting the cage down, he } opened the door, and by softly tapping the bars persuaded the birds out, } setting them free. Well, that explained the empty birdcage on the pulpit, } and then the pastor began to tell this story* } } One day Satan and Jesus were having a conversation. Satan had just come } from the Garden of Eden, and he was gloating and boasting. "Yes, sir, I } just caught the world full of people down there. Set me a trap, used bait } I knew they couldn't resist. Got 'em all!" } } "What are you going to do with them?" Jesus asked. } } Satan replied, "Oh, I'm gonna have fun! I'm gonna teach them how to marry } and divorce each other, how to hate and abuse each other, how to drink and } smoke and curse. I'm gonna teach them how to invent guns and bombs and } kill each other. I'm really gonna have fun!" } } "And what will you do when you get done with them?" Jesus asked. } } "Oh, I'll kill 'em," Satan glared proudly. } } "How much do you want for them?" Jesus asked. } } "Oh, you don't want those people. They ain't no good. Why, you'll take } them and they'll just hate you. They'll spit on you, curse you and kill } you! You don't want those people!!" } } "How much?" He asked again. } } Satan looked at Jesus and sneered, "All your tears, and all your blood." } } Jesus said, "DONE!" Then He paid the price. } } The pastor picked up the cage he opened the door and he walked from the } pulpit. Isn't it funny how simple it is for people to trash God and then } wonder why the world's going to hell. Isn't it funny how someone can say } "I believe in God" but still follow Satan (who, by the way, also } "believes" in God). } } Isn't it funny how you can send a thousand jokes through e-mail and they } spread like wildfire, but when you start sending messages regarding the } Lord, people think twice about sharing. Isn't it funny how when you go to } forward this message, you will not send it to many on your } address list because you're not sure what they believe, or what they will } think of you for sending it to them. Isn't it funny how I can be more } worried about what other people think of me than what God thinks of me? } } I pray, for everyone who sends this to their entire address book, they } will be blessed by God in a way special for them. } } } } } } } ------------------------------------------------------------ } Cindy Hasemeier, M.A. } IRB Coordinator, Department of Psychiatry } University of Illinois at Chicago } PI 328W, M/C 912 } 312.413.4563 } 312.413.4544 (fax) } }
I know Nestor wants to keep the gratuitous solicitations to a minimum, but VLSI Standards might have a valid, NIST Traceable, ISO compliant solution for you. If need be, I also have some contacts at Amray (KLA-Tencor) that might be able to assist you. Please feel free to contact me directly offline.
Regards -
Marc Helvey 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
-----Original Message----- } From: Weaver, Gary E [mailto:Gary.Weaver-at-MW.Boeing.com] Sent: Thursday, August 03, 2000 4:29 PM To: Microscopy-at-sparc5.microscopy.com
I am currently being tasked to bring my Amray 3200C SEM into compliance with ISO9000. A major issue with my supervision is the accuracy of the micron bar given with every picture - and what proceedure do I use for callibration. If you have any insight or suggestions, I would appreciate it. Thanks Gary Weaver Gary.Weaver-at-MW.Boeing.com
Again, thanks to all who have contributed ideas about this IC cross section. I think that I have it sorted out. Maybe 95% right (??). The following URL shows an annotated cross section image. The poly gate is in the central portion of the image above the substrate. I was not sure if lettering would fit or show up.
http://photoweb.net/icxsect3.htm
I have not been able to confirm or deny that the gate truly is poly. My new x-ray system won't be available for several months. It should be a Rontec UHV 133eV system. Some responses indicated that x-ray would work in analyzing the cross sectioned gate. Other responses said it would not. I guess that the proof is in the trying. I suppose that there is some limit in how small the gate is where x-ray will not work. This would be due to either confusion of the results due to x-ray dispersion in a small volume and/or limitations of spot size.
Again, thanks for the great responses. Stay tuned as more info is gained. Further work is getting under way for 0.6u EEPROM analysis. This construction uses TiW plugs.
Dear Listserver members, Sending the Bird Cage to the listserver was a mistake. I thought I was sending to a different listserver. I apologize to all. I will never do this again, but I still cannot believe the furor. Once again I apologize to all. VirginiaKriho
I picked up a big old Zeiss microscope. Care to help me understand what I have? Please send hints in private e-mail and I'll summarize later. My description is at:
I picked up a big old Zeiss microscope. Care to help me understand what I have? Please send hints in private e-mail and I'll summarize later. My description is at:
Your suggested process flow is a little different from all I am used to. First, what etches has this cross section seen? I would venture a guess that the gate is polysilicon, the layer that forms around the poly gate is a CVD (chemical vapor [or is that vapour] deposition) oxide. The next layer is a softer glass deposited and then re-flowed (or spun on). The next layer (marked poly Si), I would guess to be aluminum. The next layer (marked Al), I would guess to be CVD oxide, and the layer on top of that, (marked PSG) might be oxide or silicon nitride.
Mind you, these are all just guesses. In my small world, I have never seen aluminum deposited directly on polysilicon. (That does not mean that it is never done) Aluminum usually shows some texture in a decorated cross section. It is not usually a smooth surface. That level has the appearance I am accustomed to seeing on oxide layers (both gray tones and surface). Most processes define well regions, grow the field oxide, grow the gate oxide, put on the gates, implant source/drains (self aligned to the gates), and then build up from there. The interlevel dielectrics must be deposited, they cannot be grown, like the gate oxide and the field oxide.
I looked through my books, and found a couple with basic process descriptions:
Physical Design of CMOS Integrated Circuits using L-EDIT by John Uymura ISBN 0-534-94326-8 PWS Publishing Co. 20 Park Plaza Boston, MA 02116-4324
Microelectronic Processing An Introduction to the Manufacture of Integrated Circuits by W. Scot Ruska ISBN 0-07-054280-5 McGraw-Hill Book Company
[No financial connections with these books. I have taken several continuing education classes with John, and we have become friends from this long association]
This was a very good parable. Bill Armor, Laverne, OK ----- Original Message ----- } From: Kriho, Virginia {Vkriho-at-psych.uic.edu} To: 'Charles Meshul' {meshulc-at-ohsu.edu} ; 'Dave Yeomans' {yeomans-at-uic.edu} ; 'Dick Burry' {burry.1-at-osu.edu} ; 'Don Theyken' {dtheyken-at-mindspring.com} ; 'Dr. A. J. Tousimis' {trc-at-tousimis.com} ; 'Dr. Ed Daniels' {ewdaniels-at-earthlink.net} ; 'Dr. Hector Caruncho' {cmhector-at-usc.es} ; 'Dr. Hsi-Yuan Yang' {hyhy-at-ccms.ntu.edu.tw} ; 'Dr. Jaci Sagen' {jsagen-at-miamiproject.med.miami.edu} ; 'Dr. Marietta Issidorides' {mrissid-at-compulink.gr} ; 'Dr. P. Chien' {p-chien-at-nwu.edu} ; 'Dr. Sookja Chung' {skchung-at-hkucc.hku.hk} ; 'John Kriho' {jk-at-meadowsinfo.com} ; 'John Robinson' {robinson.21-at-osu.edu} ; 'Kathy Wolken' {wolken.1-at-osu.edu} ; 'Laura Kriho' {lkriho-at-ucar.edu} ; 'Lily Yang' {lsyang77-at-cm1.hinet.net} ; 'Lisa Adler' {horizonco-at-mindspring.com} ; 'List Server' {microscopy-at-sparc5.microscopy.com} ; 'Murat Elcin' {elcin-at-science.ankara.edu.tr} ; 'Nicholas Kriho' {idklaus1-at-uic.edu} ; 'Ralph J. Kriho' {rjkriho-at-hotmail. Sent: Friday, August 04, 2000 10:53 AM
Suprised some one from Polaroid or one of their distributors has not responded to this.
} I've always been told that the brown goop that is easy to get on your hands } while pulling apart the photo & negative of the Polaroid T55 film is a known } carcinogen and to wash your hands immediately upon contact.
According to Polaroid's MSDS (http://www.polaroid.com/service/msds/index.html) the developer in the pods of T55 is an "alkali".
If you look on page 4 of the MSDS you will see the following:
"Does not contain any chemicals listed as carcinogens by OSHA, IARC or NTP"
Guess you'll have to believe them or not.
I've gone to T53, will see how it compares.
The opinions expressed above are my own, blah blah blah yackity smackity.
michael shaffer 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 roughing pump which service our JSM6300 has began making a } rumbling noise which I might believe is bearings ... altho it could be } the pump's bearings or the motor's. Has anyone serviced this problem? } } tia and 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/ shAf, I once got a Welsh 1402 for free that was making a terrible knocking noise (also belt drive). When I rebuilt it , I found a alightly swollen phenolic vane that was sticking. A little sanding and a gasket kit and it was as good as new. An off-the-wall problem, but a possibility if the JEOL pump has other than metal vanes.
Ken Converse Quality Images third party SEM service Delta, PA
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 involved in several research projects, in two broad areas: (1) Structural characterization of viruses by cryo-EM and 3D reconstruction; (2) Study of virus assembly processes in vivo through freeze-substitution, thin sectioning and immuno-cytochemistry. Experience with one or more of these techniques would be advantageous. IMA is a well funded and rapidly expanding research institute which was established in 1995 to conduct basic and applied research in agrobiology. It is located in a modern, spacious and well-equipped building at the National University of Singapore campus. The EM lab presently has a 100kV TEM equipped with a Gatan cryo-stage and a cryo-SEM and a new 200kV LaB6 TEM dedicated to structural microscopy will be purchased shortly. 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
I will be attending M&M2000 and I am looking to visit labs and research centers that are utilizing SEM, ESEM and AFM in their studies in the fields of oil industry such as oil well cement, catalyst, corrosion and concrete. Any names, addresses will be highly appreciated. Thanks Mansour Al-Shafei Senior Lab Sci. Saudi Aramco Oil Company Lab Research & Development Center Analytical Sciences Division Advanced Instruments Unit Voice: 966-3-876-4360 E-mail: shafeima-at-aramco.com.sa
I have been asked to help a colleague with a problem he is having cutting good sections for light microscopy. He has a Reichert-Jung 2030 microtome and a new tungsten carbide microtome knife (I do not know the exact type of knife). He is trying to cut ABS with a R scale value of 105 and polycarbonate with a M scale value of 70 (R-value of ~120 according to him).
Every section he has made exhibits numerous scratch marks that are also evident in the block face. I briefly tried to face the block and also got scratch marks. I cleaned the knife edge, tried different cutting angles and speeds (manual only). I have cut this material to make good sections and block face but I used an ultramicrotome and diamond knife. But to do this for all his work is not an option.
Any suggestions as to how the sections and the block face can be improved?
Thanks so much in advance,
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
Does anyone know why freshly made lead citrate sometimes comes out yellow, and what adverse effects, if any, result? In going over past threads on the subject, it seems most people have a relatively colorless product as the end result, but a few report the yellow color as normal. After years of making up "colorless" lead citrate, mine is now yellow. I've toyed with NaOH sources and pH; I use ultra pure water, I've cleaned glasswear 'till it squeaks...no change....it's still yellow.
Sections seem to stain okay, although the stain goes bad very quickly, sometimes even on the grid itself. Since this can be a problem with colorless lead citrate, I'm not sure the yellowing is the cause. I'd sure appreciate any feedback!
Thanks, Margaret -- Margaret Dienelt Plant Pathologist
Electron Microscopy Lab FNPRU, National Arboretum ARS, USDA
B. 010A R. 238 BARC-W 10300 Baltimore Avenue Beltsville, MD. 20705
I agree with Darrell 100%. He has identified all the layers as I see them. The aluminum always show some grain boundaries after staining the section with a BOE as mentioned that you have used on these sections. Also, the aluminum would not be deposited on top of the polysilicon.
Mark
} ---------- } From: "milesd-at-us.ibm.com"-at-sparc5.microscopy.com } Sent: Friday, August 4, 2000 3:50 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: IC Structure } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Gary, } } Your suggested process flow is a little different from all I am used to. } First, what etches has this cross section seen? I would venture a } guess that the gate is polysilicon, the layer that forms around the poly } gate is a CVD (chemical vapor [or is that vapour] deposition) oxide. } The next layer is a softer glass deposited and then re-flowed (or } spun on). The next layer (marked poly Si), I would guess to be } aluminum. The next layer (marked Al), I would guess to be CVD } oxide, and the layer on top of that, (marked PSG) might be oxide or } silicon nitride. } } Mind you, these are all just guesses. In my small world, I have never } seen aluminum deposited directly on polysilicon. (That does not } mean that it is never done) Aluminum usually shows some texture in } a decorated cross section. It is not usually a smooth surface. That } level has the appearance I am accustomed to seeing on oxide } layers (both gray tones and surface). Most processes define well } regions, grow the field oxide, grow the gate oxide, put on the gates, } implant source/drains (self aligned to the gates), and then build up } from there. The interlevel dielectrics must be deposited, they cannot } be grown, like the gate oxide and the field oxide. } } I looked through my books, and found a couple with basic process } descriptions: } } Physical Design of CMOS Integrated Circuits } using L-EDIT } by John Uymura } ISBN 0-534-94326-8 } PWS Publishing Co. } 20 Park Plaza } Boston, MA 02116-4324 } } Microelectronic Processing } An Introduction to the Manufacture of Integrated } Circuits } by W. Scot Ruska } ISBN 0-07-054280-5 } McGraw-Hill Book Company } } [No financial connections with these books. I have taken } several continuing education classes with John, and we } have become friends from this long association] } } Darrell }
Specimen Preparation Engineer Northwestern University
The Electron Probe Instrumentation Center (EPIC) at Northwestern University has an immediate opening for a specimen preparation expert. EPIC is a part of the world renowned Materials Research Center (MRC) and the Department of Materials Science & Engineering at Northwestern. The EPIC facility serves over 120 users in all aspects of Scanning and Transmission Electron Microscopy. The role of the specimen preparation engineer is to assist users with their specimen preparation needs, including instruction in TEM and SEM sample preparation using IBT, FIB, PIPS, electropolishing, ultramicrotomy, cutting/grinding/polishing, vacuum evaporation etc. All microscopes in EPIC are under full service contract. Thus, the duties include training students/users, development of specialized techniques and applications, minor maintenance, record keeping and billing. A BS or technical degree in physical/biological sciences is required. The candidate must have hands-on experience in all aspects of specimen preparation as well as considerable familiarity with digital acquisition, processing and computer assisted techniques. All levels of experience will be considered. Compensation will be commensurate with experience and qualifications. Send cover letter, resume and three references to: Prof. Vinayak P. Dravid, Director EPIC Materials Science & Engineering Northwestern University, 3013A MLSB Evanston, IL 60208 E-mail: v-dravid-at-northwestern.edu Fax: (847) 491-7820 http://epic.ms.nwu.edu/epic/index.htm
Northwestern University is an Affirmative Action/Equal Opportunity Employer. Hiring is contingent upon eligibility to work in the United States.
I thank Jan for supplying the following information.
-----Original Message----- } From: Ringnalda, Jan [mailto:JRingnalda-at-FEICO.com] Sent: Monday, August 07, 2000 9:12 AM To: 'Ingber, Bruce F.'; Long, Jo; Piscopo, Irene Cc: Schaub, Daniel; Ringnalda, Jan; Rice, Trisha (CS); Booth, Steve; 'sherman-at-btny.purdue.edu'
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Folks, I requested information on a computer log-in system a week ago and received a number of replys. I am pursuing some of them including the program written by Tom Ickes which is not available at the moment but may become available soon. Also, I did install the freeware: CM Lock from FEI and it does work great as a means to control access to the TEM. However, it does not record users and use time for later billing. I am still investigating how we can record the log-ins for this purpose. What I did receive is below:
------------------------ FEI Co. (www.feico.com) has on their web site number of useful freeware software for Philips CM microscopes. One of them is "CM lock". The program runs on Win box and does what you asked for. Give them a try. I use is on my CM300 to define different levels of privileges for my users and disable microscope controls for off time.
Jerzy Gazda, Ph.D.
---------------
We have a rather good system here that does what you want and quite a lot more. It will handle several machines. It keeps track of account numbers and summaries billing information. The system was developed by Tom Ickes who was an employee here at the time. He has since left Lehigh but may be interested in setting up a system for you (for a fee, I imagine). The system is well designed in that it uses off the shelf parts, is controlled by a PC and has software that has been very well constructed.
His e-mail is: tomickes-at-cswebmail.com
Good luck, Alwyn Eades
---------------------
Debby,
I might suggest that you add a digital I/O card that will supply analog outputs from the computer used to run the TEM. These outputs could then be used for a type of interlock for turning on the HT. I would not suggest switching the HT itself. Once this was in place, with software like Visual Basic or National Instruments LabWindows or LabView you can write a routine to control the HT, mark time, and write user info to a spread sheet. I might suggest you contact National Instruments Http://www.natinst.com as they can supply all you need for this project. On another thought, if your controlling computer is a PC and can run Windows NT the password issue is much easier since it has controlled log on features built in. Not a specific answer but hope this is useful to you.
Roy Beavers
--------------
Dear Debby,
I have recently contacted Richard Benassi at BEI in Maryland. He makes the exact control mechanisms/software you are talking about. I think he's a bit pricey ($1,000 for 2 control boxes w/software), but that's just my opinion...
I too am looking to control & monitor microscopes, so if you come across something else would you be so kind as to pass the information on to me?
Thanks and good luck,
Jim
--------
If I come up with additional information, I'll send it on. 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
by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id LAA09944 for dist-Microscopy; Mon, 7 Aug 2000 11:44:50 -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 LAA09940 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Mon, 7 Aug 2000 11:44:20 -0500 (CDT) Received: from lulu.it.northwestern.edu (lulu.acns.nwu.edu [129.105.16.54]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id LAA09928 for {Microscopy-at-sparc5.microscopy.com} ; Mon, 7 Aug 2000 11:44:09 -0500 (CDT) Received: (from mailnull-at-localhost) by lulu.it.northwestern.edu (8.8.7/8.8.7) id LAA01115 for {Microscopy-at-sparc5.microscopy.com} ; Mon, 7 Aug 2000 11:38:30 -0500 (CDT) Received: from paiute (paiute.ms.nwu.edu [129.105.37.230]) by lulu.acns.nwu.edu via smap (V2.0) id xma000203; Mon, 7 Aug 00 11:36:49 -0500 Message-ID: {007701c0008d$95991440$e6256981-at-ms.northwestern.edu}
Specimen Preparation Engineer Northwestern University
The Electron Probe Instrumentation Center (EPIC) at Northwestern University has an immediate opening for a specimen preparation expert. EPIC is a part of the world renowned Materials Research Center (MRC) and the Department of Materials Science & Engineering at Northwestern. The EPIC facility serves over 120 users in all aspects of Scanning and Transmission Electron Microscopy. The role of the specimen preparation engineer is to assist users with their specimen preparation needs, including instruction in TEM and SEM sample preparation using IBT, FIB, PIPS, electropolishing, ultramicrotomy, cutting/grinding/polishing, vacuum evaporation etc. All microscopes in EPIC are under full service contract. Thus, the duties include training students/users, development of specialized techniques and applications, minor maintenance, record keeping and billing. A BS or technical degree in physical/biological sciences is required. The candidate must have hands-on experience in all aspects of specimen preparation as well as considerable familiarity with digital acquisition, processing and computer assisted techniques. All levels of experience will be considered. Compensation will be commensurate with experience and qualifications. Send cover letter, resume and three references to: Prof. Vinayak P. Dravid, Director EPIC Materials Science & Engineering Northwestern University, 3013A MLSB Evanston, IL 60208 E-mail: v-dravid-at-northwestern.edu Fax: (847) 491-7820 http://epic.ms.nwu.edu/epic/index.htm
Northwestern University is an Affirmative Action/Equal Opportunity Employer. Hiring is contingent upon eligibility to work in the United States.
Hi All, I was wondering if anyone had experience or comments on how much oxygen can safely be pumped before one has to opt for fomblin type oil in rotary pumps. Does the same limit apply to turbo pump oil, and do the pumps need to be completely cleaned for the changeover? I believe I've heard that it is unwise to try to clean a pump and reuse it, with the change of oil? Also, are there any less expensive sources for oils that can be safely used when pumping oxygen. Any comments, suggestions, or hints would be welcome.
Many Thanks,
Steve Buckingham Excellatron Solid State (770) 438 2201 All the Best,
Steve. Kratos Analytical Inc. (770) 251 6490 "Fill your bowl to the brim, and it will spill. Keep sharpening your knife, and it will blunt. Chase after money and security, and your heart will never unclench. Care about people's approval, and you will be their prisoner. Do your work, then step back... The only path to serenity." Lao-tze. Tao te Ching
Does anyone use TEM electron diffraction patterns for identifying silica in soil samples? I know that X-ray diffraction is a suitable technique, but I have been asked to find an alternative method such as TEM. If this is possible what would the sample prep entail?
Thanks in advance
Jean Howard
________________________________________________________________________ Get Your Private, Free E-mail from MSN Hotmail at http://www.hotmail.com
RMC was purchased by Ventana Medical Systems, 1-800-356-3452 or 1-800-277-4613ext3023. Greg Carter is the ultramicrotome service manager. I had him visit my lab to service our MT2B in December, 1999. Let me know if this does not work.
Sincerely, Ken
On Mon, 7 Aug 2000, Geoff Williams 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 was just informed by our Purchasing department that their RMC contact } #s no longer work. } } Is there anyone out there that can point me to the direction of the } company that might or might not still be RMC that can provide parts for } our MT2B? } } Please excuse me if this answer is common knowledge, I just can't seem } to find the information. } } TIA, } Geoff Williams } } Electron Microscope Facility Supervisor } Biology Department } Brooks Hall } Central Michigan University } Mt. Pleasant, MI 48859 } } Geoffrey.Lloyd.Williams-at-cmich.edu } 517 774-3576 } 517 774-3462 (fax) } } } }
You might also try getting your MT2B (I thought I was the only one still using one!) from Bill McGee at Microtome Services Company - 315-451-1404. He is a great guy to deal with, very reasonably priced, and has provided me with excellent service for a long time.
} } } Margaret Brannigan {brannign-at-asrr.arsusda.gov} 08/07/00 14:59 PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi,
Does anyone know why freshly made lead citrate sometimes comes out yellow, and what adverse effects, if any, result? In going over past threads on the subject, it seems most people have a relatively colorless product as the end result, but a few report the yellow color as normal. After years of making up "colorless" lead citrate, mine is now yellow. I've toyed with NaOH sources and pH; I use ultra pure water, I've cleaned glasswear 'till it squeaks...no change....it's still yellow.
Sections seem to stain okay, although the stain goes bad very quickly, sometimes even on the grid itself. Since this can be a problem with colorless lead citrate, I'm not sure the yellowing is the cause. I'd sure appreciate any feedback!
Thanks, Margaret -- Margaret Dienelt Plant Pathologist
Electron Microscopy Lab FNPRU, National Arboretum ARS, USDA
B. 010A R. 238 BARC-W 10300 Baltimore Avenue Beltsville, MD. 20705
Telephone: (301) 504-6097 Fax: (301) 504-5096
Margaret,
Lead citrate turning yellow is a new one on me. I've never heard of that peoblem. When I make lead citrate, I use a 100ml bottle that previously was used for HBSS ( I find they work better than a volumetric flask, easier to use), put the bottle on a magnetic sirrer, add lead citrate, close the bottle, let it stir for a while then had concentrated NaOH ( Fisher catalog # ss277) let it stir until solution clears, put it in refrigerator overnight (keeping it stored in refrig. at all times until use). Never turned yellow on me. I've made up my stain this way for almost 30 years and it never turned yellow and I've been able to use the stain for several months. If you have any questions, call me.
Phil Rutledge USDA/ARS voice: 410 788-4136 or 2120 e-mail:prutledge-at-ars.usda.gov fax: 410 788-4399
Yes, Although we do no do with soil sample we do identify particles from different kinds of oxides on the metal surface. With support of holly carbon film electron diffraction can be obtained from silica if the size of silica particles is around, say 100 nm or less. We have obtained electron pattern from extracted silica.
yours Sincerelly
Ping Liu
Listers:
Does anyone use TEM electron diffraction patterns for identifying silica in soil samples? I know that X-ray diffraction is a suitable technique, but I have been asked to find an alternative method such as TEM. If this is possible what would the sample prep entail?
I have a constant problem of regularly spaced wrinkles on my Spurr sections on the TEM. I stain with uranyl acetate in 50% ethanol for about 30 minutes followed by stable lead for 15 minutes.(Wrinkling is even worse with methanolic UA). I am certain the problem has something to do with staining since few wrinkles appear when I observe unstained sections. I also rarely encounter wrinkled sections with Embed 812 sections when using aqueous UA stains. Any suggestions?
} Does anyone use TEM electron diffraction patterns for identifying silica in } soil samples? I know that X-ray diffraction is a suitable technique, but I } have been asked to find an alternative method such as TEM. If this is } possible what would the sample prep entail? }
Dear Jean, I have taken selected-area ED (SAED) patterns of minerals in soil samples, but I have not yet attempted to get a structure. Of course, identi- fying a silicate of known structure from one or more zone-axis patterns is a simpler problem. The sample preparation is very easy--I just suspend the soil in water, dilute to the appropriate degree so that the soil particles
are well dispersed, put a few microliters on a carbon-formvar grid, and air-dry. When you find a particle of interest, you may have to orient the grid to get a zone axis, so you need a double-tilt or tilt-rotation stage. Once you have the proper orientation, you can ascertain the crystal form and measure the unit cell parameters and intensities. If you have an image simulation program, you can calculate the predicted intensities from a trial structure (which matches the unit cell parameters) and see what kind of agreement you get. This should work even if there is some dynamical scattering. You could try an ab initio structure determination if the cry- stal is thin enough and the voltage high enough so that dynamical scat- tering is not significant, but this is much more difficult and requires much more data. See Doug Dorset's book, Structural Electron Crystal- lography for a good overview. Good luck. Yours, Bill Tivol
Thanks for the positive feed-back and the chance for discussion.
1. If you notice the figure is labeled seed. This seed is a co-polymer of BA-B. The reason for the difference in shadow length even after cross linking with OsO4 is the heterogeneous composition. I think this is due to the polymerization rates of the two monomers. Butadiene polymerizes faster than BA so it will form particles first and the BA follows. This was then to be the seed in a core-shell latex. If you look at the third figure on the web page you will see the final latex.
2. You are right, at just the right orientation the core should look centered, but the chance of that happening frequently is small. However, if you look at the group of three particle around 11:00 you will see the middle particle having a fairly centered core.
If you have any other questions please e-mail me. Discussion is good.
Olga Shaffer
"Gerroir, Paul J" wrote:
} Hi Olga, } An impressive start to your web page! The polymer chemists/engineers here } will find these images very exciting. } I have two comments/questions. } 1. In your first example of TEM - Staining you have illustrated that a "very } short shadow indicating very little PB" It is not clear in this example of } shadowing and staining whether the system, PB/PBA is a copolymer or as I } suspect the seed referred to is PBA and the shadowing gives an indication of } the amount of shell, i.e. PB associated with the seed or core. } 2. In your second example of TEM - Staining you have pointed out the } off-center nature of the PS core. If we think of these images in two } dimensions and the latex spheres being randomly oriented why don't we see } any particles where the core appears to be more centered? If you were to } view these two-dimensional spheres from the left or right in your images } wouldn't a picture of a more centralized core result? Make a simple } three-dimensional model and try this experiment, then perhaps you can offer } me an explanation. Do you think it a little puzzling that these particles } all orient similarly? } } Regards, } Paul } } Paul J. Gerroir } Microscopy } Materials Characterization } Xerox Research Centre of Canada } 2660 Speakman Drive } Mississauga, Ontario L5K 2L1 } } Phone: (905) 823-7091, ext. 216 } FAX: (905) 822-7022 } e-mail: paul.gerroir-at-crt.xerox.com
Yes TEM electron diffraction patterns for identifying silica in soil samples can be done. One quick way is to sonicate some of the sample in water maybe with some detergent added (do different dilutions because what looks like a little bit can be a lot in the TEM.) Then take an aliquot .5-1ul and drop it onto a carbon film/holey grid. (we used to collapse our own MCE filters after lightly coating them with carbon) Allow the grid to dry, then place in scope. We used to start with the most dilute prep first and would get results off it.
good luck
} Listers:
} Does anyone use TEM electron diffraction patterns for identifying } silica in soil samples? I know that X-ray diffraction is a suitable } technique, but I have been asked to find an alternative method } such as TEM. If this is possible what would the sample prep } entail?
} Thanks in advance
} Jean Howard Jon Ekman Associate Research Specialist University of Wisconsin Milwaukee 414-229-6471
Dear Jean, I've done ED studies of soils from time to time. I have not done the sample preparation myself, but it's not too difficult. You need to disperse the soil in a liquid such as water or 2-propanol and let a drop of the suspension dry on a normal TEM grid. An ultrasonic bath is ideal for this. If the silica grains are substantially bigger than the clays, there may be fractionation, resulting in a reduced yield of silica as it can fall to the bottom of the suspension. Care is therefore needed in sampling. If the silica grains are big enough (one micron or more) you can use a double-tilt or rotation-tilt holder to align the crystallites to get a sensible zone pattern which will make interpretation easier. If the grains are v. small and you need to use a micro-diffraction technique you may find the tilting process drives you crazy as it's v. difficult to keep the grain in the beam during tilting.
Thr camera length of the microscope varies with specimen height, and specimen height varies with tilting in duoble-tilt holders. Generally, external calibration (e.g. camera length vs objective current) is not wholly satisfactory but in your case may be adequate. The projector astigmatism needs to be very well corrected if accurate d-spacings are to be obtained. A sure-fire way round this is to use an internal standard such as a poly- crystalline Al or Au film evaporated onto the carbon support film. This produces a ring pattern superimposed on the spot pattern, allowing the actual camera length in any direction to be measured directly. (Modern sputter coaters produce Au films where the grain size is too small to give sharp rings, so it has to be evaporation from a hot W wire.)
Not all silica in soils is crystalline, though, and my preferred method is to do TEM-EDX, which can resolve very small areas as the limiting factor is the minimum probe size that still gives a reasonable number of counts. It's also a lot quicker and easier.
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
A sophisticated method of soil sample analysis by high resolution TEM is thin sectioning. That requires complete sample dehydration in 100 % EtOH, gradual infiltration with LR White, (representative) suspension sampling followed by embedding in gel capsules, curing, and thin sectioning on a microtome. I recommend to collect sections on Cu / carbon lacey grids. No need to osmicate or post-stain unless you're interested in soil organisms.
Alice Dohnalkova Environmental Microbiology Battelle, PNNL MS P7-50 Richland, WA 99352 tel. (509) 372-0692 fax (509) 376-1321
-----Original Message----- } From: Jean Howard [SMTP:jmhowrd-at-hotmail.com] Sent: Monday, August 07, 2000 8:46 PM To: Microscopy-at-sparc5.microscopy.com
Listers:
Does anyone use TEM electron diffraction patterns for identifying silica in soil samples? I know that X-ray diffraction is a suitable technique, but I have been asked to find an alternative method such as TEM. If this is possible what would the sample prep entail?
Thanks in advance
Jean Howard
________________________________________________________________________ Get Your Private, Free E-mail from MSN Hotmail at http://www.hotmail.com
Excuse me folks, but isnt everyone putting the carriage before the horse? Dont you think that as scientists you need to be a bit less eager and a bit more conscientious to tell Jean Howard yes when there are more important questions to be answered?
Jean; why do you have to find an alternative method? Do you not have a diffractometer and a competent XRD person on staff? If not, why bother getting into a can of worms? Are you to analyze the soils for the regulatory crystalline silica content of 0.1% by weight for hazmat, product labelling? Why analyze it by a method that is not accepted? XRD is the accepted method for analyzing air samples for the respirable fraction by NIOSH. Almost all competent scientists that I know of analyze bulk materials with several modifications depending on the sample.
If you go and do it by TEM, you could be putting yourself in a dangerous liability situation causing question of your capabilities as a scientist. Doing it by TEM is frought with problems, and how do you propose to accurately and precisely quantify it at the 0.1% by weight level?.......you cant do it, at least not reproducibly.
The XRD method is the only way to do it if you want as close to real numbers as you can get. There are plenty of labs that will analyze it wrong for $50 a pop by XRD. I know.....I have been doing good analyses for 10 yrs, and it takes a hell of a lot more than $50 worth of work. You need to find out what the sample is composed of first, then remove components gravimetrically, with acid dissolutions or thermal treatments to render phyllosilicates amorphous. Most micas and clays have interfering peaks at the primary, secondary and tertiary quartz peaks. How do you know you arent analyzing beta or quasi metastable polymorphs of tridymite and cristobalite? You dont.
I can recommend some top notch people to verify what I have said, and who can do it by XRD better than most. Let me know. I am disappointed at how quickly so many people are responding to say it can be done by TEM without really thinking about the problems that need to be addressed first. Good way to chop your reputation up.
Lou Solebello
----- Original Message ----- } From: Ping Liu {ping.liu-at-sandvik.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, August 08, 2000 5:50 AM
Fellow Microscopists,
I am looking for 2 reliable, "clean" polyclonal (preferred) or monoclonal antibodies for use with aldehyde/detergent-treated or ethanol-treated cultured cells (HeLa). One antibody would be against vimentin, and the other against a protein that allows clear visualization of mitochondria. Any suggestions? Thank you in advance. 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
Have you tried aqueous stains for the Spurr's? I've used aq. UA/Pb and had nice staining. If you have some stain-resistant plant material, you can use KMnO4 instead of UA - it stains well.
Tamara Howard CSHL
On Tue, 8 Aug 2000, Anthony Greco 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 constant problem of regularly spaced wrinkles on my Spurr } sections on the TEM. I stain with uranyl acetate in 50% ethanol for } about 30 minutes followed by stable lead for 15 minutes.(Wrinkling is } even worse with methanolic UA). I am certain the problem has something } to do with staining since few wrinkles appear when I observe unstained } sections. I also rarely encounter wrinkled sections with Embed 812 } sections when using aqueous UA stains. Any suggestions? } } Tony Greco } } }
Hello all, Does anybody have any experience in getting 50 kV Power Supplies for Amray 1800 series electron microscopes fixed. The power supply was manufactured by CPS, a company now out of Oregon. Approximately two weeks ago we fedex'd them our supply for repair (which cost an arm and a leg), and after all this time, they're still can't even give me an approximate date when it will be completed. They haven't even diagnosed it yet. Obviously we're dead in the water with out it, and work is piling up. Does anybody know of any other options. Possibly other shops that can repair these supplies, or possibly another source for a different supply. I'm so frustrated at the fact that they didn't look at it for a week, and the fact that they have no clue when it will be done. I of all people can understand being busy, but I can't live another two weeks with the e-scope down for the count. If anybody has any ideas that would help to restore my sanity, or get my supply fixed, I would be forever indebted.
Thanks in advance, ~Jonathan Dunlap
Jonathan Dunlap Analytical Laboratory Manager Osram Sylvania Inc. 816 Lexington Avenue Warren, PA 16365 Ph: 814-726-6991 Fax: 814-726-6956
I have an aging (~15years old) Balzers Critical Point Dryer that has a burnt out heater. The dryer is type 11120 B no. 337. The heater is a plastic strip 20 cm long and 3.5cm wide which seems to have carbon or graphite laminated into the strip and connecting wires are soldered onto each end. This wraps around the chamber of the unit and was held there by heat shrink wrap. Does anyone know if you can get replacements for this heater ( in Australia would help!)? I am waiting to hear from Balzers but need to get it fixed quickly.
Regards 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
I have worked on their 30KV units in my Amray scopes. The problem is either the driver transistor or the HV generator block.
I have the schematics for the 30KV CPS unit. The HV block was about $250. The transistor was $7 from Radio Shack. I zapped the unit when the stub made hard contact with the pole piece in my 1600T. The CPS units in my 1830 and 1910 have never failed.
I found CPS to be quite helpful. I guess that this may have changed.
gary g.
At 03:25 PM 8/8/00, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
First of all, let me clarify what is being requested. The lab is interested in the regulatory crystalline silica content in soils. Currently the samples are being contracted out for XRD because there is no diffractomer or XRD scientist in house. Their results are reliable and NIOSH acceptable.
The question was whether the TEM (which is in lab) could be used for this analysis instead of purchasing a diffractometer. Let me say that I have no experience with minerals by TEM. My work has been primarily with polished metal samples. I had some doubts about using TEM for this type of work since I was not successful in finding literature on such.
I appreciate all the responses and apologize for not being more specific. No one's competency is in question here. I have received numerous suggestions but it is clear to me that the most reliable method is XRD. Should I need to speak to someone further I will not hesitate to contact you Lou.
Thank you all
Jean Howard ________________________________________________________________________ Get Your Private, Free E-mail from MSN Hotmail at http://www.hotmail.com
Provided you can obtain a binary or thresholded grain boundary image, there are a number of general image analysis programs available - free of charge - that will automatically extract the grain size. Try: * "ImageJ" http://rsb.info.nih.gov/ij/index.html * "Scion Image" http://www.scioncorp.com/frames/fr_scion_products.htm * "Image Tool" http://macorb.uthscsa.edu/dig/itdesc.html
Cheers, Paul Baggethun
} ---------- } From: Maria Luiza[SMTP:luiza-at-ppgem.faenquil.br] } Sent: Monday, July 31, 2000 7:31 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Grain size....... } } Hi. I am a studant from Brazil and I´m trying to measure the grain size of } my samples (They are Niobium 9%Tantalun). } I know there are a lot of softwares available on the internet, but I don´t } know where and if there is a specific program to analise the grain size of } metals. } Please, remember that I work at Brazil, so I don´t have any money buy a } software. We allways try to do the best we can. } } Thank you very much..... João Paulo Barros Machado..... } } Please answer this email for: jpbm-at-easygold.com.br } } }
If you are making a last minute decision as to which conferences to attend this summer/fall then take a look at ICHC 2000 "Cell Biology and Imaging Tools for the New Century". 3-8th Sept. 2000 York, UK
Take look at the fantastic Final Programme at http://www.med.ic.ac.uk/external/ichc_2000
Over 100 leading speakers in 35 symposia
York is a most beautiful medieval walled city, a great place for a vacation. You can register for the week or if close to York then come for the day!
Hello everyone; now that the corn has finally started to do something in this miserable excuse for a summer we are having, we have a grad student who would like to do some peroxidase histochemistry on a rather daunting number of samples and varieties. is there any fixation he can use to preserve both his samples and his enzyme activity? he will not be able to look at everything in a reasonable length of time. thanks, as always, in advance shea
Dr. S.Shea Miller Agriculture & AgriFood Canada Eastern Cereal and Oilseed Research Centre Rm. 2068 Neatby Building Central Experimental Farm Ottawa, Ontario Canada K1A 0C6 Phone: (613) 759-1760 Fax: (613) 759-1701 E-mail: millers-at-em.agr.ca
I have a student that would like to do TEM of diamond films that are prepared by microwave plasma CVD. The thickness of the films can be adjusted to make it electron beam transparent. The student believes that he can flake off pieces of the film.
I'm thinking maybe we could just lay the diamond film on a grid and pop it into the TEM. Is this a crazy idea? Does anyone have any suggestions?
} Excuse me folks, but isnt everyone putting the carriage before the horse? } Dont you think that as scientists you need to be a bit less eager and a bit } more conscientious to tell Jean Howard yes when there are more important } questions to be answered? }
I guess that those of us who answered thought of the question from an academic/technical viewpoint--true, at least, in my case. Yes, if there are legal questions about the methods, that is a consideration I did not think about.
} } Jean; why do you have to find an alternative method? Do you not have a } diffractometer and a competent XRD person on staff? If not, why bother } getting into a can of worms?
To characterize tens to hundreds of individual soil particles that are in the micrometer size range can be done in a few (long) sessions on the TEM. This includes EDS, ED, and imaging. I don't think that the same info can be obtained in as short a time by XRD.
} Are you to analyze the soils for the regulatory } crystalline silica content of 0.1% by weight for hazmat, product labelling?
I hadn't considered this aspect.
} } Why analyze it by a method that is not accepted? XRD is the accepted } method for analyzing air samples for the respirable fraction by NIOSH. } Almost all competent scientists that I know of analyze bulk materials with } several modifications depending on the sample.
However, to analyse many individual particles and distinguish between, e.g., sodium aluminum silicate and potassium aluminum silicate is not best done by XRD.
} } } If you go and do it by TEM, you could be putting yourself in a dangerous } liability situation causing question of your capabilities as a scientist.
As a scientist, or as a lawyer?
} } Doing it by TEM is frought with problems, and how do you propose to } accurately and precisely quantify it at the 0.1% by weight level?.......you } cant do it, at least not reproducibly.
To get the distribution of various silicates to a precision of 0.1%
would, indeed, require analysis of many thousand (million?) particles. Of course, one could, nonetheless, get such precision given enough time.
} } } The XRD method is the only way to do it if you want as close to real numbers } as you can get. There are plenty of labs that will analyze it wrong for $50 } a pop by XRD. I know.....I have been doing good analyses for 10 yrs, and it } takes a hell of a lot more than $50 worth of work. You need to find out what } the sample is composed of first, then remove components gravimetrically, } with acid dissolutions or thermal treatments to render phyllosilicates } amorphous. Most micas and clays have interfering peaks at the primary, } secondary and tertiary quartz peaks. How do you know you arent analyzing } beta or quasi metastable polymorphs of tridymite and cristobalite? You dont.
Here is where the simplicity of the TEM preparation, ability to take images, and elemental analyses of individual particles can answer these questions more quickly than just using XRD. If there is a mixture of two isomorphous silicates--say one with sodium and one with potassium but otherwise the same--I would not be optimistic about quantitating them to the 0.1% level from the ring pattern intensities, but EDS should do the job easily in a few minutes.
} } I can recommend some top notch people to verify what I have said, and who } can do it by XRD better than most. Let me know. I am disappointed at how } quickly so many people are responding to say it can be done by TEM without } really thinking about the problems that need to be addressed first. Good way } to chop your reputation up.
I look to this list as a forum for the discussion of technical points. As you said, there are often other considerations. I am happy to give technical advice where I am knowledgable and leave the decision of whether the other criteria will be met to someone else. Yours, Bill Tivol
Peroxidase is fairly resistant to glutaraldehyde or Karnovsky's fixatives, so good morphology should be possible. see JL Hall and R Sexton (1972) Cytochemical localization of peroxidase activity in root cells. Planta 108, 103-120.
Date sent: Wed, 09 Aug 2000 10:25:49 -0400 } From: "Shea Miller" {millers-at-em.agr.ca} To: microscopy-at-sparc5.microscopy.com
Off-and-on since the 1970s I've been preparing uranyl acetate for en bloc staining using maleate buffer at pH 5.2 adjusted with NaOH. However, the pKa I for maleic acid is 2.0 and the general rule for buffer selection is that the the desired pH should be within +/- one pH unit of the pKa value (p. 46 in Buffer Solutions,The Basics by Beynon and Easterby; IRL Press, 1996). Therefore, maleate buffer appears not to have a good buffering capacity at pH 5.2. Does anyone understand why maleate was chosen as bufffer for this procedure, and what I may be missing in my thinking on this subject? A current reference for this en bloc staining technique is found in Biomedical Electron Microscopy by Maunsbach and Afzelius, Academic Press, 1999.
/John/
John D. Wright, Ph.D. West Desert Test Center Dugway, UT
RMC is still part of Ventana Medical systems Inc. Feel free to contact me directly for Sales, Applications support & Service issues.
Drop by & see us at M & M
Best,
Al Coritz North American Sales Manager Ventana-RMC Microscopy Products Group 1-800-227-2155 x 2773 Direct: 520-690-2773 Fax: 520-690-2759 Cell: 520-906-3268
-----Original Message----- } From: Ken Tiekotter [mailto:tiekotte-at-up.edu] Sent: Tuesday, August 08, 2000 2:23 AM To: Geoff Williams Cc: Microscopy Listserver
RMC was purchased by Ventana Medical Systems, 1-800-356-3452 or 1-800-277-4613ext3023. Greg Carter is the ultramicrotome service manager. I had him visit my lab to service our MT2B in December, 1999. Let me know if this does not work.
Sincerely, Ken
On Mon, 7 Aug 2000, Geoff Williams 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 was just informed by our Purchasing department that their RMC contact } #s no longer work. } } Is there anyone out there that can point me to the direction of the } company that might or might not still be RMC that can provide parts for } our MT2B? } } Please excuse me if this answer is common knowledge, I just can't seem } to find the information. } } TIA, } Geoff Williams } } Electron Microscope Facility Supervisor } Biology Department } Brooks Hall } Central Michigan University } Mt. Pleasant, MI 48859 } } Geoffrey.Lloyd.Williams-at-cmich.edu } 517 774-3576 } 517 774-3462 (fax) } } } }
The MSA Education Committee is once again organizing the FREE Exhibitor Demonstrations on Tuesday, Aug. 15, 2000 at 5:00 pm. Below is a list of participants and the titles of their tutorial workshops. If you wish to attend one of these, please come to the MSA Education booth (now part of the MSA Mega-Booth) on the Exhibit Hall floor to sign up and receive a ticket allowing you to attend.
This list may not be exhaustive, so come by the booth to see what has been added!
Advanced Microscopy Techniques Corp. - Advantage HR Digital Camera System.
Allied High Tech Products, Inc. - Precision Sample Preparation for TEM, SEM and PreFIB (Multiple Sample) Thinning Using the Multiprep and MicroVision Measurement and Archiving Software.
Digital Instruments, Veeco Metrology Group - Surface Metrology Instrumentation: An Overview of Atomic Force Microscopes, Optical Interference Microscopes and Stylus Profilers.
E.A. Fischione Instruments, Inc. - Contamination-Free Specimen Preparation.
EDAX, Inc. - Quantitative Analysis of EDS Data from the Low-Vacuum SEM.
Electron Microscopy Sciences - Developments in Ultra Small Immunogold Silver Detection Systems.
Evex Analytical - Basic and Intermediate Features of the Evex Microanalysis System.
FEI Company - Precautions for Collecting EDS Data in a Low-Vac SEM.
KS Electron Technologies - Up Close with the Worlds Most Affordable TEM.
LEO Electron Microscopy, Inc. - EFTEM in Practice.
Micro Photonics, Inc. - Non-Destructive 3D Microscopy Using X-Ray Microtomography.
Microbiology/Syncroscopy - Bring Your World Into Focus with Syncroscopy.
Motic Incorporation, Ltd. - All-In-One Digital Microscope with Powerful Software.
NORAN Instruments, Inc. - Phase Identification and Orientation Mapping by EBSD.
Photon Technology International, Inc. - NovaLight: the NEW Power in Microscope Illumination.
SouthBay Technology, Inc. - Electropolishing, Tripod Polishing and Dimpling; Benefits and Limitations.
Ted Pella, Inc. - New Microwave Techniques, Accessories and Equipment.
TSL - #1 Advances in Phase Identification in the SEM. #2 Orientation Mapping in the TEM.
**************************************************************************** * 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 student who would like to do TEM on samples of clay in polystyrene. The particles are very small (around 1 nm in thickness) and are in suspension are in suspension. He believes that we could scoop up some sample onto a carbon film and examine them in an SEM. He has a chemistry paper where someone did just that. I have a TEM that has a voltage range of 100-200kV. I have never worked on polymers or a clay.
Questions: Will clay or polystyrene contaminate the TEM? What voltage range is safe? Any tricks?
By the way I do have a stage that can be liquid nitrogen cooled to minimize radiation damage.
The plant samples can be fixed in 4% paraformaldehyde and 0.5% GTA in phosphate buffer pH 7.2. I embed in paraffin and make 5-6 micron sections. I normally use the Vector Lab kit (no financial interest), peroxidase or alkaline phosphatase to do my enzyme histochemistry and it works beautifully. You can also use any fluorescent probe to label your protein of interest.
Good luck,
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:sghoshro-at-nmsu.edu http://confocal.nmsu.edu/eml
On Wed, 9 Aug 2000, Shea Miller 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; } now that the corn has finally started to do something in this miserable excuse for a summer we are having, we have a grad student who would like to do some peroxidase histochemistry on a rather daunting number of samples and varieties. } is there any fixation he can use to preserve both his samples and his enzyme activity? he will not be able to look at everything in a reasonable length of time. } thanks, as always, in advance } shea } } } } Dr. S.Shea Miller } Agriculture & AgriFood Canada } Eastern Cereal and Oilseed Research Centre } Rm. 2068 Neatby Building } Central Experimental Farm } Ottawa, Ontario } Canada K1A 0C6 } Phone: (613) 759-1760 } Fax: (613) 759-1701 } E-mail: millers-at-em.agr.ca } } }
At 11:18 AM -0600 8/9/00, Wright, John D. wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
****************** Simply put, UA does not ppt in maleate buffer. Phosphates cause hideous problems, and I've been told that cacodylate is also problematic. I side-step the whole issue and do my en bloc UA during dehydration (3%UA in 50% EtOh for 1 hr following an initial short step in 50% EtOh to remove the buffer. It works for me. Does it raise anyone's hackles? If yes, why?
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
Robin, If you put it on NaCl, you can float them off. You might be able to deposit them right onto freshly cleaved mica and float them off. You might also be able to put them onto either SiO or carbon support films directly.
If the films are too thin, they may not pop off your substrate. I had DLC films that were about 0.8 um pop off after a few days because of the stress in them. You might have to cut them down a little to help.
Carbon resists ion milling without some help in the gas. If you are interested in cross section TEM, then you will need to do low angle ion milling. I have a paper in MRS TEM book and the effects of gas compositions on ion milling. You might want to use a combination of Ne and O2.
The small angle cleavage technique worked beautifully on my DLC samples on Si. The DLC was too thin at the edge for EELS work. I would definitely try it with this if you put it on Si, GaAs, sapphire, SiC, or glass.
-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: "rgriffin-at-eng.uab.edu"-at-sparc5.microscopy.com } [mailto:"rgriffin-at-eng.uab.edu"-at-sparc5.microscopy.com] } Sent: Wednesday, August 09, 2000 11:09 AM } To: microscopy-at-sparc5.microscopy.com } Subject: TEM of diamond films } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: 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 } } } -------------------------------------------------------------- } ---------. } } } I have a student that would like to do TEM of diamond films that are } prepared by microwave plasma CVD. The thickness of the films can be } adjusted to make it electron beam transparent. The student } believes that he } can flake off pieces of the film. } } I'm thinking maybe we could just lay the diamond film on a } grid and pop it } into the TEM. Is this a crazy idea? Does anyone have any } suggestions? } } Thanks, } } Robin Griffin }
I would like to have copies of literature on the Wild M8 stereomicroscope and its accessories. Sales literature, price list, users manual, parts diagrams, service manuals,etc. are all welcome. Please contact me first to avoid duplication of your generous efforts. I will glady reimburse copying and mailing costs.
Mike Dalbey Biology Dept. University of California Santa Cruz, CA 95064
I suspect it would pop straight off if it became charged, i.e almost immediately you put the beam onto it!
One approach is to make a sticky grid, and put the film onto that. Three possible methods - 1) dip the grid into a thin solution of epoxy resin in PPO, allow all solvent to evaporate, place diamond film and cure epoxy at 60oC 2) Dissolve a pressure-sensitive adhesive from your favourite brand of sticky tape or carbon tabs. Dip grid, allow to dry, place film. 3) Coat grid with a thermoplastic solution (e.g. polystyrene) and dry. place diamond film. Bond by melting plastic on a hot-plate.
For a non-adhesive-based method you could try sandwiching the diamond film between two grids stuck together at one edge with epoxy, or use a proprietary sandwich grid (obtainable from SPI, Ted Pella, Polysciences, Agar Scientific etc.) Chris
} From: "rgriffin-at-eng.uab.edu"-at-sparc5.microscopy.com To: microscopy-at-sparc5.microscopy.com
somehow you need to glue the diamond film onto the grid, otherwise you might lose the diamond film inside the TEM.
On Wed, 9 Aug 2000 rgriffin-at-eng.uab.edu-at-sparc5.microscopy.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I have a student that would like to do TEM of diamond films that are } prepared by microwave plasma CVD. The thickness of the films can be } adjusted to make it electron beam transparent. The student believes that he } can flake off pieces of the film. } } I'm thinking maybe we could just lay the diamond film on a grid and pop it } into the TEM. Is this a crazy idea? Does anyone have any suggestions? } } Thanks, } } Robin Griffin }
I'm interested in calculating pore density using SEM image. I used several free softwares but couldn't get satisfactory results. If someone knows software or methods about calculating pore density using SEM image, please let me know.
It is not clear from your posting if your sample is a powder or a bulk specimen. The 1 nm you mention, I assume is this the clay thickness ?
We have looked at different clay materials in a polymer matrix . We normally tended to section these in a microtome (actually a cryo microtome to minimize deformation of the matrix.). We tend to use 200KeV for most of our work, but 100KeV should be OK also. The key here is to get nice thin sections because the contrast of the very thin clay particles will be better if the sections are thin. Also , could you share with me the paper (reference )you mentioned ? . We don't use a cooling holder for our samples, and contamination has not been a problem for us but then we were not doing diffraction on individual particles, we were only looking at the dispersion of the clay.
There is another possibility also which I would like to try when I get a chance. This would involve etching (plasma?) the surface of the sample to expose some of the clay and then looking at the sample either by AFM or by TEM (replicas of the etched surface). This should work on film or bulk specimens (with the clay in suspension), but I am not sure it would work OK on powder samples.
I hope this helps,
Jordi Marti Honeywell
-----Original Message----- } From: "rgriffin-at-eng.uab.edu"-at-sparc5.microscopy.com [mailto:"rgriffin-at-eng.uab.edu"-at-sparc5.microscopy.com] Sent: Wednesday, August 09, 2000 3:25 PM To: microscopy-at-sparc5.microscopy.com
I have a student who would like to do TEM on samples of clay in polystyrene. The particles are very small (around 1 nm in thickness) and are in suspension are in suspension. He believes that we could scoop up some sample onto a carbon film and examine them in an SEM. He has a chemistry paper where someone did just that. I have a TEM that has a voltage range of 100-200kV. I have never worked on polymers or a clay.
Questions: Will clay or polystyrene contaminate the TEM? What voltage range is safe? Any tricks?
By the way I do have a stage that can be liquid nitrogen cooled to minimize radiation damage.
There will be a special symposium which is dedicated to Dr. Lee Peachey, a well respected scientist and microscopist. The Symposium is number 16A on Tuesday, August 15th in Room 108. The Other Motor city: Muscle and Non-muscle Motility A Dedication to Dr. Lee Peachey
Dr. Lee Peachey uses confocal microscopy and High Voltage EM to study 3-D structural information on cell structures. The line up for the session is terrific. Dr. Saul Winegrad will chair the morning session and Dr. Clara Frazini-Armstrong will chair the afternoon session. All of the presenters are the tops in there fields. One highlight is Sir Andrew Huxley from Trinity College, Cambridge will give a talk on Microscopy of Muscle in the 19th and 20th Centuries. Dr. Huxley received the Nobel Prize in Medicine in 1963 for discoveries concerning the ionic mechanisms involved in excitation and inhibition in the peripheral and contrail portions of the nerve cell membrane.
Don't miss this session, Regards, Andy
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 wondered if you tried baking the grids and sections in a drying oven at 70 degrees C for 10 minutes or so? This will ensure that the sections will not come off the grids in any way, if that is what is happening here. (since you report that you don't seem to have this problem on unstained sections, it seems to me that your sections might be lifting a bit from the grid, causing wrinkling) If you leave the sections in the oven longer, or even overnight, I've noticed that it doesn't seem to harm them in any way. Once the sections are baked onto the grids in this manner, even a firehose directed at full spray cannot remove the sections. I might also add that heating the sections in this manner does not affect them adversely in staining. It might be the quick fix that you are looking for.
Garry
} ---------- } From: Tamara Howard[SMTP:howard-at-cshl.org] } Sent: Tuesday, August 08, 2000 2:29 PM } To: Anthony Greco } Cc: Microscopy-at-sparc5.microscopy.com } Subject: Re: TEM/Wrinkled Spurr Sections } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Have you tried aqueous stains for the Spurr's? I've used aq. UA/Pb and had } nice staining. If you have some stain-resistant plant material, you can } use KMnO4 instead of UA - it stains well. } } Tamara Howard } CSHL } } } On Tue, 8 Aug 2000, Anthony Greco 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 constant problem of regularly spaced wrinkles on my Spurr } } sections on the TEM. I stain with uranyl acetate in 50% ethanol for } } about 30 minutes followed by stable lead for 15 minutes.(Wrinkling is } } even worse with methanolic UA). I am certain the problem has something } } to do with staining since few wrinkles appear when I observe unstained } } sections. I also rarely encounter wrinkled sections with Embed 812 } } sections when using aqueous UA stains. Any suggestions? } } } } Tony Greco } } } } } } } }
New York Microscopical Society 1244 McBride Avenue West Paterson, NJ 07424
Bernard Friedman Memorial Workshop
Use of the Microscope September 16, 23, 30, October 7, 2000
A basic course on light microscopy which will cover the following topics: Theory of microscopy Kohler Illumination Diffraction Theory Contrast Methods Polarized light Phase Contrast Interference . . . . Hoffman contrast, Rheinberg, Dark-field & oblique Illumination, etc.
The workshop will consist of four consecutive Saturdays of lectures and hands on labs to cover the theoretical and practical aspects of microscopy. The course instructors include Jan Hinsch of Leica, Inc., Dennis O’Leary of Micro-Optical Methods, Mary McCann of McCann Imaging, John Reffner of SensIR Technologies, Inc. and N.Y.M.S. Instructor Don O'Leary.
WHEN: September 16, 23,30, October 7, 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: $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: Beginners and experienced users who wish to learn more about the proper use of a microscope.
HOW: Register using the form below. Limited to the first 12 registrants. Send form to Don O'Leary, 6 Chittenden Road, Fair Lawn, NJ 07410.
FURTHER INFORMATION: Call D. O'Leary (201) 797 -8849 E-mail donoleary-at-worldnet.att.net Fax (425) 988-1415
PLEASE POST ---------------------------------------------------------------------------- ------------------- Registration Form Use of the Microscope
} At 11:18 AM -0600 8/9/00, Wright, John D. wrote: } } } } } Off-and-on since the 1970s I've been preparing uranyl acetate for en bloc } } staining using maleate buffer at pH 5.2 adjusted with NaOH. However, the } } pKa I for maleic acid is 2.0 and the general rule for buffer selection is } } that the the desired pH should be within +/- one pH unit of the pKa value } } (p. 46 in Buffer Solutions,The Basics by Beynon and Easterby; IRL Press, } } 1996). Therefore, maleate buffer appears not to have a good buffering } } capacity at pH 5.2. Does anyone understand why maleate was chosen as } } bufffer for this procedure, and what I may be missing in my thinking on this } } subject? A current reference for this en bloc staining technique is found } } in Biomedical Electron Microscopy by Maunsbach and Afzelius, Academic Press, } } 1999. } } } } /John/ } } } } John D. Wright, Ph.D. } } West Desert Test Center } } Dugway, UT } } ****************** } Simply put, UA does not ppt in maleate buffer. Phosphates cause hideous } problems, and I've been told that cacodylate is also problematic. } I side-step the whole issue and do my en bloc UA during dehydration (3%UA } in 50% EtOh for 1 hr following an initial short step in 50% EtOh to remove } the buffer. It works for me. Does it raise anyone's hackles? If yes, why? } } 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
Well, my hackels are not up but I have read that long (more than a few hours) exposure to ethanol will extract cytoplasmic material even after glut. and osmium. I do agree that UA will give terrible ppt. with phosphate and I think cacodylate as well. Many rinses are needed to get the salts out. I have read that UA in distilled water works well for en bloc staining. Maleate buffer always seemed like a pain to me.
Geoff -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
} Does anyone know why freshly made lead citrate sometimes comes out } yellow, and what adverse effects, if any, result? In going over } past threads on the subject, it seems most people have a relatively } colorless product as the end result, but a few report the yellow } color as normal. After years of making up "colorless" lead citrate, } mine is now yellow. I've toyed with NaOH sources and pH; I use } ultra pure water, I've cleaned glasswear 'till it squeaks...no } change....it's still yellow. } } Sections seem to stain okay, although the stain goes bad very } quickly, sometimes even on the grid itself. Since this can be a } problem with colorless lead citrate, I'm not sure the yellowing is } the cause. I'd sure appreciate any feedback! } This happened to me a year ago, and I was equally puzzled. First I thought it was a reaction with the rubber on the plunger of the 10 ml syringe in which I store my stain (with a filter on the end it makes a great drop dispenser). I had opened a new box of *old* syringes. So I changed back to my old *new* syringes, but the newly made stain was still yellow and short lived. Same water, temperture, glassware, balance, etc. as my usual stuff. I suspected the distllled water source.
The next day a woman who has been doing TEM for ca. 35 years came in to ask if I'd ever seen yellow lead citrate. She had just goten this for the first time. She works in a different lab in a diffrent building with different water and, in fact, there should be absolutely no correlation at all. Except for phase of the moon, which theory we briefly entertained.
A few days later I successfully made up clear lead citrate. The main change I had made was to make up fresh 10N NaOH. However, the phase of the moon had changed as well.
Good luck!
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* ****************************************************************************
vendor-supplied slurry is used in lens polishing. recent polishings have resulted in scratches on lens. i have been asked to provide data with my xl40feg regarding particle sizes. am i correct in assuming that particle size distribution (in the slurry environment) is as important a criteria as simple particle diameters? can i evaluate simple particle size by "boiling off" the suspension, and sprinkling conductive tape with the remaining powder? can i "fire" the slurry into a ceramic brick, and polish the result for slurry particle distribution analysis? what would the procedure entail? thanks.
You could try the website for the American Association of Blood Banks, www.aabb.org. It is a trade organization. They may have technical leads and links.
I'm researching about the microstructure of steel, and have been usually using zet polishing and ion milling. Now I'm considering wedge polisher (Tripod) as a TEM sample preparation method, but rarely know about this.
I'd like to know whether this method can be used generally ,compared with zet polishing and ion milling. And I'd like to know advantages and disadvantages. Also, would you let me know the companies that produce wedge polisher (Tripod)?
Thanks in advace.
JungSoo.
Jung-Soo Byun School of Materials Science & Engineering College of Enineering, Seoul National University Seoul 151-742, Korea Tel : +82-2-880-7100, Fax : +82-2-885-9671 H.P. : 016-226-4358
You might consider filtering the slurry, rinsing the filtrate well. Choice of filter media is important. For you application, I would suggest a membrane filter rather than a fiberous one. ..Then dry, coat, and examine on filter -or apply filtrate to carbon tape.
Many slurrys contain more than just water and abrasive. Lubricants, suspention agents , etc. can be present which may coat the particulate if evaporative techniques are used alone.
As for firing - no. Under the correct (wrong) conditions, you may not just sinter the powder, but grain size and aglomerations may well be affected.
Woody White McDermott Technology Inc.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
vendor-supplied slurry is used in lens polishing. recent polishings have resulted in scratches on lens. i have been asked to provide data with my xl40feg regarding particle sizes. am i correct in assuming that particle size distribution (in the slurry environment) is as important a criteria as simple particle diameters? can i evaluate simple particle size by "boiling off" the suspension, and sprinkling conductive tape with the remaining powder? can i "fire" the slurry into a ceramic brick, and polish the result for slurry particle distribution analysis? what would the procedure entail? thanks.
I dont understand what is meant by x140feg. Is that an SEM? We do slurry analysis on daily basis using a TEM. Sample preparation includes to dilute slurry to about 1%vol and to follow a standard "drop grid" procedure. Please let me know if you have any further questions.
Have a nice day!
Chao Ni, PhD Rodel Inc. 451 Bellevue Road Newark, DE 19713 (302) 366-0500 ext 2812
-----Original Message----- } From: Mark Riggs [mailto:riggsm-at-svg.com] Sent: Thursday, August 10, 2000 6:55 PM To: Microscopy-at-sparc5.microscopy.com
vendor-supplied slurry is used in lens polishing. recent polishings have resulted in scratches on lens. i have been asked to provide data with my xl40feg regarding particle sizes. am i correct in assuming that particle size distribution (in the slurry environment) is as important a criteria as simple particle diameters? can i evaluate simple particle size by "boiling off" the suspension, and sprinkling conductive tape with the remaining powder? can i "fire" the slurry into a ceramic brick, and polish the result for slurry particle distribution analysis? what would the procedure entail? thanks.
Our old coater is on it's last legs so I'm looking to purchase a new carbon coater for our FESEM for EDS work. Since I inherited the old coater 'm looking for any advice (and good or bad experiences) with carbon coaters from the various manufacturers. The unit should be a bench top unit, user friendly, and as automatic as possible since it will be operated by several users.
Thanks in advance, Ron Kneisler Albemarle Corporation ron_kneisler-at-albemarle.com ****************************************************************************************** IMPORTANT NOTICE: This email message and any files transmitted with it may be confidential, may be legally privileged, and are for the intended recipient only. If obtained in error, please delete this message and confirm the deletion in an email to the sender. ******************************************************************************************
Hello everyone; after lurking for months, I have my second question in a week. Thanks to everyone who replied to my question re: fixation for enzyme histochemistry... we have decided to go with a short fixation in 1.75% glutaraldedhyde in buffer, and are keeping our fingers crossed (Kristin... I will reply in more detail to you).
Another grad student is wanting to do some in situ hybridizations on soybean and soybean seed coat. She would like to know if you can use lyophilized material.... I have only used stuff that was harvested fresh, fixed and processed through paraplast using a normal solvent dehydration, so I cannot advise her here. Any advice would be most helpful.
thanks in advance shea
Dr. S.Shea Miller Agriculture & AgriFood Canada Eastern Cereal and Oilseed Research Centre Rm. 2068 Neatby Building Central Experimental Farm Ottawa, Ontario Canada K1A 0C6 Phone: (613) 759-1760 Fax: (613) 759-1701 E-mail: millers-at-em.agr.ca
Would anyone have the web site of the "Sociedade Brasileira de Microscopia e Microanalise - SBMM"? Thanks in advance,
Adriana
Adriana Pinheiro Martinelli Rodriguez, PhD Laboratorio de Biotecnologia Vegetal CENA, Universidade de Sao Paulo Av. Centenário 303, Cx. Postal 96 13400-970, Piracicaba, SP, Brasil phone: +55-19- 429-4694 fax: +55-19- 429-4610 adriana-at-cena.usp.br http://www.cena.usp.br/labs/labbiotecveg.htm
I am trying to help a Hartford, CT high school biology teacher find microscopes for her classes. Any leads, ideas, advice for locating microscopes? I'm in Connecticut but welcome input from anywhere.
Julie Gross Dept. of Neuroscience University of CT Health Center Farmington Ave. Farmington, CT 06030 jgross-at-neuron.uchc.edu
You should look at the ProjectMicro Pages on the MSA WWW Site. There is a lot of information contained therein.
http://www.msa.microscopy.com/ProjectMicro
Caroline Schooley is the best contact for additional details/help. Her contact info is on the above page.
Nestor Your Friendly Neighborhood SysOp
} X-Sender: jgross-at-neuron.uchc.edu } Date: Fri, 11 Aug 2000 10:55:00 -0400 } To: MSA Listserver {Microscopy-at-sparc5.microscopy.com} } From: Julie Gross {jgross-at-neuron.uchc.edu} } Subject: Microscopes for High School } Mime-Version: 1.0 } Status: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
================================================================== 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
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-commserver.srrc.usda.gov
-----Original Message----- } From: Ringnalda, Jan [mailto:JRingnalda-at-FEICO.com] Sent: Friday, August 11, 2000 9:47 AM To: 'Ingber, Bruce F.'; Long, Jo; Piscopo, Irene Cc: Schaub, Daniel; Rice, Trisha (CS); Booth, Steve; 'sherman-at-btny.purdue.edu'
Ron,
We have a Bal-Tec SCD 050 Coater (6+ yrs. old) with two separate heads for carbon sputtering and gold/palladium sputtering respectively. We use carbon threads for sputtering. It has been very satisfactory for our FESEM work. The Bal-Tec unit is quite user-friendly.
------------------------------------------------------------ } Our old coater is on it's last legs so I'm looking to purchase a new carbon } coater for our FESEM for EDS work. Since I inherited the old coater 'm looking } for any advice (and good or bad experiences) with carbon coaters from the } various manufacturers. The unit should be a bench top unit, user friendly, and } as automatic as possible since it will be operated by several users. } } Thanks in advance, } Ron Kneisler } Albemarle Corporation } ron_kneisler-at-albemarle.com
As you are doing FESEM work, have you considered a high resolution deposition system? Our IBS/e Ion Beam Sputter Deposition and Etching System can act as a carbon "coater", but it can also deposit chromium, iridium etc. Using ion beam deposition allows you to controllably deposit very thin, extremely uniform films without heating the sample. Just a thought. Let me know if you would like additional information.
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.
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Our old coater is on it's last legs so I'm looking to purchase a new carbon coater for our FESEM for EDS work. Since I inherited the old coater 'm looking for any advice (and good or bad experiences) with carbon coaters from the various manufacturers. The unit should be a bench top unit, user friendly, and as automatic as possible since it will be operated by several users.
Thanks in advance, Ron Kneisler Albemarle Corporation ron_kneisler-at-albemarle.com {
South Bay Technology, Inc. is the manufacturer of the Tripod Polisher which is based on original work performed at the IBM East Fishkill facility (Anderson, Klepeis, Benedict et al). The Tripod Polisher, using the wedge polishing technique, can be used for a variety of materials. It has been used successfully for thinning semiconductors from silicon to indium phosphide, metals, ceramics, geological samples, hard disk media, composites etc. When used properly, some materials can be mechanically polished to electron transparency without the need for ion milling. Typically, a short ion milling step is required.
Jet polishing, in general, is a great method for preparing homogeneous materials if you do not have a specific area of interest. Jet polishing is a quick, effective and damage free method of preparing specimens. Of course, with a single jet polisher, you can control the area within the sample that you want to reach, but even at that, cannot very easily pick a specific spot. The Tripod Polisher provides a means to thin to a very specific area - in the case of semiconductors even to a specific sub-micron device. The main advantage of the Tripod Polisher is control. Tripod Polishing is an iterative process of polishing and visually inspecting your sample. Certain parts of the process can be automated - such as the first side polish (essentially an SEM cross section) by simply placing the Tripod on the yoke of a polishing machine. This is a nice way to do it as you can easily lift the polisher off for visual inspection and replace it within seconds. If alignment is done properly prior to 1st side polishing, first side visual inspection is minimal. The second side polish is more critical as that is where you are trying to locate your specific area of interest and progress the wedge in such a way as to maximize the thin area. Ongoing visual inspection is critical on the 2nd side or "wedge" polishing step. 2nd side polishing is generally done by hand ( although Allied High Tech sells a system that they say automates the whole process) as this allows you the most control and is the quickest way to change between the polishing and visual inspection.
The polishing is done using diamond abrasive films in a series of steps. The amount of time that you spend on any one polishing step is very short so it is important that you can quickly remove the Tripod Polisher and the diamond film to move on to the next step. This is important for a few reasons, but one of the most important is that the material you are removing at these final steps is so small, that you can over polish very quickly if you are not careful.
We have been doing Tripod Polishing here for over 10 years and have a huge base of customers as well as an extensive library of technical papers and application notes. Also, we do conduct ongoing courses in TEM specimen preparation including a specialized course in Tripod Polishing. I would be pleased to send you more detailed information if you have an interest. 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.
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Hi,
I'm researching about the microstructure of steel, and have been usually using zet polishing and ion milling. Now I'm considering wedge polisher (Tripod) as a TEM sample preparation method, but rarely know about this.
I'd like to know whether this method can be used generally ,compared with zet polishing and ion milling. And I'd like to know advantages and disadvantages. Also, would you let me know the companies that produce wedge polisher (Tripod)?
Thanks in advace.
JungSoo.
Jung-Soo Byun School of Materials Science & Engineering College of Enineering, Seoul National University Seoul 151-742, Korea Tel : +82-2-880-7100, Fax : +82-2-885-9671 H.P. : 016-226-4358
We need a nuclear LM stain for tissue sections (3-4 microns thickness) which have been processed with DAB label and embedded in epoxy. Does anyone have such a method? We would much appreciate any ideas.
We have received many questions concerning the use of Mercox for corrosion casting recently, but to provide first hand experience to those people and any others who may have an interest, we are once again sponsering the Vascular Corrosion Casting symposium at the M&M meeting next week in Philadelhia. This symposium will once again be chaired by Dr. Hossler, Dr. Aharinejad, and Dr. Lametschwandtner. It will be held Wed., Aug 16 at 8:30 am in room C122. I would urge all those who are interested to attend.
John Arnott Chairman --
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
uranyl acetate results in giving precipitates in our hands with either PBS or sodium cacodylate. For cell cultures we do the following: After osmium we do several washes with water and do the en bloc staining with 1% UA in water for 1h in the dark. We usally make a 4% UA solution and check the pH with paper before use. It should be between 4 and 5. If not we discard the solution.
Christoph Bauer University of Chicago
"Wright, John 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 } -----------------------------------------------------------------------. } } Off-and-on since the 1970s I've been preparing uranyl acetate for en bloc } staining using maleate buffer at pH 5.2 adjusted with NaOH. However, the } pKa I for maleic acid is 2.0 and the general rule for buffer selection is } that the the desired pH should be within +/- one pH unit of the pKa value } (p. 46 in Buffer Solutions,The Basics by Beynon and Easterby; IRL Press, } 1996). Therefore, maleate buffer appears not to have a good buffering } capacity at pH 5.2. Does anyone understand why maleate was chosen as } bufffer for this procedure, and what I may be missing in my thinking on this } subject? A current reference for this en bloc staining technique is found } in Biomedical Electron Microscopy by Maunsbach and Afzelius, Academic Press, } 1999. } } /John/ } } John D. Wright, Ph.D. } West Desert Test Center } Dugway, UT
If your substrate is brittle, you can also cleave it to make a small slab that you glue on a TEM grid. If the substrate is too thick, you may need to pre-thin it by mechanical polishing.
Then look with the TEM along the cleaved edge at about 45° tilt.
It's fast, it's clean, it does not require any complex tools, but of course the the transparent area is quite small!
Best regards
Philippe Buffat
} } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 that would like to do TEM of diamond films that are } } prepared by microwave plasma CVD. The thickness of the films can be } } adjusted to make it electron beam transparent. The student } believes that he } } can flake off pieces of the film. } } } } I'm thinking maybe we could just lay the diamond film on a grid and pop it } } into the TEM. Is this a crazy idea? Does anyone have any suggestions? } } } } Thanks, } } } } Robin Griffin } } } } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } 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 07710 585 401 } FAX: +44 (0) 131 653 6248 } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
__________________________________________________________________ Philippe Buffat Ecole Polytechnique Federale de Lausanne (EPFL) Centre Interdepartemental de Microscopie Electronique Address: EPFL-CIME, Batiment MX-C, CH-1015 Lausanne, Switzerland Phone: +41(21)693 29 83 Fax: +41(21)693 44 01 (Central European Time) E-mail: philippe.buffat-at-epfl.ch, WWW URL http://cimewww.epfl.ch/ ______________________________ Eudora F2.1 ___________________________
} } On Mon, 7 Aug 2000, Margaret Brannigan wrote: } } } Does anyone know why freshly made lead citrate sometimes comes out } } yellow, and what adverse effects, if any, result? } } } } Sections seem to stain okay, although the stain goes bad very } } quickly, sometimes even on the grid itself. Since this can be a } } problem with colorless lead citrate, I'm not sure the yellowing is } } the cause. I'd sure appreciate any feedback! } }
[skip]
} } A few days later I successfully made up clear lead citrate. The main } change I had made was to make up fresh 10N NaOH. However, the phase of } the moon had changed as well.
Dear Margaret and Tina, Is it possible that CO2 reacted with the old NaOH? Is PbCO3 yellow? That would explain why fresh NaOH would work. Yours, Bill Tivol
Julie, You may want to look into the possibility that a local university or business has some older scopes that they would like to donate for your efforts.
Our local microscopy society OMS (Oklahoma Microscopy Society) was graciously granted 128 microscopes that were surplus for the University of Oklahoma. We sent out notices to the public schools in the state including an application form for the microscopes. We have since placed all 128 microscopes in a variety of elementary and secondary schools all over Oklahoma. The microscopes were old, although they were in good shape and many were fine binocular scopes with 4 objectives and good sub stage condensers. The lower elementary school grades (k-4) received scopes with 2 objectives and sub stage apertures. These scopes are perfect for the lower grades because of their simplicity, although they produce nice images and are very sturdy.
We are always on the lookout for more scopes as the demand from the schools exceed the supply. Greg
Julie Gross 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 trying to help a Hartford, CT high school biology teacher find } microscopes for her classes. Any leads, ideas, advice for locating } microscopes? I'm in Connecticut but welcome input from anywhere. } } Julie Gross } Dept. of Neuroscience } University of CT Health Center } Farmington Ave. } Farmington, CT 06030 } jgross-at-neuron.uchc.edu
-- ================================================================== Greg Strout Electron Microscopist, University of Oklahoma WWW Virtual Library for Microscopy: http://www.ou.edu/research/electron/www-vl/ e-mail: gstrout-at-ou.edu Opinions expressed herein are mine and not necessarily those of the University of Oklahoma ==================================================================
We have a student working with a protein which is inside microsomes. He would like us to immunolabel the protein with a polyclonal antibody to prove that it is inside the microsome and not a contaminant from the cytoplasm. I would appreciate any suggestions from all who have attempted this or something similar.
Thanks,
Michelle Peiffer ************************************************************* Electron Microscope Facility for the Life Sciences Penn State University Biotechnology Institute 001 South Frear Lab University Park PA 16802
In the Microscopy & Microanalysis journal of Microscopy Society of America, affiliated with Brazilian Society, there are e-mail contacts for SBMM. I suggest you start with these: sbmm.ceme-at-epm.br OR: microcel-at-biomed.icb2.usp.br
Good luck,
Gib Ahlstrand
P.S. I was just in your area a few weeks ago and enjoyed the nice cool weather! Its hotter than blazes in Minnesota right now. -------------------------------------------- Responding to the message of {4.1.20000811114014.009a4820-at-mail.cena.usp.br} from "adriana-at-cena.usp.br"-at-sparc5.microscopy.com:
} Would anyone have the web site of the "Sociedade Brasileira de Microscopia } e Microanalise - SBMM"? } Thanks in advance, } } Adriana } } Adriana Pinheiro Martinelli Rodriguez, PhD } Laboratorio de Biotecnologia Vegetal } CENA, Universidade de Sao Paulo } Av. Centenário 303, Cx. Postal 96 } 13400-970, Piracicaba, SP, Brasil } phone: +55-19- 429-4694 } fax: +55-19- 429-4610 } adriana-at-cena.usp.br } http://www.cena.usp.br/labs/labbiotecveg.htm } } } } } } } .
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
Allied High Tech Products, Inc. has two different devices, the MultiPrep Precision Polishing System (Part No. 15-2000) and the TEM Wedge Polisher (Part No. 69-41000). The MultiPrep is an automated tool that is capable of thinning to samples 1 micron, as well as creating angles to 13 degrees. The TEM Wedge Polisher is a manual fixturing device that can be ste to remove material in mmicron incriments.
If you would like more information on this machine you can visit our website at www.alliedhightech.com or send me an e-mail directly.
I hope this answers your question.
Best Regards,
Armando Verdugo Laboratory Supervisor Allied High Tech Products, Inc. 2376 E. Pacifica Place Rancho Dominguez, CA 90220 (800)675-1118 US and Canada (310)635-2466 worldwide (310)762-6808 fax www.alliedhightech.com
The website address for the Sociedade Brasileira de Microscopia e Microan lise is at URL http://venus.rdc.puc-rio.br/sbme2/sbmeport.htm
It still seems to be mostly "under construction" but there is some information there.
Chuck
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It seems the northern hemisphere has collectively gone on leave - or to Philadelphia. Lets see if we can help, though its decades since I worked on starch, but those grains in seeds have not changed - I think.
Trying to fix dry seeds is near impossible and you should look at alternative methods, such as freeze etching or high resolution scanning, perhaps freeze substitution. If you can imbibe the tissue for a day or two, its still difficult. It was my notion that the GA just does not penetrate, regardless of fixation time. You may have some success by using GA at room temperature or warmer, but I doubt that. Using OsO4 you can after part processing actually see under a dissecting scope how far it has penetrated. Of course OsO4 is little use if you wish to do cytochemistry. Suggest that you use the OsO4 also at room and higher temperatures. Plant enzymes causing autolysis are fairly active in the cold and since in ice fixation times are perhaps 4x longer than at 20 degrees and maybe 8 times longer than at 37 degrees (impossible to truly quantify, but this gives the idea), shorter fixation times at higher temperatures are preferable. Do a fixation trial, at room temperature in OsO4 for say half, one and two hours and see what looks best. If the cytoplasm is dark (worse still membranes turn white) the tissue is overfixed. The granules will always tend to fall out during sectioning, but if the sections just simply fall apart, its most likely that the tissue was not fully fixed, infiltration is the less likely cause. 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 ABN: 99 724 136 560 www.proscitech.com
On Friday, August 04, 2000 11:22 PM, ABM Siddique [SMTP:siddiqueabm-at-hotmail.com] wrote: } } Dear All: } } I have been trying to study the ultra-structure (TEM) of strach granules } (in wheat and maize seeds) that contains amorphus and crystaline layers. } Unfortunately, I am unable to look at these layers under TEM even after } keeping the tissues longer time in fixative (over night in } 3%gluteraldehyde) and embedding medium (LR white/Spurs for 2 days). } } I also tried the the above procedure with isolated starch granules in low } melted agar but the result is the same - poor penetration of embedding } resin as a result I am getting broken starch granules without any } ultrastructural details. } } I would appreciate it if anybody could suggest me how to obtaine a } reasonably good ultra-structural (TEM) details of starch granules. } } With thanks, } } Siddique } } } Get Your Private, Free E-mail from MSN Hotmail at } {http://www.hotmail.com/} http://www.hotmail.com } } }
I am a postgraduate at the department of Engineering at the University of Sheffield. My samples are transparent glass melts 4cm3 with inclusions in them. I am using both transmitting light and reflecting light microscopy.
I would like to obtain a x1 mag or a 1.25 mag, objective for the Olympus CH-2 microscope, with the following features: 1. The objective lens should have a short barrel i.e. the length of the objective, from the top [ where it screws into the objective holder] to the bottom of the objective [ DT] = {30mm, -the features of my sample are fairly large so I need an objective with a short barrel, giving me a maximum depth of field. This factor is particularly important. 2. At the top where it screws into the objective holder, diameter is 20.11mm 3. with an acromat plan lens corrected for flatness of field 4. the objective will be used without a cover glass and without an immersion fluid 5. preferably the objective will be infinity corrected. I am not sure whether a standardized adjustment length would be necessary in order to allow max depth of focus, whilst using transmitting microscopy
At present the objective used on the above microscope is a x 2.5 mag, x5 mag and a x10 mag. They have the following notations MSPlan x 2.5 mag / 0.07, infinity/ - f = 180, Olympus 100621 MSPlan x 5mag / 0.13, infinity/ - f = 180 , [ this has a short barrel = 30mm ]
I would welcome any help or advice regarding obtaining the above objective lens. .
Biological electron microscopists often fall into the trap of automatically fixing everything, even when it is inappropriate to do so. I was once asked what the best fix would be for diatom frustules, to which the answer is probably to see a doctor!. But seriously, I am not quite sure why it should be thought necessary to fix starch grains either with glutaraldehyde or with osmium, or what bearing such "fixation" would have on starch grains falling out of epoxy resin sections. Let's look at some fundamentals. Starch grains are mainly crystalline or paracrystalline assemblages of glucan. The molecules are therefore in a minimum-energy configuration, which will be extremely difficult for most other molecules to enter. Unless this crystalline order is severly disrupted no significant quantity of epoxy resin will enter the grains - infiltration is essentially impossible. Glutaraldehyde and osmium will not react with starch, so it is pointless attempting to fix them unless there are traces of proteins or lipids within the grain structure that must be stabilised. I really don't know how much protein or lipid is present within the crystalline starch - I am sure you'll tell me - but the main issue here is that the embedding resin is externalized by the crystalline structure of the grains, and does not bond too well with the surface either, so that the grain readily separates from the section. I think the possibility of promoting covalent bonding of the starch grain surface to the epoxy could be a more promising avenue to pursue than experimenting with different permutations of fixatives. There was some discussion in this forum some time ago about silanes manufactured by Dow Corning that can be used to promote adhesion of various materials to epoxies and other resins.
Chris Jeffree ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 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
Dear all, In my laboratory, we study vascular endothelium with electron microscopy (Guinea pig). We tried perfusion to fix tissue but endothelium was removed. We fix them with 4% paraformaldehyde + 0,1 % Gluta + 0,1 % picric ac in phosphate buffer pH 7,4 overnight. Artery are then embedded on unicryl for immunocytochemistry. Tissue was stain with uranyl acetate and Reynolds. With this procedure cell structure isnot very evident, membrane is well defined but not very contrasted. Ac locate on all the section, not very locate in a particular place of cells. We tried other first and second antibodies but it was the same thing. What can we do? Do you have any suggestion? Thanks
SEM MASTERCLASS THE SEM APPLICATIONS COURSE 3rd to 5th October 2000 THREE DAYS $600 AUS (Not including meals and accommodation)
An opportunity for a hands-on introduction or a more advanced session, depending on your requirements, on just about every one of the major applications of modern scanning electron microscopy:
Conventional SEM - FESEM - cold stage - EDXA- cathodoluminescence - variable pressure SEM- low kV - coating techniques - film and digital recording - tricks for difficult specimens.
When? Tues-Thurs 3rd, 4th, 5th October 2000
Where? The Australian National University Electron Microscopy Unit, Canberra, Australia
Who by? Steve Chapman (Protrain, UK) and ANUEMU staff
Who for? Anyone who feels they could get more out of their time on the SEM if they had a bit more background and knew more of the possibilities. Can your machine deliver what you need with a little coaxing? Or would different equipment make a big difference to your work? For biologists or materials scientists. Participants are encouraged to bring their own samples.
What with? FOUR SEMs will be used for the course: a Hitachi 4500 FESEM, JEOL 6400 SEM, Cambridge 360, Hitachi 2250-N SEM. Accessories include three EDXA detectors, several cold stages, and a number of different SE, BSE and CL signal detectors. For equipment details, see our website, http:/www.anu.edu.au/EMU
Queries: email stowe-at-rsbs.anu.edu.au . Course Registration form also on website http://online.anu.edu.au/EMU/00chapman.htm
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
Does anyone have any experience of either/both of the Epson Expression 1600 Pro or the Minolta Dimage Multi scanners? One thing we would like to do is to scan negatives at oblique angles, rather than parallel to the film strip. My understanding is the even medium format scanners constrain the negative in a holder, whereas using a flat bed with a transmission attachment (such as the Epson) would allow rotation.
+------------------------------------------------------------------------+ | 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. Why don´t you try it with 1% aqueous UAc. Either for twenty minutes a room temperature or at 40°C for 7-10 minutes (results in more intense contrast...)
II. For staining with alcoholic UAc we apply the following procedure: 1. 20 min. 1% UAc in 70% alc. 2. wash: 1-2 drops 70% alc. 3. 1-2 drops 50% alc. 4. 3-4 drops aqua dest. 5. dry on filter paper.
You wrinkling problem might be due to a harsh change from alcohol to water, which causes damage to the sections. And, are your sections supported on the grids by a formvar or carbon film?
For Spurr´s resin I would prefer the warm, aquaeous method. Hope this works.
Greetings, Michael
} } I have a constant problem of regularly spaced wrinkles on my Spurr } sections on the TEM. I stain with uranyl acetate in 50% ethanol for } about 30 minutes followed by stable lead for 15 minutes.(Wrinkling is } even worse with methanolic UA). I am certain the problem has something } to do with staining since few wrinkles appear when I observe unstained } sections. I also rarely encounter wrinkled sections with Embed 812 } sections when using aqueous UA stains. Any suggestions? } } Tony Greco } }
I apologise if this is a little out-of-gamut for this list, but I have been trying to make digital copies of archival material which only exists as small prints on lustre paper. On a flat bed scanner, the textured surface causes objectionable specular reflections which degrade image quality. Is there a good way to avoid this?
Chris ========================================== Dr. Chris Jeffree University of Edinburgh BIOSEM - Biological Sciences Electron Microscope Facility Institute of Cell and Molecular Biology Daniel Rutherford Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JH, Scotland, UK Tel. #44 (0) 131 650 5345 FAX. #44 (0) 131 650 6563 Mobile 07710 585 401 email c.jeffree-at-ed.ac.uk =========================================
Are there problems with the system or did I somehow get deleted from the system? I haven't received any mail since last Wednesday (8/9/00). If I got deleted please re-subscribe me. Thank you.
prutledge-at-ars.usda.gov
Phil Rutledge USDA/ARS 151 Dixon Drive Chestertown, MD 21620 voice: 410.778.4136 or 2120 fax: 410.778.4399
} I apologise if this is a little out-of-gamut for this list, but I have been } trying to make digital copies of archival material which only exists } as small prints on lustre paper. On a flat bed scanner, the textured } surface causes objectionable specular reflections which degrade } image quality. Is there a good way to avoid this?
Dear Chris, I have had excellent luck with old photos by re-photographing them. The lighting has to be arranged so that there are no spurious reflections, but this is not too difficult. Using a digital camera will make scanning the photos unnecessary, and using film will give you a high-resolution negative. You will probably want to add a scale bar to the original to account for changes in magnification. You also need to take care that both the center and the edges of the print are in focus, since they can be at significantly different distances from the lens. Good luck. Yours, Bill Tivol
is there a detailed and comprehensive reference (with em pictures) to coiled bodies, pol II transcriptosomes (and pol I and pol III (?)), interchromatin granules, perichromatin fibrils, peripheral condensed chromatin, and of perinucleolar chromatin, fibillar centers, dense fibrillar component, granular component .....and other that someone knows of?????i would be eternally grateful for a lead to such a paper(s) Thanks you, marian miller
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The only EM-type ref I know of is from 1990; Raska et al. in Electron Microsc. Rec., 3:301-353. I can make a photocopy of my photocopy if you are interested, but the images are mush. it is called : "Immunocytochemistry of the Cell Nucleus".
The other refs I know of are mostly text and LM/FM, not TEM. You may have to just lookup the individual authors who work on these stuctures...I don't know if there are any sectioned images of transcriptosomes, but the others are all out there. See papers from the labs of David Spector, Robert Ochs, Marc Thiry, Daneholt, Aebi....There is a review from 1993 by Spector that talks about the regions and has refs to EM papers: "Macromolecular Domains within the Cell Nucleus" Annu. Rev. Cell Biol., 1993 9:265-315.
Hope these help! If anyone sends you the ref for a good EM review, could you please post it to the server?
Tamara Howard CSHL (IGC-land)
On Mon, 14 Aug 2000, marian miller 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 } -----------------------------------------------------------------------. } } } is there a detailed and comprehensive reference (with em pictures) to } coiled bodies, pol II transcriptosomes (and pol I and pol III (?)), } interchromatin granules, perichromatin fibrils, peripheral condensed } chromatin, and of perinucleolar chromatin, fibillar centers, dense } fibrillar component, granular component .....and other that someone } knows of?????i would be eternally grateful for a lead to such a } paper(s) Thanks you, marian miller } } }
With just one picture, you can usually retouch in Photoshop to restore the image. If you have many, then it is probably worth re-photographing them in flat light and through a polarizing filter. This should remove the reflections, but it can also will increase contrast, causing loss of detail in the very light and very dark areas of the image
I am posting this question for a colleague who is having trouble with the large size of his samples.
"I am trying to find a way to fix and embed calcified carotid endarterectomy specimens such that I will be able to obtain serial cross sections. The problem that I am encountering thus far is that I am unable to embed the 5cm specimens in a large paraffin block to allow sectioning because such large blocks either
a) do not fit on standard microtomes and therefore must be cut with sliding microtomes resulting in more distortion of the specimen
b) fall off of the blocks after 1 or 2 sections are obtained because the force of the blade against the mounted specimen shears the embedded specimen from the cassette or resin embedding block.
How can I fix and embed large specimens (~5cm long) such that I can obtain serial cross sections of the entire specimen for reconstructions without having to chop the specimen into small labeled pieces first and embed each of these pieces individually?"
Thank you ******** Cynthia J. Zeissler Physical Scientist National Institute of Standards and Technology cynthia.zeissler-at-nist.gov Tel. 301-975-3910 FAX. 301-417-1321
Cynthia: We routinely cut 3.5 to 4.0 cm paraffin blocks and cut some that exceed 5 cm in length and width on a rotary microtome. The limitation is the length of the up and down travel on your microtome. You can use a vise -type holder to hold these blocks. You can always have an adapter made if you can't purchase one that will hold large paraffin blocks. You might not want to use cassettes for this purpose but simply make a large stiff paper mold in the shape of a cube to hold the specimen like all the old original paraffin specimens were done. As far as cutting calcified blocks this may be impossible in paraffin under some circumstances if the blocks have not been decalcified first...depends on the amount of calcium. Serial sections will be a real challenge on specimens of this size and character and do not permit ready starting and stopping once commenced.
I know this may be a bit delayed (holidays you know) but maybe I can add a few points.
The micron marker is powered by the same electronics that produce magnification i.e. an error in magnification equals an error in micron marker too!
I do not know how old your instrument is or what make but this does make a considerable difference in the accuracy of magnification. It is "normal" to hope to get within 10% of the readout with an X to Y difference of less than 5%. If the following rules are not followed errors of up to 25% are quite easily obtained.
1. The sample must be a fairly flat surface
2. The sample must not be tilted
3. Turn OFF all electronic magnification and focus devices - dynamic focus, scan rotation, tilt correction
Use a 2160 lines/mm metal or TEM standard (a carbon grating replica, much cheaper and they work very well [see Spi]) to calibrate the machine but be aware that the spot size setting will have an influence upon the magnification accuracy!
The most accurate method for sample to sample consistency is to use the focus control as little as possible and use the Z control to obtain focus. In older instruments there was no zoom magnification linked to focus, so change the focus and you change the magnification. On modern instruments zoom magnification linked to focus is used in an attempt to hold the magnification correct.
After many years of testing instruments of all types, with clients on our courses there is no doubt that modern instruments are very good in relation to magnification accuracy, but some of the older machines were real dogs!
Good luck
Steve Chapman
Senior Consultant Protrain Tel & Fax - 44 (0)1280 814774 E-mail- protrain-at-emcourses.com Web Site - www.emcourses.com Protrain for SEM, TEM & EDX Training World Wide with our own full time Professional staff Courses in Australia, Canada, Europe, New Zealand, South Africa, Taiwan, UK and USA
You did not state the thickness of sections you are try to cut, this information could be important.
} From what you state, the only thing you have not tried is using a cryostat, this would give you a stable specimen without being as hard as using resin, our cryostats are routinely sectioning undecalcified bone, the 5cms. size would not pose a problem either.
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: Cynthia J. Zeissler [mailto:cynthia.zeissler-at-nist.gov] Sent: 14 August 2000 21:12 To: Microscopy-at-sparc5.microscopy.com
I am posting this question for a colleague who is having trouble with the large size of his samples.
"I am trying to find a way to fix and embed calcified carotid endarterectomy specimens such that I will be able to obtain serial cross sections. The problem that I am encountering thus far is that I am unable to embed the 5cm specimens in a large paraffin block to allow sectioning because such large blocks either
a) do not fit on standard microtomes and therefore must be cut with sliding microtomes resulting in more distortion of the specimen
b) fall off of the blocks after 1 or 2 sections are obtained because the force of the blade against the mounted specimen shears the embedded specimen from the cassette or resin embedding block.
How can I fix and embed large specimens (~5cm long) such that I can obtain serial cross sections of the entire specimen for reconstructions without having to chop the specimen into small labeled pieces first and embed each of these pieces individually?"
Thank you ******** Cynthia J. Zeissler Physical Scientist National Institute of Standards and Technology cynthia.zeissler-at-nist.gov Tel. 301-975-3910 FAX. 301-417-1321
Has anyone used the program called "Zip Shot"? I have a 1830 Amray SEM with a Noran Sigma EDS System. I would like to be able to print out my results using this program. I am not familiar with Zip Shot, it has been recommended to me. Thanks!
} } We have a AISI4340 steel sample. This sample contains iron, carbon and } nickel. We ion milled the sample. When we put the sample into our TEM, } we cannot align or focus the microscope. we are operating a JEOL100CX } microscope. } Any suggestions for imaging this sample? } Thank you Won-Sik Kim
thanks to everyone who sent me information about SBMM (Brazilian Society of Microscopy and Microanalysis)
Their current website and e-mail are:
http://www.dema.ufscar.br/sbmm/
sbmm-at-power.ufscar.br.
The website of the SBMM - Regional Chapter Rio de Janeiro is:
http://venus.rdc.puc-rio.br/sbme2/sbmeport.htm (currently under construction)
Adriana
Adriana Pinheiro Martinelli Rodriguez, PhD Laboratorio de Biotecnologia Vegetal CENA, Universidade de Sao Paulo Av. Centenário 303, Cx. Postal 96 13400-970, Piracicaba, SP, Brasil phone: +55-19- 429-4694 fax: +55-19- 429-4610 adriana-at-cena.usp.br http://www.cena.usp.br/labs/labbiotecveg.htm
Have not heard of ZipShot. Have used HyperSnap for windows as a screen capture. HS seems to be flexible and works nicely. OTOH... I have forgotten if the Sigma is running W95/8... If so have you tried ALT-PrintScreen? This will capture the displayed image to the clipboard (at the current monitor resolution). You can then paste into an image editor like Photoshop...
Woody White Mcdermott Technology Inc.
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi everyone,
Has anyone used the program called "Zip Shot"? I have a 1830 Amray SEM with a Noran Sigma EDS System. I would like to be able to print out my results using this program. I am not familiar with Zip Shot, it has been recommended to me. Thanks!
Since this material is magnetic you will need to reduce the amount of material that you put in the microscope. The first cut is to just make the 3 mm disk thinner at the start, perhaps 100 µm or less. At our lab we also use undersized (1mm or 2.3 mm) specimens and occasionally embed them in another material to make up the 3mm before we thin them. Probably the best way is to use the "window" polishing technique (see Hirsch et al.) which minimizes the amount of material while maximizing thin area. However I find it difficult to do correctly.
If you contact me offline I can put you in touch with the person at our lab who does the most of this type of work. By the way, we also order our microscopes with "extra strength" objective stigmators for working with magnetic materials.
Cheers, John Vetrano john.vetrano-at-pnl.gov
---------- } From: Won Sik Kim Sent: Tuesday, August 15, 2000 6:50 AM To: microscopy-at-sparc5.microscopy.com
} } We have a AISI4340 steel sample. This sample contains iron, carbon and } nickel. We ion milled the sample. When we put the sample into our TEM, } we cannot align or focus the microscope. we are operating a JEOL100CX } microscope. } Any suggestions for imaging this sample? } Thank you Won-Sik Kim
Does anyone have a light microscope for sale appropriate for a first year medical student? Student lives in Jacksonville FL Will attend med school in Los Angeles. Please respond off line:
Becky Garrison Pathology Supervisor ph 904-549-6237 fx 904-549-4290
I am trying to find a web or e-mail address for "Nordic" an antibody supplier in Tilburg Netherlands. My web searches haven't worked. Anyone out there deal with this supplier of immunocytochemical antibodies, etc. 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 sounds as if your sample may have become magnetized slightly. That is exactly what happened with my JEOL 1200EX-2 when I tried to examine iron particles which were slightly magnetic. I took the specimen grid and placed it into the coil I use to demagnetize my forceps with, and that solved the problem.
Franklin Bailey University of Idaho Electron Microscopy Center Moscow, ID 83844-2204 jfb-at-uidaho.edu
Reducing the size of your magnetic sample as John Vetrano has suggested is the best start. Here are some other things to consider and to try with the JEOL 100CX. (This question was last discussed on the listserver a year or so ago as I recall).
1. Be sure the sample is firmly held in the side-entry sample holder so it doesn't get pulled out by the magnetic field of the objective lens in the microscope. Sample holders with a spring-clip clamping mechanism are especially prone to this unpleasantness.
2. If you are using a double-tilt holder, check the mechanism for looseness to avoid having the sample spontaneously tilt to a position where the holder cannot be withdrawn from the microscope without scraping the polepiece or aperture holder.
3. Insert and withdraw the sample holder with the microscope in the low-magnification mode (objective lens off) to help avoid problems #1 and 2.
4. Pre-align the microscope in darkfield mode with a normal (nonmagnetic) sample before inserting the magnetic one. The available range of beam tilt correction in the 100CX microscope is much larger in the darkfield image control mode than it is in brightfield. A strongly magnetic sample will affect the beam tilts and image stigmation, but not the image focus.
5. Correct the alignments as follows: (a) Focus the image with the mechanical Z-axis translation rather than with the objective lens focus. (b) Align the beam tilts by voltage centering the image (darkfield mode to get enough tilting range). (c) Stigmate the image near its center --preferably with an unused stigmator channel so the normal one is not messed up for other users.
6. Repeat the alignment (#5) each time you tilt the sample or translate it a long way. The alignment corrections will increase a lot when the sample is tilted beyond ~5 degrees off horizontal, but should be manageable at most tilts unless the sample is too thick.
Larry Thomas larry.thomas-at-pnl.gov
From: Vetrano, John S Sent: Tuesday, August 15, 2000 10:31 AM To: 'Microscopy Listserver' Cc: 'kimwonsi-at-msu.edu' Subject: FW: AISI 4340 steel in TEM (fwd)
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html -----------------------------------------------------------------------.
Since this material is magnetic you will need to reduce the amount of material that you put in the microscope. The first cut is to just make the 3 mm disk thinner at the start, perhaps 100 µm or less. At our lab we also use undersized (1mm or 2.3 mm) specimens and occasionally embed them in another material to make up the 3mm before we thin them. Probably the best way is to use the "window" polishing technique (see Hirsch et al.) which minimizes the amount of material while maximizing thin area. However I find it difficult to do correctly.
If you contact me offline I can put you in touch with the person at our lab who does the most of this type of work. By the way, we also order our microscopes with "extra strength" objective stigmators for working with magnetic materials.
Cheers, John Vetrano john.vetrano-at-pnl.gov
---------- } From: Won Sik Kim Sent: Tuesday, August 15, 2000 6:50 AM To: microscopy-at-sparc5.microscopy.com Subject: AISI 4340 steel in TEM (fwd)
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society 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 AISI4340 steel sample. This sample contains iron, carbon and } nickel. We ion milled the sample. When we put the sample into our TEM, } we cannot align or focus the microscope. we are operating a JEOL100CX } microscope. } Any suggestions for imaging this sample? } Thank you Won-Sik Kim
Can someone advise me the appropriate temperature for regeneration of molecular seive material used in a foreline trap in a diff pump/rotary pump system on an SEM?
tia
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 following announcement is posted for: } } Roger Reyes, Group Leader } Failure Analysis Lab } International SEMATECH } 2706 Montopolis Drive } Austin, TX 78741 } (512) 356-3842 } roger.reyes-at-sematech.org } } To apply, please contact Roger directly. } ********************************************** } JOB TITLE: FA Engineer } } JOB SUMMARY: To provide critical support to the Advanced Tool Development } Facility (ATDF) and International SEMATECH's projects. } } JOB RESPONSIBILITIES: } * Continuously improve quality and accuracy of lab analysis and general } lab cycle time. } * Engineer would work on jobs that are beyond the technicians level, } providing in-depth analysis and advice on complex problems. } * Engineer would be working closely with technicians so that learning and } development occurs. } * Engineer would attend technical staff meetings. } * Some report writing and technical presentations. } * Engineer will keep pace with the leading edge technology. } } QUALIFICATIONS: } * Engineering degree. } * Experience in the semiconductor industry is preferred. } * Knowledge with databases. } * Knowledge on wet chemistries to delineate CMOS layers. } * Knowledge of Failure Analysis tool set. ( such as SEM, FIB,CVD, RIE } etc...) } * Good people skills. } } } }
Ramin Rahbari Pfizer Global Research & Development Drug Safety Evaluation 2800 Plymouth Road Ann Arbor, MI 48105 Voice (734) 622-3383 Fax (734) 622-5001 Ramin.Rahbari-at-Pfizer.com
-----Original Message----- } From: Tom Phillips [mailto:PhillipsT-at-missouri.edu] Sent: Tuesday, August 15, 2000 4:28 PM To: Microscopy-at-sparc5.microscopy.com
I am trying to find a web or e-mail address for "Nordic" an antibody supplier in Tilburg Netherlands. My web searches haven't worked. Anyone out there deal with this supplier of immunocytochemical antibodies, etc. 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 would be grateful for feedback offline from any users of the new(ish) JEOL 6700 FESEM, which appears still to be rather rare in UK.
Many thanks 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
Hello I am looking for a good source for gelatin for cryo-immunogold experiments. In the many papers on the subject I cannot find where the authors ordered the gelatin or any purity information. Any information would be appreciated. Thank you Theresa Theresa A. Fassel Core EM Unit, Mail Box MB-32 The Scripps Research Institute 10,550 N. Torrey Pines Rd. LaJolla, CA 92037-1027 tel: (858)-784-8182 fax: (858)-784-8193 tfassel-at-scripps.edu
I am looking for one each Zeiss KPl-W 10X eyepiece, catalog no. 46 40 42. Have just checked eBay with no success. does anyone have one for sale or can anyone suggest a source? Thanks, Mary Pfauth
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 one each Zeiss KPl-W 10X eyepiece, catalog no. 46 40 42. } Have just checked eBay with no success. does anyone have one for sale or } can anyone suggest a source? Thanks, Mary Pfauth
I may have a spare in the lab. I'll check tomorrow.
Please look at the Project Micro Pages on How to buy microscopes. There is alot of valuable advice there.
http://www.msa.microscopy.com/ProjectMicro
I will also be forwarding your Email to Caroline Schooley (schooley-at-mcn.org) as well as the Microscopy Listserver. Caroline will have the absolute latest information from the National Microscopy meeting which a large fraction of us from the US Microscopy Community are at this week. The Listserver members will be able to advise you on the rest of your questions, I have never used this microscope so cannot give you good advise on it's purchase.
Nestor
} } Email: jfallos-at-hotmail.com } Name: Janet Jacobson-Fallos } School: Bliss Discovery School } } Question: Hello! } } First, I need to tell you that I'm the TEACHER! :) } I haven't done anything with a microscope since college Biology 101. I am } hoping you might be able -- and willing -- to help me... } } I have been looking for quite some time to purchase a simple, yet high } quality microscope for use by highly-motivated middle school-aged } students. To find something of investment quality -- yet on a teacher's } out-of-pocket budget, I decided to look for a used Zeiss, Leica, Nikon or } Olympus. I had hoped to obtain a microscope with a 10x eyepiece and 4 } objectives -- incl. a 100x oil immersion. I've had a difficult time } finding anything under $250.00 } } I now have the opportunity to purchase an Olympus Model K ; circa 1970's?; } w/ a 10x eyepiece and 10x, 40x and 100x objectives; X,Y slide mechanism, } course and fine adjustments with locks. It has no illuminator, so I would } need to purchase one separately. The price is $140. I have seen used } illumination kits for anywhere from $40. -$200. I don't know if the 100x } objective on the microscope is designed for oil-immersion or not. (I am } currently awaiting the seller's answer to that question...) The microscope } doesn't come with any accesories -- no dust cover, case, slides, manuals, } books, etc. :( } } If you are familiar with Olympus microscopes, and hopefully the K model in } particular, I would appreciate any information/comments you might provide - } } ---------------------------------------------------------------------------
I was wondering if anyone could provide some information on tensile test fixtures for SEM's. Particularly if you have one and if you like or dislike it. I also need some manufacturers. The holder will be for a Hitachi 3200N microscope. Thanks. ______________________________________________ Roberto Garcia Senior Analyst, Metallography NC State University / Analytical Instrumentation Facility Campus Box 7531 Room 318 EGRC 1010 Main Campus Dr. Raleigh, NC 27695-7531 (919) 515-8628 (919) 515-6965 Fax rgarcia-at-unity.ncsu.edu http://spm.aif.ncsu.edu/aif http://spm.aif.ncsu.edu/asm/asm_home.htm ____________________________________________
I am looking for a copy of: Lewis J, Nelson J, Elder H: Radioimmunoassay of an antibiotic: gentamicin. Nature [New Biol] 239:1128, 1969 If anyone has this old paper sitting around please respond by email.
Thank you Karen Kelley Senior Electron Microscopist UF Biotechnology Electron Microscopy Core Lab Box 118525 Gainesville Florida 32611 lab:352-392-1184 fax: 352-846-0251 email: klv-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/staff/karenpage.html
Dear Valerie, per se your protocol looks pretty good. I have three suggestions. Fix it like that, try again perfusion fixation because it would preserve endothelium at best. Maybe you can even omit the Glut. Second, try tannic acid, this enhances contrast while preserving antigenicity. Third, try either LR-White resin or LR-Gold. Don´t cure it in the oven, use the accelerator in the cold (-20°C). I am dealing with immunoelectronmicroscopy on endothelia for over two and a half years now and we got now the breakthrough in finding a balance between structure preservation and antigenicity.
If you would like to discuss this in more detail, don´t hesitate to contact me.
michael.reiner-at-uni-koeln.de Michael Reiner University of Cologne, Germany Dept. of Anatomy I
} Valerie wrote: } Dear all, } In my laboratory, we study vascular endothelium with electron microscopy } (Guinea pig). We tried perfusion to fix tissue but endothelium was removed. } We fix them with 4% paraformaldehyde + 0,1 % Gluta + 0,1 % picric ac in } phosphate buffer pH 7,4 overnight. Artery are then embedded on unicryl for } immunocytochemistry. Tissue was stain with uranyl acetate and Reynolds. } With this procedure cell structure isnot very evident, membrane is well } defined but not very contrasted. Ac locate on all the section, not very } locate in a particular place of cells. } We tried other first and second antibodies but it was the same thing. } What can we do? } Do you have any suggestion? } Thanks } }
Michael Reiner University of Cologne, Germany Dept. of Anatomy I _______________________________________________________________________ 1.000.000 DM gewinnen - kostenlos tippen - http://millionenklick.web.de IhrName-at-web.de, 8MB Speicher, Verschluesselung - http://freemail.web.de
A well-meaning, non-microscopist friend to whom I have rented out a room in my lab decided to spiffy up the place while I was out of town. He tipped the roughing pump to a JEOL 35 up on end and stood it in the corner. When I got back from my trip, there was, of course, oil all over the floor.
The microscope was not in service at the time, but I want to place it in service as a back-up SEM some time in the future. My question:
What do I need to do (other than replace the lost oil) before starting up the pump?
In our lab we use a 275 bloom gelatin from fisher. Catalog number G8-500. We use a 2% solution. We went through many gelatins and finally was recommended to use the 275 bloom. Good luck:)
Roberto, We have a tensile stage with 500 degC heating capability which was built for us by Oxford Instruments (that division may now be part of Gatan?). The stage is for a LEICA S440 though I believe they could adapt to it for other microscopes. The system is good and has worked well. Good luck, Simon
} } } "Roberto Garcia" {rgarcia-at-unity.ncsu.edu} 17 August 2000 13:47:19 } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I was wondering if anyone could provide some information on tensile test fixtures for SEM's. Particularly if you have one and if you like or dislike it. I also need some manufacturers. The holder will be for a Hitachi 3200N microscope. Thanks. ______________________________________________ Roberto Garcia Senior Analyst, Metallography NC State University / Analytical Instrumentation Facility Campus Box 7531 Room 318 EGRC 1010 Main Campus Dr. Raleigh, NC 27695-7531 (919) 515-8628 (919) 515-6965 Fax rgarcia-at-unity.ncsu.edu http://spm.aif.ncsu.edu/aif http://spm.aif.ncsu.edu/asm/asm_home.htm ____________________________________________
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Deben Research manufacture an excellent range of micro-testing equipment for SEM chamber operation. Their details are :
Deben Research 11-15 High St Stowmarket Suffolk IP14 6QL UK Tel: +44 1728 861320 Fax: +44 1728 861350 Email: info-at-deben.co.uk WWW: 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
On Thu, 17 Aug 2000, Roberto Garcia 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 was wondering if anyone could provide some information on tensile test } fixtures for SEM's. Particularly if you have one and if you like or dislike } it. I also need some manufacturers. The holder will be for a Hitachi 3200N } microscope. Thanks. } ______________________________________________ } Roberto Garcia } Senior Analyst, Metallography } NC State University / Analytical Instrumentation Facility } Campus Box 7531 Room 318 EGRC } 1010 Main Campus Dr. } Raleigh, NC 27695-7531 } (919) 515-8628 } (919) 515-6965 Fax } rgarcia-at-unity.ncsu.edu } http://spm.aif.ncsu.edu/aif } http://spm.aif.ncsu.edu/asm/asm_home.htm } ____________________________________________ } } }
My research department is putting in a grant to ask money to buy a new set of microscopes which should work well with both bright and dark field, and coupled with color digital camera, and image analyzer. So far, we have got a quote from ZEISS for around $77K. My director wants to have the best, easy to use, easy to upgrade with great service for years to come.
Some has told us that ZEISS has the best lens quality in the world, and others said NIKON lenses are compatible to ZEISS, and its cameras are superior. Again others would recommend LEICA because it also makes excellent instruments for less money, with great service in Tampa Bay area.
Please help with your experience and knowledge in this field. Any satisfied customers? We don't have to buy the whole set from one company. We can get microscopes from one, digital camera from others.
We work most with Bone and Cartilage, paraffin, frozen, tissue culture, and plastic, if this information will have any weight on choosing which company we should go for.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } POST DOCTORAL FELLOW - ROOT/RHIZOSPHERE BIOLOGIST } $46K - $52K PLUS SUPERANNUATION } } CSIRO PLANT INDUSTRY } } Canberra Australia } } } } We are seeking a motivated Postdoctoral Fellow to join a team of } researchers in a project with the overall aim of developing a conceptual } framework for understanding the role of root/rhizosphere biology in crop } productivity. Specifically you would be investigating the impacts of } different crop management practices (e.g. conservation cropping) on root } architecture and structure, and on the in situ localisation and composition } of rhizosphere microbes. The work will involve the use of modern methods } of microscopy and bacterial identification and will also involve } collaboration with other soil biologists, agronomists and farmers. } } } The successful applicant will have a PhD and experience with root and soil } biology, preferably with field-grown plants, and technical expertise and/or } interest in the practical application of modern microscopy and } microbiological methods to the research project. } } This position is for a term of 3 years, with possible extension to 5 years. } } This post-doctoral fellowship is sponsored by the Grains Research } Development Corporation as a component of a major new initiative in Soil } Biology Research. The position is within the CSIRO Division of Plant } Industry. The Division has a staff of 700 with 60% located at its } headquarters in Canberra and other groups located at seven sites across } Australia. } } For further information contact Professor Margaret McCully (email mccully } -at-pi.csiro.au ; phone 61 2 6246 5343) or Dr John Kirkegaard (email } j.kirkegaard-at-pi.csiro.au ; phone 61 2 6246 5080) An Information Package } and Selection Documentation can be obtained by contacting the answering } service on 61 2 6246 5434 or fax 61 2 6246 5000 or email } recruit-at-pi.csiro.au This information is also available via our web site } http://www.pi.csiro.au/ } } Applicants must obtain and respond to the Selection Criteria and Duty } Statement. Address your application for the above position quoting } Reference No. PG:00082 and include details of your experience, skills and } qualifications together with the names of 3 references. Please mark } 'Confidential' and forward to: Trish Borger, Human Resources, CSIRO Plant } Industry, GPO Box 1600, Canberra ACT 2601 by Sepy. 15/ 2000 (notional). } } } } }
Several people requested copies of the full paper I presented at the Philadelphia meeting on the characterization of surface topography using AFM and SEM. I have uploaded a 720 kB "pdf" (acrobat) file containing a summary of my verbal presentation with images of the slides, for anyone interested. You can download it at {http://personal.rdu.bellsouth.net/~jcr5/MSA_paper.pdf}
I own a Zeiss, a Nikon and 2 Olympus scopes. Which is best? Depends on the application. Brand X may have the greatest optics but if the price prevents you from buying all the types of objectives needed for different approaches, then you will end up making do with the "wrong" objective and lose any real gain. The 3 manufacturers are all very similar. There are subtle advantages for each type (e.g., Nikon has the best working distance objectives). First, decide what features on objectives you need (e.g., water immersion, high NA, working distance, high UV thruput) and then buy the one you can afford.
Service is a function of your local rep and the company has very little to do with it in my experience. Zeiss owns their reps these days but it is still the local guys who you deal with.
The digital camera can come from any vendor - the best ones are probably not made my the microscope companies.
That's my prejudiced two cents worth!
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-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
Pbgrover-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 } -----------------------------------------------------------------------. } } A well-meaning, non-microscopist friend to whom I have rented out a room in } my lab decided to spiffy up the place while I was out of town. He tipped the } roughing pump to a JEOL 35 up on end and stood it in the corner. When I got } back from my trip, there was, of course, oil all over the floor. } } The microscope was not in service at the time, but I want to place it in } service as a back-up SEM some time in the future. My question: } } What do I need to do (other than replace the lost oil) before starting up the } pump? } } Paul Grover Paul, After you finish cleaning up the oil, turn the pump by hand a couple of turns just to be sure all is OK. It should be fine.
Ken Converse owner Quality Images third party SEM service Delta, PA
Good points. I am using Zeiss Axioskops despite near zero help from Zeiss. I used Olympus for years. But the newer BX series did not produce the images I need. Olympus was/is user friendly. Zeiss is user hostile as I see it. But their equipment is very good.
It takes a while to get the systems to work when there is little or no help from the maker. This is a result of their cancellation of dealers. Some Zeiss items can be found at great prices from abandoned Zeiss dealers.
gary g.
At 01:30 PM 8/18/00, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
You might also want to check your exhaust manifold and change the filter - just in case it was contaminated by some of the oil.
-- Lola Norton Research Engineer ESEM Facility EDU-114 (915) 747-8049
University of Texas at El Paso Chemistry Department Mail Code:00513 El Paso, TX 79968
} From: Ken Converse {qualityimages-at-netrax.net} } Organization: Quality Images } Reply-To: qualityimages-at-netrax.net } Date: Fri, 18 Aug 2000 20:20:34 -0700 } To: "Pbgrover-at-aol.com"-at-sparc5.microscopy.com } Cc: Microscopy-at-sparc5.microscopy.com } Subject: Re: roughing pump tipped over } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Pbgrover-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 } } -----------------------------------------------------------------------. } } } } A well-meaning, non-microscopist friend to whom I have rented out a room in } } my lab decided to spiffy up the place while I was out of town. He tipped the } } roughing pump to a JEOL 35 up on end and stood it in the corner. When I got } } back from my trip, there was, of course, oil all over the floor. } } } } The microscope was not in service at the time, but I want to place it in } } service as a back-up SEM some time in the future. My question: } } } } What do I need to do (other than replace the lost oil) before starting up the } } pump? } } } } Paul Grover } Paul, } After you finish cleaning up the oil, turn the pump by hand a couple of } turns just to be sure all is OK. It should be fine. } } Ken Converse } owner } Quality Images } third party SEM service } Delta, PA } } }
Several folks have made good points on this, so I am throwing in my two cents as well. There are many considerations for defining a "good" microscope. Personal preference (ergonmetrics, habits), price, and most importantly the task/procedures it is used for. You dont use a student microscope for R&D or vice versa.
When I consider a microscope purchase, I look only at the universals....we are scientists and use them accordingly, even in production settings. If I was a teacher, I would chose differently. My personal choice over the last 7 years is to go with the Oly BMAX line. I never cared for Oly until they got their act back together with the introduction of that line. It is a modification of Zeiss lineage. Zeiss still has the best objectives, but it is moot. Most of that difference you cannot resolve with your eyes.....just arent capable of doing so. So, I choose Oly objectives or used Zeiss objectives.
One last note. The russian scopes are incredibly enticing at their price, and they are pretty good in quality. My only problem with them is they are still built like their tanks.
Lou Solebello ----- Original Message ----- } From: Gary Gaugler {gary-at-gaugler.com} To: Tom Phillips {PhillipsT-at-missouri.edu} Cc: MSA listserver {Microscopy-at-sparc5.microscopy.com} Sent: Friday, August 18, 2000 6:13 PM
A one day TEM short course will be offered through the American Vacuum Society in Boston on Oct. 2, 2000.
For details please see: www.vacuum.org
******************************************************************* 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 ********************************************************************
I have inherited a Montage FR2 film recorder which looks to be in great shape except that I have been unable to figure out how to drive it and technical support for this product appears to be non-existent. Does anyone know how to interface the FR2 with a Mac, to generate slides from PowerPoint or Canvas?
Thanks
Gary Ward
Department of Microbiology and Molecular Genetics 214 Stafford Hall University of Vermont Burlington VT 05405 USA
I have looked at microscopes from most of the main suppliers and I have yet to find one that compares to the Leica range for quality (both optical and build) and, as far as I am concerned, ease of use and ergonomics. I use at the moment an Olympus AX which is excellent optically but is a nightamare to use. Nothing is easy to get to and every time I move my hand from the focussing or lens changer, I catch the stage even after two years of using the thing. The Zeiss microscope I had on demo was not much better. The Nikons seem quite good ( I have three of these in use)but I have not had a research one with really good optics but might go with one of them under certain circumstances i.e. financial constraints. The Leica DMR which I use for image analysis is a pleasure to use in every way and the optics are unbeatable. Richard Black
Presentation Technologies Martin Wieczorkowski 779 Palomar Avenue Sunnyvale, Ca 94086,USA
It is 6 years ago that I tried to contact the company, so I don't know if it exists any more. I purchased the Montage in 1993 from a German Company
Raebel EDV Kolbermoorer Str. 8a D-83043 Bad Aibling Tel.: ++49-806-4015 or 4016 Fax: ++49-8061-35714 email:info-at-raebel.de WWW: http://www.raebel.de
According to their homepage they sell the Montage FR2 up to now.
Cheers, Markus
At 17:40 19/08/00 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
******************************************* Dr. Markus Drechsler Friedrich-Schiller-Universitaet Jena Institut fuer Pharmazie Lehrstuhl fuer Pharmazeutische Technologie Lessingstr.8 D-07743 Jena
Two thoughts: 1) Photograph the images with very fine grain film like Tech Pan or better. You'll lose a little from the copying, but probably less than with digitizing. The negatives can then be digitized and also archived. 2) See if someone at UE has a drum scanner -- a university print shop, or some local pre-press business. Perhaps even the Art department, if they do computer art (or not).
Phil
} I apologise if this is a little out-of-gamut for this list, but I have been } trying to make digital copies of archival material which only exists } as small prints on lustre paper. On a flat bed scanner, the textured } surface causes objectionable specular reflections which degrade } image quality. Is there a good way to avoid this? } } Chris } ========================================== } Dr. Chris Jeffree } University of Edinburgh } BIOSEM - Biological Sciences Electron Microscope Facility } Institute of Cell and Molecular Biology } Daniel Rutherford Building } King's Buildings, Mayfield Road } EDINBURGH, EH9 3JH, Scotland, UK } Tel. #44 (0) 131 650 5345 } FAX. #44 (0) 131 650 6563 } Mobile 07710 585 401 } email c.jeffree-at-ed.ac.uk } =========================================
-- }}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{ 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 All, Forgive me for sounding like a cheerleader, but I want to thank the LAC gang in Philadelphia for a job very well done! The meeting was a pleasure to attend. I hope everyone is able to take a well desreved breather this week.
Next year in Long Beach.
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
Responding to the message of {v03100003b5c6b7d13bc9-at-[140.251.145.24]} from Leona Cohen-Gould {lcgould-at-mail.med.cornell.edu} : } Hi All, } Forgive me for sounding like a cheerleader, but I want to thank the LAC } gang in Philadelphia for a job very well done! The meeting was a pleasure } to attend. I hope everyone is able to take a well desreved breather this } week. } } Next year in Long Beach. }
A sentiment shared by the whole program committee for the Microscopy and Microanalysis 2000 meeting!
Plan on Long Beach for MandM 2001.
__________________ Stuart McKernan stuartm-at-tc.umn.edu 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
} Although I do not do confocal microscopy, I have been asked to, describe } it to new graduate students as part of a general lecture on microscopy. I } would like to present a Z-series in color along with a reconstructed } image. Can anyone point me to a source (preferably electronic) where I } might find such a thing. } } TIA } } Greg Erdos
Gregory W. Erdos, Ph.D. Assistant Director, Biotechnology Program PO Box 110580 University of Florida Gainesville, FL 32611
Hello All, we have a client with an old Philips EM200. The client had a large number of Mercury Batteries stored in his Refrigerator and now the supply is used-up. Does anyone have an alternate or a supplier ? The last 2 Batteries exploded. Thank you. Peter A. Stolzenberg, PESTO Inc., Philadelphia , PA 215-699-6160 FAX 215-699-5275
Microscopy listserv readers: Hildy has asked that I relay this message to the microscopy listserv, because she no longer has access to the account through which she subscribed. --John =================
} From: Hildegard H Crowley {hcrowley-at-du.edu}
} } Dear EM Netters: } } I'm interested in purchasing sterile syringe filters to be used } to rinse my grids. I have found some that has a pore size } of 0.1 microns. They have a typical fluid retention of {= 50 } lambda. The membrane is made of hydrophilic polyethersulfone. } Has anyone used these or the filter membrane? If so, do } they require priming with preboiled deionized water? } }
Donald G. Awbrey, HT(ASCP) QIHC Electron Microscopy / Image Analysis Donald Awbrey-at-hmhs.com donaldawbrey-at-hotmail.com (817)-878-5647
Honeywell in Phoenix, Arizona is looking for an experienced SEM/EDS/WDS operator to assist in the analysis of aerospace materials. Experience also desired in metallography of aerospace materials. For more information contact Juan Trillo at 602-231-2321 or reply to juan.trillo-at-honeywell.com
Harry Ekstrom Materials Laboratory (602) 231-2744 e-mail: harry.ekstrom-at-honeywell.com
Has anybody tried to download the summary of the verbal presentation at the Philadelphia meeting on the characterization of surface topography using AFM
and SEM By Dr John Russ ? For some reason all I get is a blank page. I'm not sure if it has something to do with my internet connection.
Regards 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
Dear All, We're in the process of setting up for U-Th-Pb chemical dating of monazite, and are in need of standards for Th and Pb. A particularly useful reference in this regard is that of Scherrer et al 2000 (Schweiz. Mineral. Petrol. Mitt), which summarises all kinds of stuff. And I've a couple of questions. 1) Does anyone know how to synthesise ThP2O7. Can it be done in the same way as REE orthophosphates? 2) Does anyone have any ideas on the stability under the beam of crocoite and it's suitability as a reference standard e.g., homogeneity, zoning etc. etc. Handy hints always helpful, 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: m.roberts-at-ru.ac.za SOUTH AFRICA
In a message dated 8/22/00 2:52:57 AM, willem.erasmus-at-sasol.com writes:
} Has anybody tried to download the summary of the verbal presentation at } the Philadelphia meeting on the characterization of surface topography using } AFM and SEM By Dr John Russ ? For some reason all I get is a blank page. I'm } not sure if it has something to do with my internet connection.
The paper does seem to download OK. If you are having difficulty because your browser doesn't like the ftp:// designation, try going to {http://personal.rdu.bellsouth.net/~jcr5/index.html/} and selecting the paper from there.
I was able to successfully access the website referenced by Dr. Russ. I did notice that immediately upon arriving at the site, it opened up Adobe Acrobat. Perhaps your browser does not have that plug-in.
Hope this helps,
David A. Bell Scientist Electron Microscopy Lab Millipore Corporation 80 Ashby Road Bedford, MA 01730 (781) 533-2108
We are discarding a EM-301 within the next few days. There are many new spare parts (electronic components, mechanical parts, etc.). Please contact me directly using the email address below if you need anything.
Owen
============================= Owen P. Mills Electron Optics Facility Engineer Michigan Technological University Metallurgical & Materials Engineering Rm 512 M&M Building Houghton, MI 49931 906-487-2002 906-487-2934 (FAX) opmills-at-mtu.edu
Malcolm Roberts wrote: } 2) Does anyone have any ideas on the stability under the beam of } crocoite and it's suitability as a reference standard e.g., homogeneity, } zoning etc. etc.
Dear Malcolm,
SPI uses crocoite and galena as Pb-standards in their minerals standards for X-ray microanalysis. I take this to indicate that crocoite is a suitable choice.
Best regards, Jorgen Bilde-Sorensen
***************************** J. B. Bilde-Soerensen Senior Research Scientist, Ph. D. Materials Research Department Risoe National Laboratory DK-4000 Roskilde Denmark
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Dear Microscopists
Has anybody tried to download the summary of the verbal presentation at the Philadelphia meeting on the characterization of surface topography using AFM
and SEM By Dr John Russ ? For some reason all I get is a blank page. I'm not sure if it has something to do with my internet connection.
Regards 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
by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id JAA13698 for dist-Microscopy; Tue, 22 Aug 2000 09:40:49 -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 JAA13695 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Tue, 22 Aug 2000 09:40:18 -0500 (CDT) Received: from tmpil001.tmp.allied.com (tmpil001.tmp.allied.com [198.80.19.2]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id JAA13688 for {Microscopy-at-sparc5.microscopy.com} ; Tue, 22 Aug 2000 09:40:07 -0500 (CDT) Received: from tmpnt701.allied.com (alliedsignal.com) by tmpil001.tmp.allied.com with SMTP id HAA28250 (InterLock SMTP Gateway 3.0 for {Microscopy-at-MSA.Microscopy.Com} ); Tue, 22 Aug 2000 07:34:23 -0700 Received: from 131.127.249.187 by tmpnt701.allied.com (InterScan E-Mail VirusWall NT); Tue, 22 Aug 2000 07:34:21 -0700 Received: by TMPCN190 with Internet Mail Service (5.5.2448.0) id {Q2WTNZ5W} ; Tue, 22 Aug 2000 07:34:19 -0700 Message-Id: {0380DC0B04CCD21184170008C7B1731F0194D423-at-PTBEX133}
I am looking for a new or used microtome which will work well in sectioning 0.5-5 mil polymer films and also polymer pellets. A used manual microtome would be fine. Also looking for a good bargain. Thanks!
Greetings all. I am a grad student in the psych dept at UC Berkeley. I am attempting to do HRP histochemistry on rat brain tissue. I have encountered several problems along the way. I hope someone on this list will be gracious enough to either make a suggestion or point me towards a resource which is helpful.
Briefly, my problems are these:
When I perfuse the animal via the left ventricle, frequently the lungs fill up and the brain is poorly fixed, regardless of which fixative I use. What's perplexing about this is that it seems to occur only when the fixative is added; typically the blood flush goes perfectly well. I usually don't move the 26 ga. needle when the solutions are changed. Should I be putting the needle in the aorta, as opposed to the L. ventricle? How can I do that without damaging the vessel?
I perfused a couple animals with 1% paraform/1.25% gluteraldehyde and found the brains to be mushy-- virtually unsectionable. Despite the problems with the lungs filling up & my worries about fixation, the liver appeared to be hardened, so I assumed something had happened. The glut was opened } 1 year ago, but I made the solution 48 hours before the procedure. What might have caused the brain to be so poorly fixed? Should I assume that the perfusion itself was poor, or, does the age & contact with air matter when using gluteraldehyde?
Thanks. I hope this is the appropriate list for these kinds of questions.
} Greetings all. I am a grad student in the psych dept at UC Berkeley. I am } attempting to do HRP histochemistry on rat brain tissue. I have } encountered several problems along the way. I hope someone on this list } will be gracious enough to either make a suggestion or point me towards a } resource which is helpful. } } Briefly, my problems are these: } } When I perfuse the animal via the left ventricle, frequently the lungs } fill up and the brain is poorly fixed, regardless of which } fixative I use. What's perplexing about this is that it seems to occur } only when the fixative is added; typically the blood flush goes perfectly } well. I usually don't move the 26 ga. needle when the solutions are } changed. Should I be putting the needle in the aorta, as opposed to the } L. ventricle? How can I do that without damaging the vessel? } } I perfused a couple animals with 1% paraform/1.25% gluteraldehyde and } found the brains to be mushy-- virtually unsectionable. Despite the } problems with the lungs filling up & my worries about fixation, the } liver appeared to be hardened, so I assumed something had happened. The } glut was opened } 1 year ago, but I made the solution 48 hours } before the procedure. What might have caused the brain to be so } poorly fixed? Should I assume that the perfusion itself was poor, or, does } the age & contact with air matter when using gluteraldehyde? } } Thanks. I hope this is the appropriate list for these kinds of } questions. } } Sincerely, } } bradley cooke } UC Berkeley
Bradley:
First, a 26 gauge needle is too small to deliver an adequate volume of fixative unless you are perfusing an embryo or a very young animal. However, the real problem is that you are going through the interventricular septum with the tip of the needle and perfusing the pulmonary circulation, not the systemic circulation. Thus firm lungs and mushy brain. Get a small piece of cork sheet or thin tubing. Poke the perfusion needle (20 or 18 ga.) through it so that only 2-3 mm protrudes. When entering the left vent. use the cork as a guide to prevent penetrating too far AND stay a bit to the right (your right, not the animal's) of the apex of the heart. The angle of "attack" should be about like this backslash mark. \ Also, make sure the cork does not slide on the shaft of the needle, otherwise your guide will become useless. Also, see if you can find an experienced perfuser to help you. Practice.
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 **********************************************
Hi Bradley, I haven't done whole animal perfusions in years, so I'll leave comments about that to others, but will say something about your fix solution. Your glutaraldehyde is OLD. It oxidizes over time and becomes pretty worthless as a fixative. My advice is to buy a fresh supply, preferably in small volumes. Many of the EM suppliers sell vials of 10 ml each of a variety of concentrations, sealed under nitrogen or other inert gas, so that you can open a fresh vial and have an easy dilution each time you need to make fix.
You probably need to leave the brain immersed in fix for a period of time after you've perfused it, but that will depend on how robust your antigen is....how much fixation will it tolerate?
I'm sure if you talk to the people in UC Berkely's animal facility, someone there should be able to help you get the perfusion set up properly. Its their job to make sure that animals are handled within guidelines, and they should be familiar with certain procedures.
Good luck, get fesh chemicals! 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
Hi Bradley, I haven't done whole animal perfusions in years, so I'll leave comments about that to others, but will say something about your fix solution. Your glutaraldehyde is OLD. It oxidizes over time and becomes pretty worthless as a fixative. My advice is to buy a fresh supply, preferably in small volumes. Many of the EM suppliers sell vials of 10 ml each of a variety of concentrations, sealed under nitrogen or other inert gas, so that you can open a fresh vial and have an easy dilution each time you need to make fix.
You probably need to leave the brain immersed in fix for a period of time after you've perfused it, but that will depend on how robust your antigen is....how much fixation will it tolerate?
I'm sure if you talk to the people in UC Berkely's animal facility, someone there should be able to help you get the perfusion set up properly. Its their job to make sure that animals are handled within guidelines, and they should be familiar with certain procedures.
I agree with Geoff's suggestions. We actually use a 16 gauge gavage needle and clamp it with a hemostat. This gives us good perfusion of the nervous system. I'd like to suggest a great paper that may help you:
Andrew Fix and Robert Garman, Toxicologic Pathology, vol.28, no.1, pp. 122-131, 2000. "Practical Aspects of Neuropathology: A Technical Guide for Working with the Nervous System."
Good luck, Sandy Perkins
Laboratory for Neurotoxicity Studies VA-MD Regional College of Vet Med Virginia Tech
Once you find a useful gauge of needle, use an emery stone to reduce the bevel of the tip, or get a short stub adapter of that guage and give it a slight bevel. This will prevent accidentally slicing the heart, or your finger.
A permanent method to create an insertion stop is to mix dental acrylic and build up a bead on the needle while rotating it. Continue rotating until it hardens.
Glen --
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. ************************************************************************
Experienced Confocal Microscopy/Electron Microscopy Technologist to work in Research Laboratory of Department of Pathology
The person we seek will be responsible for organization of a new facility that includes a confocal microscope and an electron microscope. The position includes overall management of the microscopy facility, user training, and user supervision. Requirements for the position include experience with light and transmission electron microscopy. This individual will oversee all aspects of specimen accession and processing, operation of the microscopes, photography, record keeping, and supervision of a technician. Knowledge of EM, biology, and pathology, as well as photographic procedures is required. The individual will work with only minimum supervision.
Position Requirements
Experience with confocal and digital imaging techniques, microinjection, visualization of living cells containing fluorescent probes, photobleaching, and fluorescence in situ hybridization.
Minimum of five years experience working in an electron microscopy laboratory with hands-on experience in tissue processing, dark room photography, and operation and maintenance of electron microscope.
Two years experience working on electron microscopy of human tissue specimens with knowledge of histology and pathology.
Excellent interpersonal and organizational skills are essential.
BachelorUs degree in science desirable.
Desirable Experience
Expertise in training in the operation of confocal microscope systems is a distinct advantage.
Familiarity with light microscopy methods, immunofluorescent staining, use of fluorescent probes for physiologic measurements and the general principles of cell biological research are critical. Significant facility with computers is desired.
Responsibilities
Serve as the technical manager of the facility and be responsible for the operation and maintenance of the confocal and EM microscope facility.
Perform routine transmission EM, including tissue processing, ultramicrotomy, and examination; do preventative maintenance on the equipment; maintain the lab, order supplies, schedule instruments, and oversee billing.
Image analysis at the light, confocal and electron microscopic levels and preparation of micrographs for publication.
Send resume to: Suzanne Mirra, M.D., Chair Department of Pathology Box 25 SUNY-Brooklyn 450 Clarkson Av. Brooklyn, NY 11203-2098
----------------- Submitted by Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
Experienced Histotechnologist to work in Neuropathology Research Laboratory of Department of Pathology
The person we seek will be responsible for maintaining a neuropathology research laboratory in the Department of Pathology at SUNY Downstate Medical Center. Knowledge of immunohistochemistry, special stains, and histopathology is required. Previous experience in handling central nervous system tissue preferred. The individual will work with only minimum supervision.
Position Requirements
Minimum of five years experience working in a laboratory with hands-on experience in tissue processing, histochemistry, immunohistochemistry, and operation of a light microscope. Experience with technical aspects of neuropathology and performing special stains including silver (Bielschowsky) and myelin stains.
Two years experience working on immunohistochemistry of CNS tissue specimens with knowledge of immunoreagents and experimental protocols.
BachelorUs degree in science desirable.
Laboratory skills including communications, ability to comply with safety and laboratory regulations, maintenance of laboratory equipment and resources, operation of computers and office equipment.
Advanced computer skills (word processing and database management) essential.
Desirable Experience
Confocal microscopy desirable.
Salary commensurate with experience
Responsibilities
Purchasing supplies and equipment, budget reports, laboratory maintenance and brain banking.
Will operate all microscopic, photographic and computer equipment, and keep accurate records of all laboratory experiments and procedures. Light and fluorescent microscopy; tissue processing for paraffin embedding, sectioning and slide stainer for immunohistochemical procedures; computer imaging (PhotoShop); general photography; and library and web searches
Familiarity with computer software for keeping records, report preparation and table/figure construction using Microsoft Office software (Word, Excel, PowerPoint) as well as Endnote and Adobe PhotoShop.
Send resume to:
Suzanne Mirra, M.D., Chair Department of Pathology Box 25 SUNY-Brooklyn 450 Clarkson Av. Brooklyn, NY 11203-2098
------------------- Submitted by Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
Experienced Electron Microscopy Technologist to work in Research Laboratory of Department of Pathology
The person we seek will be responsible for the overall operation of the EM laboratory in the Department of Pathology at SUNY Downstate Medical Center. This individual will oversee all aspects of specimen accession and processing, operation of the microscope, photography, record keeping, and supervision of a technician. Knowledge of EM, biology, and pathology, as well as photographic procedures is required. The individual will work with only minimum supervision.
Position Requirements
Minimum of five years experience working in an electron microscopy laboratory with hands-on experience in tissue processing, dark room photography, and operation and maintenance of electron microscope.
Two years experience working on electron microscopy of human tissue specimens with knowledge of histology and pathology.
BachelorUs degree in science desirable.
Laboratory management skills including effective written/verbal communication skills to interact with a diverse group, ability to comply with safety and laboratory regulations, maintenance of laboratory equipment and resources, and operation of computers and office equipment.
Desirable Experience
Previous experience in confocal microscopy highly desirable.
Previous experience in performing immunocytochemical staining and advanced computer skills usage (e.g. image analysis) is also desirable.
Salary commensurate with experience
Responsibilities
Maintain electron microscope in operating condition. Process clinical and research tissues for "thick" and "thin" sectioning. Darkroom management of photographic printing.
Send resume to: Suzanne Mirra, M.D., Chair Department of Pathology Box 25 SUNY-Brooklyn 450 Clarkson Av. Brooklyn, NY 11203-2098
----------------- Submitted by: Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
Does anyone have experience with processing lens from a dog eye in plastic? The lens was fixed in 10% NBF. I'm trying glycol methacrylate, using the JB-4 Plus kit. I trimmed the lens to 2-3 mm thick and infiltrated overnight. I'm just sectioning the first block. Areas of the lens are fractured. I'm thinking I need to try longer infiltration. But is there a better way? Thanks for any advice.
Marcia Pitzenberger Research Associate Merck & Co. WP45-251 West Point, PA 19438 (215) 652-9767 voice marcia_pitzenberger-at-merck.com
Just to make the "GA is old" point a little clearer: GA polymerises with time and higher temperatures; a nitrogen head does not change much. Frozen, GA will last an awfully long time and if stored refrigerated most people would still be happy with the product after six months and more. While in the lab storage should not be a problem, we can be sure that inappropriate storage is common. The problem is in transit, when during the summer months the GA could see some rather high temperatures during a truck or even an air journey. We have always taken some care with GA, but being a long way from the manufacturer we have adopted special precautions such as shipping with ice and mostly by air and incoming shipments are also packed in ice and refrigerated whenever not in the air. Its costly, but this assures a better product. The high purity Formaldehyde 16% does not require refrigeration, keeps for years and is an excellent fixative. I don't know how well this works in perfusion fixation, but it may well be a good alternative. 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 ABN: 99 724 136 560 www.proscitech.com
On Wednesday, August 23, 2000 4:23 AM, "lcgould-at-mail.med.cornell.edu"-at-sparc5.microscopy.com [SMTP:"lcgould-at-mail.med.cornell.edu"-at-sparc5.microscopy.com] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Hi Bradley, } I haven't done whole animal perfusions in years, so I'll leave comments } about that to others, but will say something about your fix solution. } Your glutaraldehyde is OLD. It oxidizes over time and becomes pretty } worthless as a fixative. My advice is to buy a fresh supply, preferably in } small volumes. Many of the EM suppliers sell vials of 10 ml each of a } variety of concentrations, sealed under nitrogen or other inert gas, so } that you can open a fresh vial and have an easy dilution each time you need } to make fix. } } You probably need to leave the brain immersed in fix for a period of time } after you've perfused it, but that will depend on how robust your antigen } is....how much fixation will it tolerate? } } I'm sure if you talk to the people in UC Berkely's animal facility, someone } there should be able to help you get the perfusion set up properly. Its } their job to make sure that animals are handled within guidelines, and they } should be familiar with certain procedures. } } Good luck, get fesh chemicals! } Lee } } }
Dear Bradley, Perhaps an easier solution to the danger of penetrating the wrong heart chamber is to use an over-the-needle Teflon I.V. catheter. Your animal care personnel can procure them for you, or if you have a hospital handy, their supply warehouse will have them. You can use a 20-23 gauge catheter in a rat. The sharp needle protrudes through the lumen of the catheter; you introduce the needle, bearing the catheter, into the left ventricle, just enough to penetrate the muscle wall, then slide the Teflon catheter part over the needle, deeper into the heart. With practice, I have been able to guide the catheter into the ascending aorta. The needle is then slid backwards out of the catheter and set aside. The catheter has a female hub end that will accept IV tubing, syringe hubs or injection caps, whatever your preference. Since this is a terminal procedure, you can clean the catheter and needle and reuse them. Good luck! Missy
Eleanor M. Josephson, D.V.M., Ph.D. Assistant Professor Department of Anatomy, Physiology and Pharmacology College of Veterinary Medicine 111 Greene Hall Auburn University, AL 36849-5518 Ph. (334) 844-5423 FAX (334) 844-4542 josepem-at-vetmed.auburn.edu
During the Facilities Management session at MM2000, we discussed online sign-up for instruments. At that time I could not tell anyone the specifics of the one we use. `It is free-ware and can be found at this site:
http://www.webprog.com/appoint/freetest.html
you can see it running at this site:
http://scope.biotech.ufl.edu/index.html Gregory W. Erdos, Ph.D. Assistant Director, Biotechnology Program PO Box 110580 University of Florida Gainesville, FL 32611
DEPARTMENT OF PHYSICS, UNIVERSITY OF ILLINOIS AT CHICAGO
{smaller} A postdoctoral fellow position is available at the University of Illinois at Chicago for the development and application of novel STEM/TEM techniques to the analysis of metal-oxide interfaces and grain boundaries in metals. Research in the Interface Physics Group focuses on the use of atomic resolution imaging and analytical techniques in electron microscopy, coupled with theoretical simulations, to determine the structure-property relationships at internal interfaces on the fundamental atomic scale. Other related research programs within the Interface Physics Group include ceramics, ionic conductors, high-Tc superconductors and semiconductor materials/devices, and it is expected that the successful applicant will work closely with the existing group members in these areas. =20
The experimental facilities to perform this research are comprehensive: a JEOL 2010 Field-Emission STEM/TEM featuring a 1.4=C5 probe size, "drift free" stage, high-angle annular dark-field detector (Z-contrast), Gatan Imaging Filter, Noran EDS, Gatan liquid nitrogen cooling stage, and Gatan heating stage; a VG HB601 Field-Emission dedicated STEM featuring a 2.2=C5 probe size and EDS and EELS capabilities; a JEOL 3010 conventional TEM with digital imaging capabilities and EDS; a JEOL 6320 =46ield-Emission SEM with EDS; and a JEOL JXA733 microprobe. In addition to the electron microscopes, specimen preparation facilities include a Gatan Duo-mill, Fischione precision ion-mill, SouthBay plasma cleaner and Leica Ultramicrotome. The Interface Physics Group has a Silicon Graphics R10000 Power Indigo workstation with the Molecular Simulations' Cerius 2 package incorporating the CASTEP pseudopotential code. The physics department has additional workstations and access to the UIC Convex Exemplar Supercomputer and the National Center for Supercomputing Applications at UIUC. =20
Successful candidates will be recent Ph.D. graduates in physics, metallurgy, or materials science with a sound background in the relevant materials issues and an ambition to be part of a developing program pushing at the frontiers of interface physics. Please send a resume and publication list to Professor Nigel D. Browning at the address below. Prior experience in STEM or TEM is essential. =20 However, consideration will be based on the candidates overall potential for success in the field and applicants with prior experience in related fields are encouraged to apply. Positions are for one year initially, normally renewed for a second year with possibilities existing for further years. Salary is commensurate with experience.=20 UIC is an equal opportunity employer.
Cold Spring Harbor Laboratory is seeking an experienced and responsible microscopy technician for the laboratories core microscopy facility. The individual should have practical expertise in transmission electron microscopy, confocal and widefield fluorescence microscopy, digital imaging, and microinjection. The successful candidate will be involved in designing and carrying out experimental protocols for users, training individuals in the use of various microscopes, and aligning microscopes and keeping the facility operating at an efficient and high level of productivity. Interested individuals should send their resume, including a description of their expertise and the names and addresses of 3 references to: Dr. David L. Spector, Cold Spring Harbor Laboratory, One Bungtown Road, Cold Spring Harbor, New York 11724, email: spector-at-cshl.org
At 09:45 AM 8/23/00 -0400, Greg Erdos wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I would like to second the opinion of Greg Erdos. We have been using the same calendar program and have been quite happy with it. You can see it running at our site:
http://katie.ucdavis.edu/
then click on Schedule Time.
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 would like to try to visualize ice in frozen plant leaves in a Hitachi 2460 variable pressure SEM and do not have access to a cold stage. I have heard that a block of aluminum or copper at LN temperature may make a usable substitute. Has anyone had any experience with this technique? Any and all tips would be appreciated.
TIA
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
We're considering putting a digital camera on our JEOL 1200EX STEM. Three possibles are from Gatan, AMT, and SIS. Does anyone care to comment (positively or negatively) about retrofitting an older instrument with a camera system, and recommend one system over the other?
I have seen the results of a Gatan Megaview on an older Philips 420 which was rather impressive. I have an older AMT on a 2000FX that works quite well. I saw the SIS system at M&M. I think that you will be pleased with whatever system that you buy. You may still need to use film occasionly, but the bulk of your work will use the camera. -Scott
-----Original Message----- } From: "Beverly_E_Maleeff-at-sbphrd.com"-at-sparc5.microscopy.com [mailto:"Beverly_E_Maleeff-at-sbphrd.com"-at-sparc5.microscopy.com] Sent: Wednesday, August 23, 2000 4:15 PM To: Microscopy-at-sparc5.microscopy.com
Colleagues:
We're considering putting a digital camera on our JEOL 1200EX STEM. Three possibles are from Gatan, AMT, and SIS. Does anyone care to comment (positively or negatively) about retrofitting an older instrument with a camera system, and recommend one system over the other?
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I did see this done by an application engineer when he was demonstrating a VP SEM. We really wanted to look at hydrated tissue and the only way was to freeze a metal specimen mount in liquid nitrogen and then put the sample on it as quiackly as possible. Then put it in the scope and, with the pressure as high as possible, let the vacuum "freeze-dry" the sample. If you were just wanting a quick look at hydrated material, this works fairly well. If you really want to do reproducible work with the frozen plant material, you really need a true cold stage. We do have a cryo stage and have used it for similar projects as well as for a whole host of other samples. It really is a very valuable accessory to our SEM.
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 Wednesday, August 23, 2000, Bob Wise {wise-at-vaxa.cis.uwosh.edu} wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id SAA18205 for dist-Microscopy; Wed, 23 Aug 2000 18:06:30 -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 SAA18199 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Wed, 23 Aug 2000 18:05:59 -0500 (CDT) Received: from mail.gatan.com (mail.gatan.com [205.139.92.1]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id SAA18192 for {microscopy-at-sparc5.microscopy.com} ; Wed, 23 Aug 2000 18:05:48 -0500 (CDT) Received: from connelly ([205.139.92.108]) by mail.gatan.com (8.9.3/8.8.7) with SMTP id PAA15020 for {microscopy-at-msa.microscopy.com} ; Wed, 23 Aug 2000 15:13:36 -0700 Reply-To: {tgc-at-gatan.com}
-----Original Message----- } From: tgc [mailto:tgc-at-gatan.com] Sent: Wednesday, August 23, 2000 3:43 PM To: microscopy-at-msa.microscopy.com
Hi Bob, We use the frozen block technique with a Hitachi 2250N (earlier model than yours, also fitted with Peltier and ln2-cooled stages) quite often for quick observations. It works well, but you only have a limited time (few minutes) before the ice sublimes, followed by 15-30 minutes of good stable viewing of the freeze-drying sample - if you want to examine ice distribution in situ with any accuracy at all, I dont think there is any alternative to a controllable ln2-cooled stage. Working nearer room temp with a Peltier stage at higher chamber water pressures probably wouldnt help very much either, since you would be balancing condensation/sublimation...
good luck!
Sally Stowe
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
} } } Bob Wise {wise-at-vaxa.cis.uwosh.edu} 08/24/00 03:09am } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I would like to try to visualize ice in frozen plant leaves in a Hitachi 2460 variable pressure SEM and do not have access to a cold stage. I have heard that a block of aluminum or copper at LN temperature may make a usable substitute. Has anyone had any experience with this technique? Any and all tips would be appreciated.
TIA
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
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Microscopy Core Technician.
Cold Spring Harbor Laboratory is seeking an experienced and responsible microscopy technician for the laboratories core microscopy facility. The individual should have practical expertise in transmission electron microscopy, confocal and widefield fluorescence microscopy, digital imaging, and microinjection. The successful candidate will be involved in designing and carrying out experimental protocols for users, training individuals in the use of various microscopes, and aligning microscopes and keeping the facility operating at an efficient and high level of productivity. Interested individuals should send their resume, including a description of their expertise and the names and addresses of 3 references to: Dr. David L. Spector, Cold Spring Harbor Laboratory, One Bungtown Road, Cold Spring Harbor, New York 11724, email: spector-at-cshl.org
I to was very impressed with the AMT, SIS, and Gatan cameras at the M&M 2000. I have a 1986 Hitachi 7000 STEM and would like to know if any one out there has any digital experience with this make and model. Thanks, Tim
Timothy G. Schneider Director of Electron Microscopy Department of Pathology Room 229 Jefferson Hall Thomas Jefferson University 1020 Locust St. Philadelphia Pa. 19107 215-503-4798 work 610-613-8170 cellular timothy.schneider-at-mail.tju.edu
I am looking for a introductory technical reference for the Focused Ion Beam (FIB) technique. A journal article or textbook section would be useful. Can you suggest a comprehensive source?
After consulting with Dr. Garber, we are in general agreement on his position concerning shipping and storing of GA. We believe adding cold shipping requirements are overkill, but at the same token if any of our customers feel it is necessary we are glad to oblige. We have a long history of producing highly purified GA and in forty some years have never had a problem with our guarantee of one year at RT. There are conditions that could merit special packaging and shipping and we would be glad to conform to any requests, but webelieve it to be an unnecessary expense for most people.
John Arnott Chairman
--
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
Microscope Supplies Since 1955
} } Jim Darley wrote: } } ============================================================= } } Just to make the "GA is old" point a little clearer: } } GA polymerises with time and higher temperatures; a nitrogen head does not } } change much. Frozen, GA will last an awfully long time and if stored } } refrigerated most people would still be happy with the product after six } } months } } and more. While in the lab storage should not be a problem, we can be sure } } that } } inappropriate storage is common. } } The problem is in transit, when during the summer months the GA could see } } some } } rather high temperatures during a truck or even an air journey. We have } } always } } taken some care with GA, but being a long way from the manufacturer we have } } adopted special precautions such as shipping with ice and mostly by air and } } incoming shipments are also packed in ice and refrigerated whenever not in } } the } } air. Its costly, but this assures a better product. } } (snip) } } ==============================================================
Jim Darley wrote: ============================================================= Just to make the "GA is old" point a little clearer: GA polymerises with time and higher temperatures; a nitrogen head does not change much. Frozen, GA will last an awfully long time and if stored refrigerated most people would still be happy with the product after six months and more. While in the lab storage should not be a problem, we can be sure that inappropriate storage is common. The problem is in transit, when during the summer months the GA could see some rather high temperatures during a truck or even an air journey. We have always taken some care with GA, but being a long way from the manufacturer we have adopted special precautions such as shipping with ice and mostly by air and incoming shipments are also packed in ice and refrigerated whenever not in the air. Its costly, but this assures a better product. (snip) ============================================================== Sometimes good technical people have honest differences of opinion. I am sure this is one of those instances.
I have always been led to believe that the mechanism of ageing of GA is related to the formation first of dimers and trimers of the monomer (it is an autocatalytic reaction), and that the way to ensure longest shelf life is to bring the starting purity down to levels (actually lower than I believe most EM users would need) such that one can really delay (but not indefinitely) the onset of the dimer/trimer reaction from starting. By removing the existing dimers and trimers, you remove that which starts the aging process.
When GA is purified to these levels, one can delay the onset of the aging process to such a degree, that some producers of EM glutaraldehyde guarantee their product to be good for one year at room temperature storage. SPI is one of those and as of today, I reconfirmed that the Ladd glutaraldehyde is in this category as well. Quite possibly, if not probably, there are other vendors making EM glut at that same level of purity.
But with glutaraldehyde being purified to these levels, and with these guarantees from the manufacturer for one year at room temperature, to require cold shipment in ice, or even the use of frozen gel packs (which usually are cheaper) in our opinion, is normally not required. Naturally, upon arrival, we do recommend refrigerated storage, just out of general principles, which might be more the result of "dogma" than real reasons. But we ship regularly the ampouled GA to tropical environments, unrefrigerated, and don't have any particular problems. We would discourage shipment to tropical environments via air freight, since it could end up sitting in customs for days on end but use of the courier services seems to result in no difficulties.
Now for GA being sold in screw top bottles, this would be another story. If the starting purity is good as indicated above, then refrigerated shipment should not, in general be necessary. However, we would strongly recommend the refrigerated storage upon arrival, indeed we think this should be a requirement. The guarantees for room temperature storage for one year do not extend to the product sold in the screw top bottles, only in the sealed glass ampoules. We don't know about frozen storage, but usually most users want their product in ready-to-use form and don't want to wait for something to thaw out before using.
Again, different people have different opinions about these matters, and the purpose of this posting is only to point out that there are these differences, and that we do not believe one needs to regularly incur the extra costs that would otherwise be necessary for the refrigerated shipment of GA.
Chuck
Disclaimer: SPI Supplies offers EM grade glutaraldehyde to EM laboratories worldwide and we do not refrigerate our shipments of GA and to my knowledge, we have never had a customer receive GA that has gone bad in shipment.
============================================
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 ######################## ============================================
Someone has asked what resin to use that would be clear and suitable for embedding mouse embryos. This would be for display, not for sectioning.
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 never heard of those dimmers and trimmers, just seems a change from various polymers and co-polymers. More importantly, I do not believe that Chuck's supplier has something better than vacuum distillation available to achieve highest purity. Not that I would want to get into the game of "mine is purer than yours", since I have no doubt that at production either of the three (to my knowledge) North American "refiners" produce the very best GA possible and they are comparable. The main points of this message are that: 1 For infusion and I understand cyto or immunocytochemistry the purest possible GA should be used. I have read the suggestion that a small amount of polymer may be good during conventional fixation, as this would impede osmotic shock. 2 I certainly do not agree with Chucks suggestions that GA may be stored at room temperature for a year and then used for EM fixation - regardless of initial purity. GA is not in this regard to be confused with the high purity Formaldehyde (16%), which does not require refrigeration, keeps for years and is an excellent fixative. That item come after Chuck's "snip". Hmmm. 3 The point of cold shipping that I had made was particularly pertinent to our situation were the material needs to travel a long distance. But especially during our summer months (and the USA can be just as hot) I believe that shipping GA with some ice is worth the extra cost. 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 ABN: 99 724 136 560 www.proscitech.com
On Friday, August 25, 2000 9:08 AM, Garber, Charles A. [SMTP:cgarber-at-2spi.com] wrote: } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } Jim Darley wrote: } ============================================================= } Just to make the "GA is old" point a little clearer: } GA polymerises with time and higher temperatures; a nitrogen head does not } change much. Frozen, GA will last an awfully long time and if stored } refrigerated most people would still be happy with the product after six } months } and more. While in the lab storage should not be a problem, we can be sure } that } inappropriate storage is common. } The problem is in transit, when during the summer months the GA could see } some } rather high temperatures during a truck or even an air journey. We have } always } taken some care with GA, but being a long way from the manufacturer we have } adopted special precautions such as shipping with ice and mostly by air and } incoming shipments are also packed in ice and refrigerated whenever not in } the } air. Its costly, but this assures a better product. } (snip) } ============================================================== } Sometimes good technical people have honest differences of opinion. I am } sure this is one of those instances. } } I have always been led to believe that the mechanism of ageing of GA is } related to the formation first of dimers and trimers of the monomer (it is } an autocatalytic reaction), and that the way to ensure longest shelf life is } to bring the starting purity down to levels (actually lower than I believe } most EM users would need) such that one can really delay (but not } indefinitely) the onset of the dimer/trimer reaction from starting. By } removing the existing dimers and trimers, you remove that which starts the } aging process. } } When GA is purified to these levels, one can delay the onset of the aging } process to such a degree, that some producers of EM glutaraldehyde guarantee } their product to be good for one year at room temperature storage. SPI is } one of those and as of today, I reconfirmed that the Ladd glutaraldehyde is } in this category as well. Quite possibly, if not probably, there are other } vendors making EM glut at that same level of purity. } } But with glutaraldehyde being purified to these levels, and with these } guarantees from the manufacturer for one year at room temperature, to } require cold shipment in ice, or even the use of frozen gel packs (which } usually are cheaper) in our opinion, is normally not required. Naturally, } upon arrival, we do recommend refrigerated storage, just out of general } principles, which might be more the result of "dogma" than real reasons. } But we ship regularly the ampouled GA to tropical environments, } unrefrigerated, and don't have any particular problems. We would } discourage shipment to tropical environments via air freight, since it could } end up sitting in customs for days on end but use of the courier services } seems to result in no difficulties. } } Now for GA being sold in screw top bottles, this would be another story. If } the starting purity is good as indicated above, then refrigerated shipment } should not, in general be necessary. However, we would strongly recommend } the refrigerated storage upon arrival, indeed we think this should be a } requirement. The guarantees for room temperature storage for one year do } not extend to the product sold in the screw top bottles, only in the sealed } glass ampoules. We don't know about frozen storage, but usually most users } want their product in ready-to-use form and don't want to wait for something } to thaw out before using. } } Again, different people have different opinions about these matters, and the } purpose of this posting is only to point out that there are these } differences, and that we do not believe one needs to regularly incur the } extra costs that would otherwise be necessary for the refrigerated shipment } of GA. } } Chuck } } Disclaimer: SPI Supplies offers EM grade glutaraldehyde to EM laboratories } worldwide and we do not refrigerate our shipments of GA and to my knowledge, } we have never had a customer receive GA that has gone bad in shipment. } } ============================================ } } 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 } ######################## } ============================================ } } } } } } }
Thanks to everyone who answered my query about the Topographic Measurement paper by Dr John Russ and for all the helpful suggestions. The problem seems to have fixed itself. This morning, after about 10 more attempts, I was suddenly able to download the file. I have no idea what the problem was, and neither does our IT department. It certainly wasn't the Adobe Acrobat Reader or Internet Explorer. I have learned long ago that computers are strange things, and this just proves it again !!!
Thanks Willem 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 store 25% glutaraldehyde in ampoules in the freezer compartment of a domestic fridge . It does not freeze at this temperature and is therefore ready for use very quickly.
Dave
On Thu, 24 Aug 2000 18:07:53 -0500 "Garber, Charles A." {cgarber-at-2spi.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 } -----------------------------------------------------------------------. } } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } Jim Darley wrote: } ============================================================= } Just to make the "GA is old" point a little clearer: } GA polymerises with time and higher temperatures; a nitrogen head does not } change much. Frozen, GA will last an awfully long time and if stored } refrigerated most people would still be happy with the product after six } months } and more. While in the lab storage should not be a problem, we can be sure } that } inappropriate storage is common. } The problem is in transit, when during the summer months the GA could see } some } rather high temperatures during a truck or even an air journey. We have } always } taken some care with GA, but being a long way from the manufacturer we have } adopted special precautions such as shipping with ice and mostly by air and } incoming shipments are also packed in ice and refrigerated whenever not in } the } air. Its costly, but this assures a better product. } (snip) } ============================================================== } Sometimes good technical people have honest differences of opinion. I am } sure this is one of those instances. } } I have always been led to believe that the mechanism of ageing of GA is } related to the formation first of dimers and trimers of the monomer (it is } an autocatalytic reaction), and that the way to ensure longest shelf life is } to bring the starting purity down to levels (actually lower than I believe } most EM users would need) such that one can really delay (but not } indefinitely) the onset of the dimer/trimer reaction from starting. By } removing the existing dimers and trimers, you remove that which starts the } aging process. } } When GA is purified to these levels, one can delay the onset of the aging } process to such a degree, that some producers of EM glutaraldehyde guarantee } their product to be good for one year at room temperature storage. SPI is } one of those and as of today, I reconfirmed that the Ladd glutaraldehyde is } in this category as well. Quite possibly, if not probably, there are other } vendors making EM glut at that same level of purity. } } But with glutaraldehyde being purified to these levels, and with these } guarantees from the manufacturer for one year at room temperature, to } require cold shipment in ice, or even the use of frozen gel packs (which } usually are cheaper) in our opinion, is normally not required. Naturally, } upon arrival, we do recommend refrigerated storage, just out of general } principles, which might be more the result of "dogma" than real reasons. } But we ship regularly the ampouled GA to tropical environments, } unrefrigerated, and don't have any particular problems. We would } discourage shipment to tropical environments via air freight, since it could } end up sitting in customs for days on end but use of the courier services } seems to result in no difficulties. } } Now for GA being sold in screw top bottles, this would be another story. If } the starting purity is good as indicated above, then refrigerated shipment } should not, in general be necessary. However, we would strongly recommend } the refrigerated storage upon arrival, indeed we think this should be a } requirement. The guarantees for room temperature storage for one year do } not extend to the product sold in the screw top bottles, only in the sealed } glass ampoules. We don't know about frozen storage, but usually most users } want their product in ready-to-use form and don't want to wait for something } to thaw out before using. } } Again, different people have different opinions about these matters, and the } purpose of this posting is only to point out that there are these } differences, and that we do not believe one needs to regularly incur the } extra costs that would otherwise be necessary for the refrigerated shipment } of GA. } } Chuck } } Disclaimer: SPI Supplies offers EM grade glutaraldehyde to EM laboratories } worldwide and we do not refrigerate our shipments of GA and to my knowledge, } we have never had a customer receive GA that has gone bad in shipment. } } ============================================ } } 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 } ######################## } ============================================ } } } } } } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
Is anyone familiar with the staining of silver-deposits in rat-tissues. Organs which may be involved are liver and kidneys. Is a silver-deposit within tissue possible?
Joost Bruijntjes TNO Nutrition and Food Research Zeist Holland
Exciting Opportunity for an Experienced Trasmission Electron Microscopist in Advanced Imaging and Measurement, Unilever Research, Edgewater, NJ
Position Requirements:
A full time position is immediately available for a person experienced in transmission electron microscopy of biological samples. This person will be responsible for all aspects of tissue processing, sectioning, TEM image acquisition as well as general laboratory maintenance. At least a bachelors degree in biology is required with five years of experience in bio-TEM. Previous experience in immuno-labeling, digital data acquisition and computer image processing is desirable.
Opportunity exists for the person to expand skills and capabilities in cryo-aplications (high pressure freezeing, cryo-ultramicrotomy, freze substitution/fracture etc) and environmental scanning electron microscopy.
Unilever is a leading consumer products company with world wide sales exceeding $40 billion. To support our global business, Unilever has six international research center employing over 4000 scientists. Unilever's U.S. laboratory employs over 300 scientists and is the designated headquarters for leading personal wash and skin care research. We are located on the Hudson River in Edgewater, New Jersey, within 20 minutes of midtown Manhattan with its diverse entertainment, arts and cultural resources. We offer a competitive salary, flexible benefits and excellent opportunities for both personal and professional growth. For more information about Unilever Research and Unilever visit our Web Site at http://www.unilever.com
If interested, please send resume to:
Manoj Misra Ph.D. Group Head Advanced Imaging and Measurement Unilever Research Edgewater, NJ 07020 E Mail: manoj.misra-at-unilever.com
What influence does the concentration of glutaraldehyde have on its shelf life? In the 'frig or at room temperature.
I've always thought that the higher % glut -- especially 70% -- was more likely to polymerize, and that 25% or lower had the longest shelf life. True or false?
I am assuming glutaraldehyde stored under nitrogen in sealed ampoules.
Phil
} -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } Jim Darley wrote: } ============================================================= } Just to make the "GA is old" point a little clearer: } GA polymerises with time and higher temperatures; a nitrogen head does not } change much. Frozen, GA will last an awfully long time and if stored } refrigerated most people would still be happy with the product after six } months } and more. While in the lab storage should not be a problem, we can be sure } that } inappropriate storage is common. } The problem is in transit, when during the summer months the GA could see } some } rather high temperatures during a truck or even an air journey. We have } always } taken some care with GA, but being a long way from the manufacturer we have } adopted special precautions such as shipping with ice and mostly by air and } incoming shipments are also packed in ice and refrigerated whenever not in } the } air. Its costly, but this assures a better product. } (snip) } ============================================================== } Sometimes good technical people have honest differences of opinion. I am } sure this is one of those instances. } } I have always been led to believe that the mechanism of ageing of GA is } related to the formation first of dimers and trimers of the monomer (it is } an autocatalytic reaction), and that the way to ensure longest shelf life is } to bring the starting purity down to levels (actually lower than I believe } most EM users would need) such that one can really delay (but not } indefinitely) the onset of the dimer/trimer reaction from starting. By } removing the existing dimers and trimers, you remove that which starts the } aging process. } } When GA is purified to these levels, one can delay the onset of the aging } process to such a degree, that some producers of EM glutaraldehyde guarantee } their product to be good for one year at room temperature storage. SPI is } one of those and as of today, I reconfirmed that the Ladd glutaraldehyde is } in this category as well. Quite possibly, if not probably, there are other } vendors making EM glut at that same level of purity. } } But with glutaraldehyde being purified to these levels, and with these } guarantees from the manufacturer for one year at room temperature, to } require cold shipment in ice, or even the use of frozen gel packs (which } usually are cheaper) in our opinion, is normally not required. Naturally, } upon arrival, we do recommend refrigerated storage, just out of general } principles, which might be more the result of "dogma" than real reasons. } But we ship regularly the ampouled GA to tropical environments, } unrefrigerated, and don't have any particular problems. We would } discourage shipment to tropical environments via air freight, since it could } end up sitting in customs for days on end but use of the courier services } seems to result in no difficulties. } } Now for GA being sold in screw top bottles, this would be another story. If } the starting purity is good as indicated above, then refrigerated shipment } should not, in general be necessary. However, we would strongly recommend } the refrigerated storage upon arrival, indeed we think this should be a } requirement. The guarantees for room temperature storage for one year do } not extend to the product sold in the screw top bottles, only in the sealed } glass ampoules. We don't know about frozen storage, but usually most users } want their product in ready-to-use form and don't want to wait for something } to thaw out before using. } } Again, different people have different opinions about these matters, and the } purpose of this posting is only to point out that there are these } differences, and that we do not believe one needs to regularly incur the } extra costs that would otherwise be necessary for the refrigerated shipment } of GA. } } Chuck } } Disclaimer: SPI Supplies offers EM grade glutaraldehyde to EM laboratories } worldwide and we do not refrigerate our shipments of GA and to my knowledge, } we have never had a customer receive GA that has gone bad in shipment. } } ============================================ } } 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 } ######################## } ============================================
-- }}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{ 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)
Another 2 cents worth. I have sat back and read with enjoyment all of the bantering regarding the facts and fiction on Glut and cannot believe that some people are actually arguing about issues which are so well cited in the literature and this argument is going to lead to the confusion of many researchers with no basis at all. The truth on Glut is short and simple and I do not care who is making it the facts remain the same no matter. Please see the facts below. Anyone whom would like to question these results I remain at your disposal to open my lab notebooks and results of all of the Glutaraldehyde in the industry today and its corresponding results. There are only 3 manufacturers of the final distilled product in the US and each and every one of them shows the same tendency towards temp vs time.
GLUTARALDEHYDE STORAGE ISSUES
A key detail in storage analysis for this product is that EM grade GA is usually supplied as "aqueous solution" of various concentrations and not as "pure", and the "pure iron won't rust" principle simply does not apply. This aqueous chemical system is well known to be extremely active, especially at temperatures exceeding +40 C (because of common chemical knowledge reasons, pertaining to water molecules intrinsic association tendency, on one hand, and to the particular GA_dimer polarisation of hydrating aggregates, on the other hand). As many EM users already know, GA polimerizes under the influence of traces of impurities, the process belonging to singlet-triplet activation mechanism (i.e., temperature, pH, concentration highly sensitive), and by no means is it an "autocatalytic" one. Traces of water (along with acrolein, ethanol, methanol, pyranic compounds), and a slight azeotropic tendency, lead ultimately to the formation of glutaric acid and consequently to the drastic decrease of pH, making GA improper for EM usage, as in: Hayat, M.A., "Principles And Techniques of Electron Microscopy", 2000. In those new created conditions, GA quickly reversibly turns to several polymeric species, some of them obviously detectable, some not. Alkaline artificially buffered solutions cannot prevent polymerisation either, and the process starts with a GA_trimer, as in: Fahimi, HD., et.al., "Essais de standardisation de la fixation en glutaralgehyde", J. Microsc. (Paris, France), 4: 725-736, 1965 (!!!) In time, several important kinetic stability studies were performed upon EM grade GA. For general information, and, for most of readers, as a kind reminder, we had selected conclusions made public in Histochemie, 30, 1972 (!!!), by Gillet, R., et.al., from Queen Elizabeth College, London, England: "if the absorbance"..."is plotted against storage temperature, a direct relationship is obtained between storage temperature (above 00 C) and the appearance of"... spectral ... "absorbing material".
(Please see graph attached.)
The authors wrote: "the most important storage criterion was definitely temperature. Those samples stored at -200 C showed virtually no change in"..."absorbance characteristics, even after eight months storage" ..."all samples stored at +200 C, however, showed considerable absorbance peaks"..."the effect of storage at +370 C"..."was even more pronounced." It appears very speculative and pragmatic to neglect such important facts, or categorize them as "dogmas". Certain manufacturers, who did not receive any complaints (yet) when delivering room temperature stored GA, should consider themselves extremely lucky, and should scrupulously revise their knowledge about EMS grade GA, regardless if sold in ampoules or screw cap bottles.
I look forward to hearing from anyone whom does not agree.
All of the very best Stacie Kirsch Electron Microscopy Sciences 215-646-1566
Please advice - I desperately need a source of info for comparative analysis of main existing microscope types - standard transmittance microscope,phase-contrast, interference, fluorescent, confocal, laser scanning ...in terms of their baic parameters: spatial resolution, sensitivity to light absorption, temporal resolution. Please advice a web-source or some other on-line esource if possible - in case of book/article I will not have time for ordering.
Thanks
Dmitri Lapotko,
Luikov Heat and Mass Transfer Institute Minsk Ld-at-hmti.ac.by
Not sure if this is the right forum to help me with this issue but thought I would give it a try. I have a student who wants to cut bone (non fossil) samples. She is not necessarily making thin sections but wants a cut better than can be done with a coping saw. I offered my Buehler Isomet low speed saw with a diamond cut off blade but that seem to be a bit too slow and awkward with the larger bones. Looking for ideas.
Thanks
Roy Beavers Southern Methodist University Dept. of Geological Sciences Electron Microprobe Lab P.O. Box 750395 Dallas, Tx 75275 voice: 214-768-2756 fax: 214-768-2701 E-mail: rbeavers-at-mail.smu.edu
Being an old timer, I have been around since before the advent of vials of GA. In the 70's the Merck index described a method for purifying GA. using activated charcoal.
Basically GA begins to degrade to "multimers" when stored at room temperature. The best fixation is achieved with the monomeric form, therefore the purification protocol was designed to remove the more complex forms.
Storage under nitrogen in a sealed vial slows the degradation. Lower concentration degrade more slowly. Storing at 4C is advised.
Method: Mix the concentrated liquid glutaraldehyde (25%) with 1g of activated charcoal per 100 ml of liquid. Allow to stand for 30 minutes, then vacuum filter through a Whatman #1 filter. Repeat three times. Store at 4C Repeat once a year.
I have used this method for more than 30 years and consistently obtained very satisfactory fixation. It is messy, but worth the effort.
I am helping, or trying to help, a colleague with sectioning of Drosophila legs. They are embedded in "Immunobed" (a methycrylate) or Epon (ok, Epon substitute). I don't seem to be getting a good bond between the chitin and the plastic, the tissue falls out. Any suggestions are welcome.
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 **********************************************
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Hello,
We inherited an old LKB grid stainer that has worked well for nearly two years. Now it has developed a leak.
Is there anyone who has one of these machines that they keep for spares (or to hold the door open)? I need the spindle unit for one of the pump/valve assemblies. The housing looks fine so it should be easy enough to plug in a spare unit.
For anyone who has not seen inside this machine, the above description must seem very strange. My apologies for this. My hope is that someone who has seen inside can understand my description and has the part I need.
I have already contacted Leica to see if they are still supporting this machine, so there is no need to recommend I contact them.
Thanks in advance for any help, suggestions, comments or any other e-mail you may send my way.
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
by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id PAA23100 for dist-Microscopy; Fri, 25 Aug 2000 15:55:24 -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 PAA23097 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Fri, 25 Aug 2000 15:54:53 -0500 (CDT) Received: from mail1.doit.wisc.edu (mail1.doit.wisc.edu [144.92.9.40]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id PAA23090 for {Microscopy-at-sparc5.microscopy.com} ; Fri, 25 Aug 2000 15:54:42 -0500 (CDT) Received: from [144.92.10.114] by mail1.doit.wisc.edu id PAA71884 (8.9.1/50); Fri, 25 Aug 2000 15:48:56 -0500 Mime-Version: 1.0 Message-Id: {p04320421b5cc8bee034b-at-[144.92.10.114]} In-Reply-To: {E769E1FA6D4AD311B0190008C7E65014012E44C4-at-s31xe1.systems.smu.edu} References: {E769E1FA6D4AD311B0190008C7E65014012E44C4-at-s31xe1.systems.smu.edu}
Roy,
If you don't get the answer from the microscopy group, you might try Histonet: Histonet-at-Pathology.swmed.edu to subscribe, put "subscribe" no quotes in the subject line, leave the message body blank unsubscribe the same way. Spell these correctly! They use one address for everything, so "suscribe" or "subscribe" with quotes gets sent to the list as a message, not read as a command by the serverbot. They have a bunch of folks who specialize in cutting bone, calcified and decalcified. Their archives can be searched at: http://www.histosearch.com/histonet.html
Otherwise, does someone at SMU have a diamond wire saw? Those work well for slicing intact bones. There's also a double-bladed hand bone saw that makes nice slices. Someone at the SMU med school or hospital ought to have one.
Phil
} Not sure if this is the right forum to help me with this issue but thought } I would give it a try. I have a student who wants to cut bone (non fossil) } samples. She is not necessarily making thin sections but wants a cut better } than can be done with a coping saw. I offered my Buehler Isomet low speed } saw with a diamond cut off blade but that seem to be a bit too slow and } awkward with the larger bones. Looking for ideas. } } Thanks } } Roy Beavers } Southern Methodist University } Dept. of Geological Sciences } Electron Microprobe Lab } P.O. Box 750395 } Dallas, Tx 75275 } voice: 214-768-2756 } fax: 214-768-2701 } E-mail: rbeavers-at-mail.smu.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)
If you have a machine shop cut the bone in to thin sections on a band saw sand one side smooth and super glue the bone to a disk you can mount in a lathe or a plate you can mount on a mill or grinder. Then use thin the material as thin as you want with a grinder.
Acetone will turn them loose from the plate. You can do a bunch of sections at a time on a big plate.
Embeding them before you grind them will help preserve fine structures.
Exactly how the machine shop does the work will depend on the tools they have available.
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00 - } Not sure if this is the right forum to help me with this issue but thought } I would give it a try. I have a student who wants to cut bone (non fossil) } samples. She is not necessarily making thin sections but wants a cut better } than can be done with a coping saw. I offered my Buehler Isomet low speed } saw with a diamond cut off blade but that seem to be a bit too slow and } awkward with the larger bones. Looking for ideas.
I too purified GA by charcoal filtering and even vacuum distillation; its over 25 years ago. As things were published I also played around with making thin film apertures and I know that other people have made their own filaments and even the making of a LaB6 is published in the literature, for the intrepid. Don't think that I worry about loosing some business either, that too is not one of my traits. I like to make the following observations on purifying GA, we keep cathodes for another day: Charcoal filtered GA is not as good the commercial vacuum distilled product. You should store it in a freezer (even a domestic fridge's') if its for more then a few weeks. The triple charcoal filtered product is good enough for EM fixation, if stored well. It's dubious that its good enough for cytochemistry. Charcoal is a recognized carcinogen (funny that, it used to be prescribed against diarrhea) its also expensive and a DG to ship. Several ml of GA are lost in every filtration. Any employer would cost an employee at at least 2x the hourly cost; in research its easy to forget the real world, were time is money. "Home filtered" GA is very expensive.
Different parameters may apply elsewhere, so in countries were the hourly wage is 50 cents or less, and a drum of GA fell of a truck at a leather tanning or paper towel factory (its used to soften leather and paper) and somebody is making coconut charcoal down the road . . . I would be into that and make my own purified GA too. But not in America! 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 ABN: 99 724 136 560 www.proscitech.com
On Saturday, August 26, 2000 5:35 AM, L R MELSEN [SMTP:lmelsen-at-emory.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 } -----------------------------------------------------------------------. } } } Being an old timer, I have been around since before the advent of vials } of GA. In the 70's the Merck index described a method for purifying GA. } using activated charcoal. } } Basically GA begins to degrade to "multimers" when stored at room } temperature. The best fixation is achieved with the monomeric form, } therefore the purification protocol was designed to remove the more } complex forms. } } Storage under nitrogen in a sealed vial slows the degradation. Lower } concentration degrade more slowly. Storing at 4C is advised. } } Method: } Mix the concentrated liquid glutaraldehyde (25%) with 1g of activated } charcoal per 100 ml of liquid. } Allow to stand for 30 minutes, then vacuum filter through a Whatman #1 } filter. } Repeat three times. } Store at 4C } Repeat once a year. } } I have used this method for more than 30 years and consistently obtained } very satisfactory fixation. It is messy, but worth the effort. }
..We have decided as a department to close down our transmission EM } } operation. We have a Philips EM 300 ... which we are willing to donate to someone for parts, provided they pay for disassembly and shipping to their site...
For details please contact: } } } David Kerk, Ph.D. } } } Professor and Chair } } } Department of Biology } } } Point Loma Nazarene University } } } 3900 Lomaland Dr } } } San Diego, CA 92106 } } } Ph: 619-849-2398 } } } FAX: 619-849-2598 } } } email: dkerk-at-ptloma.edu
M. Malecki, M.D., Ph.D. Director and Principal Investigator
Molecular Imaging Laboratories University of California at San Diego address: 1500 Suite Bonner Hall, 9500 Gilman Drive, La Jolla, CA 92093-0368 phone: 8588223373 fax: 8588223715 pager: 8586161420 e.mail: mmm-at-ucsd.edu e.pager: 1620024619-at-alphapage.airtouch.com www site: http://mil.ucsd.edu/ ftp site: mil1.ucsd.edu
As an alternative to charcoal purification of GA described by Dr. MELSEN, I run distillation, collect the fractions, and then select the monomeric form on the spec.
} Date: Fri, 25 Aug 2000 15:35:12 -0400 } From: L R MELSEN {lmelsen-at-emory.edu} } Reply-To: lmelsen-at-emory.edu } Organization: Emory University } X-Mailer: Mozilla 4.73 (Macintosh; I; PPC) } X-Accept-Language: en } To: MICROSCOPY {Microscopy-at-sparc5.microscopy.com} } Subject: glutaraldehyde } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
M. Malecki, M.D., Ph.D. Director and Principal Investigator
Molecular Imaging Laboratories University of California at San Diego address: 1500 Suite Bonner Hall, 9500 Gilman Drive, La Jolla, CA 92093-0368 phone: 8588223373 fax: 8588223715 pager: 8586161420 e.mail: mmm-at-ucsd.edu e.pager: 1620024619-at-alphapage.airtouch.com www site: http://mil.ucsd.edu/ ftp site: mil1.ucsd.edu
} Someone has asked what resin to use that would be clear and suitable for } embedding mouse embryos. This would be for display, not for sectioning. }
Dear Tina, How about lucite? I have no idea where to get it, but many years ago I bought a lucite cube in which a dandilion had been embedded. You could see it very clearly, and there is obviously a technique for preserving such a fragile specimen. Note that this is more along the lines of an existance theorem than a procedure. Good luck. Yours, Bill Tivol
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Tina:
The best material to use in display type applications is a polyester material. We have a material called PolyMet which is ideal. It takes a while to cure - typically overnight - but it provides a crystal clear mount. I hope this helps.
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 Tina Carvalho } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Colleagues-
Someone has asked what resin to use that would be clear and suitable for embedding mouse embryos. This would be for display, not for sectioning.
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* *************************************************************************** *
Based on recent communications it appears there is a difference of opinion on the shipping of refined Glut among the three major U.S. refiners.
Ladd and a second refiner who has not been heard from do not add ice. The third refiner does. When adding extras there is a higher cost and, though we don't always acknowledge it, these costs do get passed on to the consumer.
All three refiners know their specific product and their techniques and make their decisions on that basis. If the refiner who adds ice feels their product requires it I have great respect for that decision. They know their product and know the best way to get it to their customer.
All data and history indicates Ladd glut does not require ice. If special circumstances arise we would be glad to add ice.
We all act in good faith. If the refiner who adds ice is right and the others are wrong I will acknowledge and applaud that decision. We do add cold packaging for other chemicals like Mercox and LR White when required.
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
Hello, One day someone said that microwave oven is very dangerous to be operated in a level with the operator. Any advise, article in this matter? Thanks Mansour Al-Shafei Senior Lab Sci. Saudi Aramco Oil Company Lab Research & Development Center Analytical Sciences Division Advanced Instruments Unit Voice: 966-3-876-4360 E-mail: shafeima-at-aramco.com.sa
} Someone has asked what resin to use that would be clear and suitable for } embedding mouse embryos. This would be for display, not for sectioning. }
Dear Tina,
There is also a product called Cast N' Craft casting resin available at craft stores (Pearl, etc.) which I have had some success. You can not cast under vacuum however you get a very clear, bubble free final product. For spiders and smaller insects its been O.K but you may have difficulty with any large objects. It's only a few bucks and may be worth a shot [provided you have a few embryos to spare :)]. Good Luck.
Regards,
George R. Munzing Jr.
William Tivol {tivol-at-wadsworth.org} -at-wadsworth.org on 08/28/2000 11:42:03 AM
Sent by: tivol-at-wadsworth.org
To: microscopy-at-sparc5.microscopy.com cc:
Tina Carvalho wrote:
} Someone has asked what resin to use that would be clear and suitable for } embedding mouse embryos. This would be for display, not for sectioning. }
Dear Tina, How about lucite? I have no idea where to get it, but many years ago I bought a lucite cube in which a dandilion had been embedded. You could see it very clearly, and there is obviously a technique for preserving such a fragile specimen. Note that this is more along the lines of an existance theorem than a procedure. Good luck. Yours, Bill Tivol
{html} {font face="Times New Roman, Times"} A postdoctoral scientist is sought for a joint research program between the University of Virginia and the IBM Thomas J. Watson Research Center, centered upon real-time electron microscope studies of the evolution of epitaxial semiconductor {br} nanostructures. This position will be funded by an anticipated National Science Foundation award to establish a Materials Research Science and Engineering Center (MRSEC) at the University of Virginia. The successful applicant will be an employee of the Department of Materials Science and Engineering at the University of Virginia, but will spend the bulk of his / her time based at IBM, using the unique in-situ imaging and {br} deposition tools in the Nanoscale Materials Analysis Department in the Physical Sciences Division at the Thomas J. Watson Research Center. {br} {br} Applicants will require a PhD (or equivalent) in Materials Science, Physics or a related discipline. The successful candidate will be able to demonstrate strong background in electron microscopy, and will preferably have expertise in ultra-high vacuum techniques. A research background in electronic materials will also be a strong asset. {br} {br} Review of applications will begin on Sep 15, and will continue until the position is filled. The starting date will be in the October November 2000 timeframe. The initial appointment will be for one year, and will be extendible annually for an additional two years, depending upon continuing funding. Applications should be addressed to: Professor Robert Hull, Department of Materials Science and Engineering, University of Virginia, 116 Engineers Way, Charlottesville, Virginia 22904. Please provide a curriculum vitae, copies of up to five recent relevant publications, and contact information for three referees familiar with your work and accomplishments. {br} {br} Additional information on the Department of Materials Science and Engineering may be found at {a href="http://www.mse.virginia.edu/" eudora="autourl"} http://www.mse.virginia.edu {/a} , and on the IBM Thomas J. Watson Research Center at {a href="http://www.research.ibm.com/" eudora="autourl"} www.research.ibm.com {/a} . The University of Virginia and IBM are Equal Opportunity / Affirmative Action employers. Applicants must be able to lawfully accept employment in the United States. {br} {br} {/font} {BR} {br} {div} Robert Hull {/div} {div} Department of Materials Science and Engineering {/div} {div} University of Virginia {/div} {div} 116 Engineers Way {/div} {div} Charlottesville, VA 22904-4745 {/div} {br} {div} Tel: 804-982-5658 {/div} {div} FAX: 804-982-5660 {/div} email:hull-at-virginia.edu {/html}
Modern microwave ovens are so well shielded that you cannot get a reading with the sensitive meters that we supply. The only way you can show that the meter is working is to place it within the microwave - for a few seconds. I'd rather worry about gamma rays from outer space. 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 ABN: 99 724 136 560 www.proscitech.com
On Tuesday, August 29, 2000 9:53 PM, Shafei, Mansour A [SMTP:shafeima-at-aramco.com.sa] 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, } One day someone said that microwave oven is very dangerous to be operated in } a level with the operator. Any advise, article in this matter? } Thanks } Mansour Al-Shafei } Senior Lab Sci. } Saudi Aramco Oil Company } Lab Research & Development Center } Analytical Sciences Division } Advanced Instruments Unit } Voice: 966-3-876-4360 } E-mail: shafeima-at-aramco.com.sa } }
I am seeking advice on freeze-fracture EM. I am particularly interested in bacterial membranes and cell walls, and those who do immunolocalization on these kinds of preparations.
Please contact me OFF LIST at: 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 308 30 Convent Drive Bethesda MD 20892 ph 301-594-0025 fax 301-402-0396
We have been looking at the use of various catalysts for production carbon nanotubes. Recently we had a sample that behaved in a very unusual way. Upon focusing the electron beam on a cluster of nanotubes, the catalyst particles became fluid and ran through the tubes at about 50-100 nm/sec. I was a little surprised by the movement. The catalyst particles would stop moving within 2-5 seconds.
I have found one reference, Buffat and Borel, Phys. Rev A13, 2287(1976), that showed the melting temperature of gold nanoparticles could be reduced by 600C! for nanoparticles less than 5 nm.
In our case we have cobalt nanoparticles less than 10 nm.
My question is how high a temperature might we expect to heat the sample in the electron beam with the following parameters;
Have others seen this phenomenon? David R. Hull NASA Glenn Research Center at Lewis Field Advanced Metallics Branch Mail Stop 49-1 21000 Brookpark Road Cleveland, OH 44135
A full time position for an experienced electron microscopist is available in the laboratory of Yishi Jin at the University of California, Santa Cruz. Dr. Jin studies the cytoskeletal architecture at synapses in the nervous system of the nematode C. elegans. For more detailed information about the work being done in the lab see http://www.biology.ucsc.edu/faculty/jin.html, or consult these recent publications; Zhen et al.2000.Neuron 26:331-343 or Zhen and Jin.1999. Nature.41:371-375.
The person we seek must have prior experience in sample preparation for transmission electron microscopy of biological materials such as fixation, embedding, sectioning, staining, and photography. Proficiency in serial sectioning would be an advantage, but is not essential at the time of employment, nor is experience with C. elegans. The person selected must be highly motivated, able to work independently, and be willing to acquire new skills and techniques. A Bachelors's degree or equivalent in a biological science is desirable.
This position is funded by the Howard Hughes Medical Institute. Salary will be commensurate with experience within the range and criteria determined by HHMI.
Interested individuals should send a current CV, names, addresses, and email contact information of 3 references, and examples of previous EM work to: Dr. Yishi Jin, Department of Biology, 328 Sinsheimer Laboratories, University of California, Santa Cruz, CA 95064 by November 1, 2000. Dr. Jin's email address is jin-at-biology.ucsc.edu.
We are still using a 15 year old Link AN10000 x-ray detector. The widths of the peaks have suddenly become much wider. The zero beam or strobe used to be around 80 eV FWHM, and now is around 360 eV, with no beam in the column. To the best of my knowledge there is no light in the column, and the dewar has plenty of LN2 in it, so I am assuming the crystal is cold. Also, if I disconnect the Lemo connector (carrying the signal from the detector) the FWHM drops to about 20 eV FWHM. If anyone has any suggestions I would be very appreciative.
Thanks,
Mick Thomas ------------------------------------------------- Mick Thomas UHV-STEM Laboratory E-1 Clark Hall Cornell University Ithaca, NY 14853
A colleague is trying to use an Olympus Fluoview CLSM System on a BX50WI upright (not inverted) microscope. She is unable to obtain a decent brightfield image. Question: Has anyone developed their own set of operating directions that they would be willing to share? Unfortunately, I have no experience with CLSM and can offer no assistance to her.
Thank you.
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 ####################################################################
I have had great successs with Castoliteto make very clear, water white mounts of various natural materials. I have seen some with dandelion heads clearly defined.
Castolite can be obtained from Buehler, Ltd. 41 Waukeegan Rd. PO box 1, Lake Bluff, IL, 60044-7979, 800-283-4537
} Tina Carvalho wrote: } } } Someone has asked what resin to use that would be clear and suitable for } } embedding mouse embryos. This would be for display, not for sectioning. } } } } Dear Tina, } How about lucite? I have no idea where to get it, but many years ago } I } bought a lucite cube in which a dandilion had been embedded. You could } see it very clearly, and there is obviously a technique for preserving } such } a fragile specimen. Note that this is more along the lines of an } existance } theorem than a procedure. Good luck. } Yours, } Bill Tivol } } } } } }
If you, or people you know, have a double tilt holder that can be used for Hitachi H7000 to sale, please contact me or Dr. Soumitra Ghoshroy (505-646-3600, sghoshro-at-nmsu.edu). TIA. ---------------------------------------------------------------------------- ------------------- Prof. Jane G. Zhu Department of Physics Phone: 505-646-1933 New Mexico State University Fax: 505-646-1934 MSC 3D, P.O. Box 30001 E-mail: jzhu-at-nmsu.edu Las Cruces, NM 88003 ---------------------------------------------------------------------------- -------------------
At the recent Microscopy and Microanalysis meeting in Philadelphia, I gave a talk entitled "The Hitachi HD-2000 In Semiconductor Manufacturing Support And Research". Based on the feed back and comments I have received, I would like to restate the goals of my talk and my opinions for all that are interested.
The HD-2000 was designed as a tool for failure mode analysis and manufacturing support. It does a wonderful job in this regard. It combines traditional SEM imaging with basic STEM operation giving SEM and TEM type capabilities in the same package. Holders are compatible with the Hitachi FIB and the rapid exchange between these tools increases the ease and success rate for manufacturing support. Use is very straightforward and requires very little training for successful and productive use. The Cirent Semiconductor/Lucent technologies primary users are extremely happy with their microscope and it has increased their capabilities in support of the semiconductor fabrication lines.
Because of its dedicated STEM design and cold field emission gun, it has potential to be a successful research level microscope for imaging and spectroscopy. In my opinion, the microscope as shipped and installed at the Lucent manufacturing site in Orlando has some key shortcomings that will inhibit its implementation at this level. However, Hitachi is currently developing improved diffraction capabilities and EELS options that should address my concerns. I look forward to seeing and evaluating their improvements.
I would be glad to discuss my opinions and impressions with anyone interested.
Richard Vanfleet vanfleet-at-physics.ucf.edu 407-823-2600
Disclaimer: I am not employed by or affiliated in any way with Hitachi.
Have you changed anything around the eds setup? Plugged a new desk lamp into a mains outlet nearby? Changed the layout of the cabling? What you describe is a pretty big jump, but it may pay to check the simple things before calling in the man.
I really messed up my PC-based system a while back by connecting up an external CD writer whose mains lead went to a socket that was on a different supply circuit, thereby producing a nice big noise-capturing earth loop.
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 } -----------------------------------------------------------------------. } } } Fellow microscopists, } } We are still using a 15 year old Link AN10000 x-ray detector. The widths } of the peaks have suddenly become much wider. The zero beam or strobe used } to be around 80 eV FWHM, and now is around 360 eV, with no beam in the } column. To the best of my knowledge there is no light in the column, and } the dewar has plenty of LN2 in it, so I am assuming the crystal is } cold. Also, if I disconnect the Lemo connector (carrying the signal from } the detector) the FWHM drops to about 20 eV FWHM. If anyone has any } suggestions I would be very appreciative. } } Thanks, } } Mick Thomas } ------------------------------------------------- } Mick Thomas } UHV-STEM Laboratory } E-1 Clark Hall } Cornell University } Ithaca, NY 14853 } } Phone: 607-255-0650 } Fax: 607-255-7658 } e-mail: mgt3-at-ccmr.cornell.edu } } 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
How can we remove mucus from fresh / fixed tissue specimens - specifically fish / squid? A student has been recommended to try s-carboxy-methyl-L-cysteine but can find no information as to how to do this, e.g. concentration, time, temperature. Any information would be welcome about this or any other methods.
Thanks - Keith Ryan
_______________________ Keith Ryan (Dr) Marine Biological Association Citadel Hill Plymouth Devon PL1 2PB England
I wish to obtain the accesories for a TEM Philips EM201 listed bellow
Plate numbering device PW 6543/00 Automated exposure system PW 6542..
the parts can be used...
any help is welcome...
thanks in advance
Fernando =================================================== Fernando D. Balducci Laboratorio de Microscopia Electrónica Facultad de Ingeniería - Bioingenieria Universidad Nacional de Entre Rios Argentina e-mail: microsc-at-fi.uner.edu.ar tel: 54 43 975100 fax: 54 43 975077
} Date: Wed, 30 Aug 2000 11:50:24 +0100 } From: Keith Ryan {kpr-at-ccms.ac.uk} } To: Microscopy-at-sparc5.microscopy.com } Subject: SEM - mucus removal from specimens } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 from Plymouth UK } } How can we remove mucus from fresh / fixed tissue specimens - } specifically fish / squid? A student has been recommended to try } s-carboxy-methyl-L-cysteine but can find no information as to how to } do this, e.g. concentration, time, temperature. Any information would } be welcome about this or any other methods. } } Thanks - Keith Ryan } } } _______________________ } Keith Ryan (Dr) } Marine Biological Association } Citadel Hill } Plymouth } Devon PL1 2PB } England } } Tel. 0044 (0)1752 633279 (with answer machine) } also 0044 (0)1752 633249 } Fax. 0044 (0)1752 633102 } } e-mail: kpr-at-wpo.nerc.ac.uk
We use Sputolysin to dissolve mucus in human lung washes. You can make it yourself with 0.1 g dithiothreitol in 100 ml water. Keep this as a stock. When ready for use, dilute 1:10 with water, and use this dilution 1:1 with sample. Let sit at room temp with periodic swirling for 30-60 min. Add more and repeat if necessary. For our procedure, it thins the mucus so that we can prepare the sample for negative staining to look for viruses.
NOW, I have no idea what is in fish slime or whether this concoction will work, but it's easy enough to try. I don't know about the cysteine and will be interested to hear what you find works.
Good luck, Sara
} }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
Keith Ryan - I can relay a little tip about this but I have not studied the chemistry in detail. A group here was doing SEM on lung and airway tissues. They were prepping the tissue for SEM by fixation, dehydration in ethanol, then using hexamethyldisilazane to dry the samples. They discovered that mucus was covering the surface of the airways. They simply added a ten minute step in xylene between the last alcohol and the HMDS. The mucus disappeared. It might be worth a try. Good luck. Sincerely,
Robert J. Munn Electron Microscopy Laboratory Department of Medical Pathology University of California, Davis School of Medicine Davis, CA 95616
Dear Mick, When I had an increase in peak width in my Kevex last year, I warmed up the detector to remove any water ice that had accumulated in the bottom of the dewar or on the crystal. Turn off the bias, short out the bias plug, pour out the liquid nitrogen, then blow hot air into the dewar until everything is warm. Unfortunately, although the resolution improved 10 eV per channel, the dewar now had poor holding time, so I had to have it re-pumped. Now, everything is great. The other problem can be related to poor grounding in the pre-amp, but that caused excessive dead-time. At 04:01 PM 8/29/00 -0700, you wrote: } } Fellow microscopists, } } We are still using a 15 year old Link AN10000 x-ray detector. The widths } of the peaks have suddenly become much wider. The zero beam or strobe used } to be around 80 eV FWHM, and now is around 360 eV, with no beam in the } column. To the best of my knowledge there is no light in the column, and } the dewar has plenty of LN2 in it, so I am assuming the crystal is } cold. Also, if I disconnect the Lemo connector (carrying the signal from } the detector) the FWHM drops to about 20 eV FWHM. If anyone has any } suggestions I would be very appreciative. } } Thanks, } } Mick Thomas 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 remove mucus using a product used to clear mucus from airway and bowel in the clinical environment, it is Acetylcysteine, used at 1% concentration I believe. The only problem is that if there are other goblet cells in the tissue they will release immediately leading to yet more mucus..... It works though Simon
--------------------------- Simon C. Watkins Ph.D. MRC Path Associate Professor, Cell Biology and Physiology Director: Center for Biologic Imaging BSTS 225 University of Pittsburgh Pittsburgh PA 15261 Tel:412-648-3051 Fax:412-648-8330 http://sbic6.sbic.pitt.edu -----------------------------------
Currently we are looking at stands of DNA that have been stained with Uranyl Acetate and gold labeled, and then deposited on to Collodion film, which the strands stick to, nicely.
However, at high KeV or focused spot the film breaks easily, what I would like to do is use a carbon film, which is more robust but my problem is the DNA will not stick to this.
Perhaps someone who is more expert at DNA imaging could suggest a way to produce a more robust sample?
Many Thanks in advance
David
-- Dr. David C. Bell (612)-624-1677 AEM Manager, IT Chafac dcb-at-umn.edu 16 Shepherd Labs, University of Minnesota http://resolution.umn.edu 100 Union St S.E. Minneapolis, MN USA 55455
Have you tried collodion on carbon? Then you'd have the best of both (put the carbon on the collodion, then use the "back" of the grid). Or try some substrate other than collodion that is more stable in the beam - Butvar, for example.
However, doesn't the DNA "community" usually use carbon films? I've done DNA on carbon - just glow-disharge the carbon film within a few hours before you put the DNA on. It is more annoying than using straight plastic films, but more stable once you get to the 'scope.
Tamara Howard CSHL
On Wed, 30 Aug 2000, David Bell 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 } -----------------------------------------------------------------------. } } } } Currently we are looking at stands of DNA that have been stained } with Uranyl Acetate and gold labeled, and then deposited on to } Collodion film, which the strands stick to, nicely. } } However, at high KeV or focused spot the film breaks easily, } what I would like to do is use a carbon film, which is more robust } but my problem is the DNA will not stick to this. } } Perhaps someone who is more expert at DNA imaging could suggest } a way to produce a more robust sample? } } Many Thanks in advance } } David } } } -- } Dr. David C. Bell (612)-624-1677 } AEM Manager, IT Chafac dcb-at-umn.edu } 16 Shepherd Labs, University of Minnesota } http://resolution.umn.edu } 100 Union St S.E. Minneapolis, MN USA 55455 } }
I have been asked to identify a figurine said to be "chalkware" has anyone seen or know a definition of that term? Is there a legal definition of chalkware? Would there be a problem calling a plastic filled with 20-30% calcium carbonate - chalkware? The sample was made of gypsum (calcium sulfate) core with a calcium carbonate (whitewash) then painted which makes it what is called carnival-ware. Thanks for any help. Terry Ellis
You could try to evaporate a light coat of carbon over the whole grid after the DNA is stuck and stained. You could also try Formvar which is a little sturdier than collodion.
On Wed, 30 Aug 2000, David Bell wrote:
} Date: Wed, 30 Aug 2000 13:07:30 -0500 } From: David Bell {dcb-at-x32-135.cie.umn.edu} } To: Microscopy-at-sparc5.microscopy.com } Subject: Help Needed: Looking at DNA strands with TEM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Currently we are looking at stands of DNA that have been stained } with Uranyl Acetate and gold labeled, and then deposited on to } Collodion film, which the strands stick to, nicely. } } However, at high KeV or focused spot the film breaks easily, } what I would like to do is use a carbon film, which is more robust } but my problem is the DNA will not stick to this. } } Perhaps someone who is more expert at DNA imaging could suggest } a way to produce a more robust sample? } } Many Thanks in advance } } David } } } -- } Dr. David C. Bell (612)-624-1677 } AEM Manager, IT Chafac dcb-at-umn.edu } 16 Shepherd Labs, University of Minnesota } http://resolution.umn.edu } 100 Union St S.E. Minneapolis, MN USA 55455 } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
I am having serious problems preparing thin foils of a Mg-2.5Ag-2Nd alloy using a Fischione twin jet. I have tried two electrolytes: 5% perchloric in ethanol; 5.3g lithium chloride, 11g magnesium perchlorate, 500ml methanol and 100ml 2-butoxy-ethanol. The main problem I have seems to occur on removal from the Fischione fixture, the specimen continues to react the even the slightest amount of electrolyte-no matter what efforts I take to dilute it. Then we are left with no thin area and lots of corrosion product on the remaining thick foil.
While I haven't used the Fischione polisher, we do manufacture our own electropolisher so I have some experience to share. As our Model 550 is also capable of being used for straight chemical etching (as with HF and Bromine methanol solutions for semiconductor applications) we developed a rinser that will rinse the sample automatically immediately after the perforation is detected. The problem you have is that once a perforation is detected, even the slightest additional etching will quickly etch away the thin area surrounding the hole. If you are doing a purely electrolytic etch, then etching should stop as soon as the power is shut off. Perhaps you are also achieving some chemical etch which will continue until all of the etchant is washed away. A few things you may want to try are:
1) See if there is anyway you can rig up an automatic rinse system to the polisher. This would be ideal, but I do not know how practical it would be.
2) Try to simplify the sample holder so that electrolyte does not get trapped within the holder. If you are experiencing etching after the power is shut off, then any chemicals trapped within the holder will leak out and cause the sample to etch further. Our single jet system places the specimen on a pedestal where the electrolyte cannot become trapped. The sample is not submerged in the electrolyte so the electrolyte quickly washes away. This is also what makes our rinsing attachment easy to implement.
3) If etching still continues after removing the fixture, then you may want to try to adjust the distance between the jets and the sample. With our Model 550 you can change the shape of the dimple in the sample by moving the jet nozzle up or down. Placing the nozzle closer to the sample will provide a larger thin are and will give you additional time to stop the chemical etching before all of the thin area is etched away.
4) With our system you can easily change the LED used in the detection circuit to select a wavelength that is compatible with your material. We have been able to automatically shut off the system in some cases just prior to perforation. If you are able to do this with your system, perhaps the additional etching that occurs as you remove the sample holder will produce a sample with the appropriate thin area.
Of course, my first suggestion would be to buy one of my jet polishers! 8-)
I hope this helps to some degree. If I can be of any additional assistance, please feel free to contact me.
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 Milo Kral } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am having serious problems preparing thin foils of a Mg-2.5Ag-2Nd alloy using a Fischione twin jet. I have tried two electrolytes: 5% perchloric in ethanol; 5.3g lithium chloride, 11g magnesium perchlorate, 500ml methanol and 100ml 2-butoxy-ethanol. The main problem I have seems to occur on removal from the Fischione fixture, the specimen continues to react the even the slightest amount of electrolyte-no matter what efforts I take to dilute it. Then we are left with no thin area and lots of corrosion product on the remaining thick foil.
David, You can stabilize your collodion films with DNA on them through thermal or eb evaporation of carbon layer prior to TEM. Alternatively, you can modify carbon films through alkylamine treatment prior to spreading. Marek.
} } } } } Currently we are looking at stands of DNA that have been stained } } with Uranyl Acetate and gold labeled, and then deposited on to } } Collodion film, which the strands stick to, nicely. } } } } However, at high KeV or focused spot the film breaks easily, } } what I would like to do is use a carbon film, which is more robust } } but my problem is the DNA will not stick to this. } } } } Perhaps someone who is more expert at DNA imaging could suggest } } a way to produce a more robust sample? } } } } Many Thanks in advance } } } } David } } } } } } -- } } Dr. David C. Bell (612)-624-1677 } } AEM Manager, IT Chafac dcb-at-umn.edu } } 16 Shepherd Labs, University of Minnesota } } http://resolution.umn.edu } } 100 Union St S.E. Minneapolis, MN USA 55455 } } } } }
Marek Malecki, M.D., Ph.D. Director and Principal Investigator
Molecular Imaging Laboratories University of California at San Diego address: 1500 Suite Bonner Hall, 9500 Gilman Drive, La Jolla, CA 92093-0368 phone: 8588223373 fax: 8588223715 pager: 8586161420 e.mail: mmm-at-ucsd.edu e.pager: 1620024619-at-alphapage.airtouch.com www site: http://mil.ucsd.edu/ ftp site: mil1.ucsd.edu
I am not sure if warming will cure the problem, but is certainly worth trying. As I recall a lot of the AN10,000's had heater circuits built into the detector, with a button on the pulse processor. This may have only been the turret detectors. If present it will be less destructive than hot water.
Best wishes,
Colin
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2.
Tel: 353-1-6081820 Fax: 353-1-6770438 Email: creid-at-tcd.ie Home Page: http://www.tcd.ie/Electron_Microscope/emu/home.htm
-----Original Message----- } From: Mary Mager [mailto:mager-at-interchange.ubc.ca] Sent: Wednesday, August 30, 2000 4:37 PM To: Mick Thomas Cc: Microscopy-at-sparc5.microscopy.com
Dear Mick, When I had an increase in peak width in my Kevex last year, I warmed up the detector to remove any water ice that had accumulated in the bottom of the dewar or on the crystal. Turn off the bias, short out the bias plug, pour out the liquid nitrogen, then blow hot air into the dewar until everything is warm. Unfortunately, although the resolution improved 10 eV per channel, the dewar now had poor holding time, so I had to have it re-pumped. Now, everything is great. The other problem can be related to poor grounding in the pre-amp, but that caused excessive dead-time. At 04:01 PM 8/29/00 -0700, you wrote: } } Fellow microscopists, } } We are still using a 15 year old Link AN10000 x-ray detector. The widths } of the peaks have suddenly become much wider. The zero beam or strobe used } to be around 80 eV FWHM, and now is around 360 eV, with no beam in the } column. To the best of my knowledge there is no light in the column, and } the dewar has plenty of LN2 in it, so I am assuming the crystal is } cold. Also, if I disconnect the Lemo connector (carrying the signal from } the detector) the FWHM drops to about 20 eV FWHM. If anyone has any } suggestions I would be very appreciative. } } Thanks, } } Mick Thomas 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
probably the method with the most consistent results (and easy to perform; students do it in our EM-course) is the method by Lang and Mitani (1969). in short, put your cytochrome C stabilized DNA on the collodion film (without carbon) fix with uranylacetate/METOH and perform low angle (approx. 5 °) platinum rotation shadowing (2- 3 nanometer) followed by stationary, vertical (90°) carbon shadowing. specimens are virtually undestroyable and last "forever".
good luck! peter
************************************************** please reply to:
I knew acetylcysteine has been used for airway and stomach mucus but we never had much luck with it with intestinal mucus (rat intestines or human goblet cells cultures). Simon says "bowel" so maybe he means stomach or perhaps he has better luck with intestinal mucus also. I am not an expert with acetylcysteine so I will defer to him on that point. But I can easily believe him when he says that it causes increased mucus secretion on fresh tissue. We have found that most, if not all, of the treatments to remove or "thin" mucus are good only for biochemical preps or fixed tissues since it is especially easy to trigger a goblet cell. Even vigorous washing is enough to set them off.
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The temperature increase in your sample is not only related to the keV and e-dose you use, but also related to the thermal conductivity of your sample, it thickness, surface condition, and its surrounding materials. So I would think it is almost impossible in your case to measure experimentally or to estimate theoretically the temperature increase.
The reason for melting point decrease of any kind of nona-particles is obvious because of the tremendous surface to volume ratio. The surface Gibbs free energy should be much higher in nano-systems, and hence such systems are relatively unstable. That's why most nano-particles have to be kept in aqueous suspension and with additives.
To avoid the problem, try the following:
1. Spot size: 2 or 3 (When I drive TEM, I always first try relatively smaller spot size, usually at 2. Actually I hardly have the need to use spot size 1 on a 200kV scope. The materials I checked range from metals, intermetallic compounds, ceramics, semiconductors and polymers, either thin film or bulk or nano particles.) 2. Different keV, either higher or lower keV could be beneficial to decreasing the damage to a sample depending on which specific heating mechanism dominates. 3. Use a smaller condenser aperture. 4. Coat a very thin C film on your grids before observation.
Have a nice day!
Chao-Ying Ni Rodel Inc. 451 Bellevue Road Newark, DE 19713 (302) 366-0500 ext 2812
-----Original Message----- } From: David R Hull [mailto:David.R.Hull-at-grc.nasa.gov] Sent: Tuesday, August 29, 2000 1:22 PM To: 'Microscopy Listserver'
We have been looking at the use of various catalysts for production carbon nanotubes. Recently we had a sample that behaved in a very unusual way. Upon focusing the electron beam on a cluster of nanotubes, the catalyst particles became fluid and ran through the tubes at about 50-100 nm/sec. I was a little surprised by the movement. The catalyst particles would stop moving within 2-5 seconds.
I have found one reference, Buffat and Borel, Phys. Rev A13, 2287(1976), that showed the melting temperature of gold nanoparticles could be reduced by 600C! for nanoparticles less than 5 nm.
In our case we have cobalt nanoparticles less than 10 nm.
My question is how high a temperature might we expect to heat the sample in the electron beam with the following parameters;
Have others seen this phenomenon? David R. Hull NASA Glenn Research Center at Lewis Field Advanced Metallics Branch Mail Stop 49-1 21000 Brookpark Road Cleveland, OH 44135
The short answer is, that if conditions are right you can get significant local heating under the beam. I (and others before me) have used this technique to "melt" metal chloride crystallites and form pure metal nanoparticles inside the microscope. The key issues are how much energy you are depositing in the specimen (thickness and composition of specimen are important here) and how fast the specimen can carry the heat away. An area of the specimen that does not have good thermal contact to the grid can become very hot. I would suggest looking at Reimer "Transmission Electron Microscopy" starting about page 431 with a section called Specimen Heating.
I have tried some of these calculations before so if you run into problems I might be able to help.
Richard
At 01:21 PM 8/29/00 -0400, David R Hull wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Richard Vanfleet Assistant Professor Advanced Materials Processing and Analysis Center (AMPAC) and Department of Physics
Mail: Department of Physics University of Central Florida 4000 Central Florida Blvd. Orlando, FL 32816-2385
David R. Hull reported that catalyst particles became fluid in nanotubes under TEM examinationŠ and asked for temperature rise.
I guess that there is some lowering of the melting temperature for 10nm cobalt particles, although I do not have right now the data to compute it (in particular the interfacial energy Co-nanotube).
But about temperature rise, you have a very nice situation where inelastic interaction in the nanotubes cluster may lead to a relatively strong energy absorption, while the thermal dissipation by conductivity is low. There is only thermal radiation left for energy dissipation at the surface of the cluster and this may lead to temperature rise of several hundreds of C degrees.
We did a nice experiment (unpublished) 20 years ago with clusters a few microns in diameter containing Ag nanoparticles lying on a carbon film/copper grid. When the electron beam was focused on a cluster (about same diameter, current for "normal observation"), the cluster was instantaneously vaporized and we can follow nucleation, growth and sintering of Ag particles from nm to 50nm running on the film in the same or the next grid opening.
Considering the silver vapour pressure with temperature, this proves that the temperature was by far above 1000 K, more probably toward 1500 K. But again this strange reaction is due to
i) the size of the cluster (significant energy/electron absorption) ii) the low thermal dissipation by conductivity in the film compared to energy absorption iii) the low surface to volume ratio (= low energy dissipation by radiation / energy absorption) in these "large" clusters
It should be pointed out that for isolated nanocrystals, the situation is opposite, the surface to volume is much higher, as the radiation dissipation to energy absorption. In addition the thermal conductivity of the film becomes relatively more important. The (phonon) temperature rise is now only of a few C or tens of C even under strong irradiation.
Yours
Philippe Buffat
} Delivered-To: philippe.buffat-at-epfl.ch } X-Sender: mshull-at-popserve.grc.nasa.gov } Date: Tue, 29 Aug 2000 13:21:37 -0400 } To: "'Microscopy Listserver'" {Microscopy-at-sparc5.microscopy.com} } From: David R Hull {David.R.Hull-at-grc.nasa.gov} } Subject: TEM: Melting and Flow of Nanoparticles During TEM Observation } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 been looking at the use of various catalysts for production } carbon nanotubes. Recently we had a sample that behaved in a very } unusual way. Upon focusing the electron beam on a cluster of } nanotubes, the catalyst particles became fluid and ran through the } tubes at about 50-100 nm/sec. I was a little surprised by the } movement. The catalyst particles would stop moving within 2-5 } seconds. } } I have found one reference, Buffat and Borel, Phys. Rev A13, } 2287(1976), that showed the melting temperature of gold } nanoparticles could be reduced by 600C! for nanoparticles less than } 5 nm. } } In our case we have cobalt nanoparticles less than 10 nm. } } My question is how high a temperature might we expect to heat the } sample in the electron beam with the following parameters; } } 200 kV, LaB6, 20-30 microamp emmission current, spot size 1, 150 } micron condenser aperture. } } Have others seen this phenomenon? } David R. Hull } NASA Glenn Research Center at Lewis Field } Advanced Metallics Branch } Mail Stop 49-1 } 21000 Brookpark Road } Cleveland, OH 44135 } } (216) 433-3281 } fax (216)977- 7132 } david.r.hull-at-grc.nasa.gov
__________________________________________________________________ Philippe Buffat Ecole Polytechnique Federale de Lausanne (EPFL) Centre Interdepartemental de Microscopie Electronique Address: EPFL-CIME, Batiment MX-C, CH-1015 Lausanne, Switzerland Phone: +41(21)693 29 83 Fax: +41(21)693 44 01 (Central European Time) E-mail: philippe.buffat-at-epfl.ch, WWW URL http://cimewww.epfl.ch/ ______________________________ Eudora F2.1 ___________________________
I have a sample of albumin microspheres that need to be embedded and sectioned for TEM. They range from 2-30µm in size. Does anyone have experience embedding such a sample? If I embed them straight into a resin then they may pull out of the resin during sectioning. Any ideas? Karen Kelley Senior Electron Microscopist UF Biotechnology Electron Microscopy Core Lab Box 118525 Gainesville Florida 32611 lab:352-392-1184 fax: 352-846-0251 email: klv-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/staff/karenpage.html
We're going to be fixing and staining Xenopus cells to examine their yolk platelets, and I was hoping someone has had good experience with lipid fixation and suitable fluorescent lipophilic probes for this system. Specifically, we don't want any of our yolk platelets to float away during fixation and membrane permeabilization. I'm also curious as to which Di"X" analogs and/or Bodipy's have worked best for vitellogenin and phosvitin containing yolk. Thanks in advance.
Mike
Michael B. Ferrari
Department of Biological Sciences University of Arkansas Fayetteville, Arkansas 72701 Ph: 501-575-6372 Fax: 501-575-5349 email: ferrari-at-uark.edu http://biology.uark.edu/ferrari
A student working in my lab made ultrathin sections of polymer coated albumin microspheres either lyophilized and embedded in acrylic resin, or dehydrated from an aqueous solution and embedded in epoxy resin. I don't remember any problem with pullout, and she got reasonable looking micrographs in both preps.
Marie
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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
A Leitz Metallux II microscope, which had been stored without air conditioning for some time, recently came into my care. The objectives and other internal lenses are actually obscured by corrosion product. Before anyone gives me a hard time for letting the instrument get into this condition, let me make it quite clear that I am not the one who put it in storage, nor did I know about it until recently. I am looking for advice on how to salvage it.
I removed the objectives from the turret and disassembled one of them as far as I could - just removed the ring holding the lens in the housing. The lens is still in place but the dirt / dried corrosion is plainly exposed. I am reluctant to physically touch the glass for fear of scratching it. Flushing it with a solvent - water? - seems an appropriate first step, but I do not know what liquid to try.
Please excuse me if this is the wrong forum in which to post this query. Any suggestions are welcome. If this instrument appeared on the capital equipment manifest of any department, I could get a P.O. to get it repaired - but it does not. I'm the one who has to fix it if it is going to get fixed. Thanks in advance.
Regards, Andrew T. Werner Shaped Charge Research - Metallurgy Laboratory Schlumberger Reservoir Completions Technology Center 14910 Airline Road, Rosharon, TX 77583-1590 Voice (281) 285-5272 Fax (281) 285-5273
I think one important point needs to be made - are you sure the particles are molten? When you do a lot of TEM, you tend to forget the size and relevant time scales. A typical phonon period is about 10-10 seconds, which is similar in magnitude to the typical attempt time for single-atom diffusion. Hence it is sometimes surprising that samples sit still for long enough for one to take an image! In many cases, particularly at a free surface, what one is observing is a time average of thousands to millions of single-atom diffusion events.
Consequently what appears to be liquid-like behavior may very well be reasonably rapid solid-state diffusion. You do not need to invoke high temperatures in order to explain apparently rapid (in the microscope) motion of nanoparticles. There may be some heating (although direct displacement via radiolytic or knockon processes will be more efficient than heat), but one does not need more than a modest rise to explain the observation in most cases.
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
Hi all, I'm trying to put together a proposal to hire someone to run our JSM 840, but I'm not really sure of the salary scale for this kind of position. I would appreciate it if anyone could suggest a range or offer advice on the subject preferably to my personal e-mail at sbucks-at-charter.net Any suggestions would be appreciated. Many Thanks,
You should have no problems finding someone for a starting salary of approximately $60,000.
David Chow E-mail: david.chow-at-nrc.ca {mailto:david.chow-at-nrc.ca}
-----Original Message----- From: Buckingham, Steve [mailto:sbuckingham-at-excellatron.com] Sent: Thursday, August 31, 2000 1:47 PM To: 'Microscopy-at-MSA.Microscopy.Com' Subject: Microscopist position
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Hi all, I'm trying to put together a proposal to hire someone to run our JSM 840, but I'm not really sure of the salary scale for this kind of position. I would appreciate it if anyone could suggest a range or offer advice on the subject preferably to my personal e-mail at sbucks-at-charter.net Any suggestions would be appreciated. Many Thanks,
Is anyone writing imaging programs for the Linux OS? Or a port of NIH-Image for Linux?
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)
The tissue we were using were duodenal segments mounted in an ussing chamber, which were washed and then treated with various strains of bugs to look at mechanisms of translocation. It did get rid of the mucus, but I agree with Tom its tough to stop it coming back! Simon
-----Original Message----- } From: Tom Phillips [mailto:PhillipsT-at-missouri.edu] Sent: Thursday, August 31, 2000 8:44 AM To: Microscopy-at-sparc5.microscopy.com
I knew acetylcysteine has been used for airway and stomach mucus but we never had much luck with it with intestinal mucus (rat intestines or human goblet cells cultures). Simon says "bowel" so maybe he means stomach or perhaps he has better luck with intestinal mucus also. I am not an expert with acetylcysteine so I will defer to him on that point. But I can easily believe him when he says that it causes increased mucus secretion on fresh tissue. We have found that most, if not all, of the treatments to remove or "thin" mucus are good only for biochemical preps or fixed tissues since it is especially easy to trigger a goblet cell. Even vigorous washing is enough to set them off.
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-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
The Department of Material Science and Engineering at the University of Delaware has an immediate opening for a lab coordinator/instrument scientist to administer and help further develop the expanding electron microscopy lab in the college of engineering. The successful candidate should have an MS/PhD in MSE or related field and extensive EM laboratory experience. The primary responsibilities of the position include general maintenance of transmission and scanning microscopes (JEOL 2010FEG and 2000 FX TEM, JEOL 840 SEM) and their ancillary sample preparation equipment, user training, facility scheduling, and input on future facility development and instrument acquisition. In addition, it is anticipated that the candidate may pursue collaborative research with faculty in the college and university. The salary for the position will be commensurate with successful candidate qualifications. Applicants should send a cover letter, resume/CV, and list of 3 references to Professor Darrin J. Pochan, 301 Spencer Lab, Materials Science and Engineering, University of Delaware, Newark, DE 19716. Applications will be accepted till September 30, 2000 or until the position is filled.
I often use water first as a cleaner because often it is stuff soluble in aqueous media that is dripped on the lens and often this stuff is not soluble in organic solvents. Then I do an ethanol wash to dry and remove organic soluble stuff.
If I can add to the original question, I have had a great deal of trouble cleaning lenses and especially CCD imagers due to residual dust. I find that cotton swabs (at least those I have) leave an enormous amount of residue behind that cannot be blown off easily. Does anyone have a source of a more dust free equivalent to the cotton swab? Dave --
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Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 75 N. Eagleville Rd. U-125 Storrs, CT 06269-3125 Knecht-at-uconnvm.uconn.edu 860-486-2200 860-486-4331 (fax) home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html ************************************************************
I don't know about NIH-Image, but there are a large number of imaging programs for Linux. These include:
AVS (see www.avs.com)
Gimp (A Photoshop workalike)
VTK (The Visualization Toolkit, see www.kitware.com)
Image/J is Jave-based and runs on Linux (see http://rsb.info.nih.gov/ij/)
ImageMagick
ImgStar
There is a Mac Emulator called Executor that will allow PC users with Linux to run NIH Image. See www.cs.ubc.ca/spider/ladic/softunix.html
The site above lists a number of IP tools for UNIX, many of which will run on Linux boxes.
billo
On Thu, 31 Aug 2000, Philip Oshel 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 } -----------------------------------------------------------------------. } } } Is anyone writing imaging programs for the Linux OS? Or a port of } NIH-Image for Linux? } } 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) }
There are some very good packages out there both commercial and GNU. I will run them down for you.
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00 ----- Original Message ----- } From: "Philip Oshel" {peoshel-at-facstaff.wisc.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, August 31, 2000 2:01 PM
I have a product from Berkshire 800 242 7000 or 413 528 2602 or 2802 model LT00e166-R I got from a VCR tech. The are a Camious like swab that don't seem to leave lent. They do require more pressure on the glass but I have not had a scratch yet.
I used it to remove a very stubborn water stain on a prism that I couldn't touch with cotton and anything I had for a solvent. Isopropyl and this swab made it look easy.
Cotton has a considerable amount of oil on it as grown. Much of it is removed in processing but it still has some slight oil content that plus the fact it can pick up a static charge to a point that has to be seen to be believed make it problematic. If I was trying to use cotton so it wouldn't stick I would treat it with and anti static solution wash it in a couple of changes of hexane and use it in a high humidity environment.
A piece of lens tissue works well if you just twist it on a rough stick.\
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger
} From: "David Knecht" {knecht-at-uconn.edu} } } I often use water first as a cleaner because often it is stuff } soluble in aqueous media that is dripped on the lens and often this } stuff is not soluble in organic solvents. Then I do an ethanol wash } to dry and remove organic soluble stuff. } } If I can add to the original question, I have had a great deal of } trouble cleaning lenses and especially CCD imagers due to residual } dust. I find that cotton swabs (at least those I have) leave an } enormous amount of residue behind that cannot be blown off easily. } Does anyone have a source of a more dust free equivalent to the } cotton swab? Dave } -- }
I have a question regarding artifacts in the myelin sheath of a peripheral nerve: Samples were immersionfixed in Karnovsky´s half-strength fixative(2% Glut, 2% PFA in 0.1M Cacodylate)en bloc stained with osmiumtetroxide and UAc and embedded in Araldite. The myelin sheaths show the following appearance: 1. The fine lammellar structure is focally disturbed, the layers appear weavy, wobbly ... 2. The outer rings are stained more intense than the inner rings (to the axon). 3. The myelin sheaths are de-rounded, but this seems to be not so grave and I noted this often in other pictures published. Maybe this is "normal"?
I would appreaciate any suggestions from you all to overcome this problem. Thanks in advance! Greetings, Michael
Michael Reiner University of Cologne, Germany Dept. of Anatomy I _______________________________________________________________________ 1.000.000 DM gewinnen - kostenlos tippen - http://millionenklick.web.de IhrName-at-web.de, 8MB Speicher, Verschluesselung - http://freemail.web.de
David Knecht asked: } If I can add to the original question, I have had a great deal of } trouble cleaning lenses and especially CCD imagers due to residual } dust. I find that cotton swabs (at least those I have) leave an } enormous amount of residue behind that cannot be blown off easily. } Does anyone have a source of a more dust free equivalent to the } cotton swab?
Kodak has several application notes covering various aspects of CCD sensors available on our web site:
Specifically, application note DS 00-009 has the recommended procedure for cleaning the coverglass on our sensors. Of course, we do not speak for other manufacturers. It is always safest to check with the manufacturer of your particular sensor for the recommended cleaning procedure.
DS 00-009 is available, in Adobe Acrobat format at: http://www.kodak.com/US/en/digital/pdf/ccdCoverGlass.pdf
The engineer who cleans our light microscopes tells me that CCD coverglasses tend to attract dust because of static charges. Good housekeeping practices and the use of dust covers minimizes the need for cleaning.
Best Regards,
John Minter Eastman Kodak Company Analytical Technology Division Rochester, NY 14650-2152 Phone: (716) 722-3407 FAX: (716) 477-7781 email: john.minter-at-kodak.com
We often see some disturbance of the myelin sheaths. According to a review chapter by Friedrich and Mugnaini: "The preservation of myelin sheaths presents a special problem. The membrane lamellae often separate from one another, even in otherwise well-preserved material. On the other hand, we have also encountered specimens whose myelin sheaths were reasonably well preserved, but in which the neuronal elements were poorly fixed. Apparently the myelin sheaths differ from other tissue components in their reaction to fixation and they may require special conditions for optimal preservation. Improvements in the technology of fixation may eventually eliminate this disparity, but for the present it seems sensible to discount the condition of myelin sheaths in evaluating fixed material unless the myelin sheath itself is the main target of the investigation." He also lists a "Procedure for Myelin"..
This is from a book chapter (unfortunately, the copy I have is missing the source reference) which may be a number of years old. Perhaps someone else has some new information on this.
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
agreed. in addition, one must be aware of the difference between the melting point and the temperature of mobility (tamman temperature). the tamman temp is about 0.52 bulk melting point of a metal in degrees K. liquid-like behavior of metal particles can occur at this point. the picture is further complicated by any support interactions. in the case of nanotubes, you are dealing with a basal plane ({0001} face) of graphite on which the metal particle is nucleated. all transition metals are very sensitive to such interfacial phenomenon, which will directly impact their nucleating/wetting/spreading behavior (see some of R. T. K. Baker's controlled atmosphere electron microscopy studies, j. catal.). for instance, evaporated gold will tend to nucleate on edges of graphite, but not on the basal plane. such behavior was and still is taken advantage of using gold decoration techniques (pioneered by g. r. hennig in the 60's). cool stuff!
paul
---------------- Paul E. Anderson Catalytic and Nanostructured Materials Laboratory Department of Chemistry 102 Hurtig Hall Northeastern University Boston, MA 02115 617 373 5909 FAX 617 373 8795 paanders-at-lynx.neu.edu
} If I can add to the original question, I have had a great deal of } trouble cleaning lenses and especially CCD imagers due to residual } dust. I find that cotton swabs (at least those I have) leave an } enormous amount of residue behind that cannot be blown off easily. } Does anyone have a source of a more dust free equivalent to the } cotton swab? Dave
Photographic Solutions makes a product called "Sensor Swab" http://www.photosol.com/swab.html for use with professional digital SLR cameras to clean CCD's. They are approved by Kodak for use with their DCS Camera line. They also make a high quality optics cleaner called Eclipse.
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I would like to suggest all those people who are working on the metallurgy of Mg alloys to refer that original paper published in English. I apologize that I don't have any reprints of that paper and I don't remember the formula either, since I've left there for a long time. Regards, Zhiping
} From: Milo Kral {m.kral-at-mech.canterbury.ac.nz} } To: Zhiping Luo {zhiping_luo-at-hotmail.com} } Subject: Re: Help with TEM of Mg alloy } Date: Fri, 01 Sep 2000 07:50:04 +1200 } } Just one more thing - could you send the formula for the electrolyte } and the polishing conditions to me via e-mail? } } Thanks } Milo } } } Dear Milo, } } } } To prepare TEM thin foil samples and also the samples for optical } } metallography, please refer a paper from our group (S.Q. Zhang, } } Acta } } Metall. Sinica, 1990, Vol. 3A, p.110), which contains details for } } the electrolytes. Those electrolytes worked well for all Mg alloys we } } studied. However, they should NOT be deposited after the experiment } } since they are explosive after some time especially at higher } } temperatures. } } } } Another way is just to use the conventional ion milling. I didn't } } detect any clear beam damage by the ion milling even at room } } temperature. } } } } In Mg-based alloys there are many complex intermetallics (Luo et } } al., Scripta Metall. Mater., 1993, Vol. 28, p. 1513) and I observed } } a number of new microstructures including the new Mg-Zn-rare earth } } quasicrystals by TEM. } } } } Good luck in your work. } } } } Zhiping Luo } }
} } } From: Milo Kral {m.kral-at-mech.canterbury.ac.nz} } } } To: Microscopy-at-sparc5.microscopy.com } } } Subject: Help with TEM of Mg alloy } } } Date: Thu, 31 Aug 2000 09:36:15 +1200 } } } } } } ----------------------------------------------------------------------- - } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } } America
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Hi All, I have had a request to explore obtaining a fluorescent microscope with appropriate add-ons for doing quantitation of fluorescence. I know nothing about this and would appreciate advise on where to start looking. Would anyone who has such a system please contact me with general information about the system as well as as idea of cost. An investigator is writing a grant and would like to include such a system in the grant. Thanks, 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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J. W. Robinson Mechanical and Materials Engineering University of Windsor Windsor, Ontario N9B 3P4
Phone : 519-253-4232, ext 2598
Does someone have an address or phone number for Al Bingham of Semoptics or someone else who does service on the Nanolab SEM's? Any help would be appreciated.
It is not clear to me what you mean exactly by imaging programs. Do you mean programs for acquiring images or programs for manipulating existing images. For the later purpose, the Corel Graphics Suite (includes Draw and Photo Paint) is very powerful and I use it often rather than the Adobe Photoshop and Illustrator programs. Caveat: I have not used these programs on the Linus platfrom myself.
Larry D. Ackerman Lily & Yuh Nung Jan Laboratories Howard Hughes Medical Institute UCSF, Box 0725, Rm U226 533 Parnassus Ave. San Francisco, CA 94143
Is there anyone in Southern California who does freeze fracture/freeze etch SEM? I would like to find a collaborator or even better a service lab that I can send samples to for FF/FE work.
Gregory M. Fahy 21st Century Medicine ________________________________________________________________ YOU'RE PAYING TOO MUCH FOR THE INTERNET! Juno now offers FREE Internet Access! Try it today - there's no risk! For your FREE software, visit: http://dl.www.juno.com/get/tagj.
I have a project which needs to see the preservation of testis and kidney morphology of rats. I want to set up a couple of ultrastructural criteria for this. Can any of you provide me some info about good classical reference books or literature. Any experience or suggestions are greatly appreciated.
Regards
Gang Ning
EM Facility Medical College of Wisconsin Milwaukee, WI 53045 414-456-8141 414-456-6517 (Fax)
Dear Dr. Robinson, There is a web site for Semoptics at: http://www.magma.ca/~semoptx/ and the phone and fax number listed there is the same as before: (613) 727-1698. At 10:47 AM 9/1/00 -0400, you wrote: } Phone : 519-253-4232, ext 2598 } } Does someone have an address or phone number for Al Bingham of } Semoptics or someone else who does service on the Nanolab SEM's? Any help } would be appreciated. } } 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 Dr. Robinson, There is a web site for Semoptics at: http://www.magma.ca/~semoptx/ and the phone and fax number listed there is the same as before: (613) 727-1698. At 10:47 AM 9/1/00 -0400, you wrote: } Phone : 519-253-4232, ext 2598 } } Does someone have an address or phone number for Al Bingham of } Semoptics or someone else who does service on the Nanolab SEM's? Any } help would be appreciated. }
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 Dr. Robinson, There is a web site for Semoptics at: http://www.magma.ca/~semoptx/ and the phone and fax number listed there is the same as before: (613) 727-1698. At 10:47 AM 9/1/00 -0400, you wrote: } Phone : 519-253-4232, ext 2598 } } Does someone have an address or phone number for Al Bingham of } Semoptics or someone else who does service on the Nanolab SEM's? Any help } would be appreciated. }
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
emxray-at-uwindsor.ca-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 } -----------------------------------------------------------------------. } } J. W. Robinson } Mechanical and Materials Engineering } University of Windsor } Windsor, Ontario } N9B 3P4 } } Phone : 519-253-4232, ext 2598 } } Does someone have an address or phone number for Al Bingham of } Semoptics or someone else who does service on the Nanolab SEM's? Any help } would be appreciated.
J. W., Give Derek Saunders a call. He's in Nepean, Ontario at Electron Optics Service, Inc. e-mail eos-at-magma.ca. I believe he bought all the remaining NanoLab stock and is very familiar with their equipment
Elliptical distortion in SAED ring patterns is successfully taken into account in the newest version of the free ProcessDiffraction program. Calibration of the eccentricity and direction of axes is performed and integration goes along ellipses (as contrasted to circles). The program converts your SAED ring pattern (measured in a TEM) into and XRD-like distribution of intensity as a function of scattering vector (proportional to the radius of the ring). d-values and intensities are measured and listed. Markers show positions of known phases. And more ...
Previous problems with missing Help were also corrected for. (There was a mismatch between the name of the installed Help-file and the name the program looked for as Help.) Now the Help works fine and the Help system explains with illustrations what is the program good for and how to use it.
The program can be downloaded FREE from http://www.mfa.kfki.hu/~labar/ProcDif.htm . Steps of installation are also explained there. I suggest to uninstall previous versions and install the newest (V1.1.0) version.
I hope you will find this program useful and easy to use. Any comments, suggestions are welcome.
János
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
Sorry just checking... Tried to post something the other day but it didn't work.
---------------------------- Dr. Maxim V. Sidorov TEM Applications Specialist Philips Electron Optics, Applications Laboratory Building AAE, Achtseweg Noord 5 5600 MD Eindhoven, the Netherlands
} Hello dear microscopists! } } I have a question regarding artifacts in the myelin sheath of a peripheral nerve: } Samples were immersionfixed in Karnovsky´s half-strength fixative(2% Glut, 2% PFA in 0.1M Cacodylate)en bloc stained with osmiumtetroxide and UAc and embedded in Araldite. } The myelin sheaths show the following appearance: } 1. The fine lammellar structure is focally disturbed, the layers appear weavy, wobbly ... } 2. The outer rings are stained more intense than the inner rings (to the axon). } 3. The myelin sheaths are de-rounded, but this seems to be not so grave and I noted this often in other pictures published. Maybe this is "normal"? } } I would appreaciate any suggestions from you all to overcome this problem. } Thanks in advance! } Greetings, } Michael } } } Michael Reiner } University of Cologne, Germany } Dept. of Anatomy I
Michael Reiner University of Cologne, Germany Dept. of Anatomy I _______________________________________________________________________ 1.000.000 DM gewinnen - kostenlos tippen - http://millionenklick.web.de IhrName-at-web.de, 8MB Speicher, Verschluesselung - http://freemail.web.de
Dear All: I wrote a piece of software which I believe would be of interest to the microscopy community. It's a highly interactive program which calculates/displays the contrast transfer function of TEMs. I know that there are similar programs floating around but ctfExplorer does not only 1d but also 2d calculations/display with 2-fold and 3-fold astigmatism imposed. There are other unique features to it. All parameters (defocus, voltage, Cs, etc) can be changed interactively.
DISCLAIMER: I do work for FEI/Philips Electron Optics. However, this software is not an official product of FEI and FEI is not responsible for its distribution/support. This software is freeware.
To the best of my knowledge, the software does not have any nasty bugs. It is tested under Windows 95/98 and Windows NT4.
Please give it a try. Please direct your suggestions and comments to maxsidorov-at-bigfoot.com I hope you'll find the software useful.
Here is the link: http://clik.to/ctfexplorer
Enjoy,
Max Sidorov --- TEM Applications Specialist FEI/Philips Electron Optics Eindhoven, The Netherlands e-mail: maxsidorov-at-bigfoot.com
(Sorry again for my previous "test" post. Apparently my e-mail program was not set up correctly.)
We have just recently received and had installed a JEOL 5900LV sem and although training for this instrument is only a week away, I would like to do some basic study on the low vacuum aspect of electron microscopy. I am very familiar with high vac operation of sem's in general but lack knowledge of the techniques and uses of low vac. Any pointers to web sites that have a basic to advanced discussion of this subject would be greatly appreciated.
Peter Sterling SEM Lab, Structural Materials Labs Materials and Processes Rocketdyne Division, The Boeing Company (818) 586-1434
Open Position for an Electron Microscopist at Northwestern University Medical School
We have an opening for an Electron Microscopy Technologist in the Cell Imaging Facility of Northwestern University Medical School, hosted within the Department of Cell and Molecular Biology. This is a multi-user facility equipped with modern microscopy instruments and available for researchers of the whole university. The responsibilities of this position include: EM technical service; Supervising facility's daily activities; training users (EM, confocal and deconvolution microscopy, microinjection, and digital image analysis); maintenance of equipment; helping with developing new microscopy techniques; ordering supplies, and repairs; instrument scheduling and billing, updating web page and helping with reports. The preferred qualifications include: bachelor's degree in life sciences or the equivalent combination of education, training and experience from which comparable skills can be acquired; The candidates need to have electron microscopy and ultramicrotomy experience. Experiences with fluorescence, confocal and deconvolution microscopy, microinjection, image analysis and computer skills are very desirable. Salary is commensurate with experience and education. Please send applications to Dr. Weiming Yu, Department of Cell and Molecular Biology, Northwestern University Medical School, 303 E. Chicago Ave., Chicago, IL 60611. Detailed information about the Cell Imaging Facility can be found at http://www.basic.nwu.edu/corefacilities/cellimaging.html and http://www.cmb.nwu.edu/cif.html This is a full time position attached with full benefit package, which can be found at http://www.northwestern.edu/hr/benefits/
Weiming Yu, PhD Director, Cell Imaging Facility Northwestern University Medical School 303 E. Chicago Ave. W7-159 312/503-2841 fax:312/503-7912
Our admin people have decided to bar code all the equipment here, and need to have costs associated with the coding for security reasons. The following is a list of equipment that was either given to us, or we lost the paper trail and I'm having a hard time finding out prices for them. Prices and approximate dates of purchase, not estimated costs of present day value. If anyone has records of the same equipment and could give me a price and approximate purchase date that I could pass onto them, I would greatly appreciate it.
Edwards Coating System E306A Purchased in 1983 Reichert-Jung Ultracut E Microtome Sonicleaner Dawe Type 6443AE (rough price of a sonicator) LKB 2178 Knifemaker II Fisher slide warmer model 77 cat 12-594 Phillips EM-410 TEM
Thanks, Susan
Susan Carbyn Electron Microscopy Technician Atlantic Food and Horticulture Research Centre Agriculture and Agri-Food Canada 32 Main Street, Kentville, Nova Scotia B4N 1J5 Canada
Dear Listers, I'll be preparing samples of bacteria culture anaerobically and/or microanaerobically from sediments obtained at different bog sites in central Pa. I am looking for reference material for use with a class of graduate students. I expect to use conventional sample prep and an SEM to image Arthrobacter and Streptomyces. I am also looking for current literature on Aquaspirillum(magnetotactic bacteria). 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
Peter Sterling wrote: } I am very familiar with high vac operation of sem's in general but lack } knowledge of the techniques and uses of low vac. Any pointers to web } sites that have a basic to advanced discussion of this subject would be } greatly appreciated.
I have a few pages about EDS in LVSEM and ESEM at the address:
http://www.risoe.dk/afm/news1new.htm
Best regards, Jørgen
***************************** J. B. Bilde-Soerensen Senior Research Scientist, Ph. D. Materials Research Department Risoe National Laboratory DK-4000 Roskilde Denmark
Dear all, Out of interest (i.e. I am not in a position to buy one right now), I am interested in people's opinion on ion beam thinners on the market today.
I have used a Gatan PIPS in the past and have been very satisfied with this including many of the features including the low angle milling and the ability to mount a TV camera to watch the milling in real time (great for hole detection on transparent ceramics that defeat auto-termination systems).
I know, however, that Gatan are not the only manufacturers of ion-beam thinning equipment and I am sure that their competitors must also have some decent machines. So, please tell me what else there is on the market (since I already know plenty about the Gatan PIPS), what specifications these machines have, and about your experiences with them. Also, comparisons between different makes/models of ion beam thinner would be interesting.
Looking forward to hearing from you.
Best wishes
===== Ian MacLaren Beijing Laboratory of Electron Microscopy Chinese Academy of Sciences, P.O. Box 2724 100080 Beijing, China Email: ian.maclaren-at-physics.org / ianmaclaren-at-hotmail.com Home page: http://members.tripod.co.uk/IanMacLaren/
____________________________________________________________ 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
I am using Osmium to stain the binders in the coating layer. As you know, coated paper is composed of fibres (mechanical andchemical) and the coating layer (pigment minerals and binders). In order to try to predict how printing inks will interact with the paper surface during printing it is very important to characterise the coating layer. The quantification of the binders (styrene/butadiene latex) is thus important. How to do that?? I use to embedd paper samples in epoxy resin, take cross-section images in the SEM,BEI mode and use image analysis to quantify the binder distribution. However, it is very difficult to see any contrast between the binders in the coating layer and the epoxy resin. So, I stain the paper samples in Osmium before embedding in epoxy. The paper samples are placed in a staining chamber with a solution of 100 mg OsO4 in 10 ml water. The samples are not in contact with the solution in order to avoid structural changes in the paper cross-section. The samples are then vapour stained for 48 hrs. I get very nice results improving the contrast between the latex and the epoxy and also the mechanical (lighter) and chemical fibres. However, I have experienced a certain roughening of the paper surface due possibly to the water vapour inside the chamber. Now my question, how can I avoid this effect, Is it possible to dissolve OsO4 in another solvent?. I also tried to stain the samples with crystalline Osmium (without water), but the results are not so satisfactory.
Hi, Some time in the middle of the summer I put out a post seeking information on safe values for vacuum viewing port glass thicknesses, i.e., data on thickness of glass versus unsupported diameter for various glasses with atmospheric pressure on one side and vacuum on the other. Particular interest is in leaded glass and quartz and in small diameters in the range of 1 to 3 cm. I didn't get any responses but my guess is that a lot of list members were not monitoring the microscopy list as frequently as they normally would. So, with your indulgence, the question is reposted.
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.)
I have not used a Gatan PIPS but here at Sheffield University SERC FEGTEM Facility (http://www.shef.ac.uk/uni/academic/D-H/eee/) we have both the Leica EMRES 100 and the Technoorg Linda IV3 with an additional low energy gun. The Leica is good for general fast milling - it is completely computer controlled, with video camera viewing and end point determination, on-board timer and, in our case, Peltier cooling of the sample holder. You can just set the thing going and leave it for an hour or two. The Technoorg Linda, though not computer controlled, is more versatile, having both the facility to use a low energy ion gun and liquid nitrogen cooling to the sample holder - topping up is the problem here. Neither company has a factory in the UK (Leica's instrument is made by Balzers in Lichtenstein and Technoorg Linda are Hungarian) but both are very helpful when it comes to problem solving. Leica have a UK based rep. - not very useful in China I don't suppose! My point is that they are willing to give every possible assistance by email or phone or, if necessary, a site visit.
Hope this helps.
Kind regards,
Alan Walker
********************************************* Alan Walker Dept of Electronic and Electrical Engineering University of Sheffield Mappin Street Sheffield S1 3JD United Kingdom
} Hi! } } I am using Osmium to stain the binders in the coating layer. As you know, } coated paper is composed of fibres (mechanical andchemical) and the } coating } layer (pigment minerals and binders). In order to try to predict how } printing inks will interact with the paper surface during printing it is } very important to characterise the coating layer. The quantification of } the binders (styrene/butadiene latex) is thus important. How to do } that?? I use to embedd paper samples in epoxy resin, take } cross-section images in the SEM,BEI mode and use image analysis } to quantify the binder distribution. However, it is very difficult to see } any } contrast between the binders in the coating layer and the epoxy resin. So, } I stain the paper samples in Osmium before embedding in epoxy. The paper } samples are placed in a staining chamber with a solution of 100 mg OsO4 in } 10 ml water. The samples are not in contact with the solution in order to } avoid } structural changes in the paper cross-section. The samples are then vapour } stained for 48 hrs. I get very nice results improving the contrast } between } the latex and the epoxy and also the mechanical (lighter) and chemical } fibres. However, I have experienced a certain roughening of the paper } surface due possibly to the water vapour inside the chamber. Now my } question, how can I avoid this effect, Is it possible to dissolve OsO4 in } another } solvent?. I also tried to stain the samples with crystalline Osmium } (without water), but the results are not so satisfactory. } } Any help will be appreciated. } } Gary.
Osmium will dissolve in several organic solvents, carbon tetrachloride comes to mind.
Why dissolve it at all? I suspect that osmium crystals in a sealed container would produce enough vapor to do the job. Gentle warming would speed things up.
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 **********************************************
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Hi Ian,
I have not used a Gatan PIPS but here at Sheffield University SERC FEGTEM Facility (http://www.shef.ac.uk/uni/academic/D-H/eee/) we have both the Leica EMRES 100 and the Technoorg Linda IV3 with an additional low energy gun. The Leica is good for general fast milling - it is completely computer controlled, with video camera viewing and end point determination, on-board timer and, in our case, Peltier cooling of the sample holder. You can just set the thing going and leave it for an hour or two. The Technoorg Linda, though not computer controlled, is more versatile, having both the facility to use a low energy ion gun and liquid nitrogen cooling to the sample holder - topping up is the problem here. Neither company has a factory in the UK (Leica's instrument is made by Balzers in Lichtenstein and Technoorg Linda are Hungarian) but both are very helpful when it comes to problem solving. Leica have a UK based rep. - not very useful in China I don't suppose! My point is that they are willing to give every possible assistance by email or phone or, if necessary, a site visit.
Hope this helps.
Kind regards,
Alan Walker
********************************************* Alan Walker Dept of Electronic and Electrical Engineering University of Sheffield Mappin Street Sheffield S1 3JD United Kingdom
Have you tried using some other coating on your glass slides? Or putting the section on "Plus" slides (I use Fisher Scientific's, but I'm sure other companies have similar slides). I'm not sure the PLL is the best choice for this - I'd go with Plus slides or some "old-fashioned" type of subbing. I can send protocols if you are interested.
Good Luck!
Tamara Howard CSHL
On Wed, 6 Sep 2000, didier goux 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 Microscopist } } We persist to use Unicryl for cryo embedding (UV) while many problems. } } For in situ hybridization, the section of 3 mycron are put on polylysine } covered glasses } } this sections go away (floating) in SDS and SSC bath at 55 degres } } how could we "fixe" the sections on glasses? } } thank you for reply } } Didier } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } Didier Goux } CENTRE DE MICROSCOPIE ELECTRONIQUE } Université de CAEN Tel : (33) 02.31.56.58.13 } Campus I Fax : (33) 02.31.56.56.00 } Esplanade de la Paix 14032 CAEN CEDEX } mailto:didier.goux-at-unicaen.fr } } } } } } }
Does anyone know if the M&M 2000 proceedings have been mailed yet.?? Greg Gregory W. Erdos, Ph.D. Assistant Director, Biotechnology Program PO Box 110580 University of Florida Gainesville, FL 32611
You can probably go pretty thin with sapphire, given your small diameters and resulting pressures of less than 50 pounds. Most materials should work.
Call Melles Griot to ask about burst pressure and strength modulus for their products. That will help. Their contact information is:
E-mail: optics-at-irvine.mellesgriot.com
Call 1-800-835-2626 or 949-261-5600 and ask for the Optics Specialist
Fax 1-949-261-7790, attention Optics Specialist
Good luck,
Nathan Haese Lafayette, CA
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I think that there is old work (From Afzelius?) in which osmium tetroxide was used in some organic solvent (carbon tetrachloride or chloroform?) to fix difficult specimens (eggs ?). You could check in old texts on TEM - like Pearse's "Histological Techniques for Electron Microscopy"
Dr. A.P. Alves de Matos Anatomo-Pathology Department Curry Cabral Hospital Lisbon
----- Original Message ----- } From: Gary Dietrich Chinga {garyc-at-stud.ntnu.no} To: ERIC {biology-at-ucla.edu} Cc: {microscopy-at-sparc5.microscopy.com} Sent: Wednesday, September 06, 2000 10:33 AM
They were mailed before the mtg. Contact Bill Bailey or the Springer guy: Herb Neimerow -at- 212-460-1686
Ron Anderson, IBM, Hopewell Jct., New York, USA. anderron-at-us.ibm.com
IBM Analytical Services; http://www.chips.ibm.com/services/asg
Didier, You can use treat glass with 1% silane in acetone to make it sticky. Marek.
At 9/6/00 Wednesday01:52 PM-0700, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Marek Malecki, M.D., Ph.D. Director and Principal Investigator
Molecular Imaging Laboratories University of California at San Diego address: 1500 Suite Bonner Hall, 9500 Gilman Drive, La Jolla, CA 92093-0368 phone: 8588223373 fax: 8588223715 pager: 8586161420 e.mail: mmm-at-ucsd.edu e.pager: 1620024619-at-alphapage.airtouch.com www site: http://mil.ucsd.edu/ ftp site: mil1.ucsd.edu
Workshop Objective 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 specimen preparation technicians. This course has been updated and expanded from past workshops to include pre-FIB preparation techniques, post-FIB Plasma Trimming as well as the latest techniques available in low energy 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 October 27-28, 2000. Please contact me directly 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
Would anyone have for sale or trade-in a lens control system for JEOL's 1200EX?
Marek Malecki, M.D., Ph.D. Director and Principal Investigator
Molecular Imaging Laboratories University of California at San Diego address: 1500 Suite Bonner Hall, 9500 Gilman Drive, La Jolla, CA 92093-0368 phone: 8588223373 fax: 8588223715 pager: 8586161420 e.mail: mmm-at-ucsd.edu e.pager: 1620024619-at-alphapage.airtouch.com www site: http://mil.ucsd.edu/ ftp site: mil1.ucsd.edu
Dear Ian, I have had a VCR Group Ion Mill for several years now and have been very happy with it. It very seldom needs service, is easy for me to service if needed, does low-angle and atom thinning, has a liquid N2-cooled stage and an automatic terminator based on an ion gun, which works fine for transparent samples. My model is old now and the newer ones are computer controlled. The VCR Group is now part of South Bay Technologies. (www.southbay.com) At 09:04 AM 9/6/00 +0100, you wrote: } } Dear all, } Out of interest (i.e. I am not in a position to buy one right now), I am } interested in people's opinion on ion beam thinners on the market today. } } I have used a Gatan PIPS in the past and have been very satisfied with this } including many of the features including the low angle milling and the ability } to mount a TV camera to watch the milling in real time (great for hole } detection on transparent ceramics that defeat auto-termination systems). } } I know, however, that Gatan are not the only manufacturers of ion-beam thinning } equipment and I am sure that their competitors must also have some decent } machines. So, please tell me what else there is on the market (since I already } know plenty about the Gatan PIPS), what specifications these machines have, and } about your experiences with them. Also, comparisons between different } makes/models of ion beam thinner would be interesting. } } Looking forward to hearing from you. } } Best wishes } } ===== } Ian MacLaren } Beijing Laboratory of Electron Microscopy } Chinese Academy of Sciences, P.O. Box 2724 } 100080 Beijing, China } Email: ian.maclaren-at-physics.org / ianmaclaren-at-hotmail.com } Home page: http://members.tripod.co.uk/IanMacLaren/ } } ____________________________________________________________ } 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 } } 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
There is a Siemens Elmiskop II TEM that has been mothballed and placed on display in the lobby of a lecture hall on the University of California, Davis, campus. Several years ago we taught a course and it was used as the student scope. Once decommissioned, the power supply and ht cable were given away to an EM service company and the console was placed in the lobby as an artifact of the early days of EM on the UC campus. The lecture hall now belongs to a department that has no interest in keeping the old scope on display and would like to dispose of it. Is there any interest out there in this device either for parts or for a display item? If not, it will be buried at the landfill.
Contact me off-list if you have any interest or suggestions for this scope.
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
Try the following company to get your answers: Larson Electronic Glass (Redwood City, CA) Tel. (650)369-6734. E-mail gls2mtl-at-juno.com . I am not aware of their web site.
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax -----Original Message----- } From: donald j marshall {dmrelion-at-world.std.com} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
I was wondering the same thing. I haven't received my copy as of 6-Sept-00.
Hope they are mailed shortly. Michelle Taurino Aventis Pharmaceuticals Senior Scientist Bioimaging and Molecular Histology Tel-908-231-3357 Fax-908-231-3962 e-mail: Michelle.Taurino-at-aventis.com
-----Original Message----- } From: Greg Erdos [mailto:gwe-at-biotech.ufl.edu] Sent: Wednesday, September 06, 2000 9:47 AM To: Microscopy-at-sparc5.microscopy.com
Does anyone know if the M&M 2000 proceedings have been mailed yet.?? Greg Gregory W. Erdos, Ph.D. Assistant Director, Biotechnology Program PO Box 110580 University of Florida Gainesville, FL 32611
No one in my group has received theirs either...has *anyone* received their Proceedings yet?
Larry
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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 got mine right after returning from Philadelphia.
Randy
-----Original Message----- } From: Larry Allard [mailto:l2a-at-ornl.gov] Sent: Thursday, September 07, 2000 7:59 AM To: Microscopy-at-sparc5.microscopy.com
No one in my group has received theirs either...has *anyone* received their Proceedings yet?
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
Responding to the message of {v04210101b5dd412fa34d-at-[128.219.47.130]} from Larry Allard {l2a-at-ornl.gov} : } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } No one in my group has received theirs either...has *anyone* received } their Proceedings yet? } } Larry }
I received mine very shortly after I returned, but it is possible that I was a special case :)
Thank you all for attending, and I know next year's program commitee will look forward to seeing you next year in Long Beach CA.
__________________ Stuart McKernan stuartm-at-tc.umn.edu 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
I am preparing turf grass samples for TEM. We will be examining the crowns and lead blades for infectious agents. Any tidbits on fixation and embedding of this tissue would be appreciated!
My starting point is this protocol, which has been used for wood samples. If it is overkill, please let me know.
Fix: 2% Glutaraldehyde in 50 mM NaCacodylate pH 7.2, with light vacuum for 24 hours Wash: 3X for 1 H each RT in buffer with mild agitation PostFix: 1% OsO4 in water Overnight at 4C Wash: 3X for 1H each at RT in water with mild agitation en bloc stain: saturated UA for 30 min with mild agitation Dehydration series: 25%, 50%, 75%, 100% Acetone, each 30 min with mild agitation 100% acetone + 1% DMP for 30 min with mild agitation -- 100% acetone + 1% DMP overnight with mild agitation 100% acetone + 1% DMP overnight with mild agitation Infiltration: 33% Spurr's in Acetone + 1% DMP for 2 H min with mild agitation 66% Spurr's in Acetone + 1% DMP for 2 H min with mild agitation 100 % Spurr's for 2H with mild agitation 100 % Spurr's Overnight with mild agitation 100 % Spurr's for 8-12 H with mild agitation 100 % Spurr's for 8-12 H with mild agitation Cast in fresh resin
Thanks, Sally
Sally Burns Center for Advanced Microscopy B5 Center for Integrated Plant Studies Michigan State University East Lansing, MI 48823 (517) 355-5004 burnssal-at-msu.edu
I checked with Springer on this. They received a list of 227 names to ship Proceedings to at the end of June. Shipment occurred about August 1. It is safe to assume that not all of them have reached their destinations yet. The form you filled out in the registration bulletin said it would take 4-6 weeks
HERE'S THE IMPORTANT PART: If you signed up as a full meeting registrant AFTER June 15th (or so) and elected to have the Proceedings shipped, Springer didn't get the labels yet. I called the Meeting Manager's office and, after a short chat, got a commitment that they would ship the additional labels overnight to Springer. Herb, at Springer, committed to getting the additional proceedings shipped early next week.
Ron Anderson MSA Pres-Elect
Ron Anderson, IBM, Hopewell Jct., New York, USA. anderron-at-us.ibm.com
IBM Analytical Services; http://www.chips.ibm.com/services/asg
Available with the Center for High Resolution Electron Microscopy at Arizona State University. Position is sponsored by major chemical company and primary focus of the research will be the development and application of environmental electron microscopy to industrially relevant catalyst systems. Areas of particular interest include the study of phase transformations under reaction environments, in situ polymerization, mobility and dynamic microstructural changes. Candidate will have a Ph.D. in material science, material physics, solid state chemistry or chemical engineering, with extensive experience in catalyst characterization by transmission electron microscopy. Experience in the areas of catalyst synthesis, testing and characterization is preferred. Please submit your resume together and the names of 3 referees to: Dr. Peter A. Crozier, Industrial Associates Program, Center for Solid State Science, Arizona State University, Tempe, AZ 85287-1704, Fax (480) 965-9004, email crozier-at-asu.edu.
John C. Wheatley Lab Manager Arizona State University Center for Solid State Science PSA-213 BOX 871704 Tempe, AZ 85287-1704
} Hi - } } I have a project which needs to see the preservation of testis and } kidney morphology of rats. I want to set up a couple of ultrastructural } criteria for this. Can any of you provide me some info about good } classical reference books or literature. Any experience or suggestions } are greatly appreciated. } } Regards } } Gang Ning } } EM Facility } Medical College of Wisconsin } Milwaukee, WI 53045 } 414-456-8141 } 414-456-6517 (Fax)
See "The ultrastructure of the rat renal medulla as observed after improved fixation methods" by S-O. Bohman. J. Ultrastruc. Res 47:329-360, 1974. Beautiful EMs. This paper deals with the medulla for the most part, renal cortex is not too difficult to fix well. Any edition of "The Kidney" by Brenner and Rector (see vol. 1) should provide good references. As far as testis goes, Martin Dym was a major researcher in that field long ago (20 years or so). A lit search for his name might be productive. Or look at the list of references at the end of the chapter on male reproductive system in any good histology text (Bloom and Fawcett, Ham [when Ham was still writing it]).
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 **********************************************
Please excuse me if you've already received this message - I sent it yesterday to both servers, but received it from neither, so I think our system ate it on its way out :)
Tamara
---------- Forwarded message ----------
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I received my proceedings over 1 week ago....They were no doubt sent 3rd class so may take a number of weeks for all to be delivered depending on where you are in the country. Debby
--------------------------------------
No one in my group has received theirs either...has *anyone* received their Proceedings yet?
Larry
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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
Mine were here when I returned from the Conference.
Richard Shalvoy Arch Chemicals Cheshire, CT
-----Original Message----- } From: Larry Allard [mailto:l2a-at-ornl.gov] Sent: Thursday, September 07, 2000 8:59 AM To: Microscopy-at-sparc5.microscopy.com
No one in my group has received theirs either...has *anyone* received their Proceedings yet?
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
I'm having a bit of trouble with Formvar film stability under the TEM beam at 60 kV. The films, attached to copper or nickel grids (cleaned with dilute HCl + acetone in sonicator same day films picked up) develop holes which grow over a few minutes time until eventually the entire film more or less disintegrates, collapses. I also noticed that initially, the films are often pulled away from the rim of the grid by about one square "hole" away.
I'm using new Formvar 15/95 resin powder and new bottle of ethylene dichloride solvent, mixing to 0.3% weight to volume. I use "Rite-On" glass slides, rinsed with ethanol to clean of dust and particulates and air dried, to cast the films. The films float off easily, and look very clean under the TEM. They were dried overnight prior to inspection. I take pains to use very clean glassware for mixing the Formvar solution.
Of course, I could caarbon coat them to provide stability, but that's extra work and I don't seem to have had this kind of problem before.
I barely recall some discussion here some time ago about how Formvar has "changed" due to different process of manufacturing, or whatever.
Any clues?
Thanks in advance.
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
First, a thank you to Mary for the kind words about our ion milling system. Just a quick correction, our website is actually at www.southbaytech.com. Hopefully this will slow down the number of calls we have received asking us why we are now selling DSL service!
Unfortunately, we do not have any information yet on the website about the XLA2000 ion mill so you will need to contact me directly on that. There is some information on the Technorg-Linda Low Energy Mill.
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 Mary Mager } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Ian, I have had a VCR Group Ion Mill for several years now and have been very happy with it. It very seldom needs service, is easy for me to service if needed, does low-angle and atom thinning, has a liquid N2-cooled stage and an automatic terminator based on an ion gun, which works fine for transparent samples. My model is old now and the newer ones are computer controlled. The VCR Group is now part of South Bay Technologies. (www.southbay.com) {
we routinely use formvar at a concentration of 0.5 - 0.7 %. however, you shurely are aware that the thickness of the film critically depends on the lowering speed of the fluid (slow = thin; fast = thick). if you can manage, coat with carbon afterwards, there are no disadvantages by doing so, only advantages. good luck peter
************************************************** please reply to: peter.heimann-at-biologie.uni-bielefeld.de ("E-MAIL")
Dr. Peter Heimann Developmental Biology & Molecular Pathology; W7-107 University of Bielefeld D 33501 Bielefeld / Germany phone: xx49(0)521-106-5628 / 5627 FAX : xx49(0)521-106-5654 WEB-Site: http://www.uni-bielefeld.de/biologie/Entwicklungsbiologie WEB-Site: http://www.uni-bielefeld.de/SFB549 ******************************************************
one of the best techniques to perfuse (testis, but brain and other organs as well) is published by: Forssmann, WG et al; 1977; Anat. Rec. 188; (3); 307- 314
for excellent testis-EM look for instance after author Holstein, A.F.. I for myself use the "forssmann"-technique on mouse testis with excellent results.
good luck
peter heimann ************************************************** please reply to: peter.heimann-at-biologie.uni-bielefeld.de ("E-MAIL")
Dr. Peter Heimann Developmental Biology & Molecular Pathology; W7-107 University of Bielefeld D 33501 Bielefeld / Germany phone: xx49(0)521-106-5628 / 5627 FAX : xx49(0)521-106-5654 WEB-Site: http://www.uni-bielefeld.de/biologie/Entwicklungsbiologie WEB-Site: http://www.uni-bielefeld.de/SFB549 ******************************************************
Here in Connecticut it is often difficult to make good formvar films in the summer, due to high humidity. The films are covered with holes of varying sizes, even when made in a dry bottles and with freshly opened solvent. I think this problem is caused by condensation of water on the glass surface, which is cooled significantly during evaporation of the solvent. It doesn't take much cooling to get condensation when the ambient humidity is 90%! Perhaps you have tiny pinholes in your films that are enlarging to become visible under the beam.
Marie
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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
Position Open -- Immediately Microscopy and Microanalysis Research Assistant
An immediate position is open for an SEM microscopist at the North Carolina State University Analytical Instrumentation Facility (AIF). Duties and responsibilities include: Operation and maintenance of SEM instrumentation and sample preparation and analysis equipment; scheduling of access to and oversight of the above instrumentation; user
training and assistance; assistance with the teaching of electron microscopy laboratory classes, and analysis of a wide variety of samples. Qualifications must include a BS or MS or equivalent experience in a materials related discipline along with one or more years hands on experience with SEM or related techniques. Required skills include: extensive hands on experience with SEM and related techniques and accessories (e.g. specimen preparation and associated analytical tools). Preferred qualifications include: teaching and user training; familiarity with modern electronics; and experience with vacuum systems. Please send resume and three letters of reference to: Phil Russell, Director; Analytical Instrumentation Facility; North Carolina State University; Box 7531, Room 318A EGRC; 1010 Main Campus Drive; Raleigh, NC 27695-7531 or email PRUSSELL-at-NCSU.EDU.
North Carolina State University is an Equal Opportunity and Affirmative Action Employer. ADA Accommodations: Phil Russell, prussell-at-ncsu.edu, 919-515-7501. In its commitment to diversity and equity, North Carolina
State University seeks applications from women, minorities, and persons with disabilities
Phillip E. Russell Professor, Materials Science and Engineering Director, Analytical Instrumentation Facility Box 7531, Room 318 EGRC 1010 Main Campus Drive North Carolina State University Raleigh, NC 27695-7531
You can slightly improve stability under the beam by increasing kV, using a thinner film (and finer mesh grids), using less beam current and better still irradiating the grid at very low powers (200x) for a couple of minutes. The brightness appears great at low powers but actual beam intensity is much greater at higher powers and the lower power irradiation stabilises the film. Ultimately you can carbon coat.
BUT WHY are you and most people who really want to use plain plastic films use Formvar??? Butvar is the strongest and most stable of plastic films and is much more stable under the beam than Formvar without carbon. EM techniques (for some obvious reasons) die hard. Butvar has been around for over twenty years but people keep on going back to the old Formvar references. Disclaimer: ProSciTech does not care if you buy Butvar or Formvar. 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 ABN: 99 724 136 560 www.proscitech.com
On Friday, September 08, 2000 7:22 AM, Gib Ahlstrand [SMTP:giba-at-puccini.cdl.umn.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 Microlisters, } } I'm having a bit of trouble with Formvar film stability under the TEM beam at } 60 } kV. The films, attached to copper or nickel grids (cleaned with dilute HCl + } acetone in sonicator same day films picked up) develop holes which grow over a } } few minutes time until eventually the entire film more or less disintegrates, } } collapses. I also noticed that initially, the films are often pulled away from } } the rim of the grid by about one square "hole" away. } } I'm using new Formvar 15/95 resin powder and new bottle of ethylene dichloride } } solvent, mixing to 0.3% weight to volume. I use "Rite-On" glass slides, rinsed } } with ethanol to clean of dust and particulates and air dried, to cast the } films. } The films float off easily, and look very clean under the TEM. They were dried } } overnight prior to inspection. I take pains to use very clean glassware for } mixing the Formvar solution. } } Of course, I could caarbon coat them to provide stability, but that's extra } work } and I don't seem to have had this kind of problem before. } } I barely recall some discussion here some time ago about how Formvar has } "changed" due to different process of manufacturing, or whatever. } } Any clues? } } Thanks in advance. } } 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 }
Is coating with carbon more work than remounting a specimen after you have destroyed it with your beam. Carbon coating is part of the procedure in my lab, and students know not to stick a grid into the scope without it. We observe many samples weekly at 120kV without destroying any. Think of all the contamination you are introducing into the specimen chamber when you "burn" your film and sample.
I have a BalTec RES 100 and I am extremely pleased with it. There are quite a few options available with it. Not only have I have prepared very good TEM samples with it, but I have used it for ion polishing and ion etching of SEM samples for our High Resolution FESEM. BalTec has a number of holders for SEM applications. In addition, I can use it to sputter deposit films onto samples. The newest guns that I just had put on ours our very stable and settle down very quickly. Whatever changes they made in the software and hardware dramatically improved the operation over the first generation guns. I really like saving recipes available that I can reuse in the programming mode. Although I don't have the opportunity to use it much on glass samples, the image processing for termination is probably the most sensitive method that I have used. There are a lot of options that the user can adjust for optimizing the optical termination through image processing. Alignment of the guns is extremely easy and accurate. Further, they stay aligned. If you get the ion deflection system, they have an automatic alignment system. I bought ours because of the Faraday cup termination that they have available. However, I hardly use it. With the camera system, it is easy to watch the screen to determine when it is finished. I have the remote control/monitoring capability but have not put the system on our LAN yet. I plan to do that very soon.
If you have a FESEM in the same lab as your TEM, you should consider what this mill could do for you.
PS Let me tell you that I like the PIPS also. It is very simple to use. It has one mode that is unique to it, the ability to mill on both top and bottom of the sample from one direction coming in from the perpendicular direction to the interface of a cross section. Reza Alani of Gatan has shown many examples of beautiful samples prepared this way. But, if I were to make the decision again, I would still go with the BalTec system. It is versatile and can be used in a high tech way as well as a routine way with technicians pumping through samples. BalTec has responded to my needs and have considered things that I have said in terms of their software updates. I have experimented with some sample preparation techniques for SEM that look very promising using this mill.
Although I do not have direct experience with the other ion mills on the market, I have talked extensively with their manufacturers and they all have some nice features. The low energy guns in the Technorg-Linda system designed by Barna should be carefully considered. My advice to you is to consider what you want the mill to do in your lab with your samples and go from there. You need to weigh what features are available on each and how they would apply to your samples. You also have to weigh experience with your users to the level of competency required for each of the mills out there.
I hope this helps.
-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: Ian MacLaren [mailto:maclariz-at-yahoo.co.uk] } Sent: Wednesday, September 06, 2000 4:05 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Modern ion beam thinners } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: 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 } } } -------------------------------------------------------------- } ---------. } } } Dear all, } Out of interest (i.e. I am not in a position to buy one right } now), I am } interested in people's opinion on ion beam thinners on the } market today. } } I have used a Gatan PIPS in the past and have been very } satisfied with this } including many of the features including the low angle } milling and the ability } to mount a TV camera to watch the milling in real time (great for hole } detection on transparent ceramics that defeat } auto-termination systems). } } I know, however, that Gatan are not the only manufacturers of } ion-beam thinning } equipment and I am sure that their competitors must also have } some decent } machines. So, please tell me what else there is on the } market (since I already } know plenty about the Gatan PIPS), what specifications these } machines have, and } about your experiences with them. Also, comparisons between different } makes/models of ion beam thinner would be interesting. } } Looking forward to hearing from you. } } Best wishes } } ===== } Ian MacLaren } Beijing Laboratory of Electron Microscopy } Chinese Academy of Sciences, P.O. Box 2724 } 100080 Beijing, China } Email: ian.maclaren-at-physics.org / ianmaclaren-at-hotmail.com } Home page: http://members.tripod.co.uk/IanMacLaren/ } } ____________________________________________________________ } 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 }
----- Original Message ----- } From: {keg-at-alpha3.csd.uwm.edu} To: {keg-at-csd.uwm.edu} Sent: Friday, September 08, 2000 9:23 AM
Hello Tamara and All, An unfortunate, although interesting predicament. There are three variables that come to mind which may affect the shipping strategy: temperature change, pressure change and time. I worry that leaving the samples in methanol/acetic acid and raising the temperature would lead to denaturation of protein structure due to overfixation. If the samples could be kept acceptably cool during transport this might help, but I'm not sure of the technical difficulties (and expense) associated with overseas transport on dry ice-I believe Fed Ex has pressurized cargo (which would slow the sublimation of the dry ice). If these samples could be sent via overnight express this would probably work. Otherwise, placing the samples in a buffer solution similar to that proposed for your incubation protocol(s) for shipping could be a viable approach. Adding some sodium azide to the buffer for transport to prevent bacterial contamination would be a good idea in my opinion. I haven't tried any of these approaches but perhaps someone with more experience than I will agree/disagree and you will be able to find a quasi-reliable approach to your problem. Good Luck! Cheers, Karl Garsha ----- Original Message ----- } From: Tamara Howard {howard-at-cshl.org} To: Microscopy Listserver {Microscopy-at-sparc5.microscopy.com} ; Histology listserver {histonet-at-pathology.swmed.edu} Sent: Thursday, September 07, 2000 11:18 AM
Hello, Used equipment can be auctioned off/bidded on at www.biobid.com. It seems like a nifty idea. Regards, Karl Garsha ----- Original Message ----- } From: MICHAEL DELANNOY {delannoy-at-mail.jhmi.edu} To: {microscopy-at-sparc5.microscopy.com} Sent: Tuesday, September 05, 2000 3:21 PM
All: does anyone in the North Texas area have a carbon coater capable of handling an 8-inch wafer? We need to get a wafer coated prior to some FIB edit and ours has not returned from the vendor. Please reply to me so we don't clutter up the listserver.
I have used Sparkle Window cleaner (its the purple one!) with great success but beware that there are big differences in window cleaner brands. good luck. tom
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- 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)
A postdoctoral position is available immediately in the area of Environmental Catalysis. The position will be part of the Institute for Environmental Catalysis (http://www.iec.nwu.edu) and will involve TEM, both HREM as well as diffraction (Direct Methods) on oxide surfaces as a function of gas treatment in collaboration with other faculty at Northwestern as well as Industry, using the unique UHV-HREM at Northwestern University (see http://www.numis.nwu.edu). A strong background in TEM is required, and expertise in UHV and associated techniques (e.g. XPS) is desirable.
Send applications (by email) to: Professor L. D. Marks Department of Materials Science and Engineering mailto:ldm3-at-apollo.numis.nwu.edu
Include a CV and the names of at least two referees.
Friday, September 29, 2000 University of Wisconsin Memorial Union South 227 North Randall Avenue Madison, WI 53715 (608) 263-2600
Admission is free with MMMS Membership. MMMS Membership is $10.00 - new members will be accepted at registration
Meeting Schedule
MORNING COMBINED SESSION, Room, Union South 9:45AM - 10:30AM Registration - Coffee, Juice, Pastries will be provided.
10:30 AM Opening remarks
10:35 AM Dr. Sue Welch, UW-Madison Department of Geology and Geophysics "The Biogeochemical Cycling in the Tennyson Mine" Co-authors: Matthias Labrenz, Anne Skatvold, Gregory K. Druschel, David A. Fowle, Tamara Thomsen-Ebert and Jill Banfield
11:35 AM Dr. Will Bigelow, Professor Emeritus of Materials Engineering, University of Michigan "The current state of vacuum science in microscopy, or: why it still takes so long to pump down to an operating vacuum"
AFTERNOON BIOEMPHASIS SESSION: Room , Union South
1:30 PM Hans Ris, Professor Emeritus, UW Madison. "3D Imaging of cell structures by high resolution , low voltage , field emission SEM"
2:00 PM Daryl Meyer, UW Madison "Multiple labeling for EM: Synthesis and use of colloidal nano-particles of different metal composition and shape."
2:30 PM Judith Croxdale, UW Madison "Leaf Surfaces, Microscopy and Thievery useful in Analysis".
3:00 PM Jayne Squirrell, UW Madison "Application of multiphoton microscopy to the study of embryo development".
AFTERNOON PHYSICAL SCIENCE EMPHASIS SESSION: Room , Union South
1:30 PM Carl Loper, UW-Madison Dept of Material Science & Engineering "Interaction of Pb and Ca and Its Effect on Graphite Morphology in Cast Irons" co-authors: Junyoung Park and John Fournelle
2:00 PM Reid Cooper, UW-Madison Dept of Material Science & Engineering "Redox Reactions in Silicate Melts and Glasses"
2:30 PM John H. Perepezko, UW-Madison Dept of Material Science & Engineering " Synthesis and Initial Crystallization of Amorphous Al Alloys" coauthors: R.I. Wu, R. Hebert and Z. Dong, University of Wisconsin-Madison
3:00 PM Thomas F. Kelly, UW-Madison Dept of Materials Sceince & Engineering, and Imago Scientific Instruments Corporation, Madison, WI "Local Electrode Atom Probes: Background and Application to Microelectronic Materials"
3:45 PM CHEESE, CHEESE, & ICE CREAM (remember this is Wisconsin) RECEPTION -- UNION SOUTH
4:45 PM MEETING ADJOURNS Have a SAFE ride home
Support of this meeting has been provided by: University of Wisconsin Department of Geology & Geophysics, The Biological & Biomaterials Preparation, Imaging, & Characterization Laboratory, The Advanced Microscopy Facility, and "Microscopy Today"
Getting to Madison:
Madison is located in south central Wisconsin and is accessible via several major highways. Madison is:
- 1 1/2 hour drive from Milwaukee (via Interstate 94) - 2 1/2 hour drive from Chicago (via Interstate 90) - 4 1/2 hour drive from Minneapolis/St. Paul (via Interstate 94) - 2 hour drive from Dubuque (via US 151)
Getting to UW-Madison:
- Take I-90 or US 151 to Hwy 12/18 - Follow Hwy 12/18 exit at Park Street (Exit 261 B) - Proceed north on Park Street about four miles to Regent Street and turn left. - Proceed on Regent to Randall St. and Turn Right. (See detail map.)
Below is a message from the family of Hildy Crowley. I thought it would be of interest to the many people who subscribe to this list why she signed off recently, and so abruptly. I asked her son if it would be OK to put it here. He thought it was a good idea and that it would be what she wanted as she loved EM so much. If you think it is appropriate I will let you post it. I know that even though I never met her, your listserver brought us close and that I will miss her.
Paul Webster
HILDY CROWLEY 29 December 1935 - 7 September 2000
After a long and difficult illness Hildegard Heinrich Crowley died quietly at home on September 7. Earlier in the week she had given her grandsons, ages 2 and 4, Batman slippers. The boys showed their Oma (German for Grandma) how much higher the slippers let them jump. A few days before that she had received the Morton D. Masur Distinguished Service Award from the Microscopy Society of America because, through the years her advice also had allowed many electron microscopists to jump a little higher. With Hildy's help anyone could jump a little higher.
Hildy was born in Stuttgart, Germany. Her mother, also Hildegard, died that day. She was raised for several years by aunts and a Grandmother in North German Pomerania, now a part of Poland. Her father then remarried and she joined him again. Throughout her life she had frightening wartime dreams from those days – bombings, straffings, government repression, malnutrition, and Allied tanks arriving in her town of Eslingen.
Hildy's father, a distinguished aeronautical engineer, was one of the German scientists brought to the US after the war. Only after many months could the family join him. Years later she spoke with joy of those early teen years in America, with plenty of food, nice teachers, and wonderful friends.
A superb student, Hildy also won prizes at the County Fair for cooking and sewing, and she was an attendant to the college Homecoming Queen; beauty contests still were "in" then. After college she taught junior high school science.
In 1962 Hildy married Tom Crowley, a young medical student. In 1963 she began working as an electron-microscopy technologist at the University of Minnesota. She was a perfectionist, and EM is a perfectionist's dream and nightmare. "I love solving problems", she said, and each EM problem solved exposed ten new ones. The couple's sons, Christopher and Devin, arrived in 1965 and 1967.
After several years of raising youngsters, Hildy returned to EM at the University of Colorado Health Sciences Center, then at the National Jewish Hospital, and for the last decade, at Denver University. Armed only with a Bachelor's degree, superb intellect, and dogged determination, Hildy became internationally recognized for solving technical problems in EM, focussing in recent years on immunocytochemistry. She was especially proud of the lab's display of technical expertise in two Comparative Neurology articles (July and September 2000), on which she was a prominent co-author.
Hildy and Tom had moved to Denver for its skiing. Again the technical perfectionist, she became expert in equipment, waxes, and edge care. Her skiing continued improving well into her 60's, when she still did the easier expert runs some 35 days each year.
If Hildy was great at EM, she was even greater at teaching it. Brilliant undergraduate students came to the lab for Honors Theses. Hildy taught them biology and EM, of course, but she also supported them through romantic crises, sometimes helped them find physicians or therapists for their physical or personal problems, and advised them about their parents' illnesses or malfeasances. She also fed them delicious home-made cakes at lab meetings. She at first was miffed when someone called her the "Lab Mom", until a student said plaintively, "I want you to be our Mom. You're so much better than my own Mom". She gave students level-headed, practical, grandmotherly, every-day advice; like her ancestors from the foggy, cold Baltic, she distrusted "that touchy-feely stuff". The students went off to fine medical or dental or graduate schools, but when they visited Denver they often took her to lunch. They invited her to attend, or to be in, their weddings; one came back to ski with her. After knowing her, they all jumped a little higher.
Hildy's greatest commitment and contribution, of course, was to her family. Their loss is too large to speak of here.
Hildy learned in June 1999 that her illness would be terminal, but she wanted that kept secret from most of her friends, who, at the Portland Microscopy Society meeting, thought her gift of a limo ride to a fancy restaurant was just a madcap moment. It was a farewell.
Being Hildy, she worked with Tom to design her own simple headstone, which was erected a couple of months ago. It says, "You shall live in what you love." She loved so much, and so many, and so well. Hildy, with love, farewell. Farewell Oma, Mom, Wife, Sister, Lab Mom, Scientist, Teacher, Mentor, Friend, Colleague. Our Batman slippers fit just right. You helped us all jump higher.
-------Tom
Hildy's funeral will be at the First Universalist Church, 4101 East Hampden Avenue, Denver, at 10 AM Tuesday, September 12. A graveside interment ceremony at Fairmount Cemetery will follow.
Paul Webster, Ph.D. Associate Scientist & Director Ahmanson Advanced Electron Microscopy & Imaging Center House Ear Institute 2100 West Third St. Los Angeles, CA 90057
I had no idea who she was, but I always liked reading her postings, and her last one was just as gracious as ever.
My sympathy to her family.
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 Everybody! We have two Hitachii TEM's (H600 and H7000). We use liquid nitrogen to improve the vacuum (back trap) and to decrease contamination of the column (front trap). Recently, I have been told by the university safety officer that I can not use LN in the back trap (I and the users have to climb on a step ladder and lift LN container to fill the trap) because of the safety reasons. One of our users strained her back while filling the back trap and she has been on a sick leave for some time. They suggested to build a platform in the back of a microscope so it is more sturdy and safer. Have you ever had problems of that sort? The lab has been opened for 15 years and we have never had any accident with LN. Is the platform really safer? Will it interfere with the performance of the microscope, if it is metal one? Will performance of the scope be affected if I do not use LN in the back trap? Thanks for answers Dorota
I apologize in advance for what may well be a very sophomoric question.
I haven't paid attention to what may have been discussed regarding alternatives to Freon 113 as a final drying agent because I have scrupulously rationed my supply, ml by ml, for several years. Now I'm into my final 300 ml and must begin to search for a substitute since it has been my understanding that Freon 113 is no longer available. I only use very small volumes at a time and 4x500 ml bottles has lasted me nearly 10 years. Most of my specimens are cell layers or other small (less than 1 cubic mm) tissue samples for SEM.
Any suggestions and information about what is currently used as an alternative to critical point drying out of Freon 113 exchanged with liquid CO2 would be greatly appreciated.
Thanks,
Gerald Harrison Electron Microscope Facility Dental School Research University of Pennsylvania
We are looking for camera suggestions that will handle Low Light, Real time imaging(both for flurorescence and DIC) in the fluorescence microscope. Something in the $20K range or less for the system.
High resolution is required also. These images can be extremely dark. The user prefers also to be able to output the realtime imaging of cell growth to videotape as that has proved to be the most convenient in the past but is also interested in the "firewire" option and it's capabilities.
We realize that a compromise in some areas (likely the frame rate)will be required but high resolution during low light imaging is very important.
We would be particularly interested in hearing from others that have used a camera that would suit our application. We have commercial info for several models but would really like some user feedback.
Thanks,
Karen Rethoret York University Biology Department Toronto, Ontario 416-736-2100 x33289
I am very sad about the message that reached me today.
Unfortunatly, I knew Hildy only as a member of the listserver and by private emails. She was a source of wisdom in this community. I told her about technical problems and she repeated promptly. One day a letter from Denver reached me here in Germany with her protocol and I was so proud to receive it. Someone more or less anynomous cared for my problems, that impressed me much. I feel indeed that she was a very gentle person and I envy everybody who knew her personally.
My deeply sympathy to her family and her friends.
Michael
Michael Reiner University of Cologne, Germany Dept. of Anatomy I _______________________________________________________________________ 1.000.000 DM gewinnen - kostenlos tippen - http://millionenklick.web.de IhrName-at-web.de, 8MB Speicher, Verschluesselung - http://freemail.web.de
I have had concerns about this for some while. The difficulty isn't just with lifting something heavy (which is, most certainly, a major problem), but also that the heavy thing is inherently hazardous. If you are pouring liquid up above you, and you spill it, some will almost certainly splash on you. This is not good if the liquid is nitrogen (or any other cryogen, amongst other hazardous materials).
Using a step ladder is also not a very safe prodecure, and if you are leaning to one side to pour from a heavy dewar, it makes it all the more unstable.
I have built a wooden platform (well, I had the carpenters build it!) for the worst of our instruments. I used wood because it was much easier than designing and ordering a custom metal one, but there is no issue of interfering with the microscope. The arrangement works well. It is very simple - 2x4's and plywood, painted with floor paint. The carpenters designed it as they went along.
Another way of dealing with nitrogen filling in a safe way would be to use a pressurized tank, and fill the trap, x-ray detector or whatever through a pipe. The arrangement could be made portable. I have had in mind researching the cost of a system like this.
I wouldn't recommend an automatic fill system, though. The valves tend to stick in the open position, draining your liquid nitrogen supply and freezing (and possibly damaging) the items onto which the liquid spills. I'm talking from experience here!
Good luck!
Tony.
} Hi Everybody! } We have two Hitachii TEM's (H600 and H7000). We use liquid nitrogen } to improve the vacuum (back trap) and to decrease contamination of } the column (front trap). Recently, I have been told by the university } safety officer that I can not use LN in the back trap (I and the } users have to climb on a step ladder and lift LN container to fill } the trap) because of the safety reasons. One of our users strained } her back while filling the back trap and she has been on a sick } leave for some time. They suggested to build a platform in the back } of a microscope so it is more sturdy and safer. Have you ever had } problems of that sort? The lab has been opened for 15 years and } we have never had any accident with LN. Is the platform really safer? } Will it interfere with the performance of the microscope, if it is } metal one? Will performance of the scope be affected if I do not use } LN in the back trap? } Thanks for answers } Dorota }
** 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 **
Dr. David Kirk has a Philips 300 TEM available for donation. The recipient will pay all moving charges. If interested, please contact him directly
David Kerk, Ph.D. Professor and Chair Department of Biology Point Loma Nazarene University 3900 Lomaland Dr San Diego, CA 92106 Ph: 619-849-2398 FAX: 619-849-2598 email: dkerk-at-ptloma.edu
This is the last posting, I promise, seeking a used (of course no objection to new) Probe Current Detector (ie Faraday Cup device) for a JEOL 840 or 6400.
Anyone got one pining away in a cupboard?
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
It seems my fears were unfounded as it relates to small volume laboratory use. It is still possible to obtain Freon 113, in modest amounts, for laboratory work.
Gerald Harrison Electron Microscope Facility Dental School Research University of Pennsylvania =================================
I buy F113 fro Re-Cycle & Re-Use Industries, Mansfield, TX
Try John Metz 817.477.5207
I have about 2 gallons in stock and use maybe a quart or so a year.
gary
At 10:55 AM 9/11/00, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Dorota, I sympathize, since I have the H-800, which is the taller, 200 kV version of the H-600 and I have found that short people do have some problems filling the back trap. You are lucky you don't have the EDS, which is even higher up. My solution is to have a self-pressurized dispenser on the 50L liquid nitrogen dewar attached to a hose that can reach the back trap. Position the hose, turn on the liquid N2 slowly, close the tap when the trap is full and wait for the hose to soften before moving it. There is no reason a platform wouldn't work, so long as it is made of non-magnetic materials, such as plastic, aluminum or wood. At 01:49 PM 9/11/00 -0300, you wrote: } } Hi Everybody! } We have two Hitachii TEM's (H600 and H7000). We use liquid nitrogen } to improve the vacuum (back trap) and to decrease contamination of } the column (front trap). Recently, I have been told by the university } safety officer that I can not use LN in the back trap (I and the } users have to climb on a step ladder and lift LN container to fill } the trap) because of the safety reasons. One of our users strained } her back while filling the back trap and she has been on a sick } leave for some time. They suggested to build a platform in the back } of a microscope so it is more sturdy and safer. Have you ever had } problems of that sort? The lab has been opened for 15 years and } we have never had any accident with LN. Is the platform really safer? } Will it interfere with the performance of the microscope, if it is } metal one? Will performance of the scope be affected if I do not use } LN in the back trap? } Thanks for answers } Dorota } 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
----- Original Message ----- } } I have had concerns about this for some while. The difficulty isn't just } with lifting something heavy (which is, most certainly, a major problem), } but also that the heavy thing is inherently hazardous. If you are pouring } liquid up above you, and you spill it, some will almost certainly splash on } you. This is not good if the liquid is nitrogen (or any other cryogen, } amongst other hazardous materials). } } Using a step ladder is also not a very safe prodecure, and if you are } leaning to one side to pour from a heavy dewar, it makes it all the more } unstable. } } I have built a wooden platform (well, I had the carpenters build it!) for } the worst of our instruments. I used wood because it was much easier than } designing and ordering a custom metal one, but there is no issue of } interfering with the microscope. The arrangement works well. It is very } simple - 2x4's and plywood, painted with floor paint. The carpenters } designed it as they went along. } } Another way of dealing with nitrogen filling in a safe way would be to use } a pressurized tank, and fill the trap, x-ray detector or whatever through a } pipe. The arrangement could be made portable. I have had in mind } researching the cost of a system like this.
Building a portable LN pump would be pretty simple. You need a lid for the LN container, a wet line to the bottom of the container, a very small heating element and a vapor vent to stop the transfer.
To use the pump LN fill the tank and put the lid on with the vapor valve open. The output of the wet line is connected and the vapor valve closed and the heater turned on. A 500 Ohm resistor and 6 volts should make a good enough heater. Keep the resistor very small so it will cool rapidly. To stop the transfer open the vapor valve and turn off the heater. Once the vapor valve is open the liquid should drain back down the wet line and everything should be ready to disconnect in a minute or so. The only problem is if you get into a siphon situation. If you do that you need a way to relieve the suction.
On vapor releif and suction releif valves I would not totaly rely on something that could freeze up but I would add a breakable vapor reief valve that a sharp rap with a stick would beak and shut the system down.
I have rigged a lot of pumps that work this way using about 4 psi. The original pump I built was for pumping diesel wiht propane and if your pressure was over 4 PSI you desolved propane in the diesel and vapor locked your injectors. I have used in in a lot of situations for moving liquids. I built one that went in bung of a 55 gallon barrel and was completly sealed except for the pressure releif valve that should be open at all time when the pump is not in use.
The head on this type pump should not be very great or the transfer rate is slow, you waste a lot of vapor when you vent and the pressure can get a higher than is safe. For low head pumping 4 psi will pump 50 gallons of deisel through a 1.5 inc hose 10 or 15 minutes at a 18 inch head. or 5 or 10 minutes with a -10 inch head.
Hello, Isn't there a better alternative to Freon 113? We use ethanol for critical point drying in our lab with fine results. Freon is bad for the ozone layer. I've copied below, some information from the US Environmental Protection Agency.
C. Abiotic Effects
Freon 113 moves slowly through the lower atmosphere into the stratosphere. Photodegradation of freon 113 in the upper atmosphere releases chlorine atoms which react with ozone. Stratospheric depletion of ozone increases the amount of ultraviolet-B radiation that reaches the earth's surface (U.S. EPA 1983). Increased, surface UV radiation can adversely affect human health and the environment.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ 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 Mon, 11 Sep 2000, 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 } -----------------------------------------------------------------------. } } } I buy F113 fro Re-Cycle & Re-Use Industries, Mansfield, TX } } Try John Metz } 817.477.5207 } } I have about 2 gallons in stock and use maybe a quart or so a year. } } gary } } } At 10:55 AM 9/11/00, 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 form the Univ. of Penn., } } } } I apologize in advance for what may well be a very sophomoric } } question. } } } } I haven't paid attention to what may have been discussed } } regarding alternatives to Freon 113 as a final drying agent because I } } have scrupulously rationed my supply, ml by ml, for several years. Now } } I'm into my final 300 ml and must begin to search for a substitute since } } it has been my understanding that Freon 113 is no longer available. I } } only use very small volumes at a time and 4x500 ml bottles has lasted me } } nearly 10 years. Most of my specimens are cell layers or other small } } (less than 1 cubic mm) tissue samples for SEM. } } } } Any suggestions and information about what is currently used as } } an alternative to critical point drying out of Freon 113 exchanged with } } liquid CO2 would be greatly appreciated. } } } } Thanks, } } } } Gerald Harrison } } Electron Microscope Facility } } Dental School Research } } University of Pennsylvania } } }
I have a memory lapse-can you remind me of the reagent used as an alternative to CPD for SEM;or do you know of/recommend any safe (not too toxic) alternative method to CPD?-any reply would be appreciated
Cameron
Mr. Cameron Hind Research Scientist Advanced Technologies group Baxter R & D Europe S.C.R.L. Rue du Progrès 7 1400 Nivelles Belgium
Hildy must indeed have been quite a remarkable person. I have benefited several times over recent years from her replies and advice. I note that I currently have fifteen of her messages saved. Three of those were in connection with my own recent enquiry about folds in semi-thin resin sections. She replied so quickly to additional mailings when asked about an additional point. Her knowledge of electron microscopy and specimen processing was very wide-ranging. She will be missed by this List.
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 } -----------------------------------------------------------------------. } } } Hi Everybody! } We have two Hitachii TEM's (H600 and H7000). We use liquid nitrogen } to improve the vacuum (back trap) and to decrease contamination of } the column (front trap). Recently, I have been told by the university } safety officer that I can not use LN in the back trap (I and the } users have to climb on a step ladder and lift LN container to fill } the trap) because of the safety reasons. One of our users strained } her back while filling the back trap and she has been on a sick } leave for some time. They suggested to build a platform in the back } of a microscope so it is more sturdy and safer. Have you ever had } problems of that sort? The lab has been opened for 15 years and } we have never had any accident with LN. Is the platform really safer? } Will it interfere with the performance of the microscope, if it is } metal one? Will performance of the scope be affected if I do not use } LN in the back trap? } Thanks for answers } Dorota }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
I would add to Tony's discussion on a wooden platform, that a metal platform is not advisable in such proximity to an instrument that has both high voltage and running water. Should wire insulation crack over time due to spillage of LN2, and you get a water leak while standing on a metal platform, you won't be in a very good position! In fact, one lab in which I worked banned the use of all metal step stools in favor of wooden or plastic ones for such reasons.
On an aside, how large is the trap? Perhaps a smaller dewar for filling that particular one is in order. You could easily use a thermos sized dewar (1L) to fill that trap, especially if you are keeping it full on a regular basis.
HTH,
David Bell Scientist Electron Microscopy Lab Millipore Corporation 80 Ashby Road Bedford, MA 01730 (781) 533-2108
On Tue, 12 Sep 2000 08:53:03 -0500 Cameron Hind {hindc-at-baxter.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 Listservers, } } I have a memory lapse-can you remind me of the reagent used as an } alternative to CPD for SEM;or do you know of/recommend any safe (not too } toxic) alternative method to CPD?-any reply would be appreciated } } Cameron } } Mr. Cameron Hind } Research Scientist } Advanced Technologies group } Baxter R & D Europe S.C.R.L. } Rue du Progrès 7 } 1400 Nivelles } Belgium } } Tel:+32 67 882 511 } Fax:+32 67 217 191 } e-mail: hindc-at-baxter.com } } } } } } } } } } } } } } } } } } } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
I gather there are not alot of people out there trying to immunolabel microsomes because I only received one response last time (Thank you Jan), but I'll post the latest problem in hopes that someone may have some insight. I am attempting to immunolabel a protein inside sheep liver microsomes. Jan Leunissen suggested I combine the microsome suspension with geletin and coat the inside of an eppendorf with the geletin/microsome film then preembedd label this film.
When I finished, I had very nice geletin interspersed with large areas of indistinct matter. I think these areas are where the microsomes were, before the labelling protocol disrupted them. I processed a small amount of the geletin/microsome from the same tubes before labelling and it looked very normal: geletin interspersed with large area of microsomes. Does anyone have any ideas on how to stabilize the microsomes, or where in the world they are going?
Thank you,
Michelle Peiffer ************************************************************* Electron Microscope Facility for the Life Sciences Penn State University Biotechnology Institute 001 South Frear Lab University Park PA 16802
I guess that you are thinking of Ted Pella's Peldri. This has not been available for some years. You could try to use a solvent drying method, which for some specimens works surprisingly well.
Place a double layer of filter paper in a glass Petrie dish. Add sufficient chloroform to well saturate the filter paper. Move fixed and dehydrated specimen from ethanol onto a microscope slide placed onto the filter paper. Close Petrie dish and store in refrigerator for two days until all solvent has evaporator. Don't open Petrie dish until the dish has warmed to at least to room temperature. Mount and coat specimen etc.
Disclaimer: we do not sell glass Petrie dishes, filter papers, chloroform or refrigerators. We do sell CPD and Freeze driers! 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 ABN: 99 724 136 560 www.proscitech.com
On Tuesday, September 12, 2000 11:53 PM, Cameron Hind [SMTP:hindc-at-baxter.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 Listservers, } } I have a memory lapse-can you remind me of the reagent used as an } alternative to CPD for SEM;or do you know of/recommend any safe (not too } toxic) alternative method to CPD?-any reply would be appreciated } } Cameron } } Mr. Cameron Hind } Research Scientist } Advanced Technologies group } Baxter R & D Europe S.C.R.L. } Rue du Progres 7 } 1400 Nivelles } Belgium } } Tel:+32 67 882 511 } Fax:+32 67 217 191 } e-mail: hindc-at-baxter.com } } } } } } } } } } } } } } } } } } }
At 8:53 AM -0500 9/12/00, Cameron Hind wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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
Hi: The chemical you are looking for is called Hexamethyldisilazane (HMDS). You can find it in mist EM catalogues}
Cameron Hind wrote:
} Hi Listservers, } } I have a memory lapse-can you remind me of the reagent used as an } alternative to CPD for SEM;or do you know of/recommend any safe (not too } toxic) alternative method to CPD?-any reply would be appreciated } } Cameron } } Mr. Cameron Hind } Research Scientist } Advanced Technologies group } Baxter R & D Europe S.C.R.L. } Rue du Progrès 7 } 1400 Nivelles } Belgium } } Tel:+32 67 882 511 } Fax:+32 67 217 191 } e-mail: hindc-at-baxter.com
-- Regards, Gregory Rudomen Technical Specialist Electron Microscopy State University of New York at Stony Brook University Microscopy Imaging Center Stony Brook, NY 11794-8088 631-444-7372 Greg-at-umic.sunysb.edu ************************************************* Standard disclaimer: The opinions expressed in this communication are my own and do not necessarily reflect those of the University Microscopy Imaging Center. *************************************************
Karen, You may want to check out the Magnafire "firewire" system from Optronics. (www.optronics.com) Brett
Brett M. Connolly, Ph.D. Merck Research Laboratories Department of Pharmacology WP26A-3000 PO Box 4 West Point, PA 19486 Ph. 215-652-2501 FAX 215-652-2075 e-mail: brett_connolly-at-merck.com
} ---------- } From: Karen Rethoret[SMTP:rethoret-at-yorku.ca] } Sent: Monday, September 11, 2000 2:14 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Low Light Camera Required } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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, } } We are looking for camera suggestions that will handle Low Light, Real } time imaging(both for flurorescence and DIC) in the fluorescence } microscope. Something in the $20K range or less for the system. } } High resolution is required also. These images can be extremely dark. The } user prefers also to be able to output the realtime imaging of cell growth } } to videotape as that has proved to be the most convenient in the past but } is also interested in the "firewire" option and it's capabilities. } } We realize that a compromise in some areas (likely the frame rate)will be } required but high resolution during low light imaging is very important. } } We would be particularly interested in hearing from others that have used } a camera that would suit our application. We have commercial info for } several models but would really like some user feedback. } } Thanks, } } Karen Rethoret } York University } Biology Department } Toronto, Ontario } 416-736-2100 x33289 } } } }
Greetings, Several replies have mentioned this mysterious solvent for CPD, and that it works on some type of specimens. Could someone outline how this is used (infiltrate and then evap to air, or what?) and if there is any rhyme or reason about what kinds of specimens that work???
THanks, Tobias
} } } We use HMDS (Hexamethyldisilazane) from Sigma. } } Dave } } } On Tue, 12 Sep 2000 08:53:03 -0500 Cameron Hind } {hindc-at-baxter.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 Listservers, } } } } I have a memory lapse-can you remind me of the reagent used as an } } alternative to CPD for SEM;or do you know of/recommend any safe (not too } } toxic) alternative method to CPD?-any reply would be appreciated } } } } Cameron } } } } Mr. Cameron Hind } } Research Scientist } } Advanced Technologies group } } Baxter R & D Europe S.C.R.L. } } Rue du ProgrËs 7 } } 1400 Nivelles } } Belgium } } } } Tel:+32 67 882 511 } } Fax:+32 67 217 191 } } e-mail: hindc-at-baxter.com } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } ---------------------------------------- } Patton, David } Email: David.Patton-at-uwe.ac.uk } "University of the West of England"
Let me inform you, that the Proceedings of the "12th EUROPEAN CONGRESS ON ELECTRON MICROSCOPY" (EUREM12), Brno, Czech Republic, July 9 – 14, are available.
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The EUREM12 Proceedings consist of three printed volumes, one CD-ROM containing these three volumes (all available now), and the fourth volume (the Supplement), which will be distributed in October, 2000. The complete EUREM12 programme, accessible at the http://www.eurem2000.isibrno.cz/progra.html, brings full details about the contents of the Proceedings. The Supplement will contain the post-deadline posters, lectures of the Ernst Ruska Prize winners and the list of the participants.
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The payment has to be made in advance to the beneficiary:
Czechoslovak Society for EM, Kralovopolska 147, CZ-61264 Brno, Czech Republic, either via bank transfer as a net payment to Cs. Obchodni Banka, a.s., Sumavska 33, CZ-61140 Brno, Czech Republic, SWIFT code CEKO CZ PP BRN, account no. 08867030 or by means of a personal or company cheque issued to t
The best way for ordering the Proceedings, is using our Web Site at the http://www.eurem2000.isibrno.cz/frmvol4.html . The e-mail (eurem2000-at-isibrno.cz) or the fax (++420 5 41514337) orders are also acceptable. This offer is valid as long as the stock lasts!
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"Better" is rather tough to define--at least in my application and use. First off, I use about F-113 perhaps six times a year and use about 20cc of it initially. I dehydrate in a sequential immersion in methanol, acetone, F-113. Each immersion is accomplished in a sealed vessel. When the F-113 has too much gunk on the surface, I skim it off, put it in a salvage container and continue using the F-113 until it is too low in volume to be useful. Eventually, I have a pretty full salvage container. This is sent back to the recycle place where they clean it and re-sell the F-113. Very little is lost.
An alternative is CPD. However, considering that F-113 costs about $100 per pound and a CPD setup is perhaps $15K plus a lot of hassle, CPD is not cost effective or time effective. If I did dehydration on a regular basis, I would certainly do CPD.
Recycling and containment makes F-113 a low cost and effective approach to dehydration. And I think the method I use is environmentally friendly. There are of course ways to abuse this material. I don't think that I am doing that.
gary g.
At 10:14 PM 9/11/00, you wrote:
} Hello, } Isn't there a better alternative to Freon 113? We use ethanol for } critical point drying in our lab with fine results. Freon is bad for the } ozone layer. I've copied below, some information from the US } Environmental Protection Agency. } } C. Abiotic Effects } } Freon 113 moves slowly through the lower atmosphere into the } stratosphere. Photodegradation of freon 113 in the upper atmosphere } releases chlorine atoms which react with ozone. Stratospheric } depletion of ozone increases the amount of ultraviolet-B radiation } that reaches the earth's surface (U.S. EPA 1983). Increased, } surface UV radiation can adversely affect human health and the } environment. } } \/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ } 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 Mon, 11 Sep 2000, 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 } } -----------------------------------------------------------------------. } } } } } } I buy F113 fro Re-Cycle & Re-Use Industries, Mansfield, TX } } } } Try John Metz } } 817.477.5207 } } } } I have about 2 gallons in stock and use maybe a quart or so a year. } } } } gary } } } } } } At 10:55 AM 9/11/00, 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 form the Univ. of Penn., } } } } } } I apologize in advance for what may well be a very sophomoric } } } question. } } } } } } I haven't paid attention to what may have been discussed } } } regarding alternatives to Freon 113 as a final drying agent because I } } } have scrupulously rationed my supply, ml by ml, for several years. Now } } } I'm into my final 300 ml and must begin to search for a substitute since } } } it has been my understanding that Freon 113 is no longer available. I } } } only use very small volumes at a time and 4x500 ml bottles has lasted me } } } nearly 10 years. Most of my specimens are cell layers or other small } } } (less than 1 cubic mm) tissue samples for SEM. } } } } } } Any suggestions and information about what is currently used as } } } an alternative to critical point drying out of Freon 113 exchanged with } } } liquid CO2 would be greatly appreciated. } } } } } } Thanks, } } } } } } Gerald Harrison } } } Electron Microscope Facility } } } Dental School Research } } } University of Pennsylvania } } } } } }
I also have a Hitachi TEM (7000) and put liquid nitrogen in both the turbo molecular pump and the anticontamination trap. What I know for sure is that my Varian 880 vacuum ionization gauge usually reads about one torr better with the liquid nitrogen then with out it. As a generalization, I have very little astigmatism and would like to think that the anticontamination trap is working, but I really don't know that.
Tim
Timothy G. Schneider Director of Electron Microscopy Department of Pathology Room 229 Jefferson Hall Thomas Jefferson University 1020 Locust St. Philadelphia Pa. 19107 215-503-4798 work 610-613-8170 cellular timothy.schneider-at-mail.tju.edu
} I have a memory lapse-can you remind me of the reagent used as an } alternative to CPD for SEM
It is hexamethyldisilazane (HMDS).
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 **
Mr. Cameron Hind wrote: ============================================================= } I have a memory lapse-can you remind me of the reagent used as an } alternative to CPD for SEM;or do you know of/recommend any safe (not too } toxic) alternative method to CPD?-any reply would be appreciated ============================================================= The current material that is presently available, to my knowledge the only such material, is HMDS, or Hexamethyldisilazane, otherwise known as 1,1,1,3, 3,3 hexamethyldisilazane [CAS # 999-97-3].
More information about this material, which is offered by SPI Supplies and some of the other leading suppliers of chemicals and consumables to the EM world, can be found on URL http://www.2spi.com/catalog/chem/chem2a2.html
An opinion: Nothing has ever been developed in the way of a technique of sample preparation for SEM that is as good as critical point drying. Some years ago, Ted Pella offered a product called Pel-Dry, it was good, but not quite as good as actually doing it via CPD. HMDS, again, in my opinion, falls a bit short of what Pel-Dry used to do. But for someone without a CPD unit, wanting to prepare wet samples, the HMDS technique is lightyears better than air drying. I should also say that there are many people who use HMDS regularly, saying that for their samples, it is "good enough" either because of the lack of fine structure, or the very low magnifications they are using.
Disclaimer: SPI has offered for many years our own SPI-Chem HMDS and also a CPD unit.
Chuck
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I've been out of touch with EM of membranes and tissues for many many years, but as I recall acetone as a dehydration agent is rather harsh. After your fixation and cross-linking steps it may be that a more gently dehydration would help further preserve your plant tissue. I am definitely not up to speed on these preps and I am sure some others will respond, but I think I would try ethanol or another slightly less polar solvent for the early dehydration steps. I only say this from my (ancient) experience with acetone and its aggressive extraction behavior to plant tissue and photosynthetic membranes. Otherwise, I'd stick to your recipe.
Good Luck Brad Huggins BP Amoco
} ---------- } From: Sally Burns[SMTP:burnssal-at-pilot.msu.edu] } Sent: Thursday, September 07, 2000 9:39 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: turf grass fixation for TEM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 preparing turf grass samples for TEM. We will be examining the } crowns and lead blades for infectious agents. } Any tidbits on fixation and embedding of this tissue would be } appreciated! } } My starting point is this protocol, which has been used for wood } samples. If it is overkill, please let me know. } } Fix: 2% Glutaraldehyde in 50 mM NaCacodylate pH 7.2, with light vacuum } for 24 hours } Wash: 3X for 1 H each RT in buffer with mild agitation } PostFix: 1% OsO4 in water Overnight at 4C } Wash: 3X for 1H each at RT in water with mild agitation } en bloc stain: saturated UA for 30 min with mild agitation } Dehydration series: 25%, 50%, 75%, 100% Acetone, each 30 min with mild } agitation } 100% acetone + 1% DMP for 30 min with mild agitation } -- 100% acetone + 1% DMP overnight with mild agitation } 100% acetone + 1% DMP overnight with mild agitation } Infiltration: } 33% Spurr's in Acetone + 1% DMP for 2 H min with mild agitation } 66% Spurr's in Acetone + 1% DMP for 2 H min with mild agitation } 100 % Spurr's for 2H with mild agitation } 100 % Spurr's Overnight with mild agitation } 100 % Spurr's for 8-12 H with mild agitation } 100 % Spurr's for 8-12 H with mild agitation } Cast in fresh resin } } } Thanks, Sally } } Sally Burns } Center for Advanced Microscopy } B5 Center for Integrated Plant Studies } Michigan State University } East Lansing, MI 48823 } (517) 355-5004 } burnssal-at-msu.edu }
Further to my posting yesterday seeking a PCD for a JEOL 840, which, I am informed, is the same as that for a 6400, I am currently negotiating with a company to make me one.
Does anyone else out there want one?
It would probably ease my negotiations if there were to be more orders than just mine.
Please contact me direct.
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
Hello, Maybe you can help? I culture human preadipocytes in 35mm dishes } and then differentiate them to form lipid droplets. I have been staining } them with ORO for the past year, but have recently not been able to get the } ORO to penetrate the cells. I had been fixing the cells with formaldehyde } and calcium chloride (4g calcium chloride + 10ml of 37%formaldehyde into } ~90ml water to fix the cells for 1 hour. Then rinse with Hanks buffered } saline 2x and then stain with ORO for 15min. This has been working for 1 } year, then one day the ORO stain would not penetrate the cells any longer - } the lipid is clear, not red. What could be the problem? The formaldehyde } solution? Have you ever experienced something like this? Thank you for any } suggestions. Also, I have ordered all new fixative, CaCl2, ORO stain powder and isopropanol. The } lipid in my cells are not picking up the stain. Oh why, oh why???? I even } tried the stain on live cells without fixing and they won't stain either. Thanks for any word. } } Just for your information: I use .6g ORO in 100ml 100% isopropanol.
Georgia
Georgia Bachman NIH - Technical IRTA Gene Expression Unit 4212 N. 16th St. Phoenix, AZ 85016 Phone: 602-200-5310 or 5303.
I'm having (or trying to have) a go at doing serial ultrathin sections of Epon-embedded material. I'd like to see a long unbroken ribbon coming off the diamond knife, but after battling for a couple of hours nothing seemed to work - the sections floated off happily on their own little journeys and I wasn't going to chase them around. I tried cutting the pyramid sides with a new Weckprep blade to get them as smooth as possible, making them as parallel as railway tracks, lowering and raising the water level in the trough, and turning off vibrating things in the lab (unfortunately we're on the third floor), but nothing seemed to make any difference. I'm going home now (smiling) but would like to hear of any tips and experiences you've had that got you beautiful ribbons of sections (and any other tips for serial sectioning).
Looking forward to reading my mail tomorrow.
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 looking for a knifeblock for a Reichert OMU2 to be able to bring one of these vintage machines back on-line. Should you have this part currently gathering dust or holding open a door in your lab, please contact me.
thanks in advance
Steve
Dr. Steven Barlow EM Facility/Biology Dept. San Diego State University 5500 Campanile Drive San Diego CA 92182-4614 phone: (619) 594-4523 fax: (619) 594-5676 email: sbarlow-at-sunstroke.sdsu.edu http://www.sci.sdsu.edu/emfacility
} Date: Tue, 12 Sep 2000 18:14:08 -0600 (=?ISO-8859-1?Q?Hora_est=E1ndar_de_M=E9xico?=) } From: Mark West {mwest-at-ifcsun1.ifisiol.unam.mx} } To: Microscopy-at-sparc5.microscopy.com } Subject: serial sections } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 having (or trying to have) a go at doing serial ultrathin sections of } Epon-embedded material. I'd like to see a long unbroken ribbon coming off } the diamond knife, but after battling for a couple of hours nothing seemed } to work - the sections floated off happily on their own little journeys } and I wasn't going to chase them around. I tried cutting the pyramid sides } with a new Weckprep blade to get them as smooth as possible, making them } as parallel as railway tracks, lowering and raising the water level in the } trough, and turning off vibrating things in the lab (unfortunately we're } on the third floor), but nothing seemed to make any difference. I'm going } home now (smiling) but would like to hear of any tips and experiences } you've had that got you beautiful ribbons of sections (and any other tips } for serial sectioning). } } Looking forward to reading my mail tomorrow. } } Mark } You should trim the top and bottom of your trapezoid **on the microtome with a glass knife.**
This means that the minute scores caused on the side facet by rough trimming run parallel to the top and bottom of your block face, not perpendicular as they do when you trim by hand pushing the razor blade down from the face.
Also, as you said, the top edge and bottom edge should be parallel. But the biggest thing is not to have any scores caused by a razor blade.
If you don't know how to rough-trim with the microtome, contact me, and I'll be happy to describe it.
} } } } } } ******************************************** } 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 } ******************************************** } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
In my previous response I gave the method, though I've used Chloroform. HMDS may have a theoretical advantage, but I expect that the results would be identical. HMDS was introduced for rapid processing, particularly fast and complete dehydration. It chemically interacts with water and forms acetone. When at least the last dehydration is in HMDS, the user can be sure that dehydration is excellent even with doubtful ethanol or acetone.
During the air drying process residual water and solvent molecules exerts huge pressures onto membranes and these cause shrinkage and shriveling of any specimen. The drying mechanism: Using Peldri, drying only occurred at the Peldri/ air interface, but the specimen is held rigid in the solid Peldri material. As the Peldri slowly sublimes, more of the specimen is dried. During solvent drying the specimen is in a solvent saturated atmosphere and drying is very slow - over 48 hours. In this method there is little pressure exerted by molecules trying to pass through membranes and consequently shrinkage is minimized. Its a good alternative method and sometimes works very well. Given a choice I'll try CPD first. 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 ABN: 99 724 136 560 www.proscitech.com
On Wednesday, September 13, 2000 12:21 AM, Tobias Baskin [SMTP:BaskinT-at-missouri.edu] wrote: } } } Greetings, } Several replies have mentioned this mysterious solvent for } CPD, and that it works on some type of specimens. Could someone } outline how this is used (infiltrate and then evap to air, or what?) } and if there is any rhyme or reason about what kinds of specimens } that work??? } } THanks, } Tobias } } } } } } } We use HMDS (Hexamethyldisilazane) from Sigma. } } } } Dave } } } } } } On Tue, 12 Sep 2000 08:53:03 -0500 Cameron Hind } } {hindc-at-baxter.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 Listservers, } } } } } } I have a memory lapse-can you remind me of the reagent used as an } } } alternative to CPD for SEM;or do you know of/recommend any safe (not too } } } toxic) alternative method to CPD?-any reply would be appreciated } } } } } } Cameron } } } } } } Mr. Cameron Hind } } } Research Scientist } } } Advanced Technologies group } } } Baxter R & D Europe S.C.R.L. } } } Rue du ProgrEs 7 } } } 1400 Nivelles } } } Belgium } } } } } } Tel:+32 67 882 511 } } } Fax:+32 67 217 191 } } } e-mail: hindc-at-baxter.com } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } ---------------------------------------- } } Patton, David } } Email: David.Patton-at-uwe.ac.uk } } "University of the West of England" } } -- } _ ____ __ ____ Tobias I. Baskin } / \ / / \ / \ \ University of Missouri } / | / / \ \ \ Biological } Sciences } /_ / __ /__ \ \ \__ 109 Tucker Hall } / / / \ \ \ } Columbia, MO 65211-7400 USA } / / / \ \ \ voice: } 573-882-0173 } / /____ / \ \__/ \____ fax: 573-882-0123 } }
My name is Sorin Lazar and I am scientist in Bucharest, Romania. I have received like donation a Hitachi S 500 #01-03 scanning electron microscope. Unfortunately I don't have the operating manual. I should be grateful if someone can help me to find an Operating Manual or a copy.
Hi, Try to trim the upper and the lower edges with a glass knife. Trimming with razor blade often results in non-perfectly parallel edges. Turn the block 90 degree (right or left) trim left and right sides with perfect glass knife. Use both sides of the glass knife. I used the angle of the knife towards the block around 60 degree. When you trim your block from both sides just turn it back 90 degree - you will get perfectly parallel edges.
Your problem could be caused also by irregular air flow around microtome or some grease on the blade you have used.
Hope it will help.
Sincerely, -- Alexander Mironov Jr. Division of Tumor Biology, H4 The Netherlands Cancer Institute Plesmanlaan 121 1066 CX Amsterdam
I have read the offerings with interest and would like to make a few comments.
Firstly - when a manufacturer designs an instrument they do not fit cold traps for fun, they believe that to obtain the best performance from their instrument you need the facilities that they fit. I have worked with design teams at two manufacturers and this is the attitude that they both took!
Secondly - I have in the past, in countries where LN2 was not available, used dry ice (Solid CO2) and acetone in LN2 traps. I believe this brings the temperature down to -80deg C and it works quite well much better than no cooling at all.
Thirdly there are imaging techniques that allow us to measure the contamination rate. Place a holey film in the TEM and without LN2 in use and take a high resolution picture of a hole. Leave the beam on the specimen and repeat the picture on the same hole 20 minutes later (that is if you still have a hole). Measure the two hole widths, take the smaller from the larger and divide by 2 (2 contaminated edges) divide by the time between the two pictures and divide by the magnification. You now have your contamination rate in nm/minute. Then try this with the LN2 systems in action and you will see just how well the LN2 systems work.
The same system can be used with a SEM but use a well coated latex sphere in place of the hole. In this case the sphere will grow with time as the contamination builds round the sphere.
Many years ago (1968) Hitachi produced a paper on LN2 systems. They gist was that if you used LN2 systems for a period of time and then stopped, it took an equal period of time before the vacuum level and contamination rate fell back to the original level. Of course you could not count the time if you let the column to air.
A few more things to think about and a way of proving just how clean your microscope is.
Steve Chapman Senior Consultant Protrain For consultancy and training by professional World Wide Tel +44 1280 814774 Fax +44 1280 814007 www.emcourses.com
} I've been out of touch with EM of membranes and tissues for many many years, } but as I recall acetone as a dehydration agent is rather harsh. After your } fixation and cross-linking steps it may be that a more gently dehydration } would help further preserve your plant tissue. I am definitely not up to } speed on these preps and I am sure some others will respond, but I think I } would try ethanol or another slightly less polar solvent for the early } dehydration steps. I only say this from my (ancient) experience with } acetone and its aggressive extraction behavior to plant tissue and } photosynthetic membranes. Otherwise, I'd stick to your recipe. } } Good Luck } Brad Huggins } BP Amoco
Brad, (and all)
My experience with Ethanol is that it makes the cell walls extremely brittle and hard to section, esp in concetrations above 70% ETOH.
Time is always the problem with Plant tissue. The cell walls really like to hold on to what ever it was in last. So dehydration must proceed at a snails pace. Grass leaf blades by being very thin make the process easier but I woud imagine that the crowns would be problematic if carried too fast through dehydration.
Sincerely, Geoff Williams
Electron Microscope Facility Supervisor Biology Department Brooks Hall Central Michigan University Mt. Pleasant, MI 48859
To glue serial sections together, make a gentle adhesive by dissolving the adhesive off of a piece of Scotch tape in chloroform. Paint the upper and lower block faces with your new adhesive, and, as you cut, the sections should stick together tenaciously.
Good luck!
Dotty Sorenson Microscopy and Image Analysis Laboratory Department of Cell and Developmental Biology University if Michigan Medical School Ann Arbor, Michigan dsoren-at-umich.edu
On Tue, 12 Sep 2000, Mark West 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'm having (or trying to have) a go at doing serial ultrathin sections of } Epon-embedded material. I'd like to see a long unbroken ribbon coming off } the diamond knife, but after battling for a couple of hours nothing seemed } to work - the sections floated off happily on their own little journeys } and I wasn't going to chase them around. I tried cutting the pyramid sides } with a new Weckprep blade to get them as smooth as possible, making them } as parallel as railway tracks, lowering and raising the water level in the } trough, and turning off vibrating things in the lab (unfortunately we're } on the third floor), but nothing seemed to make any difference. I'm going } home now (smiling) but would like to hear of any tips and experiences } you've had that got you beautiful ribbons of sections (and any other tips } for serial sectioning). } } Looking forward to reading my mail tomorrow. } } 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 } ******************************************** } }
For animal tissue: 10min. in 20%,30%,50%,70%,90% and 100% ethanol (or whatever times etc you use for CPD). 2x10min. in 100% ethanl 10min. in 1:1 ethanol HMDS (or 1:2, 1:1, and 2:1) 3x10min in HMDS I put tissue on filter paper in a petri dish with lid ajar and let it evaporate in fume cupboard.
Most papers say CPD is better eg for fine hairs on leaves. I believe the first report was on very satifactory use on insects. We use it mostly on bacteria which are pretty tough anyway. I have found it disappointing on cultured cells and am planning to dust down the CPD (cryoSEM and ESEM having relegated it to an historical artifact (sic!) for most purposes in this 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 } -----------------------------------------------------------------------. } } } Greetings, } Several replies have mentioned this mysterious solvent for } CPD, and that it works on some type of specimens. Could someone } outline how this is used (infiltrate and then evap to air, or what?) } and if there is any rhyme or reason about what kinds of specimens } that work??? } } THanks, } Tobias } } } } } } } We use HMDS (Hexamethyldisilazane) from Sigma. } } } } Dave } } } } } } On Tue, 12 Sep 2000 08:53:03 -0500 Cameron Hind } } {hindc-at-baxter.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 Listservers, } } } } } } I have a memory lapse-can you remind me of the reagent used as an } } } alternative to CPD for SEM;or do you know of/recommend any safe (not too } } } toxic) alternative method to CPD?-any reply would be appreciated } } } } } } Cameron } } } } } } Mr. Cameron Hind } } } Research Scientist } } } Advanced Technologies group } } } Baxter R & D Europe S.C.R.L. } } } Rue du ProgrËs 7 } } } 1400 Nivelles } } } Belgium } } } } } } Tel:+32 67 882 511 } } } Fax:+32 67 217 191 } } } e-mail: hindc-at-baxter.com } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } ---------------------------------------- } } Patton, David } } Email: David.Patton-at-uwe.ac.uk } } "University of the West of England" } } -- } _ ____ __ ____ Tobias I. Baskin } / \ / / \ / \ \ University of Missouri } / | / / \ \ \ Biological Sciences } /_ / __ /__ \ \ \__ 109 Tucker Hall } / / / \ \ \ } Columbia, MO 65211-7400 USA } / / / \ \ \ voice: } 573-882-0173 } / /____ / \ \__/ \____ fax: 573-882-0123 }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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, You should trim the top and bottom sides of you face with a glass knife and try coating those sides with grid glue (soak double stick tape in chloroform, then remove the tape, coat with a filter paper wedge careful not to wet your face) Good Luck
Mike D.
} } } Hi, } } I'm having (or trying to have) a go at doing serial ultrathin sections of } Epon-embedded material. I'd like to see a long unbroken ribbon coming off } the diamond knife, but after battling for a couple of hours nothing seemed } to work - the sections floated off happily on their own little journeys } and I wasn't going to chase them around. I tried cutting the pyramid sides } with a new Weckprep blade to get them as smooth as possible, making them } as parallel as railway tracks, lowering and raising the water level in the } trough, and turning off vibrating things in the lab (unfortunately we're } on the third floor), but nothing seemed to make any difference. I'm going } home now (smiling) but would like to hear of any tips and experiences } you've had that got you beautiful ribbons of sections (and any other tips } for serial sectioning). } } Looking forward to reading my mail tomorrow. } } 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 } ******************************************** } }
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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, Fomvar thickness will effect stability, if you could make them slightly thicker than gray you should have no problem. If you need them really thin other than carbom coating you could try minimizing illumination/ intensity on the EM side, smaller spot size, smaller condenser aperture or lower beam current. good luck.
Mike D } } } Dear Microlisters, } } I'm having a bit of trouble with Formvar film stability under the TEM beam at 60 } kV. The films, attached to copper or nickel grids (cleaned with dilute HCl + } acetone in sonicator same day films picked up) develop holes which grow over a } few minutes time until eventually the entire film more or less disintegrates, } collapses. I also noticed that initially, the films are often pulled away from } the rim of the grid by about one square "hole" away. } } I'm using new Formvar 15/95 resin powder and new bottle of ethylene dichloride } solvent, mixing to 0.3% weight to volume. I use "Rite-On" glass slides, rinsed } with ethanol to clean of dust and particulates and air dried, to cast the films. } The films float off easily, and look very clean under the TEM. They were dried } overnight prior to inspection. I take pains to use very clean glassware for } mixing the Formvar solution. } } Of course, I could caarbon coat them to provide stability, but that's extra work } and I don't seem to have had this kind of problem before. } } I barely recall some discussion here some time ago about how Formvar has } "changed" due to different process of manufacturing, or whatever. } } Any clues? } } Thanks in advance. } } 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 } }
Rough trim the excess plastic from the specimen face with a razor blade, keeping all the tissue edges. Put the specimen chuck into the microtome arm, and put in a dry glass knife. Shave off enough plastic from the front (face) in very small increments (start with 1 um slices, decrease to 0.5 um, then 0.1 um until you get a mirror smooth face). If you must keep all the sample, then start with a very thin shaving just to get the face smooth. You should be able to tell when you are into the tissue by looking at the thin shavings. Mark the top edgeQthe one that will become the top of your trapezoid--with a magic marker on the chuck. Be sure to keep track of the plane of your sample because the clearance angle of the knife will necessitate that you return the specimen to the original position in which you trimmed the face. Rotate the chuck/specimen 90 degrees (e.g., clockwise). The edges that will become the bottom and top of your trapezoid are now vertical instead of parallel to the earth. Turn the knife holder about 30 degrees away from center (e.g., right) and approach the block carefully. Trim the left side of the block which will become the bottom of your trapezoid; this will make a smooth facet that will start with the face and go toward the microtome slanting outward at about 30 degrees. Leave the specimen in this position and turn the knife to the other side (e.g., left). Trim this side like the first; it will become the facet over the top of your trapezoid. The top and bottom should now be parallel. If they are not, you can make minute adjustments by slightly rotating the block. The purpose of this exercise it to make the scratches caused by rough trimming be parallel, not perpendicular as happens when trimming with a razor blade, to the top and bottom edges of the face.
If the face is too wide, you can trim it, either on the microtome or with a razor blade. I usually just chop the sides with a razor blade to save time; I make a trapezoid with the top edge slightly shorter than the bottom edge. However, folks with unsteady hands can make the same trapezoid by turning the block back to its original position with the magic marker at the top, then slightly rotating it right or left about 2 or 3 degrees. Trim this side with the knife still at 30 degrees away from center. Then rotate the block 2 or 3 degrees in the other direction and turn the knife 30 degrees to the opposite side. Trim the other side.
Before sectioning, turn the block back 2-3 degrees to be in the original position it was when you faced it with the mark at the top. Use a fresh knife, or even better if you have one, use a diamond. When aligning, be sure to have both the bottom edge of your block parallel to the edge of your knife as well as the plane of your block face parallel to the plane of the knife edge. To do the former, you may have to rotate your specimen very slightly. To do the latter, you may have to change the angle of your specimen in the microtome arm. These tiny adjustments may be necessary because the diamond may not be mounted in its holder exactly like the glass one you used for trimming.
Pick up the sections on slotted grids with a support film.
Good luck. Let me know if this doesn't make sense.
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
---------- Forwarded message ----------
Summary:
I am looking for a test image with control points with known positions in the warped and unwarped state and the known coefficients for a polynomial warp function to test a program I am writing. Can someone on the list help?
Details:
I am trying to write a program to correct center of gravity data for barrel distortion in the image. I am trying to implement the approach described by W. K. Pratt, Digital Image Processing, 2nd edition, pp.430-433 (1991). I imaged a rectangular lattice of squares [50 micron squares in a 100 micron square lattice] with a 5X objective. I fit the measured center of gravity coordinates to a square lattice using the points nearest the center. The difference between observed and computed values for each point shows the barrel distortion in the lens. I used the calculated value for each point as coordinate for the (unwarped) desired control point (x,y) and the observed coordinates as the coordinate for the warped control points (u,v). I'm trying to fit to the following orthogonal polynomials:
1 u1 v1 u1**2 u1v1 v1**2 1 u2 v2 u2**2 u2v2 v2**2 A = 1 u3 v3 u3**2 u3v2 v3**2 ..... 1 um vm um**2 umv2 vm**2
and the vectors u and v
u(transpose) = [u1 u2 u3 ... um] v(transpose) = [v1 u2 v3 ... vm]
I used Mathematica to compute the generalized inverse (Pseudoinverse[]) and generate the vector of coefficients, a and b by
a = PseudoInverse[A] u b = PseudoInverse[A] v
The non-unitary coefficients of a and b were much smaller than I expected and when I tried to use them to correct the center of gravity data, they did not remove the barrel distortion. I'd like a test image with known control points that produce known polynomial coefficients to test the code. Hoping that someone on the list can help....
Best Regards,
John Minter Eastman Kodak Company Analytical Technology Division Rochester, NY 14650-2152 Phone: (716) 722-3407 FAX: (716) 477-7781 email: john.minter-at-kodak.com
Hi all, Thank you all for your input regarding the typical salaries for Microscopists, it was very useful. I would now like to request that anyone who might be interested in an SEM operators position with a start-up company in the Atlanta, GA area of the USA please send me a copy of your resume and previous salary history. Although the type of position has not yet been finalized it will probably be at the entry level. Thank you to those of you who have already sent me their information, I have it on file for consideration.
Quesant Instrument Corp is looking for a customer support person whose primary duties will be: 1. Utilizing Quesant's QScopes (scanning probe microscopes) to scan samples and report back to sales prospects and customers. 2. Installation and taining of new customers. 3. Attending trade shows and demonstrating QScopes in the booth. 4. Work with marketing and R&D in testing and implementing new features.
Requirements: Advanced degree in Chemistry, Physics or Materials Science a plus but not mandatory. Must have good interpersonal skills and a profession al appearance. Must be willing to travel.
Contact Quesant for more information at 29397 Agoura Road, Suite 104 Agoura Hills, CA 91301 tel: 818-597-0311 fax: 818-991-5490 e-mail: qsales-at-quesant.com
We are closing down our EM facility. We have a Reichert Ultracut ultramicrotome available. Please make an offer, and be prepared to assume the cost of shipping to your facility.
Thanks, David Kerk
------------- David Kerk, Ph.D. Professor and Chair Department of Biology Point Loma Nazarene University 3900 Lomaland Dr San Diego, CA 92106 Ph: 619-849-2398 FAX: 619-849-2598 email: dkerk-at-ptloma.edu
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Steve, Thanks for this post. I try to get this through to our users and we faithfully use and require use of N2 on all scopes. However, there are other facilities on campus who do not use N2 or use it intermittantly. When this topic comes up again, I intend to hand them your post and ask them to determine contamination rates on their instruments to convince them of the necessity of using N2. One question that I have is that our main nitrogen trap on the SEM is above the diffusion pump to minimize backstreaming from the pump into the specimen chamber. (We also have a decontaminator close to the specimen that is cooled when we do cryo-SEM.) Now this is no doubt more of a problem with SEM than TEM due to the location of the diffusion pump. However, I wonder why most TEM manufacturers do not also use a nitrogen trap in this location, at least in the higher end scopes, in addition to the decontaminator in the specimen area. It would seem that, since contamination rate in the TEM is likely to be more critical to high resolution, it would also be used to help maintain the best possible vacuum in the column. I am aware that the majority of the contamination is produced in the TEM by interaction of the beam with the specimen thus the need for decontaminator close to the specimen. However, my understanding is that the better the vacuum, the fewer the interactions of the beam electrons with molecules in the column and the more coherent the beam....i.e. better potential resolution. Debby
Debby Sherman 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 Wednesday, September 13, 2000, Steve Chapman {PROTRAIN-at-CompuServe.COM} wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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An elegant solution indeed, Dorothy (and others suggesting similar). Just keep in mind the type of TEM for which the sections are headed, however. For high-end TEM's in the physical sciences, where FEG sources, nanoprobes and EELS analysis are quite common, the TEM specialists running the instrument can be/should be very reluctant to allow specimens which might outgas even a bit and gunk up the specimen chamber area. Your glue 'rim' around the sections would do just that, I suspect.
Should anyone be doubtful of such effects, some years ago, Don Steele (Alcan Aluminum in Kingston, Canada) did some neat serial sectioning of an Al-SiC composite. (Really! Check the MSA '91 proceedings). I seem to recall that Don had to collect the sections on formvar-coated girds (maybe even a homemade formula?). In any event, when we tried to collect X-ray maps, the film breakdown and redeposition piled up the contamination so fast as to render mapping the Al and Si distribution very difficult. Any focused beam analysis resulted in 'cones' on the section surface, rendering good EDX analysis impossible. (People used to worry about column-borne contamination in the early days of AEM. With the recent generations of instruments, specimen-borne contamination is the issue). At least our case was of a very localized phenomenon, the glue rim would result in much more widespread 'fallout'.
Finally, I always remind workshop attendees contemplating 'materials science' ultramicrotomy, to be careful regarding C-analysis on embedded materials, especially particulate. The e-beam (at least at the 120 keV of our older scope) appears to instantly result in roughly a 20-30% mass loss from any embedding resin, even at low beam intensities. This breakdown produces a thin (~10 nm) film of C on the embedded bits which does not interfere with imaging, but can be clearly seen in an EEL spectrum.
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: Dorothy Roak Sorenson [SMTP:dsoren-at-umich.edu] Sent: Wednesday, September 13, 2000 8:52 AM To: Mark West Cc: Microscopy-at-sparc5.microscopy.com Subject: Re: serial sections
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To glue serial sections together, make a gentle adhesive by dissolving the adhesive off of a piece of Scotch tape in chloroform. Paint the upper and lower block faces with your new adhesive, and, as you cut, the sections should stick together tenaciously.
Good luck!
Dotty Sorenson Microscopy and Image Analysis Laboratory Department of Cell and Developmental Biology University if Michigan Medical School Ann Arbor, Michigan dsoren-at-umich.edu
On Tue, 12 Sep 2000, Mark West 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'm having (or trying to have) a go at doing serial ultrathin sections of } Epon-embedded material. I'd like to see a long unbroken ribbon coming off } the diamond knife, but after battling for a couple of hours nothing seemed } to work - the sections floated off happily on their own little journeys } and I wasn't going to chase them around. I tried cutting the pyramid sides } with a new Weckprep blade to get them as smooth as possible, making them } as parallel as railway tracks, lowering and raising the water level in the } trough, and turning off vibrating things in the lab (unfortunately we're } on the third floor), but nothing seemed to make any difference. I'm going } home now (smiling) but would like to hear of any tips and experiences } you've had that got you beautiful ribbons of sections (and any other tips } for serial sectioning). } } Looking forward to reading my mail tomorrow. } } 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 } ******************************************** } }
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I just wanted to say I am so sorry to hear that. Her e-mails were always excellent quides for me. She doesn't know me. However, I know her very well. During the M&M meeting in Portland last year, I saw her for a minute. I was willing to meet her. I just wanted to thank her and let her know that she was also a wonderful teacher for me and for my friends, young electron microscopists in Turkey. However, I could not get a chance to do that. I was so surprised when I heard that she had retired a few months ago. Now, I learn the reason.
Thank you very much all the way from Turkey, Hildy. I will really miss you.
Gulhan
Ranan Gulhan Aktas, M.D. Trakya University Faculty of Medicine Pathology Department Edirne 22030 TURKEY
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To the List:
I ran across this in an obscure book on electron microscopy technique.
"To obtain quality serial sections for electron microscopy the following procedure must be followed for consistent results. 1) Obtain one male goat. 2) Wait for new moon after a rain storm. 3) Obtain black robes which have been treated with a Z-stat gun. 4) Sacrifice goat in the light of the new moon. 5) Collect blood. 6) Spin down blood in centrifuge and collect goat plasma and store at 4 C 7) Take piece of plastic from sample block to be cut. Pulverize into dust. 8) Mix plastic dust with goat plasma. 9) Inject plastic/goat into rabbit. 10) Wait one pay period. 11) Wait for full moon. 12) Place black robes on which have been retreated with a Z-stat gun. 13) Sacrifice rabbit which has been injected with plastic/goat antigen under the full moon. 14) Collect antiplastic/goat antibody from rabbit. 15) Place one drop of antiplastic/goat antibody in the trough of diamond knife. 16) Cut serial sections. 17) The antiplastic antisera should cause all the plastic sections to arrange themselves in the proper order. 17) This should all make anti-sense."
Bob Blystone
-------------------------------- Robert V. Blystone, Ph.D. rblyston-at-trinity.edu
Department of Biology Trinity University 715 Stadium Drive San Antonio, Texas 78212 210.999-7243 FAX 210/999-7229
Wonderful! We have a similar protocol for FISHing particularly recalcitrant mRNAs, but we use a chicken instead of a goat (not much space here so we had to scale back to a smaller organism), and the blood goes in the hybridization buffer.......
Well, thanks for the flood of suggestions about getting a ribbon. I trimmed the block sides with a glass knife (and enjoyed Mike Delannoy's suggestion of 'trimming the top and bottom sides of your face with a glass knife' - didn't know microscopy was such a painful job) and quite a decent ribbon was the result. I tried the chloroform-glue idea and that seemed to make the sections stick together more, but sometimes they gummed up a bit on the knife. The next battle is picking up the ribbon on several slot grids - I suspect that it's more practice than anything else. The method I was shown was to go underneath the ribbon at an angle with the grid, raise it until the waterline is on the top of the exposed formvar of the slot, push the ribbon onto the waterline and draw the grid up at the same time, and touch the ribbon with the hair at the point where the slot finishes to break the ribbon, and remove the slot - all easier said than done! Any ideas would be much appreciated.
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 user of our facility needs to prepare specimens to determine if sub-micrometer pores are present in plant intervascular pit membranes. The user is very concerned that critical point drying or, especially, the dehydration steps prior to CPD, may introduce 'pores' that are really artifacts. He is similarly concerned that freezing or freeze-drying his specimens might also introduce 'pores' due to ice crystal formation. Work he has already done suggests that these are valid concerns.
Has anyone sufficient experience preparing pit membranes for SEM to suggest a procedure we might follow to minimize the possibility of creating small 'pores' where none may exist?
I am interested in knowing some details the Flourescence Microscopes, their principles and applications. Please e-mail me whatever details you have available on the subject or the internet address(s) of site(s) where I can obtain such information.
Thanks, Abdulrahman
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Dear Colleagues: I have made great contacts through this listserver in the past and hope that I can again this time. I am in search of a postdoctoral fellow or very independent technician to continue a program of research using modern optical imaging methods to follow membrane trafficking in vascular endothelial cells. It is a very exciting project that cuts across the fields of cell biology, immunology, and pathology, and uses techniques from standard transmission EM to tracking GFP-labeled proteins. If you have the right background and are interested, or know of a colleague who fits these criteria, please get in touch with me. The following is the text of an ad that appeared in the August 10 issue of Nature and the September issue of Nature Cell Biology:
Postdoctoral position in Membrane Trafficking Postdoctoral position open to study membrane trafficking in endothelial cells. The ideal candidate would have experience studying regulated membrane trafficking at the cellular and molecular levels using biochemical, microscopic, and molecular biology approaches. Experience with modern imaging systems is critical. Experience with endothelial cells is a definite "plus", but not necessary. The candidate should demonstrate excellent communication skills in English, be able to work independently, as well as to interact with a lively group of productive investigators studying leukocyte-endothelial cell interactions. We are based at the Weill Medical College of Cornell University in the nicest and safest area of New York City. We interact extensively with investigators Weill as well as at The Rockefeller University and Memorial Sloan-Kettering Cancer Center, which are both just across the street. The position is fully-funded and available immediately.
Submit CV and names (and contact numbers) of three references to:
William A. Muller, MD, PhD Department of Pathology and Program in Immunology Box 69 Weill Medical College of Cornell University 1300 York Avenue New York, NY 10021 e-mail: wamuller-at-med.cornell.edu
Thank you very much for your help.
Sincerely, Bill Muller
William A. Muller, MD, PhD Associate Professor of Pathology Weill Medical College of Cornell University 1300 York Avenue New York, NY 10021
We are attempting to put a ccd camera (high resolution, large area, sensitivity = SO163) to replace films on 1200EX JEOL at a cost below $100k. Would anyone be willing to share comments on recently acquired ccd camera for TEM? Manufacturers welcome. Used ccds considered.
Marek Malecki, M.D., Ph.D. Director and Principal Investigator
Molecular Imaging Laboratories University of California at San Diego address: 1500 Suite Bonner Hall, 9500 Gilman Drive, La Jolla, CA 92093-0368 phone - office: 8588223373 phone - lab: 8585342484 fax: 8588223715 pager: 8586161420 e.mail: mmm-at-ucsd.edu e.pager: 1620024619-at-alphapage.airtouch.com www site: http://mil.ucsd.edu/ ftp site: mil1.ucsd.edu
From root Wed Sep 13 20:35:43 2000 Return-Path: {Microscopy-request-at-sparc5.microscopy.com} Received: (from daemon-at-localhost) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id UAA21870 for dist-Microscopy; Wed, 13 Sep 2000 20:31:51 -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 UAA21863 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Wed, 13 Sep 2000 20:31:21 -0500 (CDT) Received: from threedee.com (mail.threedee.com [209.83.65.10]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id UAA21854 for {microscopy-at-sparc5.microscopy.com} ; Wed, 13 Sep 2000 20:31:09 -0500 (CDT) Received: from winnie.threedee.com (winnie [192.198.5.13]) by threedee.com (8.7.5/8.7.3) with ESMTP id CAA20549 for {microscopy-at-sparc5.microscopy.com} ; Thu, 7 Feb 2036 02:11:05 -0600 Message-Id: {4.3.2.7.0.20000913200856.02dd2550-at-pc} X-Sender: jfoust-at-pc X-Mailer: QUALCOMM Windows Eudora Version 4.3.2
At 11:42 AM 9/13/00 -0500, David Kerk wrote: } We are closing down our EM facility. We have a Reichert Ultracut } ultramicrotome available. Please make an offer, and be prepared to assume } the cost of shipping to your facility.
I've got a AO / Reichert Ultracut microtome, too. It's a model 701701. It's missing the 651102 control panel, though, as well as the knife holder and object holder. It looks just like the one at: http://www.labx.com/v2/adsearch/Detail3.CFM?adnumb=54745
If I were to sell what I have, what price might it fetch?
- John
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We currently have a Bench Top Turbo carbon evaporation unit made by Denton. We are very displeased with it. We consistently get non uniform coatings that peel off of our metallographic mounts. Any suggestions on a evaporation unit that works well or any information on 'sputtering" carbon? Any other experiences out there?
Dear Mark, With coated slot grids it's easier to come down on top of your ribbon of sections. Break up your ribbon into strips which will fit over your slot first. Then hold your coated slot grid over the sections (you can see the ribbon of sections through your coating and then slowly descend onto your stip of sections. The sections adhere immediately with the first touch. This method avoids any surface tension problems between the grid and water surface which can send the grids flying away from your grid.
Good Luck. Marilyn
Mark West 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, thanks for the flood of suggestions about getting a ribbon. I } trimmed the block sides with a glass knife (and enjoyed Mike Delannoy's } suggestion of 'trimming the top and bottom sides of your face with a glass } knife' - didn't know microscopy was such a painful job) and quite a decent } ribbon was the result. I tried the chloroform-glue idea and that seemed to } make the sections stick together more, but sometimes they gummed up a bit } on the knife. The next battle is picking up the ribbon on several slot } grids - I suspect that it's more practice than anything else. The method I } was shown was to go underneath the ribbon at an angle with the grid, raise } it until the waterline is on the top of the exposed formvar of the slot, } push the ribbon onto the waterline and draw the grid up at the same time, } and touch the ribbon with the hair at the point where the slot finishes to } break the ribbon, and remove the slot - all easier said than done! Any } ideas would be much appreciated. } } 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 } ********************************************
} The next battle is picking up the ribbon on several slot } grids - I suspect that it's more practice than anything else.
Mark,
I got rather consistent results with Perfect Loop from EMS. They even describe how to pick up sections (http://www.emsdiasum.com/ems/grids/accessories.html). Nevertheless I used it in slightly different way. After step 3 I put the loop with sections in the holder (stand) in a way that I can see it looking in microtome oculars. Then I put the slot grid coated with formvar just above the loop in desired orientation and slowly lower it down until section attach to the formvar film. Then I moved the grid with attached sections to side (not up!) so that practically all water remains in the loop. Result: I got serial sections on the grid with desired orientation and without necessity to remove water (a lot of dust can go from filter papaer to the grid when you try to dry it).
I have no any interest in EMS.
Sincerely, -- Alexander Mironov Jr. Division of Tumor Biology, H4 The Netherlands Cancer Institute Plesmanlaan 121 1066 CX Amsterdam
Most modern TEMs are ion pumped (or turbo pumped) to prevent the diffision pump oil backstreaming problem. The diffusion pump is used when initially evacuating the column or gun, pumping the camera and viewing chamber and when warming up the specimen cold trap at the end of sessions. Whereas it would be good to have a N2 trap on the diffusion pump when it does pump the column the water vapour from the camera and viewing chamber (due to the films) would stall the backing pump when the N2 trap warmed up. Prior to ion pumped columns we did use a N2 trap on the TEM column diffusion pump (circa 1980). Ion pumped or turbo molecular pumped SEMs are also available.
Regards, Ron
On 13 Sep 2000 13:00:11 -0500 Debby Sherman {sherman-at-btny.purdue.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 } -----------------------------------------------------------------------. } } } Steve, } Thanks for this post. I try to get this through to our users and we faithfully use and require use of N2 on all scopes. However, there are other facilities on campus who do not use N2 or use it intermittantly. When this topic comes up again, I intend to hand them your post and ask them to determine contamination rates on their instruments to convince them of the necessity of using N2. } One question that I have is that our main nitrogen trap on the SEM is above the diffusion pump to minimize backstreaming from the pump into the specimen chamber. (We also have a decontaminator close to the specimen that is cooled when we do cryo-SEM.) Now this is no doubt more of a problem with SEM than TEM due to the location of the diffusion pump. However, I wonder why most TEM manufacturers do not also use a nitrogen trap in this location, at least in the higher end scopes, in addition to the decontaminator in the specimen area. It would seem that, since contamination rate in the TEM is likely to be more critical to high resolution, it would also be used to help maintain the best possible vacuum in the column. } I am aware that the majority of the contamination is produced in the TEM by interaction of the beam with the specimen thus the need for decontaminator close to the specimen. However, my understanding is that the better the vacuum, the fewer the interactions of the beam electrons with molecules in the column and the more coherent the beam....i.e. better potential resolution. } Debby } } Debby Sherman 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 Wednesday, September 13, 2000, Steve Chapman {PROTRAIN-at-CompuServe.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 have read the offerings with interest and would like to make a few } } comments. } } } } Firstly - when a manufacturer designs an instrument they do not fit cold } } traps for fun, they believe that to obtain the best performance from their } } instrument you need the facilities that they fit. I have worked with } } design teams at two manufacturers and this is the attitude that they both } } took! } } } } Secondly - I have in the past, in countries where LN2 was not available, } } used dry ice (Solid CO2) and acetone in LN2 traps. I believe this brings } } the temperature down to -80deg C and it works quite well much better than } } no cooling at all. } } } } Thirdly there are imaging techniques that allow us to measure the } } contamination rate. Place a holey film in the TEM and without LN2 in use } } and take a high resolution picture of a hole. Leave the beam on the } } specimen and repeat the picture on the same hole 20 minutes later (that is } } if you still have a hole). Measure the two hole widths, take the smaller } } from the larger and divide by 2 (2 contaminated edges) divide by the time } } between the two pictures and divide by the magnification. You now have } } your contamination rate in nm/minute. Then try this with the LN2 systems } } in action and you will see just how well the LN2 systems work. } } } } The same system can be used with a SEM but use a well coated latex sphere } } in place of the hole. In this case the sphere will grow with time as the } } contamination builds round the sphere. } } } } Many years ago (1968) Hitachi produced a paper on LN2 systems. They gist } } was that if you used LN2 systems for a period of time and then stopped, it } } took an equal period of time before the vacuum level and contamination rate } } fell back to the original level. Of course you could not count the time if } } you let the column to air. } } } } A few more things to think about and a way of proving just how clean your } } microscope is. } } } } Steve Chapman } } Senior Consultant Protrain } } For consultancy and training by professional World Wide } } Tel +44 1280 814774 Fax +44 1280 814007 } } www.emcourses.com } } } } } }
---------------------- 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
*Date sent: Wed, 13 Sep 2000 04:34:23 -0400 *From: Steve Chapman {PROTRAIN-at-CompuServe.COM} *Subject: LN2 and Contamination *To: American Soc {Microscopy-at-sparc5.microscopy.com}
*------------------------------------------------------------------------ *The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I understand the procedure regarding the TEM, however could you explain to what I should compare the contamination rate in the SEM.
Best regards,
Witold
* *Hi * *I have read the offerings with interest and would like to make a few *comments. * *Firstly - when a manufacturer designs an instrument they do not fit cold *traps for fun, they believe that to obtain the best performance from their *instrument you need the facilities that they fit. I have worked with *design teams at two manufacturers and this is the attitude that they both *took! * *Secondly - I have in the past, in countries where LN2 was not available, *used dry ice (Solid CO2) and acetone in LN2 traps. I believe this brings *the temperature down to -80deg C and it works quite well much better than *no cooling at all. * *Thirdly there are imaging techniques that allow us to measure the *contamination rate. Place a holey film in the TEM and without LN2 in use *and take a high resolution picture of a hole. Leave the beam on the *specimen and repeat the picture on the same hole 20 minutes later (that is *if you still have a hole). Measure the two hole widths, take the smaller *from the larger and divide by 2 (2 contaminated edges) divide by the time *between the two pictures and divide by the magnification. You now have *your contamination rate in nm/minute. Then try this with the LN2 systems *in action and you will see just how well the LN2 systems work. * *The same system can be used with a SEM but use a well coated latex sphere *in place of the hole. In this case the sphere will grow with time as the *contamination builds round the sphere. * *Many years ago (1968) Hitachi produced a paper on LN2 systems. They gist *was that if you used LN2 systems for a period of time and then stopped, it *took an equal period of time before the vacuum level and contamination rate *fell back to the original level. Of course you could not count the time if *you let the column to air. * *A few more things to think about and a way of proving just how clean your *microscope is. * *Steve Chapman *Senior Consultant Protrain *For consultancy and training by professional World Wide *Tel +44 1280 814774 Fax +44 1280 814007 *www.emcourses.com * * %%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%% Witold Zielinski, Ph.D. Warsaw University of Technology Department of Materials Science and Engineering 02-507 Warszawa, Woloska 141 POLAND
This may help? I have used a (no doubt frowned on) practice for many years for easy collection of sections:
Make up 0.2% sodium lauryl sulphate (a detergent) in distilled water. Grids are dipped in this just prior to collecting sections, then drained on filter paper before introducing under the water in the knife boat and bringing up under the sections. The sections can be manipulated along filmed slots with an eyelash before the residual fillm of water dries. You can play this game under the binocular for some minutes while watching the water evaporate. Its a bit like watching paint dry!
Sometimes, raising the water meniscus (i.e. to convex) is useful when trying to position sections prior to collection.
Too much carry-over of detergent can cause the sections to run together and bunch up (a surface tension effect of the detergent).
The main problem with this practice is that you can only do this after cutting a batch of sections. If you wish to continue cutting then it is adviseable to flush the boat with fresh water, otherwise the block face will probably "wet" if you resume cutting. I normally pipette out the water and replace it three times - that works.
Keith Ryan Marine Biological Association Plymouth, UK
Unfortunately our CPD is out of service and my bottle of PELDRI nearly empty, therefore I am forever looking for new, fast, inexpensive ways to prepare my samples.
Although I do not work with plant material I was very interested in a publication that describes the use of glycerol or triethylene glycol to infiltrate biological specimens (mostly plant material) so that they may be observed in the SEM.
("Liquid substitution: a versatile procedure for SEM specimen preparation of biological materials without drying or coating" Journal of Microscopy, Vol. 172, 3. 1993 pp195-203. Ensikat H.j. & Barthlott, W.)
.."The main objectives of these preparation methods are to stabilize the specimen, to prevent shrinkage and other artefacts during dehydration, and to render the sample electrically conductive....specimen are not dried, but their water is instead substituted for a liquid..."
If the researcher tries the method I would be very interested to hear how it worked.
Regards
Claudia
On 13 Sep 2000, at 15:50, Edward J. King wrote:
Date sent: Wed, 13 Sep 2000 15:50:14 -0600 } From: "Edward J. King" {king-at-biology.utah.edu} To: Microscopy Listserv {Microscopy-at-sparc5.microscopy.com}
I have no experience of looking at pits. Another possiblility to reduce artifacts would be to use an ESEM. I think it might be difficult to get the resolution on unfixed material with a tungsten instrument. A field emission ESEM would have the resolution at lower voltages but may still require a comparison of fixed and unfixed material. Unfixed material has to be handled carefully to avoid beam damage artifacts.
Dave
On Wed, 13 Sep 2000 15:50:14 -0600 "Edward J. King" {king-at-biology.utah.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 } -----------------------------------------------------------------------. } } } A user of our facility needs to prepare specimens to determine if } sub-micrometer pores are present in plant intervascular pit membranes. } The user is very concerned that critical point drying or, especially, } the dehydration steps prior to CPD, may introduce 'pores' that are } really artifacts. He is similarly concerned that freezing or } freeze-drying his specimens might also introduce 'pores' due to ice } crystal formation. Work he has already done suggests that these are } valid concerns. } } Has anyone sufficient experience preparing pit membranes for SEM to } suggest a procedure we might follow to minimize the possibility of } creating small 'pores' where none may exist? } } Thank you, } } Ed King }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
From root Thu Sep 14 03:51:48 2000 Return-Path: {Microscopy-request-at-sparc5.microscopy.com} Received: (from daemon-at-localhost) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id DAA22959 for dist-Microscopy; Thu, 14 Sep 2000 03:36:26 -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 DAA22955 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Thu, 14 Sep 2000 03:35:55 -0500 (CDT) Received: from haymarket.ed.ac.uk (haymarket.ed.ac.uk [129.215.128.53]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id DAA22940 for {microscopy-at-sparc5.microscopy.com} ; Thu, 14 Sep 2000 03:35:44 -0500 (CDT) Received: from srv0.bio.ed.ac.uk (srv0.bio.ed.ac.uk [129.215.136.240]) by haymarket.ed.ac.uk (8.8.7/8.8.7) with ESMTP id JAA02352 for {microscopy-at-sparc5.microscopy.com} ; Thu, 14 Sep 2000 09:29:53 +0100 (BST) Received: from BIO-SRV0/SpoolDir by srv0.bio.ed.ac.uk (Mercury 1.44); 14 Sep 00 09:29:53 +0000 Received: from SpoolDir by BIO-SRV0 (Mercury 1.44); 14 Sep 00 09:29:25 +0000
Very satisfactory results have been reported for pollen (see ref. below) but I have found it worthless on plant cells in tissue culture, for which cryo-SEM must be the way to go. HMDS may work better on densely cytoplasmic material than on very vacuolate plant tissue.
I would be interested to know what the principles of operation of HMDS are thought to be. Does anyone have specific information about this?
Chris
Chissoe,W.F.; Vezey,E.L.; Skvarla,J.J. (1994) Hexamethyldisilazane as a drying agent for pollen scanning electron- microscopy. Biotechnic & Histochemistry69, 192-198 Abstract: Use of hexamethyldisilazane (HMDS) as a final dehydrating solution provides robust, undistorted secondary electron images of a variety of angiosperm and gymnosperm pollen grains, including those considered to be susceptible to collapse in the scanning electron microscope. Ease of handling, low cost, lack of specialized equipment, minimal expenditure of time, and high rate of success are factors that favor HMDS over other drying agents for preparing pollen grains for scanning electron microscopy
} From: "Patton, David" {David.Patton-at-uwe.ac.uk} To: Tobias Baskin {BaskinT-at-missouri.edu} Copies to: Microscopy-at-sparc5.microscopy.com
I confused myself with acronyms and broadcast the result! The HMDS may be used for drying specimens for SEM as noted. I also ventured that it can be used for dehydration - wrong. For rapid, complete dehydration dimethoxypropane (DMP) is used; it converts residual water to acetone. Thanks to Tobias for spotting that. No doubt a thousand others noted the error but were too polite (?) to write. I think DMP works only well to remove residual water after prior dehydration. Cheers Jim Darley ProSciTech
On Wednesday, September 13, 2000 11:43 PM, Tobias Baskin [SMTP:BaskinT-at-missouri.edu] wrote: } Jim, } Thanks for the info. How does HMDS compare with } dimethoxypropane as a water scavenger? I use a few % of latter in } freeze substitution brews as a "cemical sieve". I have heard to folks } using DMP as one shot dehydration before embedding. I even tried it. } Worthless for our stuff. Just curious about the HMDS. } } Tobias } } } In my previous response I gave the method, though I've used Chloroform. HMDS } } may have a theoretical advantage, but I expect that the results would be } } identical. } } HMDS was introduced for rapid processing, particularly fast and complete } } dehydration. It chemically interacts with water and forms acetone. When at } } least the last dehydration is in HMDS, the user can be sure that } } dehydration is } } excellent even with doubtful ethanol or acetone. } } } } During the air drying process residual water and solvent molecules exerts } } huge } } pressures onto membranes and these cause shrinkage and shriveling of any } } specimen. } } The drying mechanism: } } Using Peldri, drying only occurred at the Peldri/ air interface, but the } } specimen is held rigid in the solid Peldri material. As the Peldri slowly } } sublimes, more of the specimen is dried. } } During solvent drying the specimen is in a solvent saturated atmosphere and } } drying is very slow - over 48 hours. In this method there is little pressure } } exerted by molecules trying to pass through membranes and consequently } } shrinkage is minimized. } } Its a good alternative method and sometimes works very well. Given a choice } } I'll try CPD first. } } 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 } } ABN: 99 724 136 560 www.proscitech.com } } } } On Wednesday, September 13, 2000 12:21 AM, Tobias Baskin } } [SMTP:BaskinT-at-missouri.edu] wrote: } } } } } } } } } Greetings, } } } Several replies have mentioned this mysterious solvent for } } } CPD, and that it works on some type of specimens. Could someone } } } outline how this is used (infiltrate and then evap to air, or what?) } } } and if there is any rhyme or reason about what kinds of specimens } } } that work??? } } } } } } THanks, } } } Tobias } } } } } } } } } } } } } } } We use HMDS (Hexamethyldisilazane) from Sigma. } } } } } } } } Dave } } } } } } } } } } } } On Tue, 12 Sep 2000 08:53:03 -0500 Cameron Hind } } } } {hindc-at-baxter.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 Listservers, } } } } } } } } } } I have a memory lapse-can you remind me of the reagent used as an } } } } } alternative to CPD for SEM;or do you know of/recommend any } } } safe (not too } } } } } toxic) alternative method to CPD?-any reply would be appreciated } } } } } } } } } } Cameron } } } } } } } } } } Mr. Cameron Hind } } } } } Research Scientist } } } } } Advanced Technologies group } } } } } Baxter R & D Europe S.C.R.L. } } } } } Rue du ProgrEs 7 } } } } } 1400 Nivelles } } } } } Belgium } } } } } } } } } } Tel:+32 67 882 511 } } } } } Fax:+32 67 217 191 } } } } } e-mail: hindc-at-baxter.com } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } ---------------------------------------- } } } } Patton, David } } } } Email: David.Patton-at-uwe.ac.uk } } } } "University of the West of England" } } } } } } -- } } } _ ____ __ ____ Tobias I. Baskin } } } / \ / / \ / \ \ University } } } of Missouri } } } / | / / \ \ \ Biological } } } Sciences } } } /_ / __ /__ \ \ \__ 109 Tucker Hall } } } / / / \ \ \ } } } Columbia, MO 65211-7400 USA } } } / / / \ \ \ voice: } } } 573-882-0173 } } } / /____ / \ \__/ \____ fax: 573-882-0123 } } } } } } } } -- } _ ____ __ ____ Tobias I. Baskin } / \ / / \ / \ \ University of Missouri } / | / / \ \ \ Biological } Sciences } /_ / __ /__ \ \ \__ 109 Tucker Hall } / / / \ \ \ } Columbia, MO 65211-7400 USA } / / / \ \ \ voice: } 573-882-0173 } / /____ / \ \__/ \____ fax: 573-882-0123
Jim, I'm not sure if it is the same, but I have used 2,2-dimethoxypropene as an intermediate agent between 100% ethanol and air drying of the sample. It works very well and there is little distortion of the surface morphology.
Franklin Bailey Electron Microscopy Center University of Idaho Moscow, ID 83844-2204
} } We currently have a Bench Top Turbo carbon evaporation unit made by Denton. } We are very displeased with it. We consistently get non uniform coatings } that peel off of our metallographic mounts. Any suggestions on a } evaporation unit that works well or any information on 'sputtering" carbon? } Any other experiences out there? }
I would look very carefully at the cleanliness of the samples before coating. I use methanol then Freon then a dry Kimwipe, and the carbon coating is tenacious on brass.
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
Your first sentence is interesting, if I may stray a bit from the actual topic. I had formed the idea that the back-streamed oil is not diff pump fluid, but rotary pump oil. But if the phenomenon disappears when a turbo or ion pump replaces the diff pump, then that kindof exonerates the rotary pump, doesn't it? Or is it just that the turbo or ion pump is better than the diff pump at preventing rotary pump vapour from diffusing backwards through it? Anybody know the definitive answer to this?
} } Most modern TEMs are ion pumped (or turbo pumped) to } prevent the diffision pump oil backstreaming problem. The } diffusion pump is used when initially evacuating the column } or gun, pumping the camera and viewing chamber and when } warming up the specimen cold trap at the end of sessions. } Whereas it would be good to have a N2 trap on the diffusion } pump when it does pump the column the water vapour from the } camera and viewing chamber (due to the films) would stall } the backing pump when the N2 trap warmed up. } Prior to ion pumped columns we did use a N2 trap on the TEM } column diffusion pump (circa 1980). } Ion pumped or turbo molecular pumped SEMs are also } available. } } Regards, } Ron }
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
In response to Tamara Howard's predicament, here is an idea to consider. One can purchase bags of phase change material from Polar Tech Industries, Inc., in Genoa, IL (60135) that can be placed into a -20oC freezer and allowed to freeze. These can be placed into a styrofoam dry ice shipping box (commercially available from VWR with an external cardboard box for shipping) and should hold your samples between -20 and 0oC during shipment. This would be your ideal temperature range for shipment, and the temperature will hold for the duration of the air flight if your box size is reasonable. The foam refrigerant material stays in the bags, so there is no mess or hazard.
For more details, contact Polar Tech at 1-800-ICE-BRIX, or 815-784-9000, or at www.polar-tech.com.
I'd like to know how this works, if you haven't already opted for another approach.
Greg Fahy, Ph.D. 21st Century Medicine
________________________________________________________________ YOU'RE PAYING TOO MUCH FOR THE INTERNET! Juno now offers FREE Internet Access! Try it today - there's no risk! For your FREE software, visit: http://dl.www.juno.com/get/tagj.
Does anyone have any information on Fourier Transform Infrared (FTIR) Spectroscopy used for organic contamination analysis? Specifically manufacturers and the appx. price for the equipment . Any information would be helpful.
Martin A. Izquierdo Analytical Laboratory Cerprobe, Inc.
Retch is right, but you could also have a poor cooling system on that pump, or just a badly designed system. In either case backstreaming of oil vapour would lead to poorly adhearing films. 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 ABN: 99 724 136 560 www.proscitech.com
On Friday, September 15, 2000 6:55 PM, Retch Sims [SMTP:r.sims-at-auckland.ac.nz] wrote: } } } } } } } We currently have a Bench Top Turbo carbon evaporation unit made by Denton. } } We are very displeased with it. We consistently get non uniform coatings } } that peel off of our metallographic mounts. Any suggestions on a } } evaporation unit that works well or any information on 'sputtering" carbon? } } Any other experiences out there? } } } } I would look very carefully at the cleanliness of the samples before } coating. } I use methanol then Freon then a dry Kimwipe, and the carbon } coating is tenacious on brass. } } 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
I plan to buy CCD camera for TEM and want to remove wet dark room. So I need good printer. I think dye sublimation printer is the best choice. But I do not know what model is the best. Is there anybody who uses dye-sub printer in EM lab? Can you suggest what model is the best? My budget is about 5000 US$ or a little more. Thank you in advance.
Best regards,
Jondo Yun Kyungnam University Division of Advanced Materials Electron Microscopy Laboratory 449 Weolyeong-dong Masan, 631-701, Korea
Jondo Depends what the end-use of your prints is going to be. The most recent 6-colour + black 1440 dpi Epson Stylus printers produce absolutely stunning A4 size prints for a lot less capital and consumables cost. At arm's length you cannot really tell the difference between the Epson inkjet prints and a dye-sub print. With your budget you could buy a couple of these printers and still have enough left to buy a top-spec laptop! Epson also produce professional spec. inkjet printers, such as the Pro 5000 which are much more expensive, but may just be within your budget. see http://www.tssphoto.com/sp/dg/news/dot_comp.html for more details and links.
However, if you are determined to buy a dye-sub type printer, check out the Fuji Pictrography printers which combine laser exposed silver halide technology with dye transfer. The results are amazing. Chris
Send reply to: "Jondo Yun À±Á¸µµ" {jdyun-at-kyungnam.ac.kr} } From: "Jondo Yun À±Á¸µµ" {jdyun-at-kyungnam.ac.kr} To: "MicroscopyListserver" {Microscopy-at-sparc5.microscopy.com}
Hi Jondo,
I have no experience of dye sublimation printers but feel I must tell you how good our Epson Stylus Photo 1270 is. Using, admittedly expensive, photo quality paper we find it beats our very expensive colour laser printer for image quality. Unless you want dozens of copies done very quickly, I would certainly give the Epson a trial - and it costs a fraction of your budget, the remainder of which could buy a lot of consumables. BTW, I do not work for Epson
Kind regards,
Alan
********************************************* Alan Walker Dept of Electronic and Electrical Engineering University of Sheffield Mappin Street Sheffield S1 3JD United Kingdom
Glad I was able to help. I now number the "we do not use LN2" excuses 1 to 10 and simply ask which a client wishes to use as there is not such a thing as a "good excuse", I have heard them all!
Ron has given you a good explanation of DP traps, uses and problems but we should discuss pumping speed and TEM contamination a little further.
A vacuum in a TEM has fast and slow pumping areas. The electron gun is a fast pumping area but small pipes and pole piece apertures restrict gas flow and produce slow pumping areas, the specimen area in a TEM for example! No matter how good the vacuum may be at the DP it is only by improving the vacuum right by the TEM specimen that we see dramatic improvements, cold traps and cold fingers right by the specimen are the best route to lowering contamination.
Contamination in the TEM deposits on the top and the bottom of the specimen thickening the material and softening the image as the structures are gradually masked. Thicker specimens mean poorer resolution, often related as resolution is equal to one tenth of the specimen thickness. Improve the vacuum and you improve the contamination rate and an area that people do not relate to, you improve the gun vacuum and high voltage stability! So a better vacuum gives us a better contamination rate and better high voltage stability these two together will improve resolution.
I note you use a cold finger in the SEM when carrying out cryo experiments, I try to use such a device as much as possible if any high resolution study is taking place. Contamination rate tests would probably indicate as much as a five fold improvement when a trap is within inches of the specimen. I have designed several circular traps to fit round the pole piece of FEG SEMs, here contamination becomes the biggest barrier to very high resolution.
Good luck with those who have not taken to the LN2 gospel, keep at it!
Steve Chapman Senior Consultant Protrain For professional training and consultancy in EM world wide Tel +44 1280 814774 Fax +44 1280 814007 www.emcourses.com
I am trying to help a local high school set up an ISI SX-30 Super III SEM. It had been sitting for several years before being moved to the school. I am unfamiliar with this scope and we are trying to do some troubleshooting. We would greatly appreciate some help.
When the [Operate] button is depressed, it should pump down the chamber/column. However, it seems as though a valve is opened to atmosphere and loads the rotary pump. We hear hissing below the chamber which seems to be coming from a nozzle attached to a solenoid. We are wondering if there should be a hose attached to the nozzle and where it might go. Or is this an electrical problem?
Also, there is a loose snap wire connector in the back above the diffusion pump. It is marked JKA6 and contains a red and a yellow wire. I could not see the wires went to.
If anyone is familiar with this instrument, we would greatly appreciate some help in getting us started. The instrument was donated to the school and there are limited funds for bringing in service engineers.
Our FTIR expert here likes the complete system by Nicolet, but also said Perkin-Elmer and Biorad are a couple of other companies that make nice systems. I'm sure I'm leaving some out, as I'm not an FTIR expert, so forgive me if your company isn't on the list! As far as pricing, he mentioned a complete system goes for about $120,00US with only one lens. You can expect to add about $15,000US for each attachment, such as ATR and micro-ATR.
Again, I'm sure some vendors can give you more pertinent information, but this should put you in the ball park.
Hope this helps,
David Bell Scientist Electron Microscopy Lab Millipore Corporation 80 Ashby Road Bedford, MA 01730 (781) 533-2108
Martin Izquierdo To: "'Microscopy-at-MSA.Microscopy.Com'" {MartinI-at-ozte {Microscopy-at-sparc5.microscopy.com} k.com} cc: Subject: Fourier Transform Infrared Spectroscopy (FTIR) 09/14/00 09:13 PM
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Does anyone have any information on Fourier Transform Infrared (FTIR) Spectroscopy used for organic contamination analysis? Specifically manufacturers and the appx. price for the equipment . Any information would be helpful.
Martin A. Izquierdo Analytical Laboratory Cerprobe, Inc.
To the best of my knowledge, there is no such thing as "sputtering carbon". But I would suggest to look into your carbon source. Try different vendors. Also, make sure your vacuum is good.
Have a nice day!
Chao-Ying Ni Rodel Inc. 451 Bellevue Road Newark, DE 19713 (302) 366-0500 ext 2812
-----Original Message----- } From: jim [mailto:jim-at-proscitech.com.au] Sent: Thursday, September 14, 2000 9:53 PM To: 'Ritchie Sims'; Ekstrom, Harry; Microscopy-at-sparc5.microscopy.com
Retch is right, but you could also have a poor cooling system on that pump, or just a badly designed system. In either case backstreaming of oil vapour would lead to poorly adhearing films. 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 ABN: 99 724 136 560 www.proscitech.com
On Friday, September 15, 2000 6:55 PM, Retch Sims [SMTP:r.sims-at-auckland.ac.nz] wrote: } } } } } } } We currently have a Bench Top Turbo carbon evaporation unit made by Denton. } } We are very displeased with it. We consistently get non uniform coatings } } that peel off of our metallographic mounts. Any suggestions on a } } evaporation unit that works well or any information on 'sputtering" carbon? } } Any other experiences out there? } } } } I would look very carefully at the cleanliness of the samples before } coating. } I use methanol then Freon then a dry Kimwipe, and the carbon } coating is tenacious on brass. } } 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
Much as I dislike cluttering up the list with complaints, I would really like to see everyone following Nestor's recommended guidelines which include posting the sender's full address, including geographical. Sometimes, obviously, we can tell from the email address but all too often we can't; and sometimes we cannot even tell the country of origin, much less the city and state. This information is especially when posting persons are seeking help with instrument problems.
I realize that full address is an option on many email programs, and I sometimes forget to choose it myself, but I urge everyone to give it more thought.
Thank you.
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 dmrelion-at-aol.com. Thank you.)
The turbo pump is simply better than the diffusion pump at minimizing back streaming. Will Bigelow in his book on Vacuum Methods in Electron Microscopy talks about this and describes the care you should take in designing a pump-down protocol to minimize the backstreaming.
Tony.
} } Hi, Ron and all } } Your first sentence is interesting, if I may stray a bit from the } actual topic. } I had formed the idea that the back-streamed oil is not diff pump } fluid, but rotary pump oil. } But if the phenomenon disappears when a turbo or ion pump replaces } the diff pump, then that kindof exonerates the rotary pump, doesn't } it? } Or is it just that the turbo or ion pump is better than the diff pump } at preventing rotary pump vapour from diffusing backwards through it? } Anybody know the definitive answer to this? } }
** 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 use a Sony Mavigraph digital printer model UP-D1500 to print images from our SEM, its been reliable for the 4 years we have had it. I don't publish (industrial use- trade secrets, industrial hygiene, quality type stuff) so the resolution limit is OK for our use. The only problem that I have had is the print paper tends to curl sometimes and won't feed unless you straighten them. I also use a Sony Mavigraph color video printer model UP-3000 to print images from our light microscopes through a video camera and analog-digital converter and its been reliable and no problems at all. They both cost about $3000 when I bought them. Terry Ellis Of course I have no interests in that company, wish I did.
Can someone provide me with information on how to obtain microscope slides which can serve as standards for measuring optical density in transmission light microscopy? We would like to use these as a staining independent standard for the imaging and image analysis of histochemically stained control and experimental slides. Are there any commercially available ones? Alternative: can someone give me "recipes" for home made ones? We were thinking of eventually using small pieces of light exposed and developed film or neutral gray filters. Has anyone tried this already?
Thanks in advance.
**************************************************** Lauran C.J.M. Oomen oomen-at-nki.nl The Netherlands Cancer Institute tel. +31 20 5121889 Digital Microscopy Facility fax. +31 20 5121893 Plesmanlaan 121 1066CX Amsterdam The Netherlands ****************************************************
} Further to my posting yesterday seeking a PCD for a JEOL 840, which, } I am informed, is the same as that for a 6400, I am currently } negotiating with a company to make me one. } } Does anyone else out there want one?
While I am not looking for a PCD myself, I have customers who may be interested. If you can let me know the company you are dealing with I would appreciate it.
Joe _________________________________________ Joe Nabity, Ph.D. JC Nabity Lithography Systems E-Beam Lithography using Commercial SEMs & STEMs PO Box 5354, Bozeman, MT 59717 USA Voice: (406) 587-0848 FAX: (406) 586-9514 E-mail: info-at-jcnabity.com Web: www.jcnabity.com
I have just had a user approach me about looking at a water-saturated sample in our Hitachi S-2460N. While we have used VP-SEM for several years to look at insulating samples, this is the first time we have been asked to look at a moist sample. I have two questions as I plunge into this.
1-What instrumental concerns are there related to using water vapor as an atmosphere? I have heard that it may sorb onto the chamber surfaces and may be slow to desorb, so our ultimate vacuum may be poor for a while afterwards. There was also a concern over possible effects of the vapor on the precision leak valve. Are there any other components that might be affected, e.g., EDS window, BSE detector, etc.? Since the water will be present as vapor at less than 2 torr and will only be used for about an hour I don't envision problems, but I would rather not be surprised.
2-What are effective ways for introducing water vapor into the SEM? I am considering partially filling a vacuum desiccator with water and hooking our vent line to it and allowing the SEM vacuum to vaporize the water. Someone suggested a moistened rag in the vent line. What other means work well?
Thank you in advance. ---------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
I am a graduate student in the Materials Engineering Department at San Jose State University in California, and need some help with my thesis research. I was wondering if anyone out there has any experience with FIB imaging of recrystallized Cu films for the purposes of grain sizing. My research requires both plan view (at various tilt angles) and cross-sectional views of blanket films. I plan to work with an outside lab that is very experienced with SEM imaging, but relatively inexperienced with FIB imaging. They just got their first system online a couple of months ago. My questions are related to sample preparation from an 8" Si wafers and suggested FIB operating parameters, tips, and tricks to get quality images with good grain-to-grain contrast. If anyone has such experience or can point me in the right direction, it would be greatly appreciated. I am also seeking a good reference book or review article that covers FIB imaging theory (especially Ga ion channeling contrast). You can respond to the listserver, or contact me directly at attz-at-pacbell.net. Thanks in advance for your time and help.
LEE FILTERS Central Way Walworth industrial Estate Andover Hampshire SP10 5AN
http://www.leefilters.co.uk/
Lee make optical density filters in polyester squares 75mmx75mm which can be cut up into small squares or circles (paper punch) and mounted under a coverslip on a microsope slide. If you want to avoid contributions to the density from glass, mountant, etc drill an array of holes in a microscope slide sized piece of aluminium sheet, and mount samples of the filters over the apertures
Kodak also make ND Wratten filters in gelatin Chris
Date sent: Fri, 15 Sep 2000 15:56:33 +0200 } From: Lauran Oomen {Oomen-at-nki.nl} To: Microscopy-at-sparc5.microscopy.com
Good theory page: http://sis.bris.ac.uk/~sd9319/spec/IR.htm
Marc Helvey Sales 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
-----Original Message----- } From: Martin Izquierdo [mailto:MartinI-at-oztek.com] Sent: Thursday, September 14, 2000 6:14 PM To: 'Microscopy-at-MSA.Microscopy.Com'
Does anyone have any information on Fourier Transform Infrared (FTIR) Spectroscopy used for organic contamination analysis? Specifically manufacturers and the appx. price for the equipment . Any information would be helpful.
Martin A. Izquierdo Analytical Laboratory Cerprobe, Inc.
The answer is that rotary pump oil is able to pass through diffusion pumps easier than turbo pumps. Michael Postek of NIST wrote on article on this. "An Approach to the Reduction of Hydrocarbon Comtamination in the Scanning Electron Microscope" Scanning Vol. 18, 269-274 (1996). One of the techniques he used to reduce oil comtamination was to create a leak in the foreline of the Diff. Pump to create viscous flow conditions that stopped the backstreaming of rotary pump oil into the Diff. pump. This stopped condensation of pump oil on his EDS detector window.
I have a box of reprints of this article in my office, since it also discussed nitrogen purging which was used in my original SEM-CLEAN product. So if any one wants one, e-mail me back and ask for the "Postek" article. Please include your mailing address.
My web site SEMCLEAN.COM has full information on XEI's anti-contamination systems. No LN or turbopump required.
Ronald Vane XEI Scientific 3124 Wessex Way Redwood City, CA 94061 (650)369-0133
-----Original Message----- } From: Ritchie Sims {r.sims-at-auckland.ac.nz} To: message to: MSA list {microscopy-at-sparc5.microscopy.com}
I'm putting in my 2 cents on this thread because we have both in our lab. This is Tektronics Phaser 450 vs. the Epson 1270. The 1270 has cheaper supplies, more variety of papers, larger format and much much higher stability in it's favor. And the 450? I have to say I still prefer the dye-subs grey tones for images over the "grittiness" of the inkjet. The dye sub print still looks more like an RC print to me. When I requested materials from Epson demonstrating the capabilities of the 1270 and 870, we got this amazing test print from them where the gradation of tones really was spectacular. None of the prints from our own photos look nearly that good on the Epson, and in side by side comparisons with the dye sub, I think the dye sub prints always look better.
I guess it boils down to price considerations, is that extra $1.50 per print + the difference in cost between printers worth it?
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------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America Mark, I sent the method below to the list some time ago. It works great. Debby Sherman
A Fool-proof Method for Mounting Serial Sections on Single Hole Grids
I did serial sectioning for years on large single hole grids using a very simple technique that made the potential problems of film thickness, wrinkles and section loss very minor. I was not the original developer of the method and do not remember who originally gave it to me. It goes as follows:
1) Have your machine shop cut some thin pieces of Plexiglas into the size of glass slides. At one end, drill about a dozen holes, roughly 5mm in diameter, in an area about the size of a formvar film cast on glass slides. These slides will serve as your template for holding your films.
2) Cast the formvar films onto glass slides using your normal method. Usually a good silver film, not gray, will work fine. I routinely used 0.2% formvar in dichloroethane when casting by immersing the slide into the solution in a small jar, etc. We now use a film caster that lets us hold the slide in the dichloroethane vapors after lowering the formvar solution level. This method tends to give you thinner films consistently so the correct solution percentage and timing would have to be redetermined.
3) Float the film off the glass slide and pick it up with the Plexiglas slide so the film covers the holes. Then draw the water out of the holes by pressing the plastic slide down onto filter paper, or using small pieces of filter paper and capillary action to draw the water out of individual holes. The films should hold nicely over the holes in the slide. Store slides until needed.
4) Next, cut your sections using a block diameter that is fairly similar to the size of the slit in the grid. Pick up the sections on UNCOATED grids by gently lowering the grid to the surface of the knife boat. I put the dull side down on the premise that the rough surface would grab the film better during step 6. The surface tension of the water will hold the sections in the grid opening. Transfer the grid to a droplet of water until you have finished sectioning. Do not invert the grid. It is important that the top of the grid (shiny side) stay dry so that the grid will float on all subsequent solutions.
5) Transfer the grid + sections + water droplet to a drop of stain. A small amount of water will be transferred but this will not interfere with staining. If you are concerned about the dilution effect, increase your staining time slightly. Allow the section to stain, then wash by transferring through a series of droplets of clean water. Continue to post-stain if desired and wash the same way. Never let the grid dry. There is minimum problem with stain precipitation if you use very clean water and transfer the grid through a sufficient number of water droplets (6-12 recommended).
6) The final step is to transfer the grid to a film suspended over the hole in a Plexiglas slide and let it dry down. The sections will now be stuck to the film with NO wrinkles and minimum breakage. When ready to view, just punch out around the grid with the tip of your forceps, grab the grid and insert into the microscope.
Believe me....the sections will still be there at the end!
I found that as long as the sections cover a substantial portion of the open area of the grid, carbon coating was not essential. I used to do 50-100 grids worth of serial sections without loosing any. The films on the plastic slides would hold for months so I could make a lot and store until needed. The method really works...do give it a try.
Debby Sherman Microscopy Center in Agriculture Dept. of Botany & Plant Pathology Purdue University West Lafayette, IN 47907
Published: Sherman, D.M. (1998) A Full-proof Method for Mounting Serial Sections on Single Hole Grids. MSA Technologist’s Forum Newsletter 16:2
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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, thanks for the flood of suggestions about getting a ribbon. I } trimmed the block sides with a glass knife (and enjoyed Mike Delannoy's } suggestion of 'trimming the top and bottom sides of your face with a glass } knife' - didn't know microscopy was such a painful job) and quite a decent } ribbon was the result. I tried the chloroform-glue idea and that seemed to } make the sections stick together more, but sometimes they gummed up a bit } on the knife. The next battle is picking up the ribbon on several slot } grids - I suspect that it's more practice than anything else. The method I } was shown was to go underneath the ribbon at an angle with the grid, raise } it until the waterline is on the top of the exposed formvar of the slot, } push the ribbon onto the waterline and draw the grid up at the same time, } and touch the ribbon with the hair at the point where the slot finishes to } break the ribbon, and remove the slot - all easier said than done! Any } ideas would be much appreciated. } } 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 } ********************************************
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I have a Kodak 8670 dye sub printer hooked up to my SEM, but it seems to me that it takes a long time to get it to print and also while it is in the printing process my computer (and hence my SEM) is locked up. Here are the details.
I have the printer (1 year old) hooked to a PC with a 333 Pentium II processor (128 MB RAM) using a parallel cable. I do get the same performance when printing from other computers so I doubt that it is anything peculiar to this one computer.
Now when I call for a printout it takes 5 minutes to get the 1MB file size print done, and almost all of the time is spent sending data to the printer - the actual printing takes a short time (30 sec). I have discussed this with Kodak Tech Support and they claim my performance is not unusual. HOWEVER, I also experienced much quicker printing times (I'm sure - I have notes, but it was a while ago) when I was first using the printer (1 minute / 1 MB of file size in the image) and this is confusing me.
My question is: how fast should this sort of printer print when using the parallel cable to feed data to it? Is 5 minutes / 1MB unusually slow? I get frustrated particularly as my computer is locked up while the data is being sent out and I cannot continue imaging until the data dump is done. I have the Win 95 spooler running of course and I don't have this problem with my HP 970 inkjet which I use normally.
I will welcome any good ideas or cold realities that apply.
The reason that you can back up a turbomolecular pump with an oil-sealed rotary vane pump and still have a system that is virtually free of oil contamination is that the compression ratio of turbo pumps varies exponentially with the square root of the molecular mass of the molecules being pumped. Thus, for nitrogen, which has M = 28, this value is about 200, whereas for a hydrocarbon molecule such as octane with M = 66 it is greater than 3000, and for a molecule of pump oil with M greater than 100 it is greater than a million. These values are for a single compression stage (rotor disk) of a turbo pump. Actual turbo pumps have from 15 to 20 such stages which act in series, so the overall value for nitrogen ends up being around 10^9, while for hydrocarbons it becomes astronimical (10^40 is often cited). Thus, if a turbo pump that is backed by a rotary vane pump is managed so that the rotor never drops below about 75% of full operating speed while the pressure in it is in the molecular flow range (below about 1 Torr is a safe figure), it is virtually impossible for oil molecules to backstream through it. These matters are all discussed in detail on pages 229 to 234 of my book 'Vacuum Methods in Electron Microscopy' (see http:/www.2spi.com/catalog/books/book48.html, and http:/pup.princeton.edu/titles/6484.html for a description) and appropriate operating protocols are are described on pp. 249 to 253 .
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
Actually, any target material, including carbon and other high purity materials, that can be used in a vacuum evaporator or DC diode coater can be utilized in an Ion Beam Sputter Deposition System.
As for carbon, TEM electron diffraction images of ion beam sputtered carbon films are amorphous. The structure of a carbon grid before the carbon film was deposited cannot be distinguished from the carbon film!
TYPICAL SPUTTER RATES
Target Å/min
Cr 8-12 W 8-10 Ta 8-10 Pt 10-12 Ir 10-12 C 2-6
DISCLAIMER: South Bay Technology produces the IBS/e Ion Beam Sputter Deposition and Etching System for high resolution microscopy applications 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 "Ni, Chao-Ying" } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Harry,
To the best of my knowledge, there is no such thing as "sputtering carbon". But I would suggest to look into your carbon source. Try different vendors. Also, make sure your vacuum is good.
Have a nice day!
Chao-Ying Ni Rodel Inc. 451 Bellevue Road Newark, DE 19713 (302) 366-0500 ext 2812 {
We currently use staff engineers to service instruments, assist in teaching classes, train users on operating equipment, and performing some minor administrative tasks (primarily in the physical sciences area). We have 12 pieces of analytical equipment and only 4 instruments on service contracts, so we rely heavily on our staff to keep our instruments running. I have performed some surveys from industry and service engineer pay scales vary approximately in the following manner:
$30's-40's: for starting engineers with little to no experience that require training mid $40's to mid $50's: for an experienced engineer mid $50's to mid $70's: for a very skilled engineer who can trouble shoot to the component level independently without any back-up help.
It would be of great help to me and my staff if you could take a few minutes and enlighten me as to your approximate salary ranges for your staff compared to their experience level, and compared to the numbers I have shown above.
Thanks very much for your time and help.
Regards, Lucille Giannuzzi
******************************************************************* 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 ********************************************************************
A colleague wishes to print "archival" quality grey scale images (micrographs) on an Epson 750 Photo printer. They have purchased CMY archival quality inks from a company called Lysonic. They are apparently unable to obtain a gray scale image but instead obtain one that is "off color". According to my colleague, the printer will not generate an image using the Black ink cartridge but only when CMY inks are used. Has anyone solved this problem (of color balance)?
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 ####################################################################
http://www.kodak.com/cgi-bin/searchKodak.cgi?searchText=neutral+density+fi lters+ will get you on a good start. I tried to send it direct and couldn't resolve your address.
Good luck Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
----- Original Message ----- } From: "Lauran Oomen" {Oomen-at-nki.nl} To: {Microscopy-at-sparc5.microscopy.com} Sent: Friday, September 15, 2000 8:56 AM
Edmund Scientific sell stepped neutral density filters with 11 discrete density steps in intervals of 0.1 OD which might be useful here, though the OD range may not be wide enough for all applications.
These filters are coated on glass 2" x 1" x 0.062" thick, cover the OD range 0.04 (92%) to 1.0 (10% transmittance). They are spectrally flat in the range 400 to 700nm. Tolerances are not stated, but they are recommended as "Ideal for applications requiring variable densities and known density values". £119 in UK, which probably translates into $119 in US.
} } Dear all, } } } } Can someone provide me with information on how to obtain microscope } } slides which can serve as standards for measuring optical density in } } transmission light microscopy? We would like to use these as a staining } } independent standard for the imaging and image analysis of } } histochemically stained control and experimental slides. Are there any } } commercially available ones? Alternative: can someone give me "recipes" } } for home made ones? We were thinking of eventually using small pieces of } } light exposed and developed film or neutral gray filters. Has anyone } } tried this already? } } } } Thanks in advance. } } } } **************************************************** } } Lauran C.J.M. Oomen oomen-at-nki.nl } } The Netherlands Cancer Institute tel. +31 20 5121889 } } Digital Microscopy Facility fax. +31 20 5121893 } } Plesmanlaan 121 } } 1066CX Amsterdam } } The Netherlands } } **************************************************** } } } } } } } } } } }
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 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
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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"
We use a Hitachi 2250-N with a variety of gases. Attaching the inlet line to a desiccator or heavy conical flask of water works fine, but the vacuum gauge calibration may need checking if you need to know pressure accurately.
If you usually run with air or nitrogen in VP conditions, you will probably find the water gives you a resolution advantage (Helium is even better).
However your user should be aware that one or two torr of water does not do a lot to keep a specimen moist unless you also use a cold stage.
good luck,
Sally
Sally Stowe EM Unit Australian National University Canberra
} } } Warren E Straszheim {wesaia-at-iastate.edu} 09/16/00 02:01am } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have just had a user approach me about looking at a water-saturated sample in our Hitachi S-2460N. While we have used VP-SEM for several years
to look at insulating samples, this is the first time we have been asked to
look at a moist sample. I have two questions as I plunge into this.
1-What instrumental concerns are there related to using water vapor as an atmosphere? I have heard that it may sorb onto the chamber surfaces and may
be slow to desorb, so our ultimate vacuum may be poor for a while afterwards. There was also a concern over possible effects of the vapor on
the precision leak valve. Are there any other components that might be affected, e.g., EDS window, BSE detector, etc.? Since the water will be present as vapor at less than 2 torr and will only be used for about an hour I don't envision problems, but I would rather not be surprised.
2-What are effective ways for introducing water vapor into the SEM? I am considering partially filling a vacuum desiccator with water and hooking our vent line to it and allowing the SEM vacuum to vaporize the water. Someone suggested a moistened rag in the vent line. What other means work well?
Thank you in advance. ---------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
FTIR is a very useful tool for analysis of organic contaminants. All FTIR systems consist of a spectrometer, or bench. The bench can be fitted with various accessories -- e.g., a beam condenser, in-compartment ATR, microscope, etc. -- that permit analysis of different states and amounts of materials. Analysis of contamination likely will require use of one or more of these sampling accessories. Manufacturers such as Nicolet, SensIR, Perkin-Elmer, Biorad, and other third-party vendors can provide specifications and application notes for each. The total system cost will depend largely on the sampling accessory and other add-ons such as cameras and databases. A standard bench and beam condenser or in-compartment ATR might cost $25K to $30K. Some benches and in-compartment accessories are sufficiently portable for field use (SensIR makes a portable diamond ATR system with integrated video that is quite nice). If your samples are routinely less than 100 microns in diameter or consist of multiple phases or layers, a microscope accessory likely will be required. A standard bench and microscope might cost $65K to $85K or more; add multiple detectors, objectives, and automated operation for QC work and the cost can exceed $125K.
I use a portable Nicolet bench coupled to a high-end Nicolet/Spectra-Tech microscope. The bench can be taken on-site for analysis of larger homogenous samples. The microscope is used in the lab for analysis of microscopic samples, including individual fibers or layer in composites. Dual detectors in the microscope cover a spectral range of 4000-450 wavenumbers for organic and many inorganic analyses, and imaging is enhanced by infinity-corrected optics, fluorescence and visible polarized illumination, and a digital/video imaging system.
Whatever you decide best suits your needs, I suggest you visit a manufacturer to test drive their equipment using your own samples. Most are only too pleased to assist.
Hope this helps, and good luck.
James Martin Principal/Research Scientist Orion Analytical, LLC PO Box 550 Williamstown, MA 01267 www.orionanalytical.com
----- Original Message ----- } From: Martin Izquierdo {MartinI-at-oztek.com} To: 'Microscopy-at-MSA.Microscopy.Com' {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, September 14, 2000 9:13 PM
Hi,
I am interested in obtaining one or more Olympus POS (vertical tue monocular with rotating stage) microscopes in good condition. Anyone have one that is no longer being used?
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 dmrelion-at-aol.com. Thank you.)
Speaking of such, check out this story at http://www.cnn.com/2000/TECH/computing/09/14/epson.image.fading.idg/ind ex.html
Ozone fades some Epson photos
Contrary to ads promising fade-resistant, long-lasting photos, color prints made with several Epson Stylus Photo ink jets turn orange in locations with heavy concentrations of ozone in the air, Epson officials confirm. The company offers to buy your printer if you're not satisfied with several recommended fixes, including using new types of paper Epson will introduce this fall. The problem came to light when some owners of Epson's Stylus Photo 870, 875DC, and 1270 reported photos they printed were turning orange.
} } John writes: } } When in doubt NEVER buy a hp printer. The HP1600 what a piece of the proverbial garbage! { {
John...you've gotta stop sugar-coating everything...just tell it like it is!
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
As part of a gradulal upgrading of facilites, we're considering putting our booking sheets online to allow users to book themselves on the TEM, SEM, confocal or LM from the comfort of their own office.
Can anyone recommend Macintosh software capable of doing this? Ideally something which can be accessed by anyone with a web browser to avoid having to install new software on everyones computer.
Any feedback as to whether this sort of system really is better than a piece of paper nailed to the wall with a pen on a string would be welcomed too.
Thank you.
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
Is there a corresponding picture for a diff pump ie why do they let RP oil through? That's assuming, of course, that the oil that builds up on eds windows IS RP oil. It is, isn't it?
cheers
rtch
} } The reason that you can back up a turbomolecular pump with an oil-sealed } rotary vane pump and still have a system that is virtually free of oil } contamination is that the compression ratio of turbo pumps varies } exponentially with the square root of the molecular mass of the molecules } being pumped. Thus, for nitrogen, which has M = 28, this value is about } 200, whereas for a hydrocarbon molecule such as octane with M = 66 it is } greater than 3000, and for a molecule of pump oil with M greater than 100 } it is greater than a million. These values are for a single compression } stage (rotor disk) of a turbo pump. Actual turbo pumps have from 15 to 20 } such stages which act in series, so the overall value for nitrogen ends up } being around 10^9, while for hydrocarbons it becomes astronimical (10^40 is } often cited). Thus, if a turbo pump that is backed by a rotary vane pump is } managed so that the rotor never drops below about 75% of full operating } speed while the pressure in it is in the molecular flow range (below about } 1 Torr is a safe figure), it is virtually impossible for oil molecules to } backstream through it. These matters are all discussed in detail on pages } 229 to 234 of my book 'Vacuum Methods in Electron Microscopy' (see } http:/www.2spi.com/catalog/books/book48.html, and } http:/pup.princeton.edu/titles/6484.html for a description) and } appropriate operating protocols are are described on pp. 249 to 253 . } } 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 } } } } 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 } } As part of a gradual upgrading of facilities, we're considering } putting our booking sheets online to allow users to book themselves } on the TEM, SEM, confocal or LM from the comfort of their own office. } } Can anyone recommend Macintosh software capable of doing this? } Ideally something which can be accessed by anyone with a web browser } to avoid having to install new software on everyones computer. } } Any feedback as to whether this sort of system really is better than } a piece of paper nailed to the wall with a pen on a string would be } welcomed too. } Andrew,
A web page that emulated the sheet nailed to the wall should work fine and the use won't have to walk to the machine to sign up. I think it is a pretty simple job to write there should be dozens of kids around that can do it. Just don't let them try to improve on the sheet of paper on the wall concept or the users will get upset.
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
Dear All has anyone any experience with Agar's melinex film for growing cells on before fixation and embedding for the TEM? I would like to find out why it is necessary to fix the film with warm fixative - doesn't this introduce some artefacts due to membrane mobility at 37 degrees? I am looking at membrane antigens! Carole Elleman
} A colleague wishes to print "archival" quality grey scale images } (micrographs) on an Epson 750 Photo printer. They have purchased CMY } archival quality inks from a company called Lysonic. They are } apparently unable to obtain a gray scale image but instead obtain one } that is "off color". According to my colleague, the printer will not } generate an image using the Black ink cartridge but only when CMY } inks are used. Has anyone solved this problem (of color balance)? }
R. Howard Berg, Ph.D. Director, Integrated Microscopy Facility, Associate Member Donald Danforth Plant Science Center/Nidus Center 893 North Warson St. Louis, MO 63141
1 MB/minute is just under 20K/second which is what we typically see with our printers. It doesn't seem to matter what printers or computers we use. We have most of our printers hooked up via ethernet connections, but even those do not seem to be much faster than those using the parallel cable. That surprises me since ethernet (10 Mbit/sec) should be able to handle almost 1000K/second (if we had the whole cable to ourselves).
It seems that there must be something about Windows that restricts throughput to 20K//sec. Perhaps someone out there knows how to speed things up. Maybe there is some Windows setting to tweak.
The fact that your speed has dropped to about 4K/second indicates that something must have changed - I don't know what. I am also surprised that your computer is tied up while printing. You might want to check out your printer properties. On the second tab (details) there is a button for spool settings. I am used to the top selection being clicked which is "Spool print jobs so program finishes printing faster", rather than the alternate choice of "print directly to printer". Our programs are ready for action in just a couple of seconds (which depends on the computer and the printing task), and other programs continue to run even while the job is spooled up. If the other box is checked, the computer will be tied up until the job is fully transmitted.
I hope some of this help.
Warren
At 12:17 PM 9/15/2000 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi All, I beg your pardon for bringing up this issue again, but when I did not need it, I did not pay attention to postings...
Now I have to include in a construction grant a price for antivibration table for FEG-ESEM XL30 and, possibly, for STEM CM12. In XL30 I have problems with vibrations at magnifications 100-200k (and sometimes starting at 50k).
Who got good results with antivibrition platforms? What type? Should only a column be isolated or a whole instrument? What is a probable price range (including installation)?
Thanks,
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: Ni, Chao-Ying [mailto:CYNi-at-rodel.com] } Sent: Friday, September 15, 2000 7:56 AM } To: Ekstrom, Harry; Microscopy-at-sparc5.microscopy.com } Cc: 'Ritchie Sims'; 'jim-at-proscitech.com.au' } Subject: RE: Carbon Evaporation Units } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Harry, } } To the best of my knowledge, there is no such thing as } "sputtering carbon". } But I would suggest to look into your carbon source. Try } different vendors. } Also, make sure your vacuum is good. } } Have a nice day! } } Chao-Ying Ni } Rodel Inc. } 451 Bellevue Road } Newark, DE 19713 } (302) 366-0500 ext 2812 } } } -----Original Message----- } } From: jim [mailto:jim-at-proscitech.com.au] } Sent: Thursday, September 14, 2000 9:53 PM } To: 'Ritchie Sims'; Ekstrom, Harry; Microscopy-at-sparc5.microscopy.com } Subject: RE: Carbon Evaporation Units } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -------------------------------------------------------------- } ---------. } } } Retch is right, but you could also have a poor cooling system } on that pump, } or } just a badly designed system. In either case backstreaming of } oil vapour } would } lead to poorly adhearing films. } 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 } ABN: 99 724 136 560 www.proscitech.com } } On Friday, September 15, 2000 6:55 PM, Retch Sims } [SMTP:r.sims-at-auckland.ac.nz] } wrote: } } } } } } } } } } } } We currently have a Bench Top Turbo carbon evaporation } unit made by } Denton. } } } We are very displeased with it. We consistently get non uniform } coatings } } } that peel off of our metallographic mounts. Any suggestions on a } } } evaporation unit that works well or any information on } 'sputtering" } carbon? } } } Any other experiences out there? } } } } } } } I would look very carefully at the cleanliness of the samples before } } coating. } } I use methanol then Freon then a dry Kimwipe, and the carbon } } coating is tenacious on brass. } } } } 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 } }
I am trying to locate a Zeiss (Oberkochen)phase-contrast condenser for their 160mm tubelength classic grey series of microscopes.
The phase-contrast condenser needs to have achromatic-aplanatic top lenses, for the phase ring inserts that I have. I am prepared to buy any condenser that i can locate. Can anyone out there help me?
(I hope that this is the right place to post this request, Nestor)
Please reply privately to me on jb_sanderson-at-yahoo.com
Regards, Jeremy Sanderson
__________________________________________________ Do You Yahoo!? Yahoo! Mail - Free email you can access from anywhere! http://mail.yahoo.com/
Basically a distillation process allows roughing pump oil to leak through the diffusion pump. The rough pump oil boils up through the top jet of the diff. pump where as a lighter weight component it may collide with another molecule and bounce up into the chamber and backstream. The heavier diffusion pump oil is less likely to lose its downward and forward momentum during a collision.
Ron Vane XEI Scientific Redwood City, CA 94061 650-369-0133
Is there a corresponding picture for a diff pump ie why do they let RP oil through? That's assuming, of course, that the oil that builds up on eds windows IS RP oil. It is, isn't it?
cheers
rtch
} } The reason that you can back up a turbomolecular pump with an oil-sealed } rotary vane pump and still have a system that is virtually free of oil } contamination is that the compression ratio of turbo pumps varies } exponentially with the square root of the molecular mass of the molecules } being pumped. Thus, for nitrogen, which has M = 28, this value is about } 200, whereas for a hydrocarbon molecule such as octane with M = 66 it is } greater than 3000, and for a molecule of pump oil with M greater than 100 } it is greater than a million. These values are for a single compression } stage (rotor disk) of a turbo pump. Actual turbo pumps have from 15 to 20 } such stages which act in series, so the overall value for nitrogen ends up } being around 10^9, while for hydrocarbons it becomes astronimical (10^40 is } often cited). Thus, if a turbo pump that is backed by a rotary vane pump is } managed so that the rotor never drops below about 75% of full operating } speed while the pressure in it is in the molecular flow range (below about } 1 Torr is a safe figure), it is virtually impossible for oil molecules to } backstream through it. These matters are all discussed in detail on pages } 229 to 234 of my book 'Vacuum Methods in Electron Microscopy' (see } http:/www.2spi.com/catalog/books/book48.html, and } http:/pup.princeton.edu/titles/6484.html for a description) and } appropriate operating protocols are are described on pp. 249 to 253 . } } 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 } } } } 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
} } My question is: how fast should this sort of printer print when using the } parallel cable to feed data to it? Is 5 minutes / 1MB unusually slow? I } get frustrated particularly as my computer is locked up while the data is } being sent out and I cannot continue imaging until the data dump is done. I } have the Win 95 spooler running of course and I don't have this problem with } my HP 970 inkjet which I use normally.
Richard,
I have the immediate predecessor to the 8670, the 8650. We have 64 meg ram in the printer. We have the ethercard connector because we were told it printed faster. There is a user sitting at the printer workstation right now printing a 14 meg image. It is taking about 35 seconds for Photoshop to process the file and export to our print server. The print server sends the image to the printer and it is taking 90 seconds to print. The NT Workstation with Photoshop is a 3 year old dual Pentium Pro with 512 meg ram. The Photoshop preferences are set to 3 scratch disks on an unused 9 gig HDD on the same machine. The NT print server is a Pentium II 400 with 256K ram. Postscript takes longer. Very large files take longer (sizes 30 meg and up). The delay occurs in Photoshop processing and between color panels once printing has started and for very large files the time can be as long as several minutes. B&W 1 meg files will print in 70 seconds.
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
Lots of antibodies don't work at LM immunocytochemistry and even more don't work at the EM level. I would like to make an informal survey. If you respond to me or the list, I will post a summary to the list.
Here are the questions:
Have you generated antibodies that work in some method (ELISA, Westerns, immunoprecipitation) but not LM immunocytochemistry?
Were they monoclonals or polyclonals?
What percentage of the antibodies you have made work at the LM level but not for EM immunocytochemistry?
If you have made antibodies against synthetic peptides, what percentage work in either LM or EM?
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)
This is going to be a *very* depressing survey.......Maybe you should include a salary survey and get all the glum stuff out of the way at one time?
Tamara Howard CSHL
On Mon, 18 Sep 2000, Tom Phillips wrote:
} Lots of antibodies don't work at LM immunocytochemistry and even more } don't work at the EM level. I would like to make an informal survey. } If you respond to me or the list, I will post a summary to the list. } } Here are the questions: } } Have you generated antibodies that work in some method (ELISA, } Westerns, immunoprecipitation) but not LM immunocytochemistry? } } Were they monoclonals or polyclonals? } } What percentage of the antibodies you have made work at the LM level } but not for EM immunocytochemistry? } } If you have made antibodies against synthetic peptides, what } percentage work in either LM or EM? } } 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) }
One more note on the oil found on EDS detector windows. It isn't always roughing pump oil. Rick Felten found through IR analysis that the plasticizer from the vacuum hose connecting the roughing pump to the diff pump was at fault. Black vacuum hose from Japan is a bad actor in this way. The Red Vacuum Hose made in the USA has less problems.
One more note on the oil found on EDS detector windows. It isn't always roughing pump oil. Rick Felten found through IR analysis that the plasticizer from the vacuum hose connecting the roughing pump to the diff pump was at fault. Black vacuum hose from Japan is a bad actor in this way. The Red Vacuum Hose made in the USA has less problems.
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Reply to: RE: % of antibodies that work in immuno Dear Tom,
Why bother? It is obvous that some antibodies will only work with a specific technique. Why this happens is still open to speculation, but it is most likely due to masking of antigenic sites. If an antibody is made that recognises a protein only when it is configured in its natural state then the antibody will not work by Western blot. It may work really well after the sample has been fixed in high aldehyde concentration (or after osmium fixation!) as long as it has been fixed in its "natural" configuration. I wonder how many really good antibodies were discarded because they did not label denatured protein on a Western?
If the fixation process is so efficient that other proteins are retained around the antigenic site, then the antibody will not work either. Maybe the antibody has been diluted in inappropriate blocking agent and does not work for EM when it worked for LM. One example I know of occurred when the researcher was using rabbit serum as a blocking agent. Everything worked well by LM but at the EM level, when he used protein A-gold, there was no signal. He diluted the gold probe in the rabbit serum which bound very efficiently to the gold and stopped it from binding to the antibody on the section.
Monoclonal antibodies are more specific than polyclonals, and anti-peptide antibodies are usually even more specific than many monoclonals. If the antigenic sequence is contained inside a molecule then is may become inaccessible after good fixation. If the antigen is a small peptide chain that can be easily dislodged, then it may be washed away duing the fixation or labeling process. Labeling success will depend on the preparation protocols used and each will be usiue for each antibody-antigen system.
All of these problems have solutions available to fix them. Saying an antibody works for one method but not another only serves to re-enforce the "black magic" aspect of our VERY sceintific discipline. All these statements really mean is that we either do not fully understand our preparation and labeling protocols, or that we have no idea what our antibodies are binding to. Regards,
Paul Webster
Paul Webster, Ph.D. Scientist II & Director Ahmanson Advanced Electron Microscopy & Imaging Center House Ear Institute 2100 West Third St. Los Angeles, CA 90057
First of all I would like to thank all of you who responded to my question on standards for measuring optical density in transmission light microscopy. As a service to the "list" (at least I hope it is) I post a summary of the responses I have got. Most of them were 'off line'. Different types of ND filters can be obtained from: 1. Edmund Industrial optics (http://www.edmundoptics.com/IOD/DisplayProduct.cfm?Productid=1945) A range of mounted or unmounted glass filters (ND 0.15-2.5) and a stepped one with 11 discrete density steps (0.1 steps from ND 0.04 to 1.0) 2. Lee Filters (http://www.leefilters.co.uk/) A range of unmounted filters (ND 0.1-0.9) and as a set of three (ND 0.3, 0.6 and 0.9) (material not specified, but might be available as resin or gelatin filters, according to the representative in the Netherlands) 3. Kodak (http://www.kodak.com/country/US/en/motion/support/h1/filtration.shtml#camfilm)
A range of unmounted filters (gelatin WRATTEN; ND 0.1-4.0) 4. Cargill Industries in NJ Not been able to find info (yet) 5. Densichron Not been able to find info (yet)
-- **************************************************** Lauran C.J.M. Oomen oomen-at-nki.nl The Netherlands Cancer Institute tel. +31 20 5121889 Digital Microscopy Facility fax. +31 20 5121893 Plesmanlaan 121 1066CX Amsterdam The Netherlands ****************************************************
Our ESEM has developed a problem with its beam. I don't know if many other instruments have this, but there is a "source" mode button which , when hit, provides a sort of view "back up the column towards the gun". It's used for saturating the filament - when properly saturated, the beam should look like a nice round to oval bright spot. Our instrument has a LaB6 source, which, because of the shape of the tip of the crystal, may produce four weaker "satellite" beams around the "main" beam - which is OK, you just want to be sure you're using the main beam. Just yesterday, and quite suddenly, this view changed somewhat - we no longer seem to have a nice oval spot, but instead a sort of cross shape, with one side smeared out into a separate, smaller beam. Signal has been dramatically reduced - I have to open up the condensor 'way more than I should, to get any kind of reasonable signal, and then the resolution is pretty degraded. I'm thinking the LaB6 has failed, perhaps with some small bit falling off one side. Previously, when one of these has died on us, it's pretty sudden and dramatic - suddenly no signal, period. Our gun vacuum is generally pretty good, but we have at least a thousand hours on this filament, so I suppose it may be on its last legs. Assuming I'm right in blaming it on the LaB6 - I can't think of any other reason for this problem - has anyone else had a filament fail in this way? I'd just like to hear of any other possible causes before I open up the gun. I suppose it's possible some little bit of crud found its way partially up the column and is causing severe astigmatism, but that sounds like a major disassembly effort, too.
F.C. Thomas MicroAnalysis Facility Bedford Institute of Oceanography Dartmouth, Nova Scotia Canada Geological Survey of Canada (Atlantic) Bedford Institute of Oceanography Dartmouth, Nova Scotia Canada B2Y 4A2
Thanks to all who responded with my question about slow printing from my Kodak dye sub printer. I have tried some things and have tracked the problem down some.
I found I can print via the parallel port to the printer just fine from a second computer (Dell 450) in the lab, using freshly reinstalled postscript drivers. A 1.2MB file prints in about 1 minute and the transfer to the printer (Progress in the Systray Printer icon box) goes in a few seconds. In turn the same file, same cable, same software, both win 95, but different computer (Compaq 333 attached to my Hitachi SEM) takes 4.5 minutes to get over the printer, before printing starts, transferring in blocks of about 60kB, 1 every 8-10 seconds. I am not logged onto the network, and have disabled my Norton anti-virus software.
This experiment rules out a lot of issues and now I'm focusing on the setup of my SEM computer to see why the Progress goes so slowly on it while it goes so much faster on the Dell. If anyone has ideas about THIS issue I'll gladly try them out. Maybe Hitachi's computer guy has some ideas.
Richard Shalvoy --------------------- original note
I have a Kodak 8670 dye sub printer hooked up to my SEM, but it seems to me that it takes a long time to get it to print and also while it is in the printing process my computer (and hence my SEM) is locked up. Here are the details.
I have the printer (1 year old) hooked to a PC with a 333 Pentium II processor (128 MB RAM) using a parallel cable. I do get the same performance when printing from other computers so I doubt that it is anything peculiar to this one computer.
Now when I call for a printout it takes 5 minutes to get the 1MB file size print done, and almost all of the time is spent sending data to the printer - the actual printing takes a short time (30 sec). I have discussed this with Kodak Tech Support and they claim my performance is not unusual. HOWEVER, I also experienced much quicker printing times (I'm sure - I have notes, but it was a while ago) when I was first using the printer (1 minute / 1 MB of file size in the image) and this is confusing me.
My question is: how fast should this sort of printer print when using the parallel cable to feed data to it? Is 5 minutes / 1MB unusually slow? I get frustrated particularly as my computer is locked up while the data is being sent out and I cannot continue imaging until the data dump is done. I have the Win 95 spooler running of course and I don't have this problem with my HP 970 inkjet which I use normally.
I will welcome any good ideas or cold realities that apply.
The XL30 FEG-ESEM is already mounted on a built-in air-supported anti-vibration system. Before you go ahead and specify (or buy) an air table, I would suggest you look very carefully at the resonances, and how they would be affected by the additional isolation. Things don't always get better - in fact, an air table could make your situation worse if the resonances are not carefully thought out.
At the very least, have a professional vibration analysis performed to show what vibrations are present at your site and how the built-in system is responding.
I don't know about the CM12 and how it is supported.
Tony Garratt-Reed.
} } Hi All, } I beg your pardon for bringing up this issue again, but } when I did not need it, I did not pay attention to postings... } } Now I have to include in a construction grant a price for } antivibration table for FEG-ESEM XL30 and, possibly, } for STEM CM12. In XL30 I have problems with vibrations } at magnifications 100-200k (and sometimes starting at 50k). } } Who got good results with antivibrition platforms? What } type? Should only a column be isolated or a whole instrument? } What is a probable price range (including installation)? } } Thanks, } } Vladimir }
** 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 **
My use of Alar was a veerrryyy looong time ago, but, my recall is that the use of the warm fixative was more to fix the cells at the same temperature as the culture medium. This is due to the fact that many of the cells contract/round up/do nasty things when exposed to cold fixative. If you are concerned about membrane mobility artifacts, I would have thought that using a fixative that is the same temperature as the culture medium would be most appropriate. However, it might be useful to compare cells fixed at 37 and 4 C for your experiment. I know that this adds another layer of complexity onto the experimental design, but if this is a major issue, then I think it should be done. Let us know how it turns out.
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals On Mon, 18 Sep 2000 11:44:12 +0100, Carole Elleman 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 has anyone any experience with Agar's melinex film for growing } cells on before fixation and embedding for the TEM? I would like to find out } why it is necessary to fix the film with warm fixative - doesn't this } introduce some artefacts due to membrane mobility at 37 degrees? I am } looking at membrane antigens! } Carole Elleman } }
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As a coincidence, one of my technologists pulled your request for survey response off the web to tell me that I was under paying him. I presume that your classification of "engineer" refers to technical degree level people. I manage a group of 6 technical people with about ten state of the art level SEM's, TEM, EMP's and XRD equipment for a very long term AFTAC contract. AFTAC HQ is at Patrick AFB. Because we reside near Silicon Valley (just West of LLNL) we must reach deeper for starting rates here where entry level housing is about $300K. We cannot compete with the high tech firms that offer stock options, signing bonuses, etc. I just lost a materials scientist who about doubled her salary at a firm in Fremont. Obviously, our starting rates are a little higher than you indicate for engineers/scientists as well as technicians with little difference for degree status. I don't think that we have started anyone in our organization lower than the mid $40's in the past couple of years. Otherwise, we work with a similar salary range for non senior level professionals without significant supervisory responsibility.
Since no one has responded to this question in the past month I will provide my perspective. I have not seen any epoxy that will adhere to chitinous structures during sectioning. If someone else has please post the information. What I typically do when sectioning the eyes is to cut ten two micrometer sections on a dry glass knife then pick up the tissue (discard the piece of surrounding epoxy) with a fine needle probe and transfer it to a drop of water on a glass microscope slide. When I have enough sections the slide is moved to a hot plate for drying and later staining with toluidine blue. This works for the eye because epoxy infiltrates the underlying tissue from the backside. The proboscis is cut away and sometimes the head is cut in half during fixation. For fixing and infiltrating legs I would expect that this technique would work if they are cut in order to provide an opening at each end for fluid transfer. Best wishes.
Larry D. Ackerman Lily & Yuh Nung Jan Laboratories Howard Hughes Medical Institute UCSF, Box 0725, Rm U226 533 Parnassus Ave. San Francisco, CA 94143
When you solve this problem, I'd be really interested to hear what it was. Printers drive me nuts, too.
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 } -----------------------------------------------------------------------. } } } Thanks to all who responded with my question about slow printing from my } Kodak dye sub printer. I have tried some things and have tracked the } problem down some. } } I found I can print via the parallel port to the printer just fine from a } second computer (Dell 450) in the lab, using freshly reinstalled postscript } drivers. A 1.2MB file prints in about 1 minute and the transfer to the } printer (Progress in the Systray Printer icon box) goes in a few seconds. } In turn the same file, same cable, same software, both win 95, but } different computer (Compaq 333 attached to my Hitachi SEM) takes 4.5 minutes } to get over the printer, before printing starts, transferring in blocks of } about 60kB, 1 every 8-10 seconds. I am not logged onto the network, and } have disabled my Norton anti-virus software. } } This experiment rules out a lot of issues and now I'm focusing on the setup } of my SEM computer to see why the Progress goes so slowly on it while it } goes so much faster on the Dell. If anyone has ideas about THIS issue I'll } gladly try them out. Maybe Hitachi's computer guy has some ideas. } } Richard Shalvoy } --------------------- } original note } } I have a Kodak 8670 dye sub printer hooked up to my SEM, but it seems to me } that it takes a long time to get it to print and also while it is in the } printing process my computer (and hence my SEM) is locked up. Here are the } details. } } I have the printer (1 year old) hooked to a PC with a 333 Pentium II } processor (128 MB RAM) using a parallel cable. I do get the same } performance when printing from other computers so I doubt that it is } anything peculiar to this one computer. } } Now when I call for a printout it takes 5 minutes to get the 1MB file size } print done, and almost all of the time is spent sending data to the printer } - the actual printing takes a short time (30 sec). I have discussed this } with Kodak Tech Support and they claim my performance is not unusual. } HOWEVER, I also experienced much quicker printing times (I'm sure - I have } notes, but it was a while ago) when I was first using the printer (1 minute } / 1 MB of file size in the image) and this is confusing me. } } My question is: how fast should this sort of printer print when using the } parallel cable to feed data to it? Is 5 minutes / 1MB unusually slow? I } get frustrated particularly as my computer is locked up while the data is } being sent out and I cannot continue imaging until the data dump is done. I } have the Win 95 spooler running of course and I don't have this problem with } my HP 970 inkjet which I use normally. } } I will welcome any good ideas or cold realities that apply. } } Richard Shalvoy } Arch Chemicals } 350 Knotter Drive } Cheshire, CT 06410 } 203-271-4394 } } 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
I work in a lab that has great success with immuno labeling on a fluoresce laser confocual level, but taking the same antibodies to TEM thin sections is generally a lost cause. My success rate is probably around 5 to 10% (and that's a lot higher then my annual raises). Tim
Timothy G. Schneider Director of Electron Microscopy Department of Pathology Room 229 Jefferson Hall Thomas Jefferson University 1020 Locust St. Philadelphia Pa. 19107 215-503-4798 work 610-613-8170 cellular timothy.schneider-at-mail.tju.edu
I am having trouble obtaining good SEM preparations of cultured endothelial cell monolayers. The monolayers show evidence of artefactual cracking along cell borders. The cracks appear whether the cells are grown on glass coverslips or transwell filters. I have tried various fixatives (buffered glutaraldehyde alone, glutaraldehyde followed by osmium and uranyl acetate), different dehydrating agents (ethanol or acetone), followed by critical point drying (CPD). I have even tried tetramethylsilane (Ted Pella) in place of CPD. The only time I get good images (i.e., no cracks) is when a small portion of the monolayer has inadvertently detached from its substrate during tissue processing. The cells on this small flap look marvelous (no cracks). I believe, in the absence of substrate adhesion, the cells on the flap shrink uniformly during dehydration when surface tension forces exert their effects. Does anyone know how to prepare cell monolayers for SEM that leaves them intact and free from artefactual cracking?
Thanks.
Alan Burns, Ph.D. Assistant Professor Cardiovascular Sciences Department of Medicine Baylor College of Medicine
This relates to the question of printing from the computer that runs the SEM.
It is my policy in our facility that computers that operate microscopes such as our Hitachi S3500N and our confocals are not used for any other purpose. Images acquired are not saved on the computer that operates the device but rather are saved to a network drive (on a server) that is mapped to the individual user. All images from all devices go to the same server. The function of the server is to distribute these images, via direct connection, http, ftp to the individual users. The computers that run the devices are thus never clogged with images and are free of any programs other than those needed to run the device. The computers run faster and have fewer failures. They are setup the same as the day they were installed except for the network connection. I have often seen computer support personnel make the mistake of assuming that they can troubleshoot a computer than runs an SEM or confocal or other high end device only to discover that these computers are integral parts of the imaging device and should only be repaired by qualified technicians familiar with the device, not the computer.
Whenever possible, I suggest isolating the computer that drives your microscope from any other function.
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
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Listers- } } Our ESEM has developed a problem with its beam. I don't know if many other } instruments have this, but there is a "source" mode button which , when } hit, provides a sort of view "back up the column towards the gun". It's } used for saturating the filament - when properly saturated, the beam should } look like a nice round to oval bright spot. Our instrument has a LaB6 } source, which, because of the shape of the tip of the crystal, may produce } four weaker "satellite" beams around the "main" beam - which is OK, you } just want to be sure you're using the main beam. } Just yesterday, and quite suddenly, this view changed somewhat - we no } longer seem to have a nice oval spot, but instead a sort of cross shape, } with one side smeared out into a separate, smaller beam. Signal has been } dramatically reduced - I have to open up the condensor 'way more than I } should, to get any kind of reasonable signal, and then the resolution is } pretty degraded. } I'm thinking the LaB6 has failed, perhaps with some small bit falling off } one side. Previously, when one of these has died on us, it's pretty sudden } and dramatic - suddenly no signal, period. Our gun vacuum is generally } pretty good, but we have at least a thousand hours on this filament, so I } suppose it may be on its last legs. } Assuming I'm right in blaming it on the LaB6 - I can't think of any other } reason for this problem - has anyone else had a filament fail in this way? } I'd just like to hear of any other possible causes before I open up the } gun. I suppose it's possible some little bit of crud found its way } partially up the column and is causing severe astigmatism, but that sounds } like a major disassembly effort, too. } } F.C. Thomas } MicroAnalysis Facility } Bedford Institute of Oceanography } Dartmouth, Nova Scotia } Canada } Geological Survey of Canada (Atlantic) } Bedford Institute of Oceanography } Dartmouth, Nova Scotia } Canada B2Y 4A2
I am trying to create a positive control for a immunohistochemistry protocal. I have lyophilized protein which I have reconstituted in PBS. Does anyone have any ideas how I can adhere this protein to a slide so that it can be stained with the normal histo protocal.
I have tried drying the solution in the oven at 37C for different lengths of time. Should I use a gelatin solution.
Frank, Sounds like on of two things - either it failed that way because of the amount of hours(as you suggested), or, the tip was brought up to "saturation" way to fast.
Gary M. Easton Scanners Corporation Third Party EM Service ----- Original Message ----- } From: Frank Thomas {thomasf-at-AGC.BIO.NS.CA} To: {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, September 19, 2000 7:25 AM
Hi Alan We also had this problem but solved it by using tannic acid and making sure the cells were never fully uncovered.
I use two plastic pipettes - as I remove the liquid from the bottom of the culture dish, I add the new liquid to the top always making sure there is a thin layer of liquid on the cells. Three fillings of the chambers at each change ensures a good exchange.
Wash: 0.1M cacodylate buffer 3x5minutes
Postfix: 1%osmium tetroxide in 0.1M cacodylate buffer 30 minutes 1% tannic acid in 0.1M cacodylate buffer 30-60 minutes 1%osmium tetroxide in 0.1M cacodylate buffer 30 minutes
Rinse: ddw
Dehydrate: 5 minutes each in 30%, 50%, 70%, 85%, 95%, 100%, 100%, 100%
CPD.
If you remove all the liquid at a change of solutions, there can be damage to cellular processes such as filopodia. The tannic acid acts as a mordant and makes a real difference to the surface of the cells. I think this method is called the OTO method (Osmium, Tannic Acid, Osmium).
I have also used Tannic Acid in the buffer wash and then just used Osmium as normal. That worked too and was less expensive on Osmium. If you have empty wells when you use the OTO method, you can take off the first osmium and put it into an adjacent well. Then after the tannic acid, put the same osmium back.
Elaine
} } I am having trouble obtaining good SEM preparations of cultured endothelial } cell monolayers. The monolayers show evidence } of artefactual cracking along cell borders. The cracks appear whether the } cells are grown on glass coverslips or transwell filters. I have tried } various } fixatives (buffered glutaraldehyde alone, glutaraldehyde followed by osmium } and uranyl acetate), different dehydrating agents (ethanol or acetone), } followed by critical point drying (CPD). I have even tried } tetramethylsilane (Ted Pella) in place of CPD. The only time I get good } images (i.e., } no cracks) is when a small portion of the monolayer has inadvertently } detached from its substrate during tissue processing. The cells on this } small flap look marvelous (no cracks). I believe, in the absence of } substrate adhesion, the cells on the flap shrink uniformly during dehydration } when surface tension forces exert their effects. Does anyone know how to } prepare cell monolayers for SEM that leaves them intact and free } from artefactual cracking? } } Thanks. } } Alan Burns, Ph.D. } Assistant Professor } Cardiovascular Sciences } Department of Medicine } Baylor College of Medicine
-- 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
It could be that the printer port on the "slow computer" could be set up in bios "wrong", i.e. as a standard printer port, with no bi-directional capability. When booting up the Compaq, hit the appropriate key to enter bios setup. Look for an "integrated peripherals" section. Then, check your printer port selection. It should be set at least to the EPP mode or the EPP+ECP(this mode gives the port DMA capabilities which further enhance high speed transfers) mode. Save, and re-boot. Win95 should automatically recognize the change and load the appropriate drivers. If that doesn't work, upgrade the Compaq's memory by 64Mb's. Win95 spools print jobs to memory(if available - if not, the hard drive). Also, check the printers settings. Make sure it is set to "Spool" rather than "Print directly to printer". Good luck!
Gary M. Easton Scanners Corporation Third Party SEM Service ----- Original Message ----- } From: Shalvoy, Richard B **CHES {RBShalvoy-at-archchemicals.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, September 19, 2000 8:50 AM
How is this really different from what others are talking about? If the computer has to talk to a printer, or in your case, to a network, the path is still external. A pristine example is when the computer saves to a local disk. Then, the disk is physically carried to some other computer and accessed at that point. A network interface requires system resources just as any other interface does. Arguably, the network can take more resources than other interfaces. It is not benign.
The method which SEM makers utilize a controlling computer is a big variable. Some use programmed data transfers while others use IRQs. The days of PIO are waning as PC OSs move to NT-type systems. PIO is not supported. Furthermore, this type of interface eats CPU bandwidth...in stark contrast to IRQ approaches. A big problem with Wintel systems is the lack of interrupts. It is an old and still aging architecture. Watch for Win to change in many ways. A DOS-less system is one of the key changes.
System management is a core function. This applies very well to computers and computer systems. A single computer shackled to a single unit is not a very useful application of resources. If the computer is so inextricably tied to the controlled system, I'd be very shy of such a system and architecture.
If a computer is "clogged" in any respect, then I would submit that these systems are poorly designed. The sore sticking point is whether these systems will be or are supported in the future. Too many computer-based products are here today, gone tomorrow. How many folks are suffering with Win3.1 or Win3.11 systems? Not much fun.
At 12:30 PM 9/19/00, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Reply to: Re: Immuno TEM Tim Schneider writes: I work in a lab that has great success with immuno labeling on a fluoresce laser confocual level, but taking the same antibodies to TEM thin sections is generally a lost cause. My success rate is probably around 5 to 10% (and that's a lot higher then my annual raises). Tim
Hi Tim,
I look forward to seeing more details about your preparation protocols before commenting on your specific case. However, in all instances it is important that general statements regarding antibody reactivity be accompanied by specific examples of protocols. Only then can we compare antibody performance.
Here is a more general reply that I hope stimulates some thought:
As an addendum to my last posting, I will make the very bold statment that if an immuno-signal is seen by light microscopy the signal will be seen by EM. How so? First look at the preparation protocol used for the LM preparation. Was it on unfixed, air dried, methanol-fixed cryostat sections? Maybe. It is a routine protocol for LM immunocytochemistry. Now look at the typical EM protocol. Good fixation to preserve morphology (maybe 4% formaldehyde and 0.1% gutaraldehyde), rapid dehydration because we do not want to wash away our antigens, finally embedding in LR White or maybe even freeze substitution into Lowicryl after rapid freezing and no chemical fixative. The morphology in the sections looks good, but where is the signal?
One possibility is that the signal cannot be generated because the sample has been so well fixed and embedded that the antigen, or the gold probe, or both cannot gain access. I bet if the same protocol used for LM was also used for the EM preparation, the signal would be there. If it could not, I would start looking for the smelly rat. Maybe the morphology would suffer a little, but as this isn't a problem for LM, why should this trouble us at the EM level? We only require the morphology to enable us to identify labeled structures.
Another possibility is that the signal really is there, but because it is in such low amount we discard it as background labeling. However, this may be the total amount of signal we will ever get on thin sections. A cryostat section is at least 10 microns thick. Dry this down to a thin smudge on the slide, make it totally accessible to antibodies by exposure to methanol and you have an 2-D sample with antigen concentrated down and totally accessible to antibody. Apply antibody, add a secondary fluorescent antibody (and maybe a bridging antibody to amplify the signal) and a very obvious result is obtained.
Take this same antibody and apply it to thin resin sections (the thinner the better to get good resolution) and there will be much less antigen present in the section (60nm v 10,000nm). Also the antibody or gold markers usually do not penetrate the section. This means that the only antigen available for binding will be the small fraction that is exposed on the surface of the section. If the antigen is present in small numbers anyway, then the specific signal will be low. If protein A-gold is used, this usually binds with antibody at a 1:1 ratio so it is possible that the specific signal for any section may be one gold particle over one structure on the whole section. This might be the result for this antibody. This is easy to check. First find out how many antigens are present in the sample and work out how many should be exposed on the section surface. Labeling efficiency (the relationship between antigen number to observed signal) is usually between 5-20% (ie. if 100 antigens are available for binding only between 5 and 20 will label). From this it is possible to get an estimate of how many antibodies will bind to the section. It is often surprising that signal is detected at all.
In conclusion: I repeat - if an antibody doesn't work, then we probably do not understand enough about the antigen to design a good labeling protocol.
Also: If the antibody works for LM it WILL work for EM. Keep preparation and labeling protocols the same for both and you will enjoy much success. I don't think that any one of my collection of 100's of antibodies works only by LM. They all also work by EM. Some do not work on resin sections and some do not work if I fix cells with glutaraldehyde, but they all label at the EM level. Some only give me one gold particle for every 20 vesicles they are supposed to label, but the particle is always over one of these vesicles. I look forward to a good discussion about this (I just finished the official writing!), but don't ask me about salaries.
Regards,
Paul Webster, Ph.D. Scientist II & Director Ahmanson Advanced Electron Microscopy & Imaging Center House Ear Institute 2100 West Third St. Los Angeles, CA 90057
Alan, problems with cells cracking when cultured as monolayers are mostly depending on artifacts induced when dehydrating or drying the samples. The most probable reason for your problem is shrinkage of the cells, which gives severe tensions between your rigid support and the fragile cells. It's very typical that you get good preservation with no shrinkage artifacts when the monolayer has lost contact with the support, as they are free to alter their volume without any hindrance from the support. In my experience, fixation with 2.5 % buffered glutar aldehyde is normally sufficient as a fixing agent. Acetone should be avoided as the rate of dehydrating is too rapid and will in may tissues induce uncontrolled shrinkage. As I don't know your exact protocol for dehydration and CP-drying, I can only suggest some hints, so what I would try is dehydrating with ethanol followed by drying from liquid CO2. I would start dehydration in a medium-low percentage, 60 - 65 % ethanol in water, and work through a series of steps with 5% increase in concentration up to 100% ethanol. Time in every step would be at least 10 - 15 minutes. Substituting the ethanol with liquid CO2 in the CPD is critical, so if you can flush the chamber with liquid gas with the chamber closed, give it at least 10 minutes before closing the unit completely. Wait at least 2 hours before another 10 minutes flush before starting the drying procedure. The temperature has to be very slowly increased, normally it takes us 20 minutes to reach the state when the pressure can be released, and this last step has also to be done very slowly, at least in terms of 30 minutes to reach normal pressure.
This would be the way I'd go for any type of a cultured monolayer, and normally works OK. Good luck! Yours sincerely
Per Hörstedt Department of Medical Biosciences Pathology Unit for Electron Microscopy University of Umeå S-90187 Umeå Sweden
Late comer to this thread, but... A few things to check: Do the 2 computers (other than the Dell's slightly faster CPU) have similar resources? That includes system memory & bus speed, open hard drive space for virtual memory, similar setup size constraints on swap file size (if any), similar background programs running, similar hard drive access speeds... Humm, can't think of others to check presently.
Also, are both parallel ports set to the same mode in the system BIOS?
If the Hitachi system is like mine, it is using a SCSI I/O to the SEM, but I don't believe that will seriously affect resources unless in use (not positive).
Given a choice I would dump Dell and/or Compaq and go for a custom, generic clone PC. I (we) had no choice on the Hitachi (Compaq), but I managed to do so for my IXRF EDS because of special circumstances - and have been very happy with the performance. I am somewhat biased (could you tell) about the proprietary stuff the bigger manufacturers add to their PCs. good luck... BTW, I am using an Epson 900, parallel I/O.
Woody White, McDermott Technology, Inc. Me: http://home.att.net/~woody.white ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thanks to all who responded with my question about slow printing from my Kodak dye sub printer. I have tried some things and have tracked the problem down some.
I found I can print via the parallel port to the printer just fine from a second computer (Dell 450) in the lab, using freshly reinstalled postscript drivers. A 1.2MB file prints in about 1 minute and the transfer to the printer (Progress in the Systray Printer icon box) goes in a few seconds. In turn the same file, same cable, same software, both win 95, but different computer (Compaq 333 attached to my Hitachi SEM) takes 4.5 minutes to get over the printer, before printing starts, transferring in blocks of about 60kB, 1 every 8-10 seconds. I am not logged onto the network, and have disabled my Norton anti-virus software.
This experiment rules out a lot of issues and now I'm focusing on the setup of my SEM computer to see why the Progress goes so slowly on it while it goes so much faster on the Dell. If anyone has ideas about THIS issue I'll gladly try them out. Maybe Hitachi's computer guy has some ideas.
Richard Shalvoy --------------------- original note
I have a Kodak 8670 dye sub printer hooked up to my SEM, but it seems to me that it takes a long time to get it to print and also while it is in the printing process my computer (and hence my SEM) is locked up. Here are the details.
I have the printer (1 year old) hooked to a PC with a 333 Pentium II processor (128 MB RAM) using a parallel cable. I do get the same performance when printing from other computers so I doubt that it is anything peculiar to this one computer.
Now when I call for a printout it takes 5 minutes to get the 1MB file size print done, and almost all of the time is spent sending data to the printer - the actual printing takes a short time (30 sec). I have discussed this with Kodak Tech Support and they claim my performance is not unusual. HOWEVER, I also experienced much quicker printing times (I'm sure - I have notes, but it was a while ago) when I was first using the printer (1 minute / 1 MB of file size in the image) and this is confusing me.
My question is: how fast should this sort of printer print when using the parallel cable to feed data to it? Is 5 minutes / 1MB unusually slow? I get frustrated particularly as my computer is locked up while the data is being sent out and I cannot continue imaging until the data dump is done. I have the Win 95 spooler running of course and I don't have this problem with my HP 970 inkjet which I use normally.
I will welcome any good ideas or cold realities that apply.
Interesting post, but I have totake issue with one statement - I've read it over and I hopeI'm not taking this out of context; please excuse me if I have:
On 19 Sep 2000, Paul Webster wrote:
Lots of snipping.....} -----
} Maybe the morphology would suffer a little, but as this isn't a } problem for LM, why should this trouble us at the EM level? We only } require the morphology to enable us to identify labeled structures. } I depend on decent morphology to tell me that I haven't cause redistribution of the antigen/mRNA/gene at some time during the processing. If the sample looks like mush but has nice label, how am I to know that the label is where it was in vivo? I realize this may be extreme, to assume that everything is going to move if not properly fixed, but I think that it critical to keep this possibility in mind when localizing anything, at any level of detection.
My $0.02 this morning;hope I haven't ticked anyone off!
Tamara Howard CSHL (NY, USA for those of you who wonder!)
We have an old LKB III microtome that needs service. I called Leica but have not heard back -- and maybe never will-- they said they weren't sure they had anyone in the Washington DC area who could work on them.
Does anyone know of someone in this area who might be able to help? THere used to be a service engineer named Patricia Cappagrossi (I think) but I can't find any numbers for her...
TIA
Margaret -- Margaret Dienelt Plant Pathologist
Electron Microscopy Lab FNPRU, National Arboretum ARS, USDA
B. 010A R. 238 BARC-W 10300 Baltimore Avenue Beltsville, MD. 20705
Paul Webster said "I don't think that any one of my collection of 100's of antibodies works only by LM. They all also work by EM. "
Wow, that is amazing. I have never heard anyone having any where near that level of success. What type of epitopes are you looking at? What tissues are they located in? I could understand it if you were routinely using polyclonal antisera against carbohydrate epitopes - my own work with mucin molecules has shown me that carbohydrates are relatively easy to localize. How many of your antisera were made against synthetic peptides?
Can you tell me what you do when you get an IgM monoclonal to make it work at the EM level? I have never succeeded with one of them at the EM level.
You don't give your success rate for antibodies at the LM level? Have you succeeded with every antibody you have tried at the LM level? I have tried numerous ones that work for westerns or immunoppt that don't work at all at the light level (chemical fixation, precipitant fixes, quick-freeze) on cryo, paraffin, resin embedded protocols with or without a host of antigen retrival. Can you give me some references to your publications that outline your EM protocols for the "hard ones" or post your protocols on the list?
I routinely use 2% paraformaldehyde as a fix but vary this when I run into failure. I routinely screen in LR White, LR Gold, and Lowicryl K4M but in some (admittedly only a small percentage) have tried other plastics and heroic procedures. I have used ultra-thin cryo sections but, that it not really practical for some tissues (e.g., seed tissues with their hard coat and softer insides). Likewise, not every tissue is ammenable to nanogold permeabilization pre-embedding approaches (e.g., seed tissue again) . What do you do when you have tried 2% paraformaldehyde fixed tissues in LR White, LR Gold and Lowicryl and it still doesn't work? I routinely use labeled secondary antibodies to avoid the limitations with protein A. In most of my cases, the antigen level should be sufficient to see if the antibody worked.
How long does it take you to determine a successful protocol for each antibody?
You say "Keep preparation and labeling protocols the same for both and you will enjoy much success." I am not sure how to translate a positive result at the LM level with an acetone fixed whole mount of tissue culture cells grown on coverslips to the EM level. Do you use acetone or other precipitant fixes at the EM level? I have actually tried that but, not surprisingly, the quality of tissue preservation was not good enough for publication?
I know this is a lot of questions but your success rate is so much higher than mine, I am quite interested in hearing the details of your protocols.
Thanks, Tom
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-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
Date sent: Tue, 19 Sep 2000 17:31:45 -0400 } From: "Spaulding, Robert F" {SpauldinRF-at-corning.com}
Hi,
A question about SEM interference. We have high electromagnetic field levels (about 5 times the Jeol recommended limits) and our SEM isn't much use above 10 000x mag (a Jeol 5410 low vacuum) - the focus is fuzzy and astigmatism hard to correct. I've had water and air pipes removed from the room, and we've been everywhere measuring fields but the fields and the poor performance don't change. Changing the room is not an option (to the people in charge of the institute!). Jeol recommended, ruling out a room change, that we fit a mu-metal shield around the column. I have contacted the company that makes the mu-metal and the thing would cost about US$6000.00 and a hassle to ship to Mexico, so I don't want to go ahead and find that we're in the same place with our performance.
Does anyone have experience with fitting a mu-metal shield to their microscope (especially a scanning EM) and resolving (or not resolving!) interference problems. I'd be very interested in hearing about your experiences with this stuff.
Thanks,
Mark
PS - the serial sections are coming along nicely (not perfect,but heading that way!), but I've still got a lot of ideas to try yet. Many thanks.
******************************************** Mark West, Unidad de Microscopia Electronica, (Electron Microscopy Unit) Instituto de Fisiologia Celular, Universidad Nacional Autonoma de Mexico, 04510 Mexico D.F.
At 08:11 PM 9/19/00 -0700, Gary Gaugler wrote: } How is this really different from what others are talking about? If the } computer has to talk to a printer, or in your case, to a network, } the path is still external. A pristine example is when the computer } saves to a local disk. Then, the disk is physically carried to } some other computer and accessed at that point. A network } interface requires system resources just as any other interface } does. Arguably, the network can take more resources than } other interfaces. It is not benign.
I disagree. Very few system chores are as demanding as printing while networking is inconsequential. Writing to a mapped drive on a 10/100 mbit card is virtually transparent. If you have a local switch then it would be entirely transparent. However, printing a 1 meg file, especially to a postscript printer, can bring your system to it's knees (as it does in this case). PS translation can easily increase a 1 meg file to many times it's original size with a corresponding hit on performance while the information is generated then spooled then sent to the printer. But that is not really the point I was trying to make. My point is that it is well to consider isolating your dedicated computers from everyday tasks such as printing and allow them to manage the device for which they were designed.
Also, we do not use removable media to transfer images from devices and prefer to save from the image acquisition device to a RAID 5 server. Removable media have too many failure points and generally require more system resources. If you start the Task Manager on your NT workstation and watch the performance hit while writing to a ZIP you can see it is considerable. Writing to a HDD or a NIC are about equivalent and much less than the ZIP. Printing locally takes a big chunk of CPU time and sending the print to a print server takes the least of all. You should also consider the printer setup. If you are sending 600 dpi files to a 300 dpi printer you are wasting system resources and gaining nothing in return so it is well to check the printer setup on the two machines our original letter writer was referring to. In my opinion, I think you should let the server do the grunt work and let the dedicated computer manage the device.
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
Frank, The other thing that can happen with a LaB6 is a rather severe accumulation of "crud" on and around the Wehnelt aperture. With the very long lifetime of the LaB6 crystal, you can accumulate significant deposit in this area. I have seen similar instances where the crystal is still OK, but the contaminants on the Wehnelt get very thick and begin to pull away from the wall of the aperture and create distortions in the emission through the Wehnelt assembly. In order avoid this I try to clean the Wehnelt cap approximately every six months. Good Luck, Brad Huggins BP Amoco, Naperville, IL
} ---------- } From: Frank Thomas[SMTP:thomasf-at-AGC.BIO.NS.CA] } Sent: Tuesday, September 19, 2000 6:25 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: ESEM beam 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 } -----------------------------------------------------------------------. } } } Listers- } } Our ESEM has developed a problem with its beam. I don't know if many } other } instruments have this, but there is a "source" mode button which , when } hit, provides a sort of view "back up the column towards the gun". It's } used for saturating the filament - when properly saturated, the beam } should } look like a nice round to oval bright spot. Our instrument has a LaB6 } source, which, because of the shape of the tip of the crystal, may produce } } four weaker "satellite" beams around the "main" beam - which is OK, you } just want to be sure you're using the main beam. } Just yesterday, and quite suddenly, this view changed somewhat - we } no } longer seem to have a nice oval spot, but instead a sort of cross shape, } with one side smeared out into a separate, smaller beam. Signal has been } dramatically reduced - I have to open up the condensor 'way more than I } should, to get any kind of reasonable signal, and then the resolution is } pretty degraded. } I'm thinking the LaB6 has failed, perhaps with some small bit } falling off } one side. Previously, when one of these has died on us, it's pretty sudden } and dramatic - suddenly no signal, period. Our gun vacuum is generally } pretty good, but we have at least a thousand hours on this filament, so I } suppose it may be on its last legs. } Assuming I'm right in blaming it on the LaB6 - I can't think of any } other } reason for this problem - has anyone else had a filament fail in this way? } I'd just like to hear of any other possible causes before I open up the } gun. I suppose it's possible some little bit of crud found its way } partially up the column and is causing severe astigmatism, but that sounds } like a major disassembly effort, too. } } F.C. Thomas } MicroAnalysis Facility } Bedford Institute of Oceanography } Dartmouth, Nova Scotia } Canada } Geological Survey of Canada (Atlantic) } Bedford Institute of Oceanography } Dartmouth, Nova Scotia } Canada B2Y 4A2 }
I am awaiting delivery of an "engineering kit" of muMetal from Advance (about $100, online order). We have some EMI from the SEM CRT which doesn't sound as bad as your situation, but does limit resolution. Since it is not practical to move the monitor far enough to mitigate the situation, I decided to try shielding. I am not yet sure how I will fit in/around the chamber , but am going to see if it will make a difference. Will let you know if it successfully eliminates the "vibes".
Woody White McDermott Technology, Inc.
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Hi,
A question about SEM interference. We have high electromagnetic field levels (about 5 times the Jeol recommended limits) and our SEM isn't much use above 10 000x mag {SNIP} ******************************************** Mark West, Unidad de Microscopia Electronica, (Electron Microscopy Unit) Instituto de Fisiologia Celular, Universidad Nacional Autonoma de Mexico, 04510 Mexico D.F.
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Reply to: Re: Immuno TEM Hi Tom,
Thanks for joining in. My posting was a good excuse to get people thinking about stuff they usually take for granted. I am surprized when people tell me about their lack of success with EM immunocytochemistry when often the problem can be solved by minor protocol modifications.
My antibodies really do all work by EM. I guess I have never really thought about why this is so, but one reason may be that I get to know the people who make them, find out as much as I can about what the antibodies label, and try to understand as much as I can about the biology of the system I work with. Some antibodies will only work on tissue culture cells that have been cultured for more than 15 days. Before this time, the antigen has not been produced in sufficent number and has not been inserted into it's functional site. I can go on with examples, but this really does not help you.
As for making IgM's work by EM, how do you store your antibodies? Do you make sure that the IgM's are not subject to freeze-thaw conditions? One round of this will break them up sufficiently to make them unuseable. Sometimes one freezing is enough if there is no cryoprotectant in the solution. Ask whoever made the antibodies if they had been frozen at least once. The surprizing thing is that for LM, even low affinity antibodies can be detected. Imagine that only about 10% of the IgM is in good shape and all the rest is totally fragmented. This will have two effects. First, the antibody dilution will have to be increased for EM detection. Second, the fragments may still bind to antigens on your section, but because they are fragments, the secondary antibody will not recognise them (know your reagents). They have competed for the specific binding sites, have won, but cannot be detected by your visualization antibody. At the EM level, this is a disaster. For LM, where there will be many more antigens present, and where amplification techniques can be used, the signal will still be detected. Know how the IgM fragments, and know what the secondary antibody recognises, and you may be on the way to understanding why the antibody does not work.
Have you ever purchased streptavidin-gold to localize biotin at the EM level? This is another tricky reagent to work with because it always seems to detect biotin at the LM level using silver enhancer. At the EM level there is never a signal. Usually, the first question I ask is what blocking agent is being used. If serum is used, then the gold probe will bind to biotin-like molecules in the serum, thus eliminating the specific label. More importantly, it seems that this probe is very unstable with the gold seperating from the protein after only short storage times. The presence of free streptavidin in the solution is not sufficient to inhibit labeling at the LM level, but it does remove almost all the EM label.
ANother example I have concerns an antibody that worked by LM but not by EM. This was an antibody that was made to a protein synthesized by endothelial cells. The solution took me a week to work out. My colleagues could see immunofluorescence labeling if they followed the protocols in published papers using the same antibody, and could even repeat the published result. Whenever they tried the antibody by EM, there was literally no signal. Now usually, it is possible to see one or two gold aprticiles on a grid that has been exposed to gold probe. After lots of thinking, and a few experiments, we found out that the specific antibody was being removed from our labeling solution. The blocking agent initially used to dilute the antibody was 10% serum. As the protein being studied was also a serum protein, the antibody was reacting to this molecule and we were getting a very good negative result (adsorption control). The authors of the published papers had never bothered to do obvious controls and were happy when they saw signal at the LM level. What were they looking at? It turns out that the cells they (and us) were using contained lots of autofluorescent organelles that were being interpreted as specific signal.
As for antibodies not working by LM. If I do not get a signal by LM, I give up the antibody as being no use to me. There is no way I would expect a signal by EM if I couldn't see it first by LM. I will usually screen antibodies first on the system I expect to use for EM labeling. Thin sections of Lowicryl-embedded material make really good LM preparations. It is possible to also test the EM immunoreagents by LM too. The silver enhancers are great for visualizing protein A-gold.
Limitations of protein A-gold, what are they? I think there are more disadvantages to using gold conjugated secondary antibodies. First, the resolution is not as good as with PAG. Second, the antibodies will deteriorate when stored. Antibody without gold will compete for binding sites and reduce the label. With PAG, one reagent, used by everyone in the lab will be less expensive and can also be easily monitored for quality. If everyone is using it successfully and one researcher has a problem, it is easy to identify where the problem is. If someone is using a secondary antibody that has not been used for 6 months, the troubleshooting procedure will be boring and time consuming. My work is mostly with mammalian cells and pathogens and I do not have many antibodies to carbohydrates. Mostly they are to membrane proteins or to other cellular components. I think I have about 6 anti-peptide antibodies that work for me.
The mention of pathogens and antibodies brings up another point. Many antibodies are made using Freund's adjuvant, a common protocol for antibody preparation. This is a homogenate of ground-up bacterial proteins. If the study involves looking at bacteria it is inevitable that some background labeling on bacteria will be observed. If we know how the antibody was prepared, then we will not be surprized by this result. If we do not know the preparation protocols, then we will spend lots of time trying out different fixation and blocking solutions, eventually giving up saying that the antibody is no good.
I hope these posts help answer some of your questions. Contact me again if there are issues I didn't address. Regards,
Paul.
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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Mark-
Are you sure that your problem is external EM interference? If it were 60Hz, then I would expect to see (at some scan rates and magnifications, anyway) a "jagginess" in the images, rather than "fuzziness". Also, if it is possible to lock the scan to the local power frequency, you could get some other recognizable effects in the image. If it is not 60Hz, but a much higher frequency, it may be getting into the 'scope circuitry and affecting the scan drives or other parts of the optics, rather than the magnetic field directly affecting the beam. A mumetal enclosure would not help this. Even if it is 60Hz, it can get in via ground loops (often accidentally set up) and would not be affected by mumetal.
Another totally different possibility is that you are blaming the wrong thing, and in fact you have a problem elsewhere, but because of the known high fields the service people have a "get out".
You say that your fields are 5 times the recommended, and that you cant use the scope above 10,000. This would imply that with the recommended fields the scope could be used up to 50,000. Is this realistic? (I don't know the specs of the 5410). If not, the implication is that there is another fault.
These are just thoughts, I know, not solutions, but hope they help.
Tony G-R
} } Hi, } } A question about SEM interference. We have high electromagnetic field } levels (about 5 times the Jeol recommended limits) and our SEM isn't much } use above 10 000x mag (a Jeol 5410 low vacuum) - the focus is fuzzy and } astigmatism hard to correct. I've had water and air pipes removed from the } room, and we've been everywhere measuring fields but the fields and the } poor performance don't change. Changing the room is not an option (to the } people in charge of the institute!). Jeol recommended, ruling out a room } change, that we fit a mu-metal shield around the column. I have contacted } the company that makes the mu-metal and the thing would cost about } US$6000.00 and a hassle to ship to Mexico, so I don't want to go ahead and } find that we're in the same place with our performance. } } Does anyone have experience with fitting a mu-metal shield to their } microscope (especially a scanning EM) and resolving (or not resolving!) } interference problems. I'd be very interested in hearing about your } experiences with this stuff. } } Thanks,
** Anthony J. Garratt-Reed ** MIT Room # 13-1027 ** 77 Massachusetts Avenue ** Cambridge, MA 02139-4307 ** USA ** ** Phone: (+) 1-617-253-4622 ** Fax: (+) 1-617-258-6478 **
Accurel Systems International Corp. has an immediate opening for a Dual Beam FIB Engineer.
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Does anyone have an old Edwards E306A (manual) vacuum evaporator mouldering in their lab? I need either a rototilt 3 electrical feedthrough or a blank plug for a size "B" hole. The same parts for the *A*306 (automatic) won't work, as Edwards changed the hole size when they changed models.
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Mark, Are you sure you have a em field problem? They usually show up as a large(assuming the frequency is low, 50-60hz) sawtooth on the displayed image. This sawtooth will change in appearance with different sem scan rates, i.e., at TV rate, the images appears to "shake" back & forth, and at the slowest visual rate, will exhibit a very well defined sawtooth. If you do have a photo CRT(probably not), the photo scan is usually synchronized to the AC line frequency, hence negating the effect of any EM interference, again, assuming your "external fields" are the same frequency. Did you shut down the SEM completely, and disconnect it from its power source when making your field checks? This is very important. If the symptoms are a fuzzy image and high astigmatism, I would check my SEM parameters to ensure that the controls are properly set for high resolution - WD, spot size, aperture size, tilt, etc.. If these are okay, look for dirt somewhere in the column, column mis-alignment, poor vacuum levels. Also make sure that the beam crossover pole pieces are inserted correctly and that the column liner tube apertures are in the correct spot. Has this problem always been there since the instrument was initially installed, or did it develop afterwards. I really don't think mu-metal will fix your problem. Good Luck!
Gary M. Easton Scanners Corporation Third Party SEM Service
----- Original Message ----- } From: "Mark West" {mwest-at-ifcsun1.ifisiol.unam.mx} To: {microscopy-at-sparc5.microscopy.com} Sent: Wednesday, September 20, 2000 12:13 PM
We have a very low success rate as well, but I am convinced that it is because we are a fee for service lab and the PI's do not want to take the time or spend the money on the numerous trials that might be required to get acceptable results. I think, that if given the opportunity, we could have a much better success rate. But we are treated as a commodity and not a research endeavor. Greg Erdos Gregory W. Erdos, Ph.D. Assistant Director, Biotechnology Program PO Box 110580 University of Florida Gainesville, FL 32611
I would be interested in heaing from anyone who has done a market survey on JEOL 8200 versus CAMECA SX100 microprobes. Which ( in the responders opinion ) has the better system for geological quantitative WDS analysis, or performs better in the SEM mode. Please give the reasons behind your preference, if at all possible. PLEASE, NO VENDOR RESPONSE.
Gary L. Lovell Phillips Petroleum Company 245a GB Bartlesville, Ok. 74004 Phone: (918)661-9691 Fax : (918)662-2047 Email: gllovel-at-ppco.com
One of our students needs a receipe for citrated saline and cannot find it in the very limited references we have available here (a materials lab where one group is doing some tissue culture). She has seen it mentioned in articles but it must be so basic (or maybe acidic) that no one tells how it is made. I hope someone can enlighten us.
Microscopy and Digital Imaging: Advances and Applications
University of Washington Seattle, WA
October 5, 2000 Room CD150, CHDD Early Education Unit 8:30 AM to 4:30 PM
A day of presentations, demonstrations and refreshments. The first in a series of opportunities to learn and share knowledge about all aspects of microscopy and digital imaging for biomedical research.
Confocal Microscope Demonstrations October 4-6th: Bio-Rad - Radiance laser scanning confocal Solamere Technologies - Spinning disk confocal
Sponsored by: The UW Center on Human Development and Disability The W.M. Keck Center for Advanced Studies in Neural Signaling The Virginia Merrill Bloedel Hearing Research Center Supported by the Bio-Rad Corporation.
Please RSVP to Glen MacDonald, glenmac-at-u.washington.edu, 616-4156, to register for attendance to the workshop, confocal demonstrations or to meet with Dr. Johnson and Dr. Danilchik. Lunch will be provided for all who pre-register by October 2nd.
Schedule: 8:30 Welcome 8:45 Glen MacDonald, UW CHDD/VMBHRC - “What is in a Digital Image, What Happens When You Enhance It?” 9:45 Coffee and snacks 10:00 Iain Johnson, Molecular Probes - “Introduction to Fluorescence and Fluorescent Probes” 11:00 Mike Danilchik, University of Oregon Health Sciences Center - “Using Confocal Microscopy to Study Early Events in Xenopus laevi Development” 12:00 Lunch 1:00 Peter Rabinovitch, UW Dept. Pathology - “ Telomere Length Measurements in Tissue Sections by Quantitative FISH Confocal Microscopy” 2:00 John Jordan, Bio-Rad - “Recent Developments in Confocal Microscopy at Bio-Rad” 2:40 George Peeters, Solamere Technology - “Introducing the Spinning Disk Confocal Microscope and its Applications ” 3:00 Coffee and snacks 3:15 Iain Johnson - “A Demonstration of Web-based Resources for Fluorescent Microscopy, Where Does the Molecular Probes’ Technical Support Staff Get Their Information?”
--
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. ************************************************************************
We have not done this for a SEM, but have mu-metal around our TEM, and it works very well. Before you go to the expense, you may want to try some soft iron as an experimental shield. It is not the best (and certainly not pretty) but it works and will tell you if the problem really is fields and not something else.
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
Are you running the scope in high vacuum mode when you have these problems? What are the operating conditions and sample characteristics when you have this trouble?
I have trouble on our Hitachi 2460N getting decent pictures above 5kx using 40Pa of He. I also re-proved the point that gold coatings are beneficial. I was looking at some silicon samples in our JEOL 840A at 10kx. I could get images without charging, but the quality was poor. The pictures looked much better with gold since the secondary signal was stronger and generally coming from an area closer to the beam.
At 10:13 AM 9/20/2000 -0600, you wrote: } Hi, } } A question about SEM interference. We have high electromagnetic field } levels (about 5 times the Jeol recommended limits) and our SEM isn't much } use above 10 000x mag (a Jeol 5410 low vacuum) - the focus is fuzzy and } astigmatism hard to correct. I've had water and air pipes removed from the } room, and we've been everywhere measuring fields but the fields and the } poor performance don't change. Changing the room is not an option (to the } people in charge of the institute!). Jeol recommended, ruling out a room } change, that we fit a mu-metal shield around the column. I have contacted } the company that makes the mu-metal and the thing would cost about } US$6000.00 and a hassle to ship to Mexico, so I don't want to go ahead and } find that we're in the same place with our performance. } } Does anyone have experience with fitting a mu-metal shield to their } microscope (especially a scanning EM) and resolving (or not resolving!) } interference problems. I'd be very interested in hearing about your } experiences with this stuff. } } Thanks, } } Mark } } PS - the serial sections are coming along nicely (not perfect,but heading } that way!), but I've still got a lot of ideas to try yet. Many thanks.
---------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
Scanning electron microscopy, x-ray analysis, and image analysis of materials Computer applications and networking
From root Wed Sep 20 17:19:34 2000 Return-Path: {Microscopy-request-at-sparc5.microscopy.com} Received: (from daemon-at-localhost) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id RAA18732 for dist-Microscopy; Wed, 20 Sep 2000 17:08:36 -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 RAA18724 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Wed, 20 Sep 2000 17:08:05 -0500 (CDT) Received: from mailhost.auckland.ac.nz (mailhost.auckland.ac.nz [130.216.1.4]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id RAA18716 for {Microscopy-at-sparc5.microscopy.com} ; Wed, 20 Sep 2000 17:07:52 -0500 (CDT) Received: from glgnov2.auckland.ac.nz (glgnov2.auckland.ac.nz [130.216.59.90]) by mailhost.auckland.ac.nz (8.9.2/8.9.2/8.9.2-ua) with ESMTP id KAA07833 for {Microscopy-at-sparc5.microscopy.com} ; Thu, 21 Sep 2000 10:01:54 +1200 (NZST) Received: from GLGNOV2/SpoolDir by glgnov2.auckland.ac.nz (Mercury 1.44); 21 Sep 00 10:07:51 +1200 Received: from SpoolDir by GLGNOV2 (Mercury 1.44); 21 Sep 00 10:07:27 +1200
Hi
Any one still awake who can tell me which ports to mount 3 x wds spectros on an 840 which has the OM?
The three spectros are
STE/RAP PET/LIF PET/LIF
The three ports on the head of the OM aren't the same, which leads me to suspect that the positioning may not be arbitrary.
Please express your answer numbering the ports clockwise from the operator, bird's-eye view, 1, 2, 3.
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
Does anyone out there know of a SEM/TEM facility in the DFW area that is willing to rent time on their machine? .
I'm posting this for a colleague who has been using both SEM and TEM facilities on a pay for use basis for forever. Unfortunately those facilities are under new management who are reluctant to give outside users access to their equipment. So he is looking for alternatives. Thanks in advance for your input.
Email: laniven-at-julian.uwo.ca Name: Leigh Ann School: UWO Question: What is GFP? How do you convert a microscope for birefringence and what type of microscopy is this?
Listers, Anyone out there know of a good(hopefully easy!) way to remove carbon tracking deposits on a ceramic electon gun? Thanks in advance. Gary M. Easton Scanners Corporation
Date sent: Wed, 20 Sep 2000 10:13:03 -0600 (Hora estándar de México) } From: Mark West {mwest-at-ifcsun1.ifisiol.unam.mx} To: microscopy-at-sparc5.microscopy.com
} I found I can print via the parallel port to the printer just fine from a } second computer (Dell 450) in the lab, using freshly reinstalled postscript } drivers. A 1.2MB file prints in about 1 minute and the transfer to the } printer (Progress in the Systray Printer icon box) goes in a few seconds. } In turn the same file, same cable, same software, both win 95, but } different computer (Compaq 333 attached to my Hitachi SEM) takes 4.5 minutes } to get over the printer, before printing starts, transferring in blocks of } about 60kB, 1 every 8-10 seconds. I am not logged onto the network, and } have disabled my Norton anti-virus software. } } This experiment rules out a lot of issues and now I'm focusing on the setup } of my SEM computer to see why the Progress goes so slowly on it while it } goes so much faster on the Dell. If anyone has ideas about THIS issue I'll } gladly try them out. Maybe Hitachi's computer guy has some ideas.
Basically, you are using the wrong kind of OS and hardware for trying to do two things at once. "Go unix :-("
It seems that the key feature here is that the computer is also trying to control the SEM at the same time. Presumambly there is a resources bottlenck because the computer is desperately trying to do too many things in too many little chunks too often? If you've checked out the printing side and that's OK then maybe take a look at how the computer is talking to the microscope. At what rate does the computer normally try to "converse" with the microscope for example. If you used to be able to run the microscope and simultaneously print in reasonable time when the system was first installed, then perhaps this microscope polling rate (or whatever) has been subsequently increased excessively. Too fast a rate could cause an excessive slowdown when you try to get the computer to do something else intensive such as printing. Will your microscope software allow you to find out how often the computer talks to the microscope and allow you to change it? You might only need to slow down the rate of conversation with the microscope slightly to get a big improvement in both operations. Some experimentation might allow you to find an acceptable balance between printing time and microscope responsiveness.
Just a thought.
} } Richard Shalvoy } --------------------- } original note } } I have a Kodak 8670 dye sub printer hooked up to my SEM, but it seems to me } that it takes a long time to get it to print and also while it is in the } printing process my computer (and hence my SEM) is locked up. Here are the } details. } } I have the printer (1 year old) hooked to a PC with a 333 Pentium II } processor (128 MB RAM) using a parallel cable. I do get the same } performance when printing from other computers so I doubt that it is } anything peculiar to this one computer. } } Now when I call for a printout it takes 5 minutes to get the 1MB file size } print done, and almost all of the time is spent sending data to the printer } - the actual printing takes a short time (30 sec). I have discussed this } with Kodak Tech Support and they claim my performance is not unusual. } HOWEVER, I also experienced much quicker printing times (I'm sure - I have } notes, but it was a while ago) when I was first using the printer (1 minute } / 1 MB of file size in the image) and this is confusing me. } } My question is: how fast should this sort of printer print when using the } parallel cable to feed data to it? Is 5 minutes / 1MB unusually slow? I } get frustrated particularly as my computer is locked up while the data is } being sent out and I cannot continue imaging until the data dump is done. I } have the Win 95 spooler running of course and I don't have this problem with } my HP 970 inkjet which I use normally. } } I will welcome any good ideas or cold realities that apply. } } Richard Shalvoy } Arch Chemicals } 350 Knotter Drive } Cheshire, CT 06410 } 203-271-4394
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/
Before heading down an expensive path with the mu-metal, check out a few basic things first. The power source and step-down transformer would be easy to verify if they are correct.
Make sure the power is wired with a ground that runs all the way back to the distribution panel and is not grounded to the local disconnect box which would be a "conduit" ground. This type of ground is a good source
for noise to the microscope. Also, have a look at the step-down transformer. This could be incorrectly wired too.
I would imagine with a 5410 you could roll it down the hall, plug it in and see if it changes anything.
Bob Roberts EM Lab Services, Inc. 2409 S. Rural Rd Suite C Tempe, Arizona 85282 (480) 967-3946
I am after a supplier (preferably in Australia) for some doncaster dishes. These are glass petri dishes / watchglass dishes containing raised, concentric circles. Thanks Garry
Dr. Garry Rosewarne Plant Industry CSIRO, Black Mountain
On Wed, 20 Sep 2000 20:26:39 -0500, laniven-at-julian.uwo.ca 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 | -----------------------------------------------------------------------.
| | Email: laniven-at-julian.uwo.ca | Name: Leigh Ann | School: UWO | Question: What is GFP? How do you convert a microscope for birefringence | and what type of microscopy is this? |
Hi Leigh Ann - I can answer the first part of your question (to some extent), but I'm not very sure about the second part. GFP = Green Fluorescent Protein ; it's a protein that naturally fluoresces, and I believe that it was first discovered as a natural product from some marine animal (name ?). What I know of GFP is that it's gaining usage as a fluorescent marker. I believe it's been used as a marker for determining the location of X compound in a cell when viewed by fluorescent microscopy, but I'm sure that other usages have been cited in other research papers. As for the second part ... someone in this list may correct me, but I believe that birefringence is correlated with polarized light. You'll definitely need to consult with someone else in this list to get more information. Good luck getting more answers. Nelson Conti
_______________________________________________________ Say Bye to Slow Internet! http://www.home.com/xinbox/signup.html
I assume that the tracks are on a ceramic insulator which is not glazed, I am not sure what might happen if a glazed surface is abraded.
Alumina powder made up to a paste in Alcohol is good for polishing out tracks on high voltage insulators (guns and accelerators). If it is really bad then grit blasting with Alumina powder may be needed. Make sure you don't damage seals or other surfaces and that you wash thoroughly to remove all traces of the powder. (That's wash the gun of course, you should not get it all over yourself.) The insulator should be gently baked out in a (clean) vacuum oven to remove the final traces of solvent, but you may get away with baking out gently in air and pumping out for a long time before applying the voltage.
Beware of the limitations imposed by other materials used in the constuction of the component, epoxy resins etc.
Good luck, Ron
On Wed, 20 Sep 2000 20:15:50 -0500 "Gary M. Easton" {gary.easton-at-scannerscorp.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 } -----------------------------------------------------------------------. } } } Listers, Anyone out there know of a good(hopefully easy!) way to } remove carbon tracking deposits on a ceramic electon gun? Thanks in } advance. Gary M. Easton Scanners Corporation } } }
---------------------- 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 dryers we're using are a couple of Polaron E-3000 CPD's, and they are working perfectly. The main advantage is that you manually control inlet- outlet- and evacuation valves, which means you can totally control the substitution of dehydration agent when the chamber is closed. Temperature is controlled with ordinary tap water, controlled by an ordinary thermostat-mixer. They also have a large viewing window that makes it very easy to visualize everything that goes on inside the chamber, i.e. for example the level of liquid CO2 which is very important when flushing the chamber. The chamber has a large volume that can contain many and/or large samples. I've had a workshop making some sets of specimen carriers for different types of specimen, so for example when working with biopsies, we can prepare 24 at the same time. We've used them for many years now and I find them highly recommendable. Yours sincerely
Per Hörstedt Department of Medical Biosciences Pathology Unit for Electron Microscopy University of Umeå S-90187 Umeå Sweden
Building test shields from 0.30 cent per pound iron is a good test for how well MU metal will work.. If you have an Ag Engineering department you might be able to borrow it from them at no cost except on favor to be announced at a later date. .30 cent iron my be less expensive.
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00 ----- Original Message ----- } From: {"bobrob-at-uswest.net"-at-sparc5.microscopy.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, September 21, 2000 12:10 AM
HCL works great as long as you protect the other (metal) elements.
Earl
"Gary M. Easton" 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 } -----------------------------------------------------------------------. } } Listers, Anyone out there know of a good(hopefully easy!) way to } remove carbon tracking deposits on a ceramic electon gun? Thanks in } advance. Gary M. Easton Scanners Corporation
-----Original Message----- } From: Granville Sent: Thursday, September 21, 2000 1:51 PM To: Luc
Hi Ann, GFP is the Green Fluorescence Protein. Jellyfish Aequorea produces bioluminescence from two proteins, aequorin, which is a Ca++ binding protein that produces blue light. GFP emits green light by absorption of the aequorin. GFP was discovered in1962 by O. Shimomura. In 1992 D.Prasher and colleagues sequenced it. Now there are variants of GFP. GFP is a 238 amino acid protein. Absorption at 395 nm and 475 nm, emission at 508 nm. It can expressed in a cell/tissue /animal with along another protein of interest. It is, therefore, a reporter or label. It can be used to report gene expression or gene transfer, for a highly specific label for protein localization, quantification or dynamics. greetings Werner
Margaret, You may call Norman J. Woodside, an ex-LKB employee, and he may help you. His address and phone and fax numbers: NJW Supplies 7 Tanglewood Rd. Catonsville, MD 21228 Phone: 410-744-9574; Fax 410-744-1580 Thanks! Pat
Birefringence is observed under a light microscope by putting a polarizer before the specimen (and generally before the condenser) and a similar device called the analyser between the specimen and the eyepiece: the analyser is usually set at 90^ to the polarizer. If the crystal (or even a piece of stretched plastic or a blob of liquid crystal) has different refractive indices in different directions, the polarized light passing through the specimen gets divided into two out-of-phase components which are recombined in the analyzer to give pretty interference colours, from which one can also make quantitative deductions. The people that use this technique most are geologists and mineralogists looking at thin polished rock sections, but we plastics people also look at polymers in this way. I'm not sure if it's used much in bio/med applications.
A good book to explain the physics and applications of the technique is:
Wood, Elizabeth Armstrong Crystals and light.
while there are many books which concentrate more on the setup of the microscope. Here's a selection out of our library catalogue:
Hartshorne, N.H. Crystals and the polarising microscope.
Polarized light microscopy by Walter C. McCrone, Lucy B.McCrone and John Gustav Delly
The polarizing microscope by A. F. Hallimond Hallimond A.F.
+------------------------------------------------------------------------+ | 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 | +------------------------------------------------------------------------+
} Email: laniven-at-julian.uwo.ca } Name: Leigh Ann } School: UWO } Question: What is GFP? How do you convert a microscope for birefringence } and what type of microscopy is this?
Tamara Howard writes: } Maybe the morphology would suffer a little, but as this isn't a } problem for LM, why should this trouble us at the EM level? We only } require the morphology to enable us to identify labeled structures. } I depend on decent morphology to tell me that I haven't cause redistribution of the antigen/mRNA/gene at some time during the processing. If the sample looks like mush but has nice label, how am I to know that the label is where it was in vivo? I realize this may be extreme, to assume that everything is going to move if not properly fixed, but I think that it critical to keep this possibility in mind when localizing anything, at any level of detection.
I seem to have caught someone's attention with my very controvertial post. First let me address Tamara's question, which also covers part of an issue brought up by Tom Phillips. You take exception that I should advocate the use of poorly fixed material for EM localization. Yet, at the LM level this same poor fixation is considered a routine part of the protocol. How many people who perform LM immunolocalizations routinely freeze the tissue without aldehyde fixation or cryoprotection and then fix the sections with acetone or methanol, air-drying them before or after this treatment? Who would conside that they are working with a) badly fixed material?, b) material where the target antigens have either been removed or relocated? My guess is that very few people have even given this a thought. [If you think that performing LM immunochtochemistry using aldehyde fixed material followed by detergent permiabilization (without drying the sample) may I direct you to Hannah, Weiss & Huttner 1998 Differential extraction of proteins from paraformaldehyde-fixed cells: Lessons from synaptophysin and other membrane proteins Methods (a companion to Meth. Enzymol.) vol16 pp170-181)].
These issues about morphology are never taken into consideration at the LM level. Why then should we decide it is important to have good morphology at the EM level? One reason is that we are attempting perfection and feel that good morphology is aesthetically pleasing. More importantly we need to have sufficient contrast information within the sections to obtain a colocalization with recognizable subcellular structures (and hopefully see the antigen location at sites where it exists in vivo).
If we are able to perform multiple labeling experiments, where we can use one antibody to identify specific organelles, then absolute morphology is not important. We can disrupt our cells so that they become bags of loosely packed organelles and our antibodies have total accessibility to antigens. We can even partially purify these organelles and work with them as a pellet or adsorbed onto specimen grids. We will loose spatial information if we do this, but we will get a signal.
Similarly, if the only way we can get a signal by LM is by air drying and acetone fixing, then use the same protocol for EM. The protocol will be easy (pre-embedding label) but the morphology will be awful. Remember, this is what light microscopists accept as good morphology! Once the label system has been established to work by EM (ie all the reagents work to give a result), then it will be possible to start modifying the protocol to improve morphology. Perhaps fixing the sample in 2% formaldehyde for 30 seconds, followed by a short homogenation (improve membrane preservation and control cellular disruption) is the answer. Maybe a fixation in 5% glutaraldehyde in a low osmolality buffer, so that the cells swell, is the answer. The exact recipe will depend on what is known about the antigen. If it is a membrane protein, is the epitope deeply embedded inside the membrane? If so, will this require membrane disruption (freeze-thaw, detergent?) or will it be sufficient to just remove the cytoplasm? Each system has to be approached as a completely new project and each will have an answer. Knowing as much as we can about the antigen is our first step to success, allowing ourselves to work with samples that do not have "text-book" quality morphology is the next step. Being totally open about how we are willing to prepare samples and look at our results, completes the process. My protocols will not help many people with their own particular immunolabeling problems but my experience might. So will the experience of all the other researchers who have faced similar problems. Many different immunolabeling protocols are published but, not surprizingly, are not broadcast in technical journals. The best protocols are hidden in scientific papers where the approach was unique and led to an interesting discovery about a biological system. As I said, the methods are unique to each system. There is little point in publishing lists for someone to follow without thinking.
We have to face the fact that whatever method we use to prepare our sample, each will have limitations. Our role is to try to make these limitations work in our favor. Knowing what the limitations are for each preparation protocol makes our job a little easier.
Do you want my replies to Tom Phillips posted here, or should I reply to him off-line? I am sure there are many people already bored by this discussion.
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
At 3:04 PM -0400 9/20/00, Greg Erdos wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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
At 8:26 PM -0500 9/20/00, laniven-at-julian.uwo.ca () wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Leigh GFP stand for green fluorescent protein. It is a naturally fluoescent protein, originall isolated from a jelly fish and nowavailable as a construct that you can insert into your protein of choice using routine molecular biology techniques. It obviates the need for immuno-labelling. In order to see it, you need a UV light source that emits at/around 488nm and the appropriate fluorescence filter sets. The ones designed for FITC work just fine. YOu'd need to add a fair number of components to your 'scope...if you can convert it. Your local microscope sales/service person will be able to gie you the details.
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'm sorry, but I think questions like yours aren't fair. You may get a number of very public gripes from people who may have a good point OR are getting bad results because of their poor skills. Respectable vendors are defenceless against these things because if they rebutted all complaints it could look like they were petty or argumentative or had something to hide. Consider the Golden Rule.
I can remember seeing a respected vendor raked over the coals by a TEM operator who claimed that the TEM in question was unstable. It turns out that he liked bright screens (easier to focus) and recorded all of his pictures of biological tissue specimens with the largest beam size and the beam at crossover.
It's best that you conduct your own comparisons, using your own bread-and-butter specimens. If you are just at the info gathering stage, it would have been MUCH better, in my opinion, to REQUIRE private responses to you instead of having them all go the listserver. You've already required vendors to remain silent!
Ron
Ron Anderson, IBM, Hopewell Jct., New York, USA. anderron-at-us.ibm.com
IBM Analytical Services; http://www.chips.ibm.com/services/asg
"Gary Lovell" {gllovel-at-ppco.com} on 09/20/2000 04:22:52 PM
To: Microscopy-at-sparc5.microscopy.com cc:
I would be interested in heaing from anyone who has done a market survey on JEOL 8200 versus CAMECA SX100 microprobes. Which ( in the responders opinion ) has the better system for geological quantitative WDS analysis, or performs better in the SEM mode. Please give the reasons behind your preference, if at all possible. PLEASE, NO VENDOR RESPONSE.
Gary L. Lovell Phillips Petroleum Company 245a GB Bartlesville, Ok. 74004 Phone: (918)661-9691 Fax : (918)662-2047 Email: gllovel-at-ppco.com
Greetings, Paul Webster asks: Why then should we decide it is important to have good morphology at the EM level?
One answer is that the questions at light and electron levels are distinct. At the light level, we might interested in finding out what tissue expresses a particular antigen, whereas at the electron level, we might be wondering whether it is in the outer or the inner membrane of the mitochondrion. In the latter case, if we can't distinguish outer from inner membrane, let alone a mitochondrion from a lysosome, we are in trouble.
Greetings, Paul Webster asks: Why then should we decide it is important to have good morphology at the EM level? snip snip
One answer is that the questions at light and electron levels are distinct. At the light level, we might interested in finding out what tissue expresses a particular antigen, whereas at the electron level, we might be wondering whether it is in the outer or the inner membrane of the mitochondrion. In the latter case, if we can't distinguish outer from inner membrane, let alone a mitochondrion from a lysosome, we are in trouble.
Years ago while doing lithography using a JEOL 840A, we observed definite* line frequency interference in some of the patterns. The source of the noise was traced to a power bus running in the ceiling to another lab, which could not easily be changed.
JEOL provided a mu-metal shield on a trial basis that was specifically made for the 840. As I recall, their price (~10 years ago) was $6k US. We could quantitatively measure the change in the patterns and found that the interference was reduced by about a factor of 3. Note that this shield surrounded just the lower part of the column, i.e., it extended just above the sample chamber.
With that said, I will now add that I agree with the other responses that suggest the external noise may not be your primary problem. In my experience, I have not seen that line frequency interference causes "fuzzy" focus and astigmatism problems. Since the SEM scan is (or should be) in sync with the line frequency, line frequency interference problems can show up as localized distortions in the image position, depending on the scan frequency.
There are so many possible reasons for fuzzy focus and astigmatism problems (as other responses have pointed out), that it is hard to suggest where you should start. I think you could get better suggestions if you could describe the recent history of the SEM related to its performance. For example, has the SEM always been in its present location and has it always had this problem? If it was moved to the room with the interference, did it work properly before this? What is the best performance that has been seen on your 5410? Have you contacted other 5410 users to check on their experiences? (JEOL should have a user list. If not, I have two customers with 5400's that may be able to offer some guidance.) Do you have an experienced SEM operator who can work with your SEM and diagnose the problems? (I know you have JEOL working on it, but while many SEM technicians - from all SEM vendors - are very good, there are always those who are just learning or just don't really care. Also, as pointed out previously, since your room is out of spec, they have an easy excuse and may not really be trying to solve the problem.)
Joe _________________________________________ Joe Nabity, Ph.D. JC Nabity Lithography Systems E-Beam Lithography using Commercial SEMs & STEMs PO Box 5354, Bozeman, MT 59717 USA Voice: (406) 587-0848 FAX: (406) 586-9514 E-mail: info-at-jcnabity.com Web: www.jcnabity.com
(*When doing lithography, you can often measure the frequency of the interference in the patterns. Not that it really helps you, but if you are interested, example lithography patterns with line frequency noise problems can be seen at "www.jcnabity.com/pictures.htm" under the Diagnostic Images heading.)
To 1 litre with deionised water, pH to 7.8 with hydrochloric acid.
Hope this helps.
Lesley Weston.
On Wed, 20 Sep 2000, Ronald LHerault wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } One of our students needs a receipe for citrated saline and cannot find it } in the very limited references we have available here (a materials lab } where one group is doing some tissue culture). She has seen it mentioned } in articles but it must be so basic (or maybe acidic) that no one tells } how it is made. I hope someone can enlighten us. } } Thanks. } } Ron } lherault-at-bu.edu } } } }
I must have missed the original question, but Birefringence is the difference between the two refractive indices displayed by an anisotropic substance under polarized light observation.
Phil Aviles Forensic Microanalyst
} -----Original Message----- } From: Nelson Conti [SMTP:NelsonC51-at-excite.com] } Sent: Thursday, September 21, 2000 12:50 AM } To: Leigh Ann } Cc: Microscopy-at-sparc5.microscopy.com } Subject: Re: What is GFP? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 Wed, 20 Sep 2000 20:26:39 -0500, laniven-at-julian.uwo.ca 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 } | } -----------------------------------------------------------------------. } } | } | Email: laniven-at-julian.uwo.ca } | Name: Leigh Ann } | School: UWO } | Question: What is GFP? How do you convert a microscope for } birefringence } | and what type of microscopy is this? } | } } } Hi Leigh Ann - } I can answer the first part of your question (to some } extent), but I'm not } very sure about the second part. } GFP = Green Fluorescent Protein ; it's a protein that } naturally } fluoresces, and I believe that it was first discovered as a natural } product } from some marine animal (name ?). What I know of GFP is that it's gaining } usage as a fluorescent marker. I believe it's been used as a marker for } determining the location of X compound in a cell when viewed by } fluorescent } microscopy, but I'm sure that other usages have been cited in other } research } papers. } As for the second part ... someone in this list may correct } me, but I } believe that birefringence is correlated with polarized light. You'll } definitely need to consult with someone else in this list to get more } information. } Good luck getting more answers. } Nelson Conti } } } } } } _______________________________________________________ } Say Bye to Slow Internet! } http://www.home.com/xinbox/signup.html }
Dear Listers, I am seeking input from those of you who use an automated critical point dryer. The chamber on all of those I've considered is much smaller than that of the Polaron E3000 which I have used for many years. What are the advantages of automated / semi-automated critical point dryers? 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, I am looking for a supplier of Mitsubishi thermal dye-sub paper (with ribbon) catalog CK 200sa for printer. I was supplied by Kipp and Sons, apparently now out of business. Preferable that supplier be close to DC Baltimore region. Thank You.
Hi All, I have a picky question that I think would be good for all of us to be clear on, so I'll ask. Can someone define the difference between formaldehyde and formalin? My impression has always been that formalin is to formaldehyde as Kleenex is to tissues (aka a type of brand). I have just come across a protocol that specifically says "formalin (not formaldehyde)". Thanks for your help, Kristen
Kristen A. Lennon Department of Plant Pathology 351 Bessey Hall Iowa State University Ames, IA 50011 email: kalen-at-iastate.edu
Observing birefringence is a very useful means for identifying and locating optically anisotropic materials, i.e. crystalline and polymeric materials. For example, oxalates in tissues would display obvious birefringence, whereas the rest of the tissue would not and would appear black with the polarizer and analyzer inserted (termed "crossed nicols"). See the response below from Olley for setup, but I would recommend the purchase of a polarizing (transmitted light) microscope rather than converting something. The "pretty interference colors" are termed birefringence and should be viewed without the use of the condenser lens. Conoscopic light can be used, however, for such things as the determination of the orientations of the maximum and minimum refractive indices, the magnitude of the anisotropy, and the orientation of the optic axes. These are best determined by observing the interference figure that forms above the objective lens with the nicols crossed and a Bertrand lens. A retardation plate (gypsum, mica, or quartz wedge) is often inserted between the sample and the analyzer. If you don't have a Bertrand lens, simply remove the ocular to view the figure (be sure that the condenser inserted).
All this seems totally unrelated to GFP. You can put a UV source on a polarizing light microscope if necessary.
Donald Miser
} -----Original Message----- } From: Robert H. Olley [SMTP:r.h.olley-at-reading.ac.uk] } Sent: Thursday, September 21, 2000 9:14 AM } To: laniven-at-julian.uwo.ca } Cc: Microscopy-at-sparc5.microscopy.com } Subject: Re: What is GFP? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Leigh Ann, } } Birefringence is observed under a light microscope by putting a polarizer } before the specimen (and generally before the condenser) and a similar } device called the analyser between the specimen and the eyepiece: the } analyser is usually set at 90^ to the polarizer. If the crystal (or even } a piece of stretched plastic or a blob of liquid crystal) has different } refractive indices in different directions, the polarized light passing } through the specimen gets divided into two out-of-phase components which } are recombined in the analyzer to give pretty interference colours, from } which one can also make quantitative deductions. The people that use this } technique most are geologists and mineralogists looking at thin polished } rock sections, but we plastics people also look at polymers in this way. } I'm not sure if it's used much in bio/med applications. } } A good book to explain the physics and applications of the technique is: } } Wood, Elizabeth Armstrong } Crystals and light. } } while there are many books which concentrate more on the setup of the } microscope. Here's a selection out of our library catalogue: } } Hartshorne, N.H. } Crystals and the polarising microscope. } } Polarized light microscopy by Walter C. McCrone, Lucy B.McCrone } and John Gustav Delly } } The polarizing microscope by A. F. Hallimond } Hallimond A.F. } } } +------------------------------------------------------------------------+ } | 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 } | } } +------------------------------------------------------------------------+ } } } Email: laniven-at-julian.uwo.ca } } Name: Leigh Ann } } School: UWO } } Question: What is GFP? How do you convert a microscope for birefringence } } and what type of microscopy is this?
Hello everyone on the list who is doing Immuno-EM! Dear Greg, Paul, Tom and Tim
I have read your contributions with great interest and smiled everytime. Ok, since my bible in Immunoelectronmicroscopy is the outstanding book of G.Griffiths ("Fine structure immunocytochemistry", 1993), and as I am trying to do good IEM for over three years, I do agree with all of you.
First, let me state that it is particulary difficult to enable monoclonal antibodies for EM immunoprocedures. In fact, I was not able to run a good EM-Immuno with a monoclonal one (but the polyclonals do work.). And I agree with you, most of the (monoclonal) antibodies provided today are very well suited for Western blotting and sometimes even for LM because they were fitted for degenerative conditions applied in these techniques. I am always aware of that approximately 90% of the epitopes are gone, due to embedding and sectioning.
Second, many collegues who do have quite satisfactorily results at LM ask me why, the signal at EM is not worth looking at, to poor. Many of them are not aware of the dimensions we are talking about, a few milimetres in square and a section thickness of 50-70 nms is little, and fixation and post-processing leads to a loss of antigenicity.
Third, if you do have an antibody running well in IEM, use it. There are, in fact too few AB suitable for IEM. I am wondering why there isn´t any company already which is offering AB´s for IEM which recognize their epitope after aldehyde degeneration/fixation and acrylic plastic embedding.
At least, counting your signal should help. Sometimes you don´t recognize specific signals because you only see almost nothing at a magnification of x20,000 or more. If you start counting more fields, you are often surprised by the high specifity, althoug it´s very weak.
So, my two cents to this topic are: Try different antibodies from different Labs made against the same epitope! If it works, you should always use it as reference marker. And don´t forget: IEM does take so much more time and spirit as LM, this business is more tricky.
The best wishes for the IEM´s on this list.
Michael Reiner
PS: In memoriam Hildy Crowley, I already do miss her reply to this topic. _______________________________________________________________________ 1.000.000 DM gewinnen - kostenlos tippen - http://millionenklick.web.de IhrName-at-web.de, 8MB Speicher, Verschluesselung - http://freemail.web.de
Formaldehyde is a gas which may be dissolved in water.
A 40% solution of formaldehyde dissolved in water is called Formalin. Normally, various additives are put into the 40% solution of formaldehyde in order to stabilize it (I believe that traces of methanol may be added). Because of impurities, Formalin is usually considered unsuitable for electron microscopy studies. Confusion often arises when people refer to percentages of these solutions. For example, a 10% solution of formaldehyde is quite different than a 10% solution of Formalin ((10% versus 4%, respectively).
Hope this answers your question.
} I have a picky question that I think would be good for all of us to } be clear on, } so I'll ask. Can someone define the difference between formaldehyde and } formalin? My impression has always been that formalin is to formaldehyde as } Kleenex is to tissues (aka a type of brand). I have just come across } a protocol } that specifically says "formalin (not formaldehyde)".
#################################################################### 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 ####################################################################
} Hi All, } I have a picky question that I think would be good for all of us to be clear on, } so I'll ask. Can someone define the difference between formaldehyde and } formalin? My impression has always been that formalin is to formaldehyde as } Kleenex is to tissues (aka a type of brand). I have just come across a protocol } that specifically says "formalin (not formaldehyde)". } Thanks for your help, } Kristen } } Kristen A. Lennon } Department of Plant Pathology } 351 Bessey Hall } Iowa State University } Ames, IA 50011 } email: } kalen-at-iastate.edu
Hi Kristen:
Formaldehyde is a gas, when bubbled through water the solution becomes saturated to about 37-40%. Methanol is usually added as a stabilizer. Formalin is the name often given to a 1:9 dilution of the "formaldehyde" solution, it is about 3.7-4% formaldehyde. The nomenclature is not exact, to put it mildly. To know exactly the person who wrote the protocol means, you will probably have to ask him/her. I don't know how you could have formalin without formaldehyde!
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 concur with everything John said and point out that one also commonly sees 3.7% formalin in protocols. This is the "same" as 4% usually since it depends on whether you mean % w/w or % w/v (i can't remember which is which).
Also, besides the impurities, formalin is often bought in gallon or barrel sizes so it can sit around a long time and form polymers not present in freshly depolymerized paraformaldehyde.
So the words should never be taken as synonomous.
Tom
} } } Formaldehyde is a gas which may be dissolved in water. } } A 40% solution of formaldehyde dissolved in water is called } Formalin. Normally, various additives are put into the 40% solution } of formaldehyde in order to stabilize it (I believe that traces of } methanol may be added). Because of impurities, Formalin is usually } considered unsuitable for electron microscopy studies. Confusion } often arises when people refer to percentages of these solutions. } For example, a 10% solution of formaldehyde is quite different than } a 10% solution of Formalin ((10% versus 4%, respectively). } } Hope this answers your question. } } } I have a picky question that I think would be good for all of us to } } be clear on, } } so I'll ask. Can someone define the difference between formaldehyde and } } formalin? My impression has always been that formalin is to formaldehyde as } } Kleenex is to tissues (aka a type of brand). I have just come } } across a protocol } } that specifically says "formalin (not formaldehyde)". } } #################################################################### } 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 } ####################################################################
-- 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)
Formaldehyde is a gas. An aqueous solution saturated with the gas is called formalin. This saturated solution is called 100% formalin, which contains approximately 37% formaldehyde. Thus, a solution of 10% formalin is very different in strength from a solution that is 10% formaldehyde. Thus, an instruction may say to use "10% formalin (not formaldehyde)" to make sure that you indeed are using the correct strength.
DL
Kristen A. Lennon 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 have a picky question that I think would be good for all of us to be clear on, } so I'll ask. Can someone define the difference between formaldehyde and } formalin? My impression has always been that formalin is to formaldehyde as } Kleenex is to tissues (aka a type of brand). I have just come across a protocol } that specifically says "formalin (not formaldehyde)". } Thanks for your help, } Kristen } } Kristen A. Lennon } Department of Plant Pathology } 351 Bessey Hall } Iowa State University } Ames, IA 50011 } email: } kalen-at-iastate.edu
-- ____________________________________________________________________ Donald L. Lovett voice: 609-771-2876 Assoc. Professor fax: 609-637-5118 Dept. of Biology e-mail: lovett -at- tcnj.edu The College of New Jersey P.O. Box 7718 www.tcnj.edu/~scholars/lovett.html Ewing, NJ 08628-0718
Kristen: Formalin is commonly used to describe a mixture of37% to 38% formaldehyde in water (this reagent grade chemical is also called formaldehyde solution). This solution of dissolved formaldehyde gas generally contains some small amount of methanol in it also. Now for the good part! Traditionally this reagent grade mixture of the 37% or so formaldehyde in water is considered a 100%formalin concentration (the maximum concentration range you can buy commercially). A traditional 10% formalin solution would then contain about 3.7% to 3.8% aldehyde concentration. Many E-M folks prefer to make their formaldehyde solutions from the polymer paraformaldehyde--in part I think believing that there is less methanol in the final product yet folks have had success using both kinds in the right application. I hope this helps to clarify the common usage of these terms.
Dear Listers, For preembedment EM-ISH, is a two-step detection method more sensitive than a single-step method? Specifically, I am planning to detect digoxigenin-labeled RNA probes with mouse anti-dig antibody, followed by anti-mouse gold-conjugated secondary antibody (then enhancing ultrasmall gold particles). Or would direct detection of the dig probes with an anti-dig gold-conjugated antibody lend any advantage? One review attributes higher sensitivity for two-steps, but I think this might be restricted to postembedment methods only (J. Histochem. Cytochem. 45:481-92,1997).
If a two-step method is used, would gold-conjugated antibodies (against biotin probes) have advantages over streptavidin-gold? As recently discussed on this list, unconjugated streptavidin will compete with the gold-labeled streptavidin, thus reducing signal strength. Nanoprobes makes covalently bound gold antibodies, but I have seen a higher background with these than with noncovalently bound antibodies, at least in cultured rat neurons.
Finally, will glutaraldehyde levels greater than 0.05% tend to inhibit hybridization? Rat glioma monolayers have been labeled with EM-ISH by Punnonen, et al., using 4% paraformaldehyde and 0.05% glut for 30 min. (J. Histochem. Cytochem. 47:99-112, 1999). However, this protocol included 0.1% saponin permeabilization for 30 min. Their probes were up to 850 bp and "partially hydrolyzed" while my probes are only 50 bp long. I have immunolabeled neurons fixed in 2% glut with no permeabilization; so if the antibodies can make it into the cells, then the smaller RNA probes should too, right?
I know that target abundance is an issue here. I plan to include positive controls for abundant RNAs (e.g., beta-actin), and I will consider Tyrimide Signal Amplification, if necessary.
Thank you for your helpful comments on any of these questions. I will be happy to elaborate if this inquiry is too brief.
Michael Plociniak Research Technician Albert Einstein College of Medicine Neuroscience Dept. Rose Kennedy Center, 529 Bronx, NY 10461 (718) 430-3509 plocinia-at-aecom.yu.edu
Win95 has a utility named system monitor (Start | Programs | Accessories | System Tools |). I don't believe it's installed by default. You may have to add it through the custom install section. It could help you rule out the physical disk, CPU, and RAM as bottlenecks. You might compare the charts using your different printers. It's a "watch it in real time" affair though you might be able to use {Alt-Print Screen} to get a snapshot for comparison or baselining (it does work, I tried it : ) ).
It contains counters to monitor your CPU, physical disk, virtual memory, and network connections. A knowledgeable person in your computer department should be able to help you sort which counters might be useful and what they mean. Or you could play with different counters. Unfortunately the help portion of the utility is the stuff of legends.
I'd probably start with Kernel Processor Usage %, File System Bytes Written/Second & Dirty Data (This sounds like the amount of queued data), and Memory Manager Disk Cache Size & Page Faults.
Hope it helps,
Chuck ------------------------------------- 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
Please keep the discussion coming! I have found it extremely informative and helpful, as I am attempting pre-embedding anti-GFP immunolabelling with nanogold and silver enhancement. ImmunoEm is so important now that many people have localized their GFP transfected proteins by confocal. They want to know what are the fluorescent blobs they cannot quite resolve. Before I attempt any immunoEm with anyone, they must do a fixation series of formaldehyde and glutaraldehyde dilutions to see where they lose their fluorescent signal. We attempt to get the best fixation possible, yet penetration of the ab is an issue too. It is always a compromise. There are many ways to skin a cat, and only time and effort will hopefully give you an answer. JoAnn Buchanan Stanford University School of Medicine Stanford, CA
} I have a picky question that I think would be good for all of us to be clear on, } so I'll ask. Can someone define the difference between formaldehyde and } formalin? My impression has always been that formalin is to formaldehyde as } Kleenex is to tissues (aka a type of brand). I have just come across a protocol } that specifically says "formalin (not formaldehyde)". } Thanks for your help, } Kristen
As I have understood it, formalin is the (fairly concentrated) aqueous solution, formaldehyde is the actual pure compound HCHO.
Not that I've ever used, the stuff, though.
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
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Reply to: Re: Immuno TEM Tobias Baskin writes: One answer is that the questions at light and electron levels are distinct. At the light level, we might interested in finding out what tissue expresses a particular antigen, whereas at the electron level, we might be wondering whether it is in the outer or the inner membrane of the mitochondrion. In the latter case, if we can't distinguish outer from inner membrane, let alone a mitochondrion from a lysosome, we are in trouble.
Hi Tobias,
I agree totally. The level of acceptance for poor morphology will depend on the result expected. Again, each system will have its own protocol for getting that particular result. I am not encouraging everyone to go out and prepare samples with such poor morphology that any result they get will be meaningless. This would be irresponsible for me to advocate and a waste of time for anyone who was foolish enough to follow this advice.
Rather, I suggest that if it is possible to label purified (or semi-purified) sub-cellular particles, then do that. If you are looking for antigens on the outside of mitochondria, perform a pre-embedding labeling on purified fractions. As the mitochondria are so recognizable, it is not even necessary to get very pure fractions. If the antigen is inside the mitochondria, then sectioning is probably the best way. But even then, if the profile that is labeled is so obviously a mitichondion, then it doesn't matter if the cristae are not perfectly preserved (unless there is extraction or redistribution of antigen). If specific membrane markers are available, structural morpholgy is even less important. Co-localizing the marker with the test antibody on a membrane or organelle fragment is often very acceptable. Sometimes semi-purified fractions can be obtained just by breaking open cells to let out the antigens we are not interested in (unless the antigens are cytoplasmic of course). Gently lysed cells have wonderful contrast and it is possible to recognize a great number of the intracellular organelles if they have been fixed correctly. The added accessibility obtained by removing the cytoplasm makes it possible to reach antigens that are usually covered. I know an investigator who labeled the lumen of the RER with a very specific antibody. He couldn't do this on well fixed, heathly cells as the antibody could not gain access to the luminal antigens. He solved his problem by noticing that some of the cells in the pellet had obviously died before being fixed, and had really poor morphology. However, the RER was easily recognizable and had lots of gold particles in the swollen lumen. Is this unacceptable because the cells were not perfectly preserved? The journal reveiwers didn't think so.
I think that immunocytochemistry is a very special branch of EM that is almost impossible to provide as a service. I know that this is what is being asked of EM labs all over the world and I know it will be impossible to change this. However, it is our responsibility to educate our colleagues and to allow them to become involved in the discovery process of how their antibodies work. I have no good solution to this, but do know that if someone is involved in their own specimen preparation and data collection, their work will progress much faster. If new EM users are taught why particular approaches should be applied instead of being given one protocol to apply exactly as written, pretty neat ideas originate from their work.
We must stop being the "black box" of science. Regards,
Paul Webster.
Reccommended reading: G. Griffiths 1993 Fine Structure Immunocytochemistry. Springer Verlag, Heidelberg & Berlin {http://www.amazon.com/exec/obidos/ASIN/038754805X/qid%3D969583271/102-2138449-1518557}
Discalimer: I have no financial interest in Amazon or Springer. I have co-authored papers with Griffiths and we teach immunocytochemical methods together.
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
Tobias Baskin 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, } Paul Webster asks: Why then should we decide it is } important to have good morphology } at the EM level? snip snip } } One answer is that the questions at light and electron levels } are distinct. At the light level, we might interested in finding out } what tissue expresses a particular antigen, whereas at the electron } level, we might be wondering whether it is in the outer or the inner } membrane of the mitochondrion. In the latter case, if we can't } distinguish outer from inner membrane, let alone a mitochondrion from } a lysosome, we are in trouble. } } Tobias } -- } _ ____ __ ____ Tobias I. Baskin } / \ / / \ / \ \ University of Missouri } / | / / \ \ \ Biological Sciences } /_ / __ /__ \ \ \__ 109 Tucker Hall } / / / \ \ \ } Columbia, MO 65211-7400 USA } / / / \ \ \ voice: } 573-882-0173 } / /____ / \ \__/ \____ fax: 573-882-0123 } } } } RFC822 header } ----------------------------------- } } Received: from ultra5.microscopy.com [206.69.208.10] by mailhouse.hei.org } with ESMTP } (SMTPD32-6.00) id A14011010214; Thu, 21 Sep 2000 17:01:04 -0700 } Received: (from daemon-at-localhost) } by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id LAA02943 } for dist-Microscopy; Thu, 21 Sep 2000 11:06:33 -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 LAA02940 } for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Thu, 21 Sep 2000 } 11:06:03 -0500 (CDT) } Received: from umc-mail01.missouri.edu (umc-mail01.missouri.edu } [128.206.10.216]) } by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id LAA02932 } for {Microscopy-at-sparc5.microscopy.com} ; Thu, 21 Sep 2000 11:05:52 -0500 (CDT) } Received: from [128.206.81.167] (houndbyte.biosci.missouri.edu } [128.206.81.167]) by umc-mail01.missouri.edu with SMTP (Microsoft Exchange Internet Mail } Service Version 5.5.2650.21) } id RL53XF67; Thu, 21 Sep 2000 10:59:59 -0500 } Mime-Version: 1.0 } X-Sender: baskint-at-pop.email.missouri.edu } Message-Id: {p04320412b5efe08d6ca7-at-[128.206.81.167]} } Date: Thu, 21 Sep 2000 11:00:03 -0500 } To: Microscopy-at-sparc5.microscopy.com } From: Tobias Baskin {BaskinT-at-missouri.edu} } Subject: Re: Immuno TEM } Content-Type: text/plain; charset="us-ascii" ; format="flowed" } Errors-to: Microscopy-request-at-sparc5.microscopy.com } X-RCPT-TO: {pwebster-at-hei.org} } X-UIDL: 257555211 } Status: U }
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Another thing that can be critical is service from the local reps in your area. This varies enormously around the globe and can also vary between different regions of a country. Ask others in your area what the service has been like over the last few years. If you've got a complex piece of machinery you'll appreciate good service, especially as it ages.
Rosemary
Rosemary White Microscopy Centre CSIRO Plant Industry GPO Box 1600 Canberra, ACT 2601 Australia
Dear all, we have received a SEM Topcon SM510 and we would like to upgrade this system with LaB6 filament. It is clear that the system is almost ready for this upgrade but we have a lot of problems to obtain information from Topcon in Europe (our SEM is in Italy). We need help to know the ion pump necessary to upgrade our system, how to install it and the procedures to convert the SEM for LaB6 filament use. We need also one complete copy of Topcon SM510 manual in order to clearly understand how to correctly work with LaB6 filament.
Hi All, Thanks to all of you who responded to my query. It was a question that had been sitting in the back of my mind for several years (since the time a mentor of mine told me that formalin was a "safer" alternative to formaldehyde as my fellow students and I proceeded to be pretty much covered with it while doing some mass field fixes). When that protocol hit, it was a good excuse to settle it once and for all! Thanks, Kristen
Kristen A. Lennon Department of Plant Pathology 351 Bessey Hall Iowa State University Ames, IA 50011 email: kalen-at-iastate.edu
I am writing in response to Paul Webster's recent description of gold labeling the lumen of RER by way of lyseing cells and isolating the resulting membranes and vesicles in a fraction. The basic question I have is, at what point was the fraction exposed to the primary antibody and the secondary antibodie that would have been conjugated to colloidal gold? Did these steps occur before any fixation? Was the fraction fixed, dehydrated, embedded, thin sectioned and then exposed to the primary and secondary antibodies? Or was a entirely different procedure used.
For the past 9 years I have been working with Plasmodium falciparum during its 48 hour asexual life cycle in human erythrocytes. I have been able to isolate various membranous structures which we believe contain proteins that are of interest to us. We have had only marginal success in labeling said structures in lightly fixed, L. R. White embedded sections. I might be able to adept your mentioned RER fractionation technique to meet my needs. Thanks in advance for any help, Tim
Timothy G. Schneider Director of Electron Microscopy Department of Pathology Room 229 Jefferson Hall Thomas Jefferson University 1020 Locust St. Philadelphia Pa. 19107 215-503-4798 work 610-613-8170 cellular timothy.schneider-at-mail.tju.edu
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Reply to: RE: TEM in-situ hybridization
Michael Plociniak wrote: } } Dear Listers, } For preembedment EM-ISH, is a two-step detection method more sensitive than } a single-step method? Specifically, I am planning to detect } digoxigenin-labeled RNA probes with mouse anti-dig antibody, followed by } anti-mouse gold-conjugated secondary antibody (then enhancing ultrasmall } gold particles). Or would direct detection of the dig probes with an } anti-dig gold-conjugated antibody lend any advantage? One review } attributes higher sensitivity for two-steps, but I think this might be } restricted to postembedment methods only (J. Histochem. Cytochem. } 45:481-92,1997).
Hi Michael,
As I have been getting lots of off-line e-mail encouraging me to continue writing, I will attempt to offer my thoughts on your question for all to see. As you know, in situ hybridization is just another form of affinity localization labeling protocol. In this instance the probe is a nucleic acid sequence instead of an antibody. Much the same rules apply. I certainly don't know all the answers but can share some of what I have learned the hard way. I have embedded my comments in your text. :
Two step, sequential detection methods (primary label, followed by secondary with attached visualization probe) are usually more sensitive. This was first observed by Coons in the 1950's, and he is generally considered to be the father of immunoctyochemistry. This sequential labeling can be applied to any labeling system you want to use. You must be careful to differentiate between signal amplification and sensitivity. What you (and all of use) are looking for is to maximize the amount of target that reacts with affinity marker (increasing sensitivity). Signal amplification refers to the process of making the reacted agents more visible. } } If a two-step method is used, would gold-conjugated antibodies (against } biotin probes) have advantages over streptavidin-gold? As recently } discussed on this list, unconjugated streptavidin will compete with the } gold-labeled streptavidin, thus reducing signal strength. Nanoprobes makes } covalently bound gold antibodies, but I have seen a higher background with } these than with noncovalently bound antibodies, at least in cultured rat } neurons.
If you are using antibodies to digoxigenin, then a secondary antibody-gold probe should be sufficient. In your system this would be mouse anti-digoxigenin followed by anti-mouse-gold. Remeber that if you choose to use nanoprobes, it might be difficult to later perform multiple labeling experiments. If you want to look for biotin labeled nucleic acid probes try using anti-biotin antibodies and cold water fish skin gelatin as a blocking agent - not serum. } } Finally, will glutaraldehyde levels greater than 0.05% tend to inhibit } hybridization? Rat glioma monolayers have been labeled with EM-ISH by } Punnonen, et al., using 4% paraformaldehyde and 0.05% glut for 30 min. (J. } Histochem. Cytochem. 47:99-112, 1999). However, this protocol included } 0.1% saponin permeabilization for 30 min. Their probes were up to 850 bp } and "partially hydrolyzed" while my probes are only 50 bp long. I have } immunolabeled neurons fixed in 2% glut with no permeabilization; so if the } antibodies can make it into the cells, then the smaller RNA probes should } too, right? } The only answer to this question is what you will tell us when you have performed the experiment. Each system has its own unique qualitites and sensitivity to fixatives will be one of these variables. If it bothers you to include glutaraldehyde then leave it out. Fix with a stong glutaraldehyde solution after you have performed your hybridization and silver enhancing steps.
Watch out for the mistake that many people make in assuming that immunoreagents are inert objects and that their size is the only property to be taken into account. As someone who mailed me said: immunoreagents are biological molecles and not chemicals. They are charged molecules and have many complex, variable interactive and binding properties that we cannot begin to know. Try out your protocol and see if it works. If is does then the question about your RNA probe is answered.
} I know that target abundance is an issue here. I plan to include positive } controls for abundant RNAs (e.g., beta-actin), and I will consider Tyrimide } Signal Amplification, if necessary.
One possible step you could put in is to first try out your whole preparation and immunolabeling protocol at the LM level. Use the same reagents and same preparation protocols as you plan to use for EM and expose to silver enhancment for a longer time so that you can see the signal by LM. If you see a signal then you will be confident that you will see a signal when you go to examine the EM experiment. You might even try to find a way of even preparing the LM sample that incorporates the resin embedding so that you can control for possible digestion of the silver signal by osmium tetroxide.
Regards,
Paul Webster
} } Thank you for your helpful comments on any of these questions. I will be } happy to elaborate if this inquiry is too brief. } } } Michael Plociniak } Research Technician } Albert Einstein College of Medicine } Neuroscience Dept. } Rose Kennedy Center, 529 } Bronx, NY 10461 } (718) 430-3509 } plocinia-at-aecom.yu.edu } } transmission and scanning electron microscopy } 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 } } } } RFC822 header } ----------------------------------- } } Received: from ultra5.microscopy.com [206.69.208.10] by mailhouse.hei.org } with ESMTP } (SMTPD32-6.00) id AA6F3AA023A; Thu, 21 Sep 2000 23:21:35 -0700 } Received: (from daemon-at-localhost) } by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id SAA04892 } for dist-Microscopy; Thu, 21 Sep 2000 18:10:29 -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 SAA04889 } for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Thu, 21 Sep 2000 } 18:09:59 -0500 (CDT) } Received: from post.aecom.yu.edu (post.aecom.yu.edu [129.98.1.4]) } by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id SAA04882 } for {Microscopy-at-sparc5.microscopy.com} ; Thu, 21 Sep 2000 18:09:47 -0500 (CDT) } Received: from amca (amca.aecom.yu.edu [129.98.90.216]) } by post.aecom.yu.edu (8.9.3/8.9.3) with SMTP id TAA05342 } for {Microscopy-at-MSA.Microscopy.com} ; Thu, 21 Sep 2000 19:03:54 -0400 (EDT) } Message-Id: {3.0.6.32.20000921185842.0079b8e0-at-mailserver.aecom.yu.edu} } X-Sender: plocinia-at-mailserver.aecom.yu.edu (Unverified) } X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.6 (32) } Date: Thu, 21 Sep 2000 18:58:42 -0400 } To: Microscopy-at-sparc5.microscopy.com } From: Michael Plociniak {plocinia-at-aecom.yu.edu} } Subject: TEM in-situ hybridization } 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: 257555229 } Status: U }
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Daniela: I have a TOPCON 500/510 wet SEM and I have the manual although I don't know how to get a copy to you. I use the tungsten filament, don't have theLaB6 filament. You might ask the people I have repair our SEM, they all used to work for TOPCON and they have parts for them.
Chuck Humphreys Image Control Inc. P.O. Box 720596 Orlando, FL 32876-0596 U.S.A. email : cchumph-at-ibm.net
Terry Ellis Hallmark Cards Inc. 2501 McGee Kansas City, MO 64141 U.S.A. email: tellis2-at-hallmark.com
THE NORTH CAROLINA SOCIETY FOR MICROSCOPY AND MICROBEAM ANALYSIS
present the
NINETEENTH ANNUAL SYMPOSIUM ON ADVANCES IN MICROSCOPY
"Molecules, Cells & Microchips"
Coastline Convention Center, Wilmington, North Carolina
October 13 - 15, 2000
FINAL ANNOUNCEMENT!
The Nineteenth Annual Symposium, sponsored by the North Carolina Society for Microscopy and Microbeam Analysis (NCSMMA), has been planned with a theme of "Molecules, Cells & Microchips" Continuing with the tradition of the symposium, the guest lecturers are composed of both nationally and internationally distinguished scientists.
Speakers who have agreed to participate (so far) this year include:
* Ian Anderson (ORNL, Oak Ridge, TN)Spectrum Imaging: Microanalysis for a New Millennium * Ken Downing (University of California, Berkeley, CA) High Resolution Protein Structure * Jan Ho (Johns Hopkins University, Baltimore, MD) Scanning Probe Microscopy in Biology * Stefan Jeglinski (4pi Analysis Inc.) Open Source software as a Business Model for a Microscopy Company? * Dan Kiehart (Duke University Medical Center, Durham NC) Flies 'R Us * Richard Palmer (Duke Univ. NC) Recent Developments in Infrared Imaging and Microscopy * Peter So (MIT Cambridge, MA) New Microscopy Instrumentation for Biomedical Research * Rich Superfine (UNC, Chapel Hill, NC)Remote Nanomanipulation: from Viruses to Nanotubes
* The meeting has several purposes, not the least of which is to draw attention of the scientific community to emerging developments in the practical and basic research aspects of exciting new fields, and to bring people together from diverse disciplines to discuss how innovative techniques will be relevant to the future direction of microscopy and microprobe analysis. In particular, this year, special emphasis will be placed on how recent advances in electron, optical and probe microscopies have resulted in new knowledge that has benefited microscopy in general and are having a significant impact in the biological and physical sciences. The symposium also offers an opportunity for interested participants including students to submit abstracts of related studies for poster display.
* 3 special WORKSHOPS/TUTORIALS will be offered at NO ADDITIONAL CHARGE to participants in the Symposium: (a) Immuno/Microwave EM Techniques, (Cindy Hastings, VA Little Rock AR) (b) Atomic Force Microscopy (Jan Ho) (c) Digital Imaging Methods (John Mackenzie, North Carolina State Univ). These are practical, introductory sessions and no previous experience or knowledge is necessary.
Registration Fees, Hotel rates
The $90 ($100 on site) per person and $50 for students ($60 on site). Registration fee includes: symposium attendance and materials, Saturday lunch, breaks, and Friday and Saturday evening meals. Additional Friday evening tickets are available for Adults - $20; Children 10 years of age and under - $10. Additional Saturday evening tickets are available for Adults - $20; Children 10 years of age and under - $10. There is a $15 fee for all cancellations.
COAST LINE INN special rates $80/room/night (single or double). Tel: 1-800 617-7732
For questions or further information on Registration, please telephone Betty Gooch, Duke University Medical Center: (919) 286-0411 x 6508 or email: b.gooch-at-cellbio.duke edu
Website: http://152.3.167.174/NCAnnSymp2000.html
Or call Peter Ingram/Ann LeFurgey (919) 660-2671 email: p.ingram-at-cellbio.duke.edu
Today's question is, how will PMMA react in the PIPS? I have a thin film of Ti/Cu/Ti grown on a substrate of (1 1/2um thick) PMMA . My plan is to do a regular film to film x-section, dimple down to about 10um and finish it in the PIPS...I'm just not sure how PMMA mills?
If anyone has done this sort of sampIe I would love to hear about it. As always any suggestions are gladly received.
Looking for a TEM sample Prep technician at SEMATECH in Austin TX
SEMATECH is a research and development consortium working on advanced materials and processes for semiconductor manufacturing.
JOB RESPONSIBILITIES: Prepare samples, both cross sectional and plan view, of semiconductor material and devices for TEM analysis. Using flat-polishing, wedge-polishing, mechanical dimpling, ion milling, Focused Ion Beam milling and additional techniques as needed. Maintain lab equipment in working order, maintain detailed records, lab safety, housekeeping, order supplier and perform additional tasks as needed. QUALIFICATIONS: Two year technical degree, Superior fine motor skills and near vision, Ability to work with limited supervision and direction, Ability to multitask effectively, Strong organizational skills Ability to work with limited supervision and direction, Experience with sample polishing and the semiconductor industry is preferred Experience with FIB and TEM prep greatly preferred
Interested parties should send resumes to:
Carolyn Gondran 2706 Montopolis Dr. Austin TX 78741
} we have received a SEM Topcon SM510 and we would like to } upgrade this system with LaB6 filament. } It is clear that the system is almost ready for this } upgrade but we have a lot of problems to obtain } information from Topcon in Europe (our SEM is in Italy). } We need help to know the ion pump necessary to upgrade our } system, ...
I believe an ion pump is definitely necessary ... LaB6 emitters are extremely susceptable and unstable with respect to contamination. For adding an ion pump, you will need (1) the ion pump itself and its power supply ... (2) and you'll need add a port near the gun for the ion pump, and an isolation valve for isolating the gun from the rest of the vacuum system while the ion pump is working. For LaB6 emitters, (1) you'll likely need modify your power supply for the voltage/current requirements for heating the emitter, and (2) you'll most likely need a different Wehnelt. (...hmmmmm? ... have I forgot anything? ....)
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 am interested in finding out more about the Oklahoma City, OK Ugly Bug contest which is sponsored by the Microscopy Society there. Thanks. Barbara Hodges bbddhodges-at-aol.com
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With regard to my question on the model of dye sub printer for the EM photos, eighteen people have sent me replys. Most of them suggested to use Epson printer, 870, 900 or 1270 not only due to the price, but also due to the image quality. Dr. John Mackenzie also kindly recommended to use both of Epson 900 for the resolution and 870 for the archiving because of fading problem of 900. But some of them wrote that dye sub is better than ink jet printer. They were using Codonics, Fuji Pictography, Tektronics, or Kodak 8670 dye sub printers.
I am thinking to buy Epson 900 and small Kodak dye sub printer 4270. 4270 is not so expensive, and prints only 4 inch wide photos, good for EM photo only. Many of its users are for passport size personal photographs. I will see how good is resolution of 4270.
Epson 900 sprays 3 picoliters of color dots, smallest one that I've ever known, but 10 picoliters of grey-scale dots, somewhat large one. Epson 870 sprays 4 picoliters of color dots, but unknown for grey scale. I do not know how much difference 3 and 4 picoliters dots would make. Epson 870 is photo version and has various capability of photography. Epson 900 is for office color printing. If the difference is insignificant, 870 would be much better. Any comments?
Thank all of you who have kindly posted suggestions and comments.
Best regards,
Jondo Yun Kyungnam University Division of Advanced Materials Electron Microscopy Laboratory 449 Weolyeong-dong Masan, 631-701, Korea
Gary M. Easton 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 } -----------------------------------------------------------------------. } } Listers, Anyone out there know of a good(hopefully easy!) way to } remove carbon tracking deposits on a ceramic electon gun? Thanks in } advance. Gary M. Easton Scanners Corporation Gary, The concentrated HCl is great but the metal parts become a real problem. Try Bon Ami if the surface is glazed. It shouldn't scatch. If it's not glazed then be more aggressive with Comet or Ajax.
Bon Ami also works wonders on a yellowed specimen chamber if you can work with just the chamber in a sink. It beats acetone or alcohol hands-down.
} } I am interested in finding out more about the Oklahoma City, OK Ugly Bug } contest which is sponsored by the Microscopy Society there. Thanks. Barbara } Hodges bbddhodges-at-aol.com
Barbara -
The Oklahoma Microscopy Society website is http://www.ou.edu/research/electron/oms/, and the popular Ugly Bug contest has its own (very entertaining) site: http://www.ou.edu/research/electron/oms/uglybug/ Paige Johnson {paige.l.johnson-at-iolok.com} chairs the OSM outreach program.
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
Postdoctoral Positions in Materials Science University of Kentucky
Two post-doctoral researcher positions are available at the University of Kentucky, as detailed below. The research programs will make extensive use of the university's Electron Microscopy Facility, which houses two SEMs, two TEMs and one AFM. Most of the research will be conducted on a JEOL 2010F Field Emission TEM outfitted with an Oxford energy dispersive x-ray spectrometer (EDS), Gatan electron energy loss imaging filter and acquisition hardware and software for spectrum imaging.
1. The first position will be in the area of carbon-based materials under the auspices of the NSF-sponsored Center for Advanced Carbon Materials. The structure and chemistry of chemically-modified carbon nanotubes and metal-doped diamond-like carbon films will be investigated and correlated with physical property measurements. Most experimentation will involve the use of high-resolution electron microscopy (HREM) and electron energy loss spectroscopy (EELS).
2. The second position will be in the area of nanocrystalline oxide materials for sensor applications. In collaboration with the Chemistry Department at the University of Kentucky, we are studying the crystallization kinetics of metal-oxide precursors which have been designed at the molecular level to facilitate low-temperature synthesis routes. Phase and particle analysis of various oxide materials will be assessed quantitatively by TEM and XRD. Dopant segregation and the effects on dielectric behavior will also be studied.
Qualified candidates will have Ph.D. in Materials Science, Physics or any related field and practical experience in analytical electron microscopy. The salary will be commensurate with qualifications and experience. Please send applications, including two references to:
Professor Elizabeth Dickey Department of Chemical and Materials Engineering University of Kentucky A254 ASTeCC Bldg. Lexington, KY 40506-0286 ecdickey-at-engr.uky.edu
The University of Kentucky is an equal opportunity employer.
Could you tell me if agarose is PAS positive? I stained a dinoflagellate using agarose embedding technique and it was negative, however, others in my field tell me it should have stained positive. Can you clarify?
Sue Tyler, Biologist MD Dept. of Natural Resources 904 S. Morris Street Oxford, MD 21654 410-226-5193
Does anybody knows an efficient and quick way to get spare parts for Edwards Auto 306 coater?. It seems that EPROM in the control unit is dead and the instruments is out of control (after switching on the display diodes light randomly). An official Edwards' representative in Poland told us that it would take months for them to get the needed parts from the Edwards headquarters.
Leszek Kepinski
Institute of Low Temperature and Structure Research, Polish Academy of Sciences, P.O.Box 1410, 50-950 Wroclaw, Poland
Does anybody know an efficient and quick way to get spare parts for Edwards Auto 306 coater?. It seems that EPROM in the control unit is dead and the instruments is out of control (after switching on the display diodes light randomly). An official Edwards' representative in Poland told us that it would take months for them to get the needed parts from the Edwards headquarters.
Leszek Kepinski
Institute of Low Temperature and Structure Research, Polish Academy of Sciences, P.O.Box 1410, 50-950 Wroclaw, Poland
Greetings, I have PAS stained samples embedded in agarose and then in plastic and there was never an agarose reaction.
However, this info is simply anecdotal. I have no idea is PAS is "supposed" to stain agarose, and I don't do PAS "properly" (just perioidic acid and then Schiff's). Did you have a positive control? Sometmes the Schiff's reagent goes bad. I recall placing a drop on a piece of white tissue paper and if it went pink in a moment or two then the reagent was still ok.
Hope this helps even a little, Tobias
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On Wednesday, September 20th, there was an accident at the New York-Presbyterian Hospital that might be of some interest to members of the list. It was to me since my wife works just outside the accident area.
For those that get the New York Times, it was on page B12; it was on page 2 of the New York Daily News.
In summary, a supervisor was killed by asphyxiation while working alone in a poorly ventilated trailer located outside the hospital. He was part of a team installing a new MRI at the hospital. For reasons not clear, one of the liquid nitrogen tanks connected by a valve to the room started leaking, filling the trailer space and causing the asphyxiation. Six others were treated for dizziness and other minor injuries. Two of the six injuries resulted from attempting to rescue the supervisor as they went into the room before it had been ventilated. At that time, it was not known that the room had been filled with nitrogen and that the oxygen had been displaced.
I cannot determine if the trailer was a temporary facility or part of the new installation that was being done. The nitrogen tanks were outside the trailer and the trailer where the supervisor was working was on the order of 15 x 40. It is not clear from the reports why the supervisor was working alone or how long he had been working in the trailer. Also, there is no indication of whether the trailer was part of the installation or a temporary facility.
Just thought you all would be interested.
Tony Mitchell, Ph. D. 7 East Willow Street Beacon, NY 12508
I have watched the replies to your question, and have noted that for the most part they reflect my experience. Perhaps you could try to begin your dehydration at very low %, e.g. 30% or even 10% EtOH, and do a more gradual (10,20,30,40,50,60,70,80,90,90,100,100,100) series. This has worked for some of the cultured cells I have prepared for SEM over the years. Other little nuances have included a very slow bleed in for the initial CO2 step in the CPD, and then watch the flow during the process. A final issue is watching the temperature the CPD cycles at for final drying. Sometimes the CPD tends to run a little hot, and that can also affect the final results. Hope one or more of these are of use.
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals On Tue, 19 Sep 2000 14:16:25 -0500, Alan Burns 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 having trouble obtaining good SEM preparations of cultured endothelial } cell monolayers. The monolayers show evidence } of artefactual cracking along cell borders. The cracks appear whether the } cells are grown on glass coverslips or transwell filters. I have tried } various } fixatives (buffered glutaraldehyde alone, glutaraldehyde followed by osmium } and uranyl acetate), different dehydrating agents (ethanol or acetone), } followed by critical point drying (CPD). I have even tried } tetramethylsilane (Ted Pella) in place of CPD. The only time I get good } images (i.e., } no cracks) is when a small portion of the monolayer has inadvertently } detached from its substrate during tissue processing. The cells on this } small flap look marvelous (no cracks). I believe, in the absence of } substrate adhesion, the cells on the flap shrink uniformly during dehydration } when surface tension forces exert their effects. Does anyone know how to } prepare cell monolayers for SEM that leaves them intact and free } from artefactual cracking? } } Thanks. } } Alan Burns, Ph.D. } Assistant Professor } Cardiovascular Sciences } Department of Medicine } Baylor College of Medicine }
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RSSL's Microscopy Laboratory has an extremely varied and busy working environment and its members of staff are committed to providing a first class microscopy service. A major proportion of work concerns Foreign Body Analysis, largely for the pharmaceutical and food sectors, for which the Laboratory is a market leader and enjoys a good reputation amongst its customers. Light and scanning electron microscopy imaging and Research and Development accounts for most of the remainder of the work. In house techniques available include LM (using different contrasting methods), SEM, cryo-SEM, X-ray microanalysis, FT-IR microspectroscopy, image analysis and freeze fracture/etching; TEM is undertaken off site.
Due to expansion and subsequent staff development, an opportunity has arisen in to assist in the Foreign Body Identification service that runs from the Microscopy Laboratory. The position also supports ongoing research projects, particularly in both chocolate and sugar confectionery project areas. Other tasks include provision of support in maintenance of laboratory equipment including the electron and light microscopes, digital imaging and dark room facilities. On occasions, work into the evenings and at the weekends might be required and this post carries a requirement to share in a rota to undertake out of hours analyses as part of RSSL's comprehensive Emergency Response Service.
We are looking for a candidate who is educated to degree level or equivalent with a minimum of 4 years microscopy experience in food science, biology or materials science. Although training will be given, experience in X-ray microanalysis of bulk samples and/or of FT-IR microspectroscopy is highly desirable. Experience or knowledge of general equipment maintenance and a willingness to become involved in this; be an excellent team player with the ability to work independently; be customer focussed and able to work under pressure, as well as capable of providing written reports of long and short term projects.
If you are a self starter, looking for an interesting and challenging role and believe you can make a positive contribution to our already successful team, we would be interested to hear from you.
Please send your CV with a covering letter including your salary expectations to Jane Bienkowski, Human Resources Officer, Reading Scientific Services Lld, Lord Zuckerman Research Centre, PO Box 234, Whiteknights, Reading, RG6 6LA or e:mail jane.a.bienkowski-at-rssl.co.uk or fax to 0118 986 8932.
Hi Phil, We have odd bitz for the above. Let me know the dimensions of the feed thro and I'll have a rummage in the box. Chris Smith, IACR-Rothamsted, UK. chris.smith-at-bbsrc.ac.uk
Take the chance to learn about the developments in scanning electro microscopy! A course on "Scanning Electron Microscopy - Imaging and Microanalysis", SEM 2000, will be given at Chalmers University of Technology, Gothenburg, Sweden, October 17 - 19, 2000.
This course will be given in collaboration with four microscope manufacturers (Hitachi, Jeol, Leo, Philips/Fei) and suppliers of equipment for EDX, WDX, EBSP and CL.
The aim of this 3-day intensive course is to give a theoretical background in the morning sessions and experimental insights in the afternoons. The lectures and the demonstrations will be given by application specialists from the different companies representad at the course, and also by researchers working with different applications in SEM. The demonstrations and lab classes will be carried out on equipment brought to Chalmers specifically for this course.
Detailed information about SEM 2000, including course programme and registration form, is posted on: http://fy.chalmers.se/microscopy This web-site will be continuously updated until the course starts.
Looking forward to seeing you at SEM 2000!
Birgitta Castedal, Lena Falk and Mats Halvarsson Course Organisers
____________________________
Associate Professor Lena Falk Department of Experimental Physics, Chalmers University of Technology, SE-412 96 Göteborg, SWEDEN
Your description does not fit the classic EM fields interference pattern, as other responses had pointed out. The high level of the stray fields in the room is bad, yet it will not necessary cause a problem. Your problem could be resulted from a combination of conditions. JEOL, however, has a reason for recommending the mu-metal shield. You can perform a quick and easy test to see whether external electromagnetic fields are responsible for the problem. Assuming that the SEM column is clean (especially the final aperture!), and focusing circuit operates properly. 1. Obtain an image in continuous LIVE (no integration!) mode on the monitor screen with contrast enough vertical features, so distortion of the vertical lines is clearly visible. Magnification may vary, but you will need at least several 1000s. This distortion (if present), will be a sawtooth pattern, with the period smaller at the slowest beam scan speed, larger at higher speeds, and becoming a single "wave period" at the fastest speed which is a TV rate. The later will be an image "floating" back and forth horizontally on the screen. If the described pattern is not obtainable, the problem is unlikely to be caused by stray AC fields. 2. Change accelerating voltage at least 50%. Focus the image. Make sure that you have exactly the same sample feature in the image. Do not change any other settings. If the described distortion is caused by external electromagnetic fields, the amplitude of the sawtooth pattern (slow scan) or the amplitude of "floating" (TV mode) will change in approximately reverse proportion to the accelerating voltage value, i.e. at 10 kV will be twice of what it was at 20 kV, and so forth. 3. If the distortion will stay the same, then the source of your problem could be, for example, the mechanical vibrations. Those are caused by AC electric motors (vacuum pumps, air compressors, elevators, etc.) most of the time, vibration frequencies being harmonics of the AC line frequency. So the distortion will look very similar, except its amplitude will not be a function of accelerating voltage. Also, interference affecting the electronic circuits (ground loops, for instance), may look the same and will not be a function of the accelerating voltage. 4. Ground loops are the most frequent problem in my experience. Their influence could be inconsistent, which makes the problem difficult to identify. For example: the AC line conduit was used for the ground. Everything was fine until big air-conditioning unit was installed behind the wall. Now, a strong 60Hz AC field is measured every time when aircond. motor kicks in. Then expensive mu-metal shield was installed, and... made no difference. The reason: new unit was connected to the same power line. Existing ground loop was OK before due to low power consumption of the SEM. It is greatly increased now, and ground loop badly affects electronics of the SEM. The SEM column itself, at the same time, was sufficiently shielding external fields. 5. In any case, try self-made soft Iron shield before spending big $$ on it.
Good luck.
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax -----Original Message----- } From: Mark West {mwest-at-ifcsun1.ifisiol.unam.mx} To: microscopy-at-sparc5.microscopy.com {microscopy-at-sparc5.microscopy.com}
Hi Again, I received many replies to my question of late regarding the composition of formalin vs. formaldehyde. All of them were pretty consistent except for the most recent which touched on a very important, related issue, safety. As I think that it is particularly important, I'm forwarding to the whole list (with the permission of the author - who's name I've withheld). Be careful out there. Kristen
"Kristen,
I didn't follow the thread on this topic and what has been said. However, the comment about being "pretty much covered with it while doing some mass field fixes" caught my eye. In case no one else mentioned it, exposure to formaldehyde is thought by some pulmonology specialists, especially those researching Idiopathic Pulmonary Fibrosis (IPF), suspect that formaldehyde exposure may trigger the disease. IPF is fatal and has no treatment except lung transplant. I pass this on as one who has experienced the type of exposure you describe, the diagnosis of IPF, and the double lung transplant that is something that very, very few people are fortunate enough to get. And even with a lung transplant, the five year life expectancy is only 48%. PLEASE, please be very careful with lab chemicals and teach those you work with to do likewise.
} My exposure to formalin/formaldehyde occurred during my undergraduate and } graduate years, not only during standard class preparations but also in } the field when we went collecting algae at freshwater and more so marine } habitats. It was there when we were exposed to a lot of it; I remember my } fingers getting "fixed", my eyes watering and stinging, you know the } feeling. The fact is that the disease occurred about 20 years later. Yes } the story has a "happy" ending, so far, but for reasons other than getting } a rare double lung transplant and doing fine after 3 years. But that is } another story! Yes, many people who handle chemicals in the lab don't get } IPF, but don't take the attitude that nothing can happen to you. There } are many other ways that chemical exposure can damage your } health. Remember, take care of your body as it is the only one you get } and damn few people get spare parts! :-) } } You may pass this along if you want, if it helps one person it's worth it."
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I have a problem with Digital Micrograph 2.1 related to image file sizes.
Sometimes DM saves an image at 1.2MB while at other times (in fact, the very next image) may be saved at 1MB or even less. Unfortunately, when we do image analysis on these images the measurements will be off since the number of pixels varies. I have tried saving in various formats and nothing seems to change the events.
What gives here? Why does the program save in such differing file sizes and so randomly? I can not figure out any logic to this....... It is driving me crazzzzzzzzzzzzzzzyyyyyyy.
Thank you,
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 ####################################################################
A response directed to Paul Webster, c/o this invaluable and open forum:
I would like to continue with the topic of immunogold labeling for TEM, relating to your suggestion:
"One possible step you could put in is to first try out your whole preparation and immunolabeling protocol at the LM level. Use the same reagents and same preparation protocols as you plan to use for EM and expose to silver enhancment for a longer time so that you can see the signal by LM. If you see a signal then you will be confident that you will see a signal when you go to examine the EM experiment."
How reliable is LM for predicting TEM immunogold labeling results?
I have tried this approach before, with mixed results. It sounds like a great idea, but in attempting to "modify" the EM protocol into a LM pilot experiment, it becomes a completely different beast. During primary fixation, EM requires glutaraldehyde to preserve ultrastructure. You suggested that glut not be used until AFTER immunolabeling, but won't the damage to the fine structure already be done? I was under the impression that glut "postfixation" is intended primarily to crosslink gold antibodies in place after they have bound to primary antibody.
Also, silver and gold enhancement reagents come in LM and EM formulae; usually, EM versions are less acidic. Does a longer LM enhancement accurately predict the action of a brief EM enhancement? Perhaps EM formulae can be used to directly monitor enhancement with the light microscope. However, because optimal EM enhancement is invisible by LM, one will need to empirically determine EM enhancement times for each cell system. Doesn't this argue against spending too much time with LM experiments, when EM also includes osmium, uranyl acetate, dehydration, and resin infiltration?
I still use pilot LM experiments, but more for their complimentary benefit, rather than for their predictive value.
Routinely, I see a discouraging signal to noise ratio at the LM level. However, at the EM level noise is restricted to the first couple of resin sections of a monolayer culture, and subsequent sections contain beautiful results. I interpret this as nonspecific binding of antibodies to the poly-L-lysine and lamin-coated glass substrate. In this example, routine LM (DIC optics without optical sectioning) does not accurately predict EM performance.
Sincerely,
Michael Plociniak Research Technician Albert Einstein College of Medicine Neuroscience Dept. Rose Kennedy Center, 529 Bronx, NY 10461 (718) 430-3509 plocinia-at-aecom.yu.edu
Thanks everyone who responded to my request for labs in the DFW area that would consider "renting" space on their scopes. I've sent the information on to the researcher who is looking for space and he and I were impressed by the many places that were gracious enough to consider letting outsiders use their equipment.
Whenever I use Schiffs reagents for student lab classes I test it first by adding just a couple of drops of neutral buffered formalin (the diluted form = 3.8% formaldehyde) to about 10ml in a clear tube and they can then watch the colour develop over a few seconds. If was also 'water white' to begin with and with no hint of pink we use it. It hasn't failed me so far and the students get to see a dramatic colour change before they do the method.
Hope this helps?
At 08:30 2000-09-25 -0500, Tobias Baskin wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Med vänliga hälsningar/With best regards
Gareth
"A wise person is certain of few things - a fool of everything"
"The foolish reject what they see, not what they think; the wise reject what they think, not what they see." Huang Po
Gareth Morgan MPhil MSc FIBMS, Institutionen för Mikrobiologi, Patologi och Immunologi(IMPI), H5, Karolinska Institutet, Avdelningar för biomedicinsk laboratorievetenskap och biomedicinska ämnen, Box 12 773, S 11 296, Stockholm Sverige
In celebration of the acquisition of a new JEOL 3010 transmission electron microscope with environmental cell capabilities, the Naval Research Laboratory Stennis Space Center will be conducting a day long symposium on November 2, 2000. Outstanding microscopists from around the country will join local experts to discuss marine environmental and materials science applications of TEM. On Friday morning, November 3, participants are invited to visit informally with NRL researchers in the new Microscopy Facility.
In addition to the opportunities for exploring and sharing insights during the period of formal presentations, transportation will be provided Thursday evening to the French Quarter in New Orleans. Here participants may enjoy interacting with each other in a setting famous for its music, its food, and its stimulating atmosphere.
We are also strongly soliciting contributed papers. Although a primary area of emphasis is expected to be environmental cell and in-situ microscopy, submissions of a wide variety of ‘frontier’ microscopy results are welcome!
Invited speakers include: Larry Allard - Oak Ridge National Laboratory -- “Aberration Correction for High Resolution Imaging and Advanced Analytical Electron Microscopy†Frances Ross - IBM Watson Research Center Renu Sharma - Arizona State University Judith Yang - University of Pittsburgh
Other speakers planning to participate include: Tyrone Daulton – NRL -- “Application of High-Resolution and In-Situ TEM to Solving Problems in Diverse Fields of Study†Yoko Furukawa – NRL -- “Ultrastructure of Aquatic Sediments and its Effect on Early Diagenesis†Dawn Lavoie – NRL -- “Preparation Techniques and Quantitative Characterization of Clay Sediments using TEM†Brenda Little – NRL -- “Environmental Electron Microscopy Investigations of Microbiologically Influenced Corrosion†Eric Stach – NCEM -- “In-situ TEM of Dislocations in Thin Filmsâ€
This symposium is co-sponsored by the Southeastern Microscopy Society.
We anticipate there will be approximately 16 presentations at the Symposium. Arrangements are underway to develop a special section of Microscopy and Microanalysis devoted to the papers presented. Finished manuscripts are due October 15. The total number of participants will be limited to a maximum of 60.
REGISTRATION Please register early by completing and returning the form below; the registration deadline is October 13.
Sincerely, your organizers: Bhakta Rath, Co-Chair - Naval Research Laboratory Eric Stach, Co-Chair - National Center for Electron Microscopy, Lawrence Berkeley National Laboratory Matthew Hulbert, Coordinator - Research Dynamics
______I wish transportation to the French Quarter Thursday evening.
______I do not wish transportation to the French Quarter.
If you wish to give a presentation, please provide its title below.
Return registration information to Matthew Hulbert. E-mail is preferred -- matthew_hulbert-at-hotmail.com If you choose to use US mail, the address is 1332 Pembroke Drive, West Chester, PA 19380.
Most standard histology books will give a procedure for digestion of a sample as negative control. If you need the procedure please let me know and I will e-mail it to you.
Pamela A Marcum Histology/Microscopy Product Development Manager Polysciences, Inc.
-----Original Message----- } From: Tobias Baskin [mailto:BaskinT-at-missouri.edu] Sent: Monday, September 25, 2000 9:30 AM To: Microscopy-at-sparc5.microscopy.com Cc: STYLER-at-dnr.state.md.us
Greetings, I have PAS stained samples embedded in agarose and then in plastic and there was never an agarose reaction.
However, this info is simply anecdotal. I have no idea is PAS is "supposed" to stain agarose, and I don't do PAS "properly" (just perioidic acid and then Schiff's). Did you have a positive control? Sometmes the Schiff's reagent goes bad. I recall placing a drop on a piece of white tissue paper and if it went pink in a moment or two then the reagent was still ok.
Hope this helps even a little, Tobias
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
In celebration of the acquisition of a new JEOL 3010 transmission electron microscope with environmental cell capabilities, the Naval Research Laboratory Stennis Space Center will be conducting a day long symposium on November 2, 2000. Outstanding microscopists from around the country will join local experts to discuss marine environmental and materials science applications of TEM. On Friday morning, November 3, participants are invited to visit informally with NRL researchers in the new Microscopy Facility.
In addition to the opportunities for exploring and sharing insights during the period of formal presentations, transportation will be provided Thursday evening to the French Quarter in New Orleans. Here participants may enjoy interacting with each other in a setting famous for its music, its food, and its stimulating atmosphere.
We are also strongly soliciting contributed papers. Although a primary area of emphasis is expected to be environmental cell and in-situ microscopy, submissions of a wide variety of ëfrontierí microscopy results are welcome!
EARLY REGISTRATION WOULD BE GREATLY APPRECIATED!!!
Invited speakers include: Larry Allard - Oak Ridge National Laboratory -- ìAberration Correction for High Resolution Imaging and Advanced Analytical Electron Microscopyî Frances Ross - IBM Watson Research Center Renu Sharma - Arizona State University Judith Yang - University of Pittsburgh Other speakers planning to participate include: Tyrone Daulton ñ NRL -- ìApplication of High-Resolution and In-Situ TEM to Solving Problems in Diverse Fields of Studyî Yoko Furukawa ñ NRL -- ìUltrastructure of Aquatic Sediments and its Effect on Early Diagenesisî Dawn Lavoie ñ NRL -- ìPreparation Techniques and Quantitative Characterization of Clay Sediments using TEMî Brenda Little ñ NRL -- ìEnvironmental Electron Microscopy Investigations of Microbiologically Influenced Corrosionî Eric Stach ñ NCEM -- ìIn-situ TEM of Dislocations in Thin Filmsî
This symposium is co-sponsored by the Southeastern Microscopy Society.
We anticipate there will be approximately 16 presentations at the Symposium. Arrangements are underway to develop a special section of Microscopy and Microanalysis devoted to the papers presented. Finished manuscripts are due October 15. The total number of participants will be limited to a maximum of 60.
REGISTRATION Please register early by completing and returning the form below; the registration deadline is October 13.
Sincerely, your organizers: Bhakta Rath, Co-Chair - Naval Research Laboratory Eric Stach, Co-Chair - National Center for Electron Microscopy, Lawrence Berkeley National Laboratory Matthew Hulbert, Coordinator - Research Dynamics
REGISTRATION INFORMATION Name: ______________________________________________________ Organization: ______________________________________________ Address: ___________________________________________________
___________________________________________________________ e-mail Address: ____________________________________________ __I wish transportation to the French Quarter Thursday evening. __I do not wish transportation to the French Quarter. If you wish to give a presentation, please provide its title below. Return registration information to Matthew Hulbert. E-mail is preferred -- matthew_hulbert-at-hotmail.com If you choose to use US mail, the address is 1332 Pembroke Drive, West Chester, PA 19380. _________________________________________________________________________ Get Your Private, Free E-mail from MSN Hotmail at http://www.hotmail.com.
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Have any other microscopists had problems with Peltier coolers failing on water cooled CCD cameras?
In my facility I have three such cameras all plumbed into the microscope water system and connected, by the camera manufacturer, into wall sockets. On one of the three cameras I have had the Peltier cooler fail four times in two years. After the latest failure the manufacturer asked if the camera had been run without water. I told him not intentionally but when we have power cuts and the power is restored, the camera comes back on but the microscope (and water) does not. The camera can therefore run for some time without water cooling. Power failures are not uncommon at this university!
Many e-mails to the manufacturer have resulted in no response to my questions about reliability of the Peltier coolers. The manufacturer is treating the failure as an out of warranty issue but I believe there has been something wrong with this camera since it was installed.
Any experiences of listservers with Peltier cooler failures would be appreciated. Please e-mail me directly.
Regards
Alan W Nicholls, PhD Electron Microscopy Service Director Research Resources Center - East (M/C 337) Room 100 Science and Engineering South Building The University of Illinois at Chicago 845 West Taylor St Chicago, IL 60607-7058
Alan W Nicholls, PhD Electron Microscopy Service Director Research Resources Center - East (M/C 337) Room 100 Science and Engineering South Building The University of Illinois at Chicago 845 West Taylor St Chicago, IL 60607-7058
Just this morning we were discussing the same problem with our Gatan CCD cameras on our HD-2000, one of which is retractable and used for TEM imaging, and the other of which is at the end of the GIF for energy-filtered imaging. These cameras are connected in series for the water flow, and have very tiny water lines which can easily clog, even with recirculated water. There is no provision by the manufacturer to provide for a safety cut-off of the Peltier cooler in the event of loss of water flow. We have recently experienced a burn-out of the TEM camera due to a clog in the line, which resulted in 2 weeks of downtime for repair (nicely done for free by Gatan), but still no response to queries about some sort of protection. So we have decided to find an appropriate water flow sensor, and to install it in an appropriate place in the water line, and to connect it to the camera controllers to provide a power cut-off when the water flow diminishes. I'll post the details of the solution when it is completed.
Larry
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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 am having trouble processing bacteria in liquid culture for TEM/SEM. Too much of sample is being lost because I only have a small table top microfuge. The culture won't spin down into a hard enough pellet. Does anyone have a different protocol for processing bacteria in liquid culture? Please reply to capri-at-zianet.com Kristine Fambrough
Dear Alan, My experience with Peltiers does not involve a CCD camera, but it may be instructive nonetheless. When we wanted to provide a constant temperature for the chemicals in the darkroom--which required sometimes heating the water and at other times cooling it--we decided that Peltiers were the best solution. We bought a commercial thermoregulating unit which sent current of the appropriate polarity through the Peltiers to adjust the temperature to the desired setpoint. The Peltiers failed at an expensive rate, and we found that the thermoregulator was such that it provided a specific current, which it turned on and off in a duty cycle (the greater the temperature difference, the greater fraction of time the current was on). The sudden changes in current were the cause of the Peltiers' demise. Our shop designed and built a regulator which operated in such a way that the current was not turned on and off, but was proportional to the temperature difference, and the Peltiers worked properly for many years. We were also warned that moisture in the Peltiers could lead to an electrochemical process which caused corrosion and subsequent failure. Check that the controller does not vary the current rapidly and that the Peltiers are protected from condensation in your CCD setup.
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
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Perform all technical operations necessary to produce specimens for viewing in the scanning and transmission electron microscopes. Will work independently and report to the EM Manager in performing daily responsibilities according to established procedures. Focus of the job is on the application of skills and techniques to assist users with the extremely varied research projects. Duties include preparation of specimens for TEM and SEM, autoradiography, immunoelectronmicroscopy, serial sectioning, shadowing, negative stating, particle counting and SEM. Produce prints and publication ready layout. Periodic instrument maintenance, billing operations, maintain logs, supply inventories, purchase records. Attend workshops and maintain current knowledge of topics related to EM and Center experimentation.
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John, for all high resolution work and image analysis, I always collect images as a ".gat" (with a Digital Micrograph 3.3.1 on our system, I get 2.1 MB file). I have similar problem with exporting files to users not having DM (not able to open gatan files); either tiff or jpeg files are only about half the original size. I get better resolution if I go through the "File/Export" option rather than just "File/Save As/". Does anyone have a more elegant way? Thanks, Alice.
Alice Dohnalkova Environmental Microbiology Battelle, PNNL MS P7-50 Richland, WA 99352 (509) 372-0692
} From: John J. Bozzola [mailto:bozzola-at-siu.edu] Sent: Monday, September 25, 2000 3:40 PM To: microscopy-at-sparc5.microscopy.com
I have a problem with Digital Micrograph 2.1 related to image file sizes.
Sometimes DM saves an image at 1.2MB while at other times (in fact, the very next image) may be saved at 1MB or even less. Unfortunately, when we do image analysis on these images the measurements will be off since the number of pixels varies. I have tried saving in various formats and nothing seems to change the events.
What gives here? Why does the program save in such differing file sizes and so randomly? I can not figure out any logic to this....... It is driving me crazzzzzzzzzzzzzzzyyyyyyy.
Thank you,
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 ####################################################################
Kristine There was a thread on this a few months ago, the concensus of which was that for TEM most people advocated solidifying a spun- down bacterial suspension using agar or agarose before getting too far into the process. You might want to consider whether this should be done pre-fix or post-fix. I would advocate the latter. Anyway, this produces a chunk of material that can be handled through teh solution changes more conveniently than a cell suspension. However, many people found that a pellet solidifed in this way would infiltrate rather poorly if left in the bottom of the microfuge tube / eppendorff, etc. The best solution is probably to detach it and perhaps chop it up a little to minimize specimen size, providing reagents and solvents free access to all surfaces. Best wishes Chris Jeffree
} From: "sghoshro-at-nmsu.edu"-at-sparc5.microscopy.com Date sent: Tue, 26 Sep 2000 09:21:58 -0600 (MDT) To: microscopy-at-sparc5.microscopy.com
The original question by Tom Phillips has stirred up a great discussion on this list. Even though we're getting a little away from the original subject, I wanted to throw in my views with respect to the issue of antibodies working in LM and not in EM. I'm afraid it will get kind of lengthy, but I hope it is useful.
When immunodetection at the LM level works, but not at the EM level, I prefer to follow a logical procedure to find where the bottle necks are. Basically there may be two differences in the approaches: 1. the reagents used, 2. the specimen preparation method.
To make things simple, let us assume the labeling at the LM level works reliably. Then we are left with a situation where we need to know whether the antigens (epitopes) are still recognizable and available in the EM specimen and whether the reagents for EM detection will do the job they are supposed to do. How do we get an answer to these questions?
The first thing one should do is to check the reactivity of the reagents for EM with the primary antibody of interest. This does not necessarily have to be checked on the specimen. It is even better not to do that, because there are too many other parameters different from the LM approach. A simple dot-spot test can do the trick and give a conclusive answer in about two hours. In practice: a dilution series of primary antibody (monoclonal or affinity purified) is applied to a strip of nitro-cellulose. The strip is blocked and incubated with the immunogold reagent. If applicable (with small particle conjugates) the strip can be silver enhanced. If antigen is available, the dot-spot test can even be used to get a rough idea about the influence of fixatives. A dilution series of antigen (or an enriched cell lysate) is applied to the nitro-cellulose strip, the strip can be treated with fixative (exactly the same concentrations, time and temperature as intended to be used for specimen fixation), then the strip is washed with glycine, blocked, incubated with primary antibody, gold conjugate and silver enhanced. A comparison of different strips (fixed with several different fixatives and unfixed) gives somewhat of an indication whether a particular fixative is worthwhile to try. It is not an absolute test, as the environment of the antigen may play a role in the effect of fixation as well (cross-linking, densely packed structures). Should anyone need information how to do this in practice, please get in touch with us. If antigen is not available, you could use cryostat sections or even paraffin sections that worked at the LM level and run the same test. Make sure that the gold and silver enhancement reagents being tested here are the same reagents that will be used for EM labeling.
What about the influence of specimen preparation? To some extent the issue of fixation has been addressed above. If reactivity of the immunogold and silver enhancement reagents had been tested fine and EM specimens had been prepared the same way as for LM (only thinner of course, at least in the end), the approach should work. Unfortunately, the specimen preparation for EM often differs a lot from the one for LM. So let us start with the EM sample preparations that are similar to those for LM, because logically those should be the most suited, at least from the point of view of antigen preservation and presentation. In ultrathin cryosections the influence of the embedding material is avoided, so antigens will not be as masked as they are in resin or acrylate embedded specimens. I don't know if there still is the same consensus about the validity of this technique as there was some years ago, but we always found that cryo ultra microtomy was the approach to most likely give a positive result.
Another approach, that has been addressed in a few postings recently, is to use pre-embedding labeling. In the time period before ultra small gold conjugates became available, pre-embedding immunolabeling with the classic gold conjugates was only feasible after harsh detergent treatments which were often applied during chemical fixation. As a result much of the cytoplasm and organelles were either destroyed or washed out. The state of ultrastructural preservation was far from ideal and this has imprinted the pre-embedding approach with a negative image for a long time. However, with the ultra small gold conjugates becoming available, harsh detergent treatments of the sample as a trade off for immunoreagents to penetrate were no longer necessary. Even only a pre-treatment with sodium borohydride, which was applied to quench aldehyde groups, was sufficient to make some types of single cells accessible for ultra small gold reagents.
I don't want to state that these two approaches would be the only ones I would consider useful. But they are good steps to start with. A great advantage of the combination of ultrathin cryosections and ultra small gold reagents is that these reagents penetrate with greater ease into the section’s interior, compared to larger gold conjugates, detecting not just antigens exposed on the surface.
For me cryosectioning also worked with plant material like soy beans, but then again they were not the dormant dry seeds, these were sprouts. I am sure they are a lot easier to cut than the dry ones. Teaching workshops at botany departments I have learned how hard it is to get even an acceptable level of ultrastructural preservation and good immunolabeling, although I got the impression that high pressure freezing can be a very good starting point.
To be able to use ultra thin cryosections you need a cryo ultra microtome. For pre-embedding you don't need a lot of fancy equipment. The results some of us are getting with pre-embedding immunolabeling are really very encouraging, especially with Fab2 and single Fab fragment-ultra small gold reagents and efficient and homogeneous silver enhancement reagents being available. I don’t think I should be the one doing a lot of talking here since it is not my work. But I hope that researchers who have done this or are experienced with pre-embedding will jump into the discussion. Hong, why don't you tell us? And yes, I realize, useless for internal antigens in plant tissue. But for cultured cells and animal tissue it can be great.
I wholeheartedly agree with Paul Webster that it is a must for an investigator to try to understand the ins and outs of a method and that ‘we must stop being the "black box" of science’. That is the only way to understand the results and to make progress. When I was still associated with the University in Utrecht I experienced very often that non-EM people have a tendency to look upon EM as a way to make the picture to fit the story. As a sort of confirmation of the biochemical or molecular data they already have fixed in their minds. You're lucky (!) if you can confirm their beliefs, and they're happy to have a nice demonstration supporting their ideas. IEM is a scientific method that not just adds up to the results that are there, but my own experiences convinced me that it may provide results that have been overlooked with other, biochemical or whatever analysis methods. I learned something from my PhD professor that I still value very much: "don't try to get the results that you or somebody else wants you to get, but try to understand the results you're actually getting".
In spite of the fact that most of you will have fallen asleep by now as a result of this lengthy posting, I would like to make a few more comments.
Don't compare apples with pears! Use the same reagents for LM and EM. Protocols for LM and EM should be comparable as much as possible, including the use of buffers, blocking reagents, dilutions.
I am reluctant about using cell fractionation in some instances: you might only get to see what you want to see. This may degrade the labeling to a confirmation of a mental picture, and prevent you to find labeling in places you didn't expect before and which may not be present in your fraction-specimen anymore. And also, if you already know where the antigen is, why would you bother to do immuno EM? But often this so-called knowing is just an idea based on data acquired up to that point in time. Immuno EM may reveal additional data that shed a new light on the question.
Secondary antibody conjugates vs. protein A: theoretically secondary antibodies give less resolution, true. Is there anyone who actually made the comparison and found one to be different from the other in that respect? I mean not in molecular labeling: molecules or viruses attached to a film on grid and labeled for specific epitopes, but labeling in or on a section?
To end this message: unfortunately, sometimes it happens that even if you've checked carefully and think it should work, sometimes it doesn't. Those cases are rare and although they are most annoying they are also the most interesting cases from a technological/methodological point of view, because they can teach us something we didn't understand before.
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} From: John J. Bozzola, bozzola-at-siu.edu
{I have a problem with Digital Micrograph 2.1 related to image file sizes.
{Sometimes DM saves an image at 1.2MB while at other times (in fact, {the very next image) may be saved at 1MB or even less. Unfortunately, {when we do image analysis on these images the measurements will be {off since the number of pixels varies. I have tried saving in various {formats and nothing seems to change the events.
{What gives here? Why does the program save in such differing file {sizes and so randomly? I can not figure out any logic to this.......
Fun with Digital Micrograph :-(
Actually in DM2.X and lower when you "Save As.." you are in effect saving a screen resolution image and the size of the image you save is dependent on how much you have zoomed the image on the screen. So if you zoomed out once and the image was 512X512 on screen (with a 1024 camera) then your image would get saved as a 512X512 image and you lost half of your resolution. What I always did was to save the image in DM format first without touching anything in the image (contrast, brightness, zoom, etc). That way I knew I had a pristine image that had not been messed up in anyway and I could go back to that when needed. In order to save a tiff or whatever, I would make sure that my zoom was 1:1 and then I would save it in the format needed.
In DM3.X they modified that slightly. If you hit "Save as.." and save it in DM format it is OK, if you "Save as.." in any other format it saves just as I mentioned above. However, they now have an export menu item that saves the image in original resolution in whatever file type you want. I always tell my students to ignore the "Save as.." feature and just use export. Gatan has also removed some of the bugs in the auto save feature. If you have the extra $$$$$$$$$ it is probably worth upgrading to DM3.X just note that they changed their DM image format somewhat but new DM can read old DM images but not the other way around.
Norm Olson
******************************************* Norm Olson Senior Research Electron Microscopist Department of Biological Sciences Lilly Hall of Life Sciences Purdue University West Lafayette, IN 47907
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Hi, Alan
} we have power cuts and the power is restored, the camera comes back on but } the microscope (and water) does not. The camera can therefore run for some } time without water cooling. Power failures are not uncommon at this } university! }
I think that all equipment like this should be set up so that when the power goes off, the equipment stays off when the power comes on again.
Devices to acheive this are common in the world of electricians, they call them "zero-voltage switches" (at least down here they do).
Get your electrician to install one for the mains socket from which the camera derives its power.
Mine have saved me from complications many times.
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
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Reply to: RE: Can LM predict immunogold TEM? Hi Michael,
I read your comments with some dismay. I may have misunderstood, but the impression I get is that you feel there will be such large differences between LM labeling and EM labeling that you don't think it worthwhile to check your antibodies by LM first. If there were such differences then the experiment should not be performed in the first place. The LM result should be the same as the one you see by EM. I would not expect anything less.
One point of my previous posting was to suggest that immunolabeling experiments can be carefully designed so that LM screening is possible before moving onto the EM level. I think that the reason my antibodies work for EM is that I first tested them by LM to make sure I would get a result.
I do not think that glutaraldehyde is a prerequisite for good morphology, or that samples have to be treated with osmium tetroxide, uranyl acetate either. What I was suggesting is that it should be possible to screen samples by LM before moving onto the EM level. This will confirm that immunoreagents will react with the sample, will give an idea of the best dilutions to use, and will give information on the relative number and ditribution of antigens. All this should be possible using the same samples for LM and EM.
As an example, suppose I wanted to look for a mast cell antigen in nasal mucosa. I know from my biology lessons that there will be few mast cells in this tissue so I know I will have to search for them before I can label them. Now I make a choice between pre- and post-embedding labeling. I know that the antigen I want is going to be inside the granules of the mast cells, which are inside the tissue. For this reason, I decide that pre-embedding labeling may not be the best approach. I choose to label sections. Now, I have to decide whether I want to cryosection or to embed in resin. As I am lucky enough to have a cryoultramicrotome in my lab, and as I know from experience that I usually get higher sensitivity on cryosections, I choose to prepare the samples this way. If I want to perform immunolabeling on the samples, I will choose not to fix in glutaradehyde, which gives the tissues an autofluorescent glow (and which can also be a useful counterstain, when used properly). I therefore choose to fix with formaldehyde (which, btw, is a gas and which when dissolved in water becomes formalin, and is actually converted to methylene glycol CH2O + H2O = HO CH2 OH). I also know that at concentrations above 2% the methylene glycol begins to polymerize to form polyoxymethylene glycol, and that these polymers are what react with and crosslink proteins in biological material. My choice of fixative might then be an initial immersion fix in 4% buffered formaldehyde for perhaps 1 hr, followed by a second immersion fix in 8% buffered formaldehyde for 24 hr. I would make sure not to use phosphate buffered saline, but something with a better buffering capacity (such as 100mM phosphate buffer).
Once fixed, I would cut off a small portion of the tissue (carefully so as not to crush the sample), infiltrate with sucrose and prepare cryosections, mount them onto glass coverslips and label them with my antibody and a fluorescent secondary. I could also visualize the bound antibody by reacting with protein A gold and then silver enhancing the bound gold. I have no concern about whether the silver enhancer will work at the EM level because I know I will be able to easily detect the 5nm protein A-gold at the magnifications I use.
Once I had found a part of the tissue containing the mast cells, I would then trim down the block, cool the ultramicrotome and cut thin sections for EM. These I would label with antibody and protein A-gold, confident that the sections contained mast cells and that the immunoreagents would be working.
If I found that the antibody gave unusual results, suggesting either a migration or extraction of antigen, I would take part of the tissue still in aldehyde and embed it in Lowicryl. For this I have the choice of PLT (progressive lowering of temperature) or freeze substitution. As I know that freeze substitution generally gives increased sensitivity, I would probably use that. I would cryoprotect small pieces of sample in sucrose (above 1.6M) and freeze by immersion in liquid nitrogen (only because I do not have a high pressure freezer), and put the frozen blocks in cold methanol (to increase contrast I may add 1% osmium tetroxide or uranyl acetate to the substitution medium). While still cold, I would gradually replace the methanol with unpolymerized Lowicryl until I was in 100% resin and could polymerize it by exposure to UV light, still at low temperature.
The sectioning step would be the same as above. I would first prepare sections for LM to screen the tissue for mast cells and for immunolabeling. Once I had both, I would thin section and look at the sample by EM. If this routine is kept standard then the level of success will increase as the level of frustration drops.
As for your comment that you see background in the first few sections of your EM labeling, I presume that this is outside the cells. It is possible that this background is also there in your LM preparations but because it is only in a thin layer, the LM may not have the sensitivity to register it. If you are getting a high signal to noise ration at the LM level but not by EM, it would be a good idea to figure out why this is. The result may affect the way you interpret your EM pictures. I notice that many antibodies when tested by LM appear to be very specific with no background, but when used at the EM level, are unusable because of the large amount of background they produce. If the LM is carefully performed and the results examined before they have been exposed to the process I have called "Photoshop purification of antibodies", the background can be picked up there too. Only with very few antibodies have I noticed that no background is detected by LM or by Western blotting, but that it comes up in the EM. Usually this is a result of the high concentrations I use the antibodies at, compared to what is used for other detection emthods. The offending signal, which is still specific, has been diluted out. This background can be eliminated by careful preparation and purification of antibodies.
By all means expose the labeled sections to a fixation step after the labeling protocol is complete. It is such a small part of the process as to be negligible. I would be interested, however, in any quantitative results that actually show this step to retain gold particles when compared with labeling protocols that leave it out.
Regards,
Paul Webster.
Disclaimer: Other than a satisfied user, I have no affiliation with or interest in Lowicryl resins or Adobe Photoshop. All opinions are my own and are freely given. How they are used is not my responsibility.
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
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 } -----------------------------------------------------------------------. } } } A response directed to Paul Webster, c/o this invaluable and open forum: } } I would like to continue with the topic of immunogold labeling for TEM, } relating to your suggestion: } } "One possible step you could put in is to first try out your whole } preparation and immunolabeling protocol at the LM level. Use the same } reagents and same preparation protocols as you plan to use for EM and } expose to silver enhancment for a longer time so that you can see the } signal by LM. If you see a signal then you will be confident that you will } see a signal when you go to examine the EM experiment." } } How reliable is LM for predicting TEM immunogold labeling results? } } I have tried this approach before, with mixed results. It sounds like a } great idea, but in attempting to "modify" the EM protocol into a LM pilot } experiment, it becomes a completely different beast. During primary } fixation, EM requires glutaraldehyde to preserve ultrastructure. You } suggested that glut not be used until AFTER immunolabeling, but won't the } damage to the fine structure already be done? I was under the impression } that glut "postfixation" is intended primarily to crosslink gold antibodies } in place after they have bound to primary antibody. } } Also, silver and gold enhancement reagents come in LM and EM formulae; } usually, EM versions are less acidic. Does a longer LM enhancement } accurately predict the action of a brief EM enhancement? Perhaps EM } formulae can be used to directly monitor enhancement with the light } microscope. However, because optimal EM enhancement is invisible by LM, } one will need to empirically determine EM enhancement times for each cell } system. Doesn't this argue against spending too much time with LM } experiments, when EM also includes osmium, uranyl acetate, dehydration, and } resin infiltration? } } I still use pilot LM experiments, but more for their complimentary benefit, } rather than for their predictive value. } } Routinely, I see a discouraging signal to noise ratio at the LM level. } However, at the EM level noise is restricted to the first couple of resin } sections of a monolayer culture, and subsequent sections contain beautiful } results. I interpret this as nonspecific binding of antibodies to the } poly-L-lysine and lamin-coated glass substrate. In this example, routine } LM (DIC optics without optical sectioning) does not accurately predict EM } performance. } } Sincerely, } } Michael Plociniak } Research Technician } Albert Einstein College of Medicine } Neuroscience Dept. } Rose Kennedy Center, 529 } Bronx, NY 10461 } (718) 430-3509 } plocinia-at-aecom.yu.edu } } transmission and scanning electron microscopy } } } } } RFC822 header } ----------------------------------- } } Received: from ultra5.microscopy.com [206.69.208.10] by mailhouse.hei.org } with ESMTP } (SMTPD32-6.00) id AC422BAD01DC; Tue, 26 Sep 2000 11:34:42 -0700 } Received: (from daemon-at-localhost) } by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id SAA03897 } for dist-Microscopy; Mon, 25 Sep 2000 18:47:37 -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 SAA03893 } for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Mon, 25 Sep 2000 } 18:47:07 -0500 (CDT) } Received: from post.aecom.yu.edu (post.aecom.yu.edu [129.98.1.4]) } by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id SAA03886 } for {Microscopy-at-sparc5.microscopy.com} ; Mon, 25 Sep 2000 18:46:53 -0500 (CDT) } Received: from amca (amca.aecom.yu.edu [129.98.90.216]) } by post.aecom.yu.edu (8.9.3/8.9.3) with SMTP id TAA26413 } for {Microscopy-at-MSA.Microscopy.com} ; Mon, 25 Sep 2000 19:40:58 -0400 (EDT) } Message-Id: {3.0.6.32.20000925193749.0094c470-at-mailserver.aecom.yu.edu} } X-Sender: plocinia-at-mailserver.aecom.yu.edu } X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.6 (32) } Date: Mon, 25 Sep 2000 19:37:49 -0400 } To: Microscopy-at-sparc5.microscopy.com } From: Michael Plociniak {plocinia-at-aecom.yu.edu} } Subject: Can LM predict immunogold TEM? } 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: 257555315 } 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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I need some help. If anyone out there has a Philips 420 TEM, I would appreciate some info on hooking up the main transformer (i.e. from the installation manual or anything else that I might use), our electrian cann't figure out the wiring for hooking it up. Any help would be greatly appreciated. Thank You, Chuck
Hello, I have just changed the Ar tank that is used on my PIPS. I now have a hugh amount of contamination from the grid on my TEM samples. The only thing that has changed is the tank. I made sure the guns are aligned and the flow adjusted properly.
Has anyone else had this happen after changing the tank and is there some advice for me?
I have a bunch of microslide storage boxes, the plastic kind that hold 100 glass slides. They have been used, but are in good condition . I don't need them any more, let me know if you would like some or away they go.
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
Mark: I can't comment on a mu-metal shield, but have used field-cancelling systems on SEMs and FIBs to great effect. I can't remember what we paid for ours, so I don't know if such a system would cheaper than the shield. But if you're curious try this URL: http://www.spicerconsulting.com/. This company is in the UK, but I think they have people in the Americas.
Mark West 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, } } A question about SEM interference. We have high electromagnetic field } levels (about 5 times the Jeol recommended limits) and our SEM isn't much } use above 10 000x mag (a Jeol 5410 low vacuum) - the focus is fuzzy and } astigmatism hard to correct. I've had water and air pipes removed from the } room, and we've been everywhere measuring fields but the fields and the } poor performance don't change. Changing the room is not an option (to the } people in charge of the institute!). Jeol recommended, ruling out a room } change, that we fit a mu-metal shield around the column. I have contacted } the company that makes the mu-metal and the thing would cost about } US$6000.00 and a hassle to ship to Mexico, so I don't want to go ahead and } find that we're in the same place with our performance. } } Does anyone have experience with fitting a mu-metal shield to their } microscope (especially a scanning EM) and resolving (or not resolving!) } interference problems. I'd be very interested in hearing about your } experiences with this stuff. } } Thanks, } } Mark } } PS - the serial sections are coming along nicely (not perfect,but heading } that way!), but I've still got a lot of ideas to try yet. Many thanks. } } ******************************************** } 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 } ********************************************
In one of his recent postings, Dr. Webster suggested using cell fractionation for immuno-localization. This pointed to an alternative path to the usual route I take for immunolabeling. I of course understand that in some circumstances, this approach might be the only way to acquire valid data. However, on the other end of the spectrum, there are also situations, perhaps a majority, in which the localization is only relevant when the cellular and tissue organization is intact. I work a lot with brain tissue, in which the membrane integrity is essential for identifying different neuronal elements, and the labeling technique is often intended to help figure out the relationship among those elements. In this case, cell fractionation is certainly not suitable, and even in intact tissue, the morphology can only be compromised to a certain point. Inconveniently, this point may well be beyond the threshold that allows antigen survival or antibody access.
Fortunately, technology has advanced so much thanks to the relentless efforts of many. With the availability of ultrasmall gold conjugates and silver enhancement reagents for EM, the detectability of antigens in fixed, wet tissue with excellent ultrastructure is no longer unreachable. I routinely use 4% paraformaldehyde plus 0.2% glutaraldehyde to perfuse brain and then post fix it with the same fixative for 1 hour. The ultrastructure quality obtained with this fixation is such that there is no severe dilation of the ER and Golgi, very little swelling of mitochondia, and most membranes are still intact. Yet, with this morphology, I have labeled epitopes in the cytoplasm, nuclei, both inside and outside of Golgi and ER, and on both sides of the cell membranes. Too bad one is not allowed to post images on this listserver.
There are two stages involved in the final preservation of ultrastructure during pre-embedding immuno-labeling. The initial fixation determines the starting point of ultrastructure quality and is limited by the tolerance level of the antigens. Subsequently, the immunolabeling procedure itself is responsible for preventing deterioration of the ultrastructure. The latter is closely related with reagents to be used, and has been the focus of reagent improvement for the last few years. Using a single Fab conjugate allows less harsh permeablization of the tissue. A near neutral pH silver enhancement reagent reduces the harm of pre-osmium enhancement on morphology.
Will it work at the EM level if it does at the LM level? Pre-embedding labeling with ultrasmall gold conjugates and silver enhancement has definitely made it easier to carry LM success to the EM level.
Thank you.
Hong
============= Hong Yi Emory Neurology Microscopy Core Laboratory Emory University School of Medicine 6215 Woodruff Memorial Research Building 1639 Pierce Drive Atlanta, GA 30322 Tel: (404) 727-8692 Fax: (404) 727-3157 Email: hyi-at-emory.edu
.. I am just curious: What's the good thing about DM? "Everbody" seems to be complaining about this "image format"(?). - What's the advantage of DM compared to TIFF?
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Have you thought about filtering your bacteria on polycarbonate filters? This method would obviously only work for the SEM.
There was a thread a while ago and I think I copied it from the archives where I searched for the the keyword "filters". Quite a few people were discussing how to prepare bacteria for SEM and nearly all suggested filters.
A typical protocol was: filtering suspension of bacteria through 0.2 um polycarboante filter, remove filter from housing and place into 2.5% glutaraldehyde in buffer, follow with buffer washes, optional post fixation with OsO4, gradient alcohol series (3 x 100 % EtOH) and then either 3 washes in HMDS (hexamethyldisilizane) followed by air drying or critical point drying.
I am working with leukemia cell cultures and pellet the cells for TEM (my cells are easier to pellet) and filter them on polycarbonate filters for SEM.
Regards
Claudia Dr. C. Hayward-Costa School of Life Sciences Kingston University Penrhyn Road, Kingston upon Thames Surrey KT1 2EE, UK 44(0)208 547 2000 x 2240 Email: c.hayward-at-kingston.ac.uk
I am looking at Field Emission SEMs with a view to a lasting and meaningful relationship, and am reminded of the problem of verifying manufacturer's claims about resolution. I dimly recall having been here before, but it is such a long time since I had the luxury of considering SEM purchase that it has become buried in my deep subconscious, alongside other strongly repressed issues like full costs recovery, the Oedipus complex, resentment of academic payscales and Bo Derek.
The more I reflect on resolution the more it looks like a can of worms. For example, it has recently become apparent that some FESEMs can "resolve" 1nm gold on a featureless carbon film under conditions where the manufacturer is only claiming a lesser resolution. This situation must be analogous to the situation where 15nm gold can be imaged by reflection light microscopy, or stars are imaged by eye or telescope. In other words the individual point sources of light (or secondaries) are seen, but are represented by the imaging system as objects larger than their true size. Most manufacturers seem to use gold particles on carbon or mag tape as resolution test objects. I have reservations about this, partly because of the effect described above, which will result from the massive contrast between gold and carbon, but also because it is not a realistic test of performance in the context of low-kV operation and light-element materials and biological samples.
I would therefore be grateful for your experience and advice on some of the following issues:
How should resolution be defined?
Is there a rigorous way of measuring the resolution achieved in a given SEM image?
Is there a rigorous way of establishing the fundamental resolving power of the SEM imaging system? (I think these two questions are different)
What would you recommend as a test object for testing the practical resolution of the sytem in imaging of light-element samples?
Consider the following conditions:
Field width on specimen = 1µm Magnification = 200k at display width of 200mm (8") At a digital image size of 1024x768 pixels (typical basic FESEM display resolution) the pixel size is therefore about 1nm Presumably at this magnification even if the SEM is resolving 1nm (let's assume it is) the display system is inadequate to demonstrate the fact.
Question: How many pixels would be required to image and resolve the 1nm data in this image? The answer to this question is presumably influenced by the criteria employed for establishing the resolution (point to point? peak width at half height? etc) and presumably dictates either the minimum magnification or the minimum digital resolution necessary to test resolution at the approximately 1nm current limit of FEGSEM performance.
I would be grateful for your comments 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
My thanks to everyone for their suggestions about how best to troubleshoot my slow dye sub printer. I can't say that I have gotten a conclusion yet, but I have learned enough to see that taking the printer off the SEM's computer is my best bet. It prints fine from every computer I have tried EXCEPT the SEM one and after some problems I've had with that one, it seems wise to separate this function from that system. That done, all is fine.
Thanks again to all here who were so generous of their time and ideas.
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A very basic point that is almost always overlooked when discussing SEM "resolution," is the difference between "resolution" and "resolving power."
An SEM might have a resolving power of 1nm or so based on the observation of gaps between gold particles on a featureless carbon film, as you point out. Fine--if your job is to look at gold on C.
For the actual 3-D samples most everyone looks at, EVERY micrograph has a DIFFERENT resolution, closer to 10nm, because of secondaries generated by back scattered electrons, edge effects, ... etc. If you want resolutions closer to the instrument's resolving power you have to look at thin TEM specimens without sharp edges and angles.
In our semiconductor lab we have several FESEMs advertised to have resolutions of 1nm that can't resolve 4 and 5 nm layers in a film stack. There is no question that these instruments are superior to non-FESEMs and that today's SEMs are feature-filled and wonderfully reliable--just don't expect 1nm resolution in real images and remember the difference between resolution and resolving power.
So if every instrument has one "resolving power" and a different "resolution" for each picture, to answer your question: the only test of "resolution" for you is to take your bread-and-butter samples to each manufacturer and compare the resulting images. There isn't any theoretical formula that will provide a more meaningful answer.
Ron
Ron Anderson, IBM, Hopewell Jct., New York, USA. anderron-at-us.ibm.com
IBM Analytical Services; http://www.chips.ibm.com/services/asg
We haven't changed the Ar tank yet, so I have not observed the problem you describe. However, we do get Cu contamination periodically in the PIPS due to buildup of contamination on the bottom surface of the shutter (facing the sample). This is easily fixed by removing the shutter and cleaning it with very rough, 320 grit or coarser, grinding paper. The timing may be coincidental with the change of Ar tank, but just in case check that the presure is set correctly in the tank regulator (8 psi) and the gas control valves; a higher pressure may have increased the Ar in the beam and the sputtering from the shutter to the sample. The most common source of contamination from the grid is the grid itself: grid thickness varies, and when it is larger than nominal the grid "shadows" the sample at low milling angles sputtering grid material onto the sample. I hope this helps.
Augusto Morrone Seagate Technology Augusto_A_Morrone-at-notes.seagate.com
"Leah Wajdyk" {leah.wajdyk-at-onsemi.com} on 09/26/2000 05:55:59 PM
To: Microscopy-at-sparc5.microscopy.com cc:
Hello, I have just changed the Ar tank that is used on my PIPS. I now have a hugh amount of contamination from the grid on my TEM samples. The only thing that has changed is the tank. I made sure the guns are aligned and the flow adjusted properly.
Has anyone else had this happen after changing the tank and is there some advice for me?
There is a group here that would like to use digital microscopy to characterize sizes of vesicles obtained by fractionation of myocytes. The problem is to get them to stick to glass coverslips in a non-selective way. Because we had it on hand, we tried poly-L-lysine and letting the vesicles settle overnight at 4C. The proportion of material that remained on the coverslips was very small. I have a few questions: 1) Is it possible that different populations (with different membrane proteins etc.) would adhere differentially to the positively-charged surface? 2) Does anyone have experience with different substrates (e.g. Cel-Tak, silane, albumin) that would be more appropriate. 3) Should we abandon this approach and use the more reliable (?) negative stain?EM route.(I worry about shrinkage artifacts due to air-drying and heavy-metal stains) Any advice will be gratefully considered and references appreciated. Thanks in advance. 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
An additional consideration for any low angle ion milling is to change to Mo grids. They have a much lower sputter rate. They are also thinner, allowing a much lower milling angle on the grid side of the specimen. Using 1 mm diameter grids, and with the feature of interest cnetered in the hole, we routinely mill as low as 3 degrees into the grid hole. They are particularly suitable for specimens which require extended low angle milling, on the order of 2-3 hours.
Phil
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
The Digital Micrograph (DM) format is not a image file format for say in the way that TIFF, PICT, JPEG, GIF, etc. are file formats (some of which use lossy compression). Digital Micrograph, in addition to saving the image (with no reduction in data), saves a lot of experimental header information, for instance calibrations, images size, keywords, magnification, electron energy, TEM operator name, image annotations etc. Many of these can be entered in the Object Info dialog. Also, DM can save multiple images in a single DM file.
Tyrone Daulton
Gunnar Glasser wrote:
} Hi, } } .. I am just curious: What's the good thing about DM? "Everbody" seems to } be complaining about this "image format"(?). - What's the advantage of DM } compared to TIFF? } } Dipl.Ing.(FH) G.Glasser } Elektronenmikroskopie } Max Planck Institut fuer Polymerforschung } Ackermannweg 10 } 55128 Mainz, GERMANY } phone: ++49 +6131 379195 } fax : ++49 +6131 379100 } web: http://www.mpip-mainz.mpg.de/~glasser } } Disclaimer: The above statement is mine alone and does not implicitly represent the position of the MPI-P or the MPG!
-- _____________________________________________________ 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
The Mid-Atlantic Microbeam Analysis Society and the NIST Surface and Microanalysis Science Division will be meeting at the National Institute of Standards and Technology Gaithersburg, MD on
Thursday, October 19, 2000 10:15 am to 3:00 pm NIST Administration Building Lecture Room D
10:30 am Energy-Filtered TEM Jim Bentley Oak Ridge National Laboratory 2000-2001 MAS Tour Speaker
1:00 pm Electron Microprobe Dating of Minerals John Hanchar Department of Earth and Environmental Sciences The George Washington University
For more information, contact John Henry Scott 301-975-4981, johnhenry.scott-at-nist.gov, or Ryna Marinenko 301-975-3901, ryna.marinenko-at-nist.gov Don't forget to bring your MAMAS dues ($5.00)
Hope to see you there, -- John Henry
John Henry J. Scott Bldg 222/Rm A113 NIST Microanalysis Research Group 100 Bureau Drive Stop 8371 (301) 975-4981; (301) 417-1321 FAX Gaithersburg, MD 20899-8371
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
{Subject: .... again: Digital Micrograph??? {From: Gunnar Glasser, glasser-at-mpip-mainz.mpg.de {.. I am just curious: What's the good thing about DM? "Everbody" seems to {be complaining about this "image format"(?). - What's the advantage of DM {compared to TIFF?
It is a proprietary format, no other common program can use it. In an earlier post I say that I save everything in Digital Micrograph format and that is my archival image. If you have properly calibrated the mag on your camera and you spend time putting in keywords, etc, these become a permanent part of the image and you can always go back and get that image. I always try to be as verbose as I can with information. For example, my image name may be: 000927rhinovirus.1 which has today's date and a short sample name. This also allows me to sort files by a name that can be chronologically based. In the tags and keywords I have the pixel size (based on mag), sample name, collaborator's name, sample prep conditions, etc., etc. You can also measure items automatically without having to calculate in your mind that x pixels in the image equals y nm in the sample. You cannot do that with a TIFF or whatever. After I have saved my image in DM format, then I will resave it again, if I want to, in any other format but I always have the untouched original to fall back to.
Norm Olson
******************************************* Norm Olson Senior Research Electron Microscopist Department of Biological Sciences Lilly Hall of Life Sciences Purdue University West Lafayette, IN 47907
by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id JAA09576 for dist-Microscopy; Wed, 27 Sep 2000 09:53:32 -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 JAA09572 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Wed, 27 Sep 2000 09:53:02 -0500 (CDT) 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 JAA09565 for {Microscopy-at-sparc5.microscopy.com} ; Wed, 27 Sep 2000 09:52:46 -0500 (CDT) 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 PAA21211; Wed, 27 Sep 2000 15:46:23 +0100 (BST)
Claudia, do you notice any difference in your cell cultures between HMDS and CPD?
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 Kristine } } Have you thought about filtering your bacteria on polycarbonate } filters? This method would obviously only work for the SEM. } } There was a thread a while ago and I think I copied it from the } archives where I searched for the the keyword "filters". Quite a few } people were discussing how to prepare bacteria for SEM and } nearly all suggested filters. } } A typical protocol was: filtering suspension of bacteria through 0.2 } um polycarboante filter, remove filter from housing and place into } 2.5% glutaraldehyde in buffer, follow with buffer washes, optional } post fixation with OsO4, gradient alcohol series (3 x 100 % EtOH) } and then either 3 washes in HMDS (hexamethyldisilizane) followed } by air drying or critical point drying. } } I am working with leukemia cell cultures and pellet the cells for } TEM (my cells are easier to pellet) and filter them on polycarbonate } filters for SEM. } } Regards } } Claudia } Dr. C. Hayward-Costa } School of Life Sciences } Kingston University } Penrhyn Road, Kingston upon Thames } Surrey KT1 2EE, UK } 44(0)208 547 2000 x 2240 } Email: c.hayward-at-kingston.ac.uk }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
The encoding of the image information in DM is the TIFF format, with the extra header info listed below. It is possible on files containing single images to strip the header info and open them with Photoshop.
Craig Wall
*%*#*%*#*%*#*%*#*%*#*%*#*%*# Disclaimer: The above statements are mine alone and do not implicitly represent the position of the Goodyear Tire & Rubber Co. Craig G. Wall, Ph.D. The Goodyear Tire & Rubber Company 142 Goodyear Boulevard Akron, Ohio 44305 (330)796-2335 (330)796-3304 FAX
The Digital Micrograph (DM) format is not a image file format for say in the way that TIFF, PICT, JPEG, GIF, etc. are file formats (some of which use lossy compression). Digital Micrograph, in addition to saving the image (with no reduction in data), saves a lot of experimental header information, for instance calibrations, images size, keywords, magnification, electron energy, TEM operator name, image annotations etc. Many of these can be entered in the Object Info dialog. Also, DM can save multiple images in a single DM file.
Tyrone Daulton
Gunnar Glasser wrote:
} Hi, } } .. I am just curious: What's the good thing about DM? "Everbody" seems to } be complaining about this "image format"(?). - What's the advantage of DM } compared to TIFF? } } Dipl.Ing.(FH) G.Glasser } Elektronenmikroskopie } Max Planck Institut fuer Polymerforschung } Ackermannweg 10 } 55128 Mainz, GERMANY } phone: ++49 +6131 379195 } fax : ++49 +6131 379100 } web: http://www.mpip-mainz.mpg.de/~glasser } } Disclaimer: The above statement is mine alone and does not implicitly represent the position of the MPI-P or the MPG!
-- _____________________________________________________ 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
Hi, anybody has experiences on small angle(or 90 degree) cleavage method of making cross section TEM samples with MgO substrates? Any information would be appreciated.
---------- } From: Norm Olson {nho-at-bilbo.bio.purdue.edu} } To: MSA {Microscopy-at-sparc5.microscopy.com} } Subject: Fwd: .... again: Digital Micrograph??? } Date: September 27, 2000 11:35 AM }
} } } {Subject: .... again: Digital Micrograph??? } {From: Gunnar Glasser, glasser-at-mpip-mainz.mpg.de } {.. I am just curious: What's the good thing about DM? "Everbody" seems } to } {be complaining about this "image format"(?). - What's the advantage of } DM } {compared to TIFF? } } It is a proprietary format, no other common program can use it. In an } earlier post I say that I save everything in Digital Micrograph format } and that is my archival image. If you have properly calibrated the mag } on your camera and you spend time putting in keywords, etc, these become } a permanent part of the image and you can always go back and get that } image. I always try to be as verbose as I can with information. For } example, my image name may be: } 000927rhinovirus.1 which has today's date and a short sample name. ................After I have saved my image in DM format, then } I will resave it again, if I want to, in any other format but I always } have the untouched original to fall back to. } } Norm Olson }
For what it's worth, our NORAN Voyager system has its own proprietary image format called a .grey (theoretically readable only by Voyager systems), which also incorporates important information such as pixel size, magnification, date, and whatever other notes you wish to include. I'm told by one of my colleagues that with a little creative backstripping of headers, and a foreknowledge of the size of the file, you can open these things in Photoshop, or CorelDraw or whatever. So far I haven't really had to do anything like that - our images all get stored both as .greys, and as .tiffs. With the Voyager software, you can always make a new .tiff from a .grey, but you can't make a .grey from a .tiff. These proprietary image formats can be a bit of a pain sometimes, but at least they do allow the inclusion of all that extraneous information. In our lab, when a job is finished, the client gets a CD containing all their images in both formats and I make a duplicate for myself, as an archive. Theoretically, the client can't look at the .greys. The client's CD then becomes an off-site archive copy for me, should mine turn out to be a dud (hasn't happened yet, but I remain pessimistic). If the client is from "outside", there may be ownership/confidentiality issues involved, so I always check to see if they mind me keeping an archive for them. So far, they've all been happy to have me keep a copy, just in case. But then, I'm not doing any work for military or spy agencies, or even big corporations, or anybody like that.
F.C. Thomas Geological Survey of Canada (Atlantic) Bedford Institute of Oceanography Dartmouth, Nova Scotia Canada B2Y 4A2
"William F. Tivol" wrote: } My experience with Peltiers does not involve a CCD camera, but it may be } instructive nonetheless. When we wanted to provide a constant temperature for } the chemicals in the darkroom--which required sometimes heating the water and at } other times cooling it--we decided that Peltiers were the best solution. We } bought a commercial thermoregulating unit which sent current of the appropriate } polarity through the Peltiers to adjust the temperature to the desired setpoint. } The Peltiers failed at an expensive rate, and we found that the thermoregulator } was such that it provided a specific current, which it turned on and off in a } duty cycle (the greater the temperature difference, the greater fraction of time } the current was on). The sudden changes in current were the cause of the } Peltiers' demise.
Sorry, but that seems not to be the case.
http://www.tellurex.com/resource/txfaq.htm
"Can I use pulse-width modulation to control my Peltier device if I keep the voltage at VMax or below? Yes, and this is one of the most electrically-efficient ways to control voltage to your device—although you must observe some precautions."
And they suggest 2kHz PWM frequency.
I think your temperature regulator supplied too much voltage or current to the peltier, causing overheating and destruction. Just a guess.
} We were also warned that moisture in the Peltiers could lead to an } electrochemical process which caused corrosion and subsequent failure.
This is valid. Water inside peltier will cause both mechanical trouble (freezing inside peltier), and corrosion problems etc..
To the original poster: I _really_ think you should think of some way to make certain that there either is water flow _always_ to the peltier, or that power to peltier (and camera?) is switched off at power/flow failure. The latter (switch off) should be easier. The peltier _WILL_ die from overheating if there is not sufficient cooling. It might kill your camera by overheating it as well.
If you use just water (not glycol or something) as coolant in recirculators, perhaps you could add redundancy by adding a couple of magnetic valves to the peltier cooling lines so that while there is mains current, the valves lets recirculator water to flow through peltier, and otherwise let water from water mains to flow through peltier and to sink. Something like this:
water mains } ----------- valve1 -----o | o--------} peltier in | recirculator out } ------ valve2 -----o
recirculator in {------- valve3 -----o | o-------- { peltier out | sink {------------------ valve4 -----o
Where: valve1: normally open (NO = conducts when coil NOT powered) valve2: normally closed (NC = conducts when coil IS powered) valve3: normally closed valve4: normally open
Just connect all valve coils in parallel and to circuit below.
The idea is to switch on the system (ie. recirculator flow) with the button. With relay switched on, contacts 1 and 2 are closed and power goes to valves. Next, when mains power goes off, relay is nolonger switched on, and no power goes to valve coils (thus mains water cooling). User has to press button again to put relay on - return of mains power won't do this.
Total cost for valves etc. about 100usd or less.
Of course, if you are using glycol or some other chemical, it might well be that regulations forbid you from mixing piping of water mains with your glycol piping, because of danger of contaminating water mains.
STILL, I really think it would be electrically trivial to make certain that the camera does NOT switch on when there is no coolant flow (or simpler yet, no mains - just use circuit above with camera connected instead of valve coils). A simple pressure or flow switch at coolant line, and power to camera via the pressure/flow switch will be easy.. This should be pretty fool-proof protection for the camera in _any_ case of coolant flow failure. I think sooner or later you _will_ have coolant failure without mains failure (recirculator pump failure or something), and this should be anticipated in your design. Of course, you could as well add flow/pressure switch NOcontact in series with relaycoil in circuit above.
} Date: Wed, 27 Sep 2000 10:04:02 -0500 } From: Andrew Ochalski {aochalsk-at-science.uottawa.ca} } To: Microscopy-at-sparc5.microscopy.com } Subject: Sticking vesicles to glass coverslips } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 Listmembers, } } There is a group here that would like to use digital microscopy to characterize sizes of } vesicles obtained by fractionation of myocytes. The problem is to get them to stick to } glass coverslips in a non-selective way. Because we had it on hand, we tried poly-L-lysine and } letting the vesicles settle overnight at 4C. The proportion of material that remained } on the coverslips was very small. I have a few questions: } 1) Is it possible that different populations (with different membrane proteins etc.) would adhere } differentially to the positively-charged surface? } 2) Does anyone have experience with different substrates (e.g. Cel-Tak, silane, albumin) that would } be more appropriate. } 3) Should we abandon this approach and use the more reliable (?) negative stain?EM route.(I worry } about shrinkage artifacts due to air-drying and heavy-metal stains) } Any advice will be gratefully considered and references appreciated. Thanks in advance. } 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 } Not sure if this will help, but it's worth a try. Many years ago, we did SEM of red blood cells and needed to have them stuck to a substrate. We treated glass coverslips with collagen, added the RBCs, and glued them down with glutaraldehyde. They stuck very well.
Coverslips Chip one edge of a cover slip (we used 9 x 9 mm) in a specific corner so that you will be able to know which side is up if it gets flipped over. _______ | | | | | c |______|
You can do this by holding the cover slip between thumb and forefinger of one hand and gently dragging the tips of an old pair of EM forceps across the edge at a downward angle. We had about 70% sucess without breaking.
Acetone clean cover slips and let air dry. Put them into a moist chamber (Petri dish with wet filter paper) and add a large drop of 1% aqueous collagen. Note the orientation of your chip mark on the cover slip. Don't let the drop flow over the sides, but just sit there and rounded up.
Incubate 30 min.
Remove drop of collagen with Pasteur pipette for future use.
Wash coverslips well with dH2O. Can use right away or store dried for future use.
Add membrane prep, cells, etc., without letting drop run over the sides. (Surface tension will allow you to cover the glass with a rounded-up drop.) Do not add too much sample; leave enough room to add more fluid later. Let sit 30 min.
Then add to the suspension a drop of concentrated glutaraldehyde. We added 50 ul of 25% glut in PBS to the cell suspension without letting the drop of suspension flow over the coverslip edge. Do this very slowly so as not to disturb the stuck material. (The glut is a di-aldehyde that links things together; in your case, hopefully, the material to the cover slip. Immunologists use this method to link antigens or antibodies to solid substrates.)
Let sit another 30 min, then GENTLY wash off the suspension. (We then critical point dried for SEM, but you can do your routine prep from here.)
Collagen This stuff comes as a flaky powder. Weigh out 0.1 gram and add to 10 ml dH-at-O. Stir overnight in the cold (cold to minimize bacterial growth). Filter through Whatman or any kind of filter paper to remove undissolved chunks. Keep refrigerated. Can autoclave if long term storage desired.
Good luck.
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
Department of Cellular & Structural Biology The University of Texas Health Science Center at San Antonio
The Department of Cellular and Structural Biology is recruiting faculty for tenure track positions at the Assistant, Associate and Full Professor level. The Department is particularly interested in recruiting qualified applicants with interests in cell and molecular biology, aging, cancer, immunology or neurobiology, and who utilize optical imaging approaches as a main research tool. The ability/willingness to teach in one of the Department's professional and/or graduate courses is also required.
Interested individuals should submit their curriculum vitae, a statement of research and teaching interests and experience, and three letters of recommendation to:
Dr. Ellen Kraig Chair, Cellular & Structural Biology Faculty Search Committee The University of Texas Health Science Center 7703 Floyd Curl Drive Mail Code 7762 San Antonio, Texas 78229-3900
Applications received prior to December 1, 2000 will be considered.
Further information about the Department and its optical imaging facility can be found at: http://www.uthscsa.edu/csb/csbhome.html
The University of Texas is an Equal Employment Opportunity/Affirmative Action Employer.
Dear sirs Respectfuly,i beg to let you now that i am a 26 year old Iranian researcher.Since 1993 to present i have been ivolved in research activities.I am the director of an Iranian research institute .One of the main activities of whichis to operate departments of electron microscopes.Also i am responsible electron microscope department of the Isfahan university of medical scienceis(one of the most reputed universities of Iran).Meanwhile i am in close relations with other electron microscope departments all through the country. Unfortunately hoewer, in Iran ,in spite of the existence of a great number of electron microscopes,the science of electron microscopy is not very developed and most of the electron microscope departments are eather inactive or have little activities.Therefore i intend to stablish the first center for development of electron microscopy technology in Iran by the assistance of the private sector.In this connection i will very much appreciate receiving assistance from your center.I intend to pass a complementary course in the field of electron microscopy at your center and then developing the above mentioned center in one of the cities or international free zones of Iran under support and assistance of your center .The programs of this center will include accepting electron microscopy students from Iran or your country, instruction of applied electron microscopy,instruction of electron microscopy courses at different Iranian universities,helding electron microscopy workshops,operating electron microscopy departments,supplying electron microscopy equipments and consumption electron microscopy materials,performing different electron microscopy techniques,praviding expanded electron microscopy services to scientific researche, industrial and medical diagnosis departments. With regard to the acceptance that receives the activities of our institute in the field of electron microscopy it seems that in the future the activities of the center for development of electron microscopy technology will be welcomed in a similar way by both public and private sectors. Therfore i would like to ask you to advise me in the above mentional fields.port of my executive and research cactivities are as follows: 1.Director of Isfahan's Sinapooyesh institute of research . 2.Responsibility of electron microscope department of Isfahan university of medical scienceis. 3.Responsibility of processing system of electron microscope department at Shahid Beheshti university of medical scienceisof Tehran. 4.Membership of the Iranian national standard commitee. 5.Iam the first provider of nanotechnology in Iran. 6.Receiving the appreciation medallion of the twelve th Kharazmi research festival in Iran. 7.Recieving the first prize of the national young researchers festival in Iran. 8.Reseiving the special prize at the national congress of the security of communications of Iran. 9.Instruction of electron microscopy courses at MSc and phD levels in reputed universitesof Iran. 10.Participaton in several national and international research congresses and presenting several scientific articles and activities. Meanwhile,from 1993 to 1995 i have studied in the field of medical diagnosis and then from 1995 to 1998 i have learnt electron microscopy techniques at Isfahan university of medical scienceis .since 1998 to present i am pursuing my studies in the field of electronic engineering. Thank you very much in advance for your sincer assistance and advice in this regard. sincerely yours Iman moradi
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There is No advantage to digital micrograph over other formats. It is a format supported by Gatan by force feeding it to users of their software. You can jump through some hoops and get it to export images in other formats though.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ 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 Wed, 27 Sep 2000, Gunnar Glasser 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 just curious: What's the good thing about DM? "Everbody" seems to } be complaining about this "image format"(?). - What's the advantage of DM } compared to TIFF? } } } Dipl.Ing.(FH) G.Glasser } Elektronenmikroskopie } Max Planck Institut fuer Polymerforschung } Ackermannweg 10 } 55128 Mainz, GERMANY } phone: ++49 +6131 379195 } fax : ++49 +6131 379100 } web: http://www.mpip-mainz.mpg.de/~glasser } } Disclaimer: The above statement is mine alone and does not implicitly represent the position of the MPI-P or the MPG! } }
Do any of you have experience with the GATAN DigiScan hardware and Digital Micrograph for converting a non-digital sem to a digital sem? We have a JEOL T200 we are considering for conversion. Your responses are greatly appreciated.
Sincerely, Ken Tiekotter, Adjunct Professor The University of Portland Department of Biology 5000 N. Willamette Blvd. Portland, OR 97203
Tyrone is correct in saying that some of the file formats in circulation (for example JPEG) use lossy compression and one should be careful with them. TIFF, however, is not necessarily one of them. TIFF usually allows to select compression options, which can include lossless compression (LZW). Also, TIFF is a flexible format where additional information can be stored in so-called "tags" (hence the name: Tagged Image File Format). There are public tags (like image size, etc), but each software can implement private tags, which are just ignored by other software. That way it is possible to incorporate additional information into the file itself without making it proprietary. That's the strategy we have pursued. TIFF, apparently like DM, can also store multiple pages in one file. Other formats, like BMP, only encode the actual image data and one needs additional mechanisms like a database to keep track of other elements like keywords.
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: Tyrone L. Daulton [mailto:tdaulton-at-nrlssc.navy.mil] Sent: Wednesday, September 27, 2000 8:24 AM To: Microscopy-at-sparc5.microscopy.com
The Digital Micrograph (DM) format is not a image file format for say in the way that TIFF, PICT, JPEG, GIF, etc. are file formats (some of which use lossy compression). Digital Micrograph, in addition to saving the image (with no reduction in data), saves a lot of experimental header information, for instance calibrations, images size, keywords, magnification, electron energy, TEM operator name, image annotations etc. Many of these can be entered in the Object Info dialog. Also, DM can save multiple images in a single DM file.
Tyrone Daulton
Gunnar Glasser wrote:
} Hi, } } .. I am just curious: What's the good thing about DM? "Everbody" seems to } be complaining about this "image format"(?). - What's the advantage of DM } compared to TIFF? } } Dipl.Ing.(FH) G.Glasser } Elektronenmikroskopie } Max Planck Institut fuer Polymerforschung } Ackermannweg 10 } 55128 Mainz, GERMANY } phone: ++49 +6131 379195 } fax : ++49 +6131 379100 } web: http://www.mpip-mainz.mpg.de/~glasser } } Disclaimer: The above statement is mine alone and does not implicitly represent the position of the MPI-P or the MPG!
-- _____________________________________________________ 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
The prices I have heard for field canceling systems has ranged from ~US$15k to $30k. A list of suppliers is available at "www.jcnabity.com/links.htm#Magnetic Field Cancellation".
Joe _________________________________________ Joe Nabity, Ph.D. JC Nabity Lithography Systems E-Beam Lithography using Commercial SEMs & STEMs PO Box 5354, Bozeman, MT 59717 USA Voice: (406) 587-0848 FAX: (406) 586-9514 E-mail: info-at-jcnabity.com Web: www.jcnabity.com
In a message dated 9/27/2000 3:30:27 AM Mountain Daylight Time, r-holdford-at-ti.com writes:
} Mark: I can't comment on a mu-metal shield, but have used field-cancelling } systems on SEMs and FIBs to great effect. I can't remember what we paid for } ours, so I don't know if such a system would cheaper than the shield. But } if } you're curious try this URL: http://www.spicerconsulting.com/. This } company } is in the UK, but I think they have people in the Americas. }
I have surplus a LKB Ultratome V microtome in full working order. This would be available free to an academic institution (you pay shipping) or for a reasonable cost (offers?) to a commercial lab. It is an older instrument but still produces excellent thin sections at room temperature. We have replaced it with a new instrument with cryogenic capability.
Obviously, the closer you are to Cambridge, Massachusetts, the easier shipping would be!
Contact me directly if you have interest or any questions.
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: (+) 1-617-258-6478 **
I am interested in synchronising Hela cells and collecting samples in each stage of Mitosis. Does anybody have a protocol and the times at which each stage is reached after release from G1?
Ken Blight Senior Scientific Officer Electron Microscopy Unit Imperial Cancer Research Fund London England
In an early posting I wrongly suggested that the Ar pressure in the PIPS should be set at 8psi:
"...The timing may be coincidental with the change of Ar tank, but just in case check that the presure is set correctly in the tank regulator (8 psi) and the gas control valves; a higher pressure may have increased the Ar .."
Neal Zimmermann, who works in my lab, corrected me: the pressure should be set to 25psi as stated in the Gatan PIPS manual. Fortunately we had it set right, just my memory lapsed.
Augusto
Augusto Morrone Seagate Technology Augusto_A_Morrone-at-notes.seagate.com
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Reply to: RE: Immuno LM/EM Hi Jan,
Thanks for joining in. I was beginning to feel a bit lonely out here. It is interesting, however, to read the many off-line relplies I have been getting about this posting. Obviously many people are struggling with there attempts to understand the many different choices available for immunolabeling cells and tissues. I thank you for the support you have given.
I agree with your posting almost entirely and hope that we might have made our point by now. I have a few extra points to make which will probably end this thread for good (mostly from boredom, I guess).
First, I think we both agree that we should have reliable immunoreagents. I have preferences in the reagents I use, but I know there are other choices available. Use whatever works.
You express a reluctance to using cell fractions, something that someone off-line also disagreed with. I am not saying use poor morphology to get a result, I suggest using the best morphology that gives the result. In some instances the use of purified fractions of organelles has provided important results. Perhaps the number of specialized organelles in a cell system is so low that it is impossible to find sufficient profiles of them in the thin sections we use. If we can positively identify these organelles by some other unique marker, then it is a simple task to section the fraction and apply two antibodies (one being the identifier, one being the experimental). This gives the information we look for and thus allow us to continue, hopefully to label sections of whole cells at a later date. This is just part of the data collection process for the whole project. Again, use what works.
I was being negative about secondary marker antibodies to add a little spice to my posting. I know that many people use these markers but I did want them to question their reliability. There is a tendency to condemn the primary antibody without thinking there may be other causes for signal to be absent. Maybe there is a need for careful monitoring of the gold probes during storage.
I do not know of one published work that has extensively examined the effect of storage on colloidal gold probes (same antigen system, same preparation protocols, antibody-gold compared with protein A-gold, systematic counting of gold particles etc.). If anyone knows of such a study, I would be interested to hear of it. I often use unconjugated secondary antibodies as a bridge for protein A-binding. It is clear that this extra step increases sensitivity and also adds more gold particles to the section (supported by published results from Jan Slot, I think). Although the gold is not so closely associated with the structures I expect to label, I still get results from this sort of labeling.
The problems I have with the use of pre-embedding techniques are the same I had when I first used them in the 1980's (and my morphology was pretty good in those days too). It was always difficult to quantify the result because I could never be completely sure that all the antigens had been totally accessible to the antibodies or visualization reagents. This problem may not be due to size of the particles used, or with permiabilization (which do play a role), but may also be due to other factors that cannot be controlled for, such as biological activity within whole mounts. I am also reluctant to use pre-embedding labeling as a routine protocol because I cannot yet see a way to easily perform multiple labeling experiments. I look forward to being enlightened.
The possible lack of total accessibilty will also make it difficult to prove a negative result (how can I be sure that the vesicle lumen not labeled is because the antigen is not there, and not because the antibody or gold probe is not there?). I know these things are also problems with on-section labeling, but the methods for on section labeling are a little better understood. For this problem I would prepare thinner sections or count the gold particles. It may not improve things but I would be sure there is some possibility for labeling to occur. Limitations are easier to live with when we understand them a little more. Having siad all that, I realize that for some systems, pre-embedding is the way to go. If it works, use it.
I strongly support your advice that we should let the result drive our work and accept that what we find may not fit in with the favorite theory of the lab. The protocols, reagents and equipment available to us for the study of morphology and immunocytochemistry are the most sophisticated we have ever had. For this reason we have the ability to provide completely unambiguous quantitative results that are difficult to ignore. Learning to stand by them is our next step.
Finally, in an attempt to spice things up and to keep this thread going in some form, here are some extra comments to consider. Remember my question many years ago on this listserver when I asked if EM was a science or an art? All of the inital replies were that EM is an art. Only when I asked why we published in scientific journals did I get a few slightly more scientific replies. They didn't convince me too much though. I still think that many EM people see themselves as artists able to perform wonderful things (i.e. miracles) with complicated machines. We are wrong to think this. We are also wrong to think that we are able to provide every researcher who walks through the door with the result they need to finish their grant/paper/report. With the tools available, we can provide unbiased results that will have biological meaning only in the context of what else is known about the system. For immunocytochemical experiments at least, we should really be active participants in the whole project. Non-microscopists need to read this stuff and think about what they expect from EM too.
Best 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 SAA15014 for dist-Microscopy; Thu, 28 Sep 2000 18:30:21 -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 SAA15011 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Thu, 28 Sep 2000 18:29:50 -0500 (CDT) 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 SAA15004 for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 28 Sep 2000 18:29:39 -0500 (CDT) X-Sender: zaluzec-at-ultra5.microscopy.com Message-Id: {v03130301b5f983159703-at-[206.69.208.21]} Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
CENTRAL INSTITUTE FOR THE DEAF, St. Louis, Missouri
announces the following:
New Frontiers in the Amelioration of Hearing Loss
A Conference in Honor of James D. Miller
CID is now accepting poster submissions and registration for this conference in St. Louis, MO, March 22-25, 2001.
The conference will offer two expert seminar tracks, one for cellular and molecular biologists and one for scientists studying applied technologies and methods for the rehabilitation of children and adults with hearing loss. The speakers include some of the most active and progressive researchers working today.
In addition to serving as a forum for cutting-edge research, the conference is designed to bridge the information gap between basic and applied research related to hearing and deafness. Scientists from different tracks will be provided with helpful information during a primer session held before each expert seminar track. Social sessions will bring together diverse professionals to foster new perspectives and ideas. Attendees will have the opportunity to tour new CID campus facilities, including the Fay and Carl Simons Center for Biology of Hearing and Deafness and CID's new "quiet" oral school.
For poster guidelines, preliminary program and registration materials, please contact Sarah Uffman, CID Research Department, 4560 Clayton Avenue, St. Louis, MO 63110, or visit our web site at www.cid.wustl.edu { {http://www.cid.wustl.edu/} http://www.cid.wustl.edu/} .
Jaclynn M. Lett, Research Assistant
Faye and Carl Simons Center for the Biology of Hearing and Deafness Central Institute for the Deaf 4560 Clayton Avenue St. Louis, MO 63110
Mitosis is quite short, so if you get the cells reasonably synchronized (they pass through mitosis in a Gaussian distribution), you can probably catch most if not all stages if you time your collection right.
See our paper in Methods in Enzymology for a synchronization protocol:
Brady, R.C, M.J. Schibler and F. Cabral. 1986. Isolation of mitotic spindles from mammalian cells. Meth. Enzymol. 134: 217-225.
Matthew J. Schibler Ph.D. UCLA Brain Research Institute 1524A Gonda (Goldschmied) Center for Neuroscience and Genetics Los Angeles, CA 90095-1761
-----Original Message----- } From: ken blight [mailto:blight-at-icrf.icnet.uk] Sent: Thursday, September 28, 2000 8:16 AM To: EM information site
I am interested in synchronising Hela cells and collecting samples in each stage of Mitosis. Does anybody have a protocol and the times at which each stage is reached after release from G1?
Ken Blight Senior Scientific Officer Electron Microscopy Unit Imperial Cancer Research Fund London England
The answer I gave Kristine was purely theoretically - because I read about it.
Unfortunately our CPD is out of service since years and so far I have not used HMDS because I still have a few mls of PELDRI II.
Sorry, I can't give you an answer.
Regards
Claudia
Dr. C. Hayward-Costa School of Life Sciences Kingston University Penrhyn Road, Kingston upon Thames Surrey KT1 2EE, UK 44(0)208 547 2000 x 2240 Email: c.hayward-at-kingston.ac.uk
My apologies for using network bandwidth for this, but I can't possibly respond to all the e-mail and phone messages individually. However, the microtome I offered yesterday has already found a good home where it will be productively used by students. Many thanks to all of you who responded.
Tony.
** Anthony J. Garratt-Reed ** MIT Room # 13-1027 ** 77 Massachusetts Avenue ** Cambridge, MA 02139-4307 ** USA ** ** Phone: (+) 1-617-253-4622 ** Fax: (+) 1-617-258-6478 **
Some colleagues are trying to fix plant tissue for LM and possibly confocal microscopy using ETOH and acetic acid--they asked me if I knew what % of glycerol to include to keep the samples from being too brittle. Does anyone know a good protocol for glycerol fixation? They don't want to go the glutaraldehyde &/or osmium route. I remember seeing something about glycerol fixation in an ancient book on plant microtechnique but the book disappeared long ago...
Thanks for bailing me out again, Margaret -- Margaret Dienelt Plant Pathologist
Electron Microscopy Lab FNPRU, National Arboretum ARS, USDA
B. 010A R. 238 BARC-W 10300 Baltimore Avenue Beltsville, MD. 20705
Thanks to all who replied to my query on staff salaries.
To summarize the findings: The ranges that I reported for industry are similar for university facilities, although the universities tend to be in the lower end of each range. The people in higher cost of living regions don't seem to be making that much more than those living in other regions. The actualy salary offerings vary with duties required and experience (as they should), so one should not use just the numbers to compare salaries.
The (sad) bottom line is: Administrators (the person(s) above the lab director) think that staff salaries are adequate.
Thanks, Lucille
******************************************************************* 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 ********************************************************************
I am looking for a replacement part for an LKB Huxley ultramicrotome Mark 2. Does anyone know of an LKB Dealer or possibly have in their lab a spare Suspension Strip Hinge (LKB Part No. W52271-30)? This piece is made of polypropylene rod with 2 thin indented areas in the rod.
Thanks a lot for all the responses. However, it seems that not everbody has the same opinion regarding the possibility of cleaving samples with MgO substrate for x-section TEM. Does anybody have sucessful experices on this? Thanks a lot.
} } } Margaret Brannigan {brannign-at-asrr.arsusda.gov} 09/29 8:31 AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi,
Some colleagues are trying to fix plant tissue for LM and possibly confocal microscopy using ETOH and acetic acid--they asked me if I knew what % of glycerol to include to keep the samples from being too brittle. Does anyone know a good protocol for glycerol fixation? They don't want to go the glutaraldehyde &/or osmium route. I remember seeing something about glycerol fixation in an ancient book on plant microtechnique but the book disappeared long ago...
Thanks for bailing me out again, Margaret -- Margaret Dienelt Plant Pathologist
Electron Microscopy Lab FNPRU, National Arboretum ARS, USDA
B. 010A R. 238 BARC-W 10300 Baltimore Avenue Beltsville, MD. 20705
On Fri, 29 Sep 2000, Margaret Brannigan wrote: } Hi, } } Some colleagues are trying to fix plant tissue for LM and possibly } confocal microscopy using ETOH and acetic acid--they asked me if I } knew what % of glycerol to include to keep the samples from being too } brittle. Does anyone know a good protocol for glycerol fixation?
Steve Ruzin's Plant Microtechnique book gives a brief mention of softening dried specimens in "10% glycerin-ethanol-water; probably 10% glycerol in 50% EtOH" and gives the reference as Peterson et al, 1989(Comparative leaf anatomy of the annual Muhlenbergia. Nord. j. Bot. 8:575-583)
Lots of people have asked me for results of the survey. My initial numbers below were obtained from an industry (not university) inquery. These numbers are higher than what my staff is actually receiving: (an I am therefore trying to fix this).
$30's-40's: for starting engineers with little to no experience that require training mid $40's to mid $50's: for an experienced engineer mid $50's to mid $70's: for a very skilled engineer who can trouble shoot to the component level independently without any back-up help.
Summary of Responses: (I have removed any reference to individual names of people/locations, etc.)
Based on what I have seen, and as reported by the ACS, your ranges are quite acceptable.
} From what I have deduced from my commercial EM engineers, I think you're right on the money. We don't have in house engineers that do this kind of servicing. The in house guys get the middle range that you listed.
Your figures are pretty much on the mark. What you might consider is how technicians (not engineers) fit into the picture. They can be at pay scales $10K-$20K less overall yet still do a very good job. Plus, they tend to stay on the premises much longer than engineers, who seem to seek greater rewards more frequently.
Your numbers look good to me, though I would add supervisory responsibilities and lab management for someone in the upper range.
I am employed at XXX and my job is to service the electron microscopes on campus. I repair at the component level and have not required manufacturer help. I have many instruments I am responsible for. I worked 11+ years for YYY in service and have been here over 4. My salary is low 50's. Even though my job does not require me to do so I work on other things as well. I put a new motor in a print processor today. I often work on microtomes. I know I saved a department about 6K a couple weeks ago by replacing a component in their power supply. After inspection ZZZ would have sold them a new power supply. I fixed a Horse treadmill last year. I love it here. A whole lot of fun.
These salaries seem to be in line (maybe a little higher) with what universities pay technical persons doing these kinds of jobs. In my case, I reluctantly transitioned from industry after 19 years and went from 70's to low 40's. My observation is this, if you are trying to find good people with experience then most likely you would need to offer salaries near the upper end of these ranges. Keep in mind that technology changes, and even experienced people need training and interaction with vendors. I believe this investment will pay off as you are able in time to eliminate expensive service contracts on all instruments. Salary is just part of the package that will attract good people. I believe a good technical person will flourish in the academic environment if they have a good salary, are free from benefit concerns, and have the resources to do the job well.
As a coincidence, one of my technologists pulled your request for survey response off the web to tell me that I was under paying him. I presume that your classification of "engineer" refers to technical degree level people. I manage a group of 6 technical people with about ten state of the art level SEM's, TEM, EMP's and XRD equipment for a very long term contract. Because we reside near Silicon Valley we must reach deeper for starting rates here where entry level housing is about $300K. We cannot compete with the high tech firms that offer stock options, signing bonuses, etc. I just lost a materials scientist who about doubled her salary at a firm in Fremont. Obviously, our starting rates are a little higher than you indicate for engineers/scientists as well as technicians with little difference for degree status. I don't think that we have started anyone in our organization lower than the mid $40's in the past couple of years. Otherwise, we work with a similar salary range for non senior level professionals without significant supervisory responsibility.
We have service contracts on all machines so much of our staff is engaged in teaching etc.. and minor maintenance. In any case the $s sound about the same for us as well... except the third category for which we do not have anyone. But benefits (health, 403b, facilities etc..) are of great interest to most which varies so widely across institutions
I get 65K, and probably should get a hell-of-alot more. But then again, I am such a push-over.
In our Characterization Facility most of the user training and class teaching is done by PhD staffmembers with the title "research associate" or "senior research associate" (both faculty rank, non-tenure track positions). At many universities the analogous staff are called "assistant research professor" or "associate research professor". Some training is done by "junior" or "senior" level "scientists" with bachelors or masters degrees; these are "civil service" staff and members of the state workers union. Our instrument service and some administrative tasks are done by the above people, or by our technician, altogether ranging from bachelors to PhD degrees in various sciences (materials science, physics, chemistry, geology). I'm not sure how this translates to your "staff engineers". Our pay ranges have always struck me as pitifully low, relative to what our staff could make in analogous positions in industry. Here they are: PhD staff: low $40k's to low $60k's including very experienced people throughout the range. bachelors to masters staff: low $30k's to high $40k's. All of these people are capable of independent troubleshooting. Most, however, are not electrical or mechanical engineers. Thus much of our service is contracted to the instrument companies. We are funded almost entirely by our revenue, so down time is very detrimental and in many cases it makes no sense to try to service things ourselves at the level of the guts of electronics. Of course we do our own troubleshooting to identify the problematic instrument component, but in most cases we do not service things in the sense of an electrical engineer. No doubt it is difficult to make comparisons given the diversity of scenarios at the various universities plus industry. I applaud you for attempting.
we have three people who work in the areas you have discussed. They train students and faculty to use the instruments, and they maintain eleven microscopes and sample prep equipment. They also perform administrative tasks. They all have more than 15 years experience with microscopes. They are in the $50K to $60K salary range. Because of their many years of service and the high quality of their work, I consider their pay to be at least 15% lower than it should be. If possible, I would be interested in the results of the other responses.
I am looking for a replacement part for an LKB Huxley ultramicrotome Mark2. Does anyone know of an LKB Dealer or possibly have in their lab a spare Suspension Strip Hinge (LKB Part No. W52271-30)? This piece is made of polypropylene rod with 2 thin indented areas in the rod.
Betty Loraamm Univ. of South Florida Dept. of Biology SCA 110 4202 E. Fowler Ave Tampa, FL 33620-5150 (813)974-2676 Fax:(813)974-3263
My posting to this list server from a few weeks ago stating our problems with the Denton Bench Top Turbo may have been taken out of context. Denton is resolving our problems with the carbon coater. There customer service has been excellent. I want to offer my apologies for any misrepresentation I may have caused them.
Harry Ekstrom Materials Laboratory (602) 231-2744 e-mail: harry.ekstrom-at-honeywell.com
I have been carrying our quantitative analysis of a lead titanate like oxide using a Microspec WDX-2A-3PC on a scanning electron microscope. I have modified the normal experimental procedure and the reproducability improved from 3-5% to 1%. Are there any further modifications of experimental procedure that might further improve the reproducability by another factor of 2?
I am using a compound standard similar to all of the unknowns. All samples are polished well. Count times are long enough to get statistics well better than 1% for each of the 5 elements. Data is collected from each element on standard and then on sample, and thus beam current in the SEM is stable to better than 1%.
I am talking about reproducability of the k ratios being accurate to better than 1%, not about the accuracy of the measured composition. I think the compound standard will produce the best accuracy one can do.
Hello everyone, I am an undergraduate at Miami University, and could use some protocol advise to best achieve the results I am looking for. I have already obtained a protocol for general drosohpila SEM sample preparation, but my specific area of interest is the eye. I was wondering what the best method for acquiring a clean cross-section would be. If you have done this, or could give me some helpful hints it would be greatly appreciated. Thanks, Thomas Litzinger
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Can anybody suggest a supplier from whom I can buy some 0.001" diameter Pt wire? Please include e-mail address if possible.
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
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Sorry all but it looks like I screwed up again. Over the weekend I was editing the filtering software trying to increase the SPAM/Junk Mail rejection capabilities..... Well, I manaaged to stop all mail to the List which means I was 100% effective in filtering the Junk Mail!! ;-)
In any event, I think things are back on track, and working again. If you tried to post anything between Oct 1 and today you should consider it as being permanently lost in the Ether..
Just resubmit them...
BTW, thanks to all those people who started sending me Email saying " I haven't gotten any mail in the last few days" it started me looking for problems. I guess it's good to know you all miss your Daily Microscopy Email-Fix. Almost sounds addictive. (Hmmm... too bad I can't bottle it an make a few $$)
I have an application for separating various size particles using Thin Layer Chromatography. Question, has anyone prepped the polyester backing, with silica gel coating for SEM?
I have a couple hypothetical protocols I could try but I thought I would send this question out to the masses, to see if anyone has had experience examining TLC plates in the SEM (regluar SEM not an ESEM or VPSEM).
Thanks in Advance, Geoff Williams
Electron Microscope Facility Supervisor Biology Department Brooks Hall Central Michigan University Mt. Pleasant, MI 48859
A colleague on campus is having a problem with their ISI Alpha 9 SEM.
They have diagnosed a failure of the high voltage power supply. Specifically, the gun side of the multi-tap power supply has been fried.
So, they are looking for the power supply for the Alpha 9 or an ISI Mini SEM. If anyone has such a component (or even the whole SEM) for sale or donation, please contact me.
Thank you.
John B.
#################################################################### John J. Bozzola, Ph.D., Director IMAGE (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
} The Chemical Technology Division of Argonne National Laboratory has an } opening for a staff microscopist to perform transmission electron } microscopy in support of the nuclear fuel cycle, waste management, and } other programs as required. This person will also develop new processes } and/or equipment and other technology to support the programs, and } interface with sponsors to maintain existing support and develop new } initiatives. } } This position requires a Ph.D. in chemistry, materials science, or } related discipline and 2-5 years of experience (or equivalent). } } For further information or to send resume, contact David Chamberlain by } electronic mail : chamberlain-at-cmt.anl.gov. } }
Any comments on which would make a better detector, Ge or Si ? only PGT sells Ge detectors and I'm aware of the decrease in resolution with Ge detector. Could someone lists or details the benefits of having a Ge detector as oppose to a Si one.
Sincerely
N.V.Vo
N.V.Vo, Ph.D. Intermagnetics General Corporation 450 Duane Avenue Schenectady, New York 12304 Phone: 518 346 1414 x 3107 Fax: 518 346 6080 E-mail: nvo-at-igc.com www.igc.com
Simon I am not a diatom expert, but I know someone who is. Put your question to Professor David Mann at the Royal Botanic Garden Edinburgh. He eats, sleeps and breathes diatoms (maybe I exaggerate a little!). His email address is d.mann-at-rbge.org.uk
good hunting Chris Jeffree
} From: "s_woodland-at-madasafish.com"-at-sparc5.microscopy.com Date sent: Wed, 4 Oct 2000 10:04:39 -0500 To: Microscopy-at-sparc5.microscopy.com
Faculty Position Department of Materials Science and Engineering LEHIGH UNIVERSITY
Lehigh seeks to fill a tenure-track position, at the level of associate or assistant professor, in Materials Science and Engineering. Research interests in any area of materials will be considered but the person selected will have a major commitment to the use of electron microscopy. The Materials Department at Lehigh has research strengths in areas of ceramics, metals, polymers and electronic materials. Details can be found from the faculty pages at the departmental web site: http://www.lehigh.edu/~inmatsci/inmatsci.html. It will be considered an advantage if the research interests of the applicant are related to existing research interests in such a way that collaborative work is promoted. Lehigh runs an outstanding electron microscopy facility. Favorable consideration will be given to candidates whose research will involve substantial use of electron microscopy, especially when the microscopy is innovative rather than routine. Lehigh is committed to recruiting, retaining and tenuring women and members of minority groups. Please submit an application by December 15, 2000, to Sharon Coe, Department of Materials Science and Engineering, Lehigh University, 5 East Packer Avenue, Bethlehem PA 18015-3195, USA (slc6-at-lehigh.edu). To discuss the post contact Alwyn Eades (jae5-at-lehigh.edu) at the same address.
================================================================== Dr. 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 Minnesota Microscopy Society will be holding it's annual Fall Kick-Off event on Thursday, October 12, 2000
This will follow the traditional format of a dinner followed by a talk, which this year is:
"Nature's Jewels and Scientific Tools: Confessions of a Career Diatomist"
by Mark B. Edlund from the Science museum of Minnesota.
Abstract:
Diatoms are single-celled or colonial micro-organisms that possess a cell wall of ornamented opaline silica (biologically-produced glass) and are commonly referred to as the golden-brown algae. It is the wall ornamentation and shape that are used to identify the nearly 25,000 species of living diatoms. Diatoms were "discovered" within the first hundred years of microscopy; Leeuwenhoek himself may be credited with one of the earliest illustrations (1703). Since that time, diatoms have been the jewels, the postage stamps, and the tools of amateur and professional diatomists. Microscopical societies burgeoned in the latter half of the 19th century with members actively exchanging diatomaceous material, slides, and methods and discussing publications and exsicattae. Diatoms figured strongly in the development of light microscopy as test organisms for resolving power; even today, many microscopists have an Amphipleura pellucida slide in the drawer. Transmission electron microscopy revealed much-needed knowledge on the cytology and mechanism of cell wall formation in diatoms, but it has been the scanning electron microscope that has truly changed the field. Ultrastructural details seen in the SEM coupled with "rediscovery" of beautiful cytological work done in the late 1800's and early 1900's radically changed our modern understanding of diatom systematics. Lastly, diatoms have proved to be excellent indicators of water quality and environmental change. Whereas descriptive and biodiversity studies are still common, applied methods now dominate the current scientific efforts.
Speaker: Mark B. Edlund, Research Scientist Science Museum of Minnesota
Mark Edlund has a PhD in Natural Resources from the University of Michigan, and has been specializing in diatoms for the last ten years. Besides field work in the United States, he has done work at Lake Baikal in Siberia, and recently in Mongolia.
Program: 5:30-6:00 PM Social hour 6:00-7:00 PM Dinner 7:00-7:15 PM Business meeting 7:15-8:15 PM Speaker
During the social hour, wine, cheese and crackers will be served. The cost of the meal is $20 per person.
Location: Cherrywood Room, 2nd Floor, St. Paul Students Cente,r St Paul Campus, University of Minnesota, 2017 Buford Ave., St. Paul
Make Your Reservations Today
An accurate head count for the dinner is an absolute necessity. Contact Stuart McKernan by Friday, October 6, to make your reservation (612-624-6009, or stuartm-at-tc.umn.edu).
__________________ Stuart McKernan stuartm-at-tc.umn.edu 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
You might try a swimming pool supply store. Look for Diatomaceous Earth filter media. As to preparation... Techniques will depend on the type of microscopy employed. If using a high vacuum electron microscope, an electrically conductive film will need to be applied to the diatoms which have been (typically) sprinkled onto double faced tape. The tape will not have to be conductive (if it is - so much the better) if the whole mount is conductively coated.
Woody White McDermott Technology, Inc. http://www.mtiresearch.com/
Is there a problem with the server? I haven't received any messages in the last couple of days. I checked with our server and everything seems to be O.K. Did my address get taken off by accident? In case it was my address is: prutledge-at-ars.usda.gov Thanks,
I would try the two following companies as possible sources of fine platinum wire:
GoodFellow, P.O. Box 628, Doylkestown, PA 18901-9975 FAX: 1-800-283-2020
Refining Systems Inc., P. O. Box 72446, Las Vegas, NV 89170 FAX: 702-368-0933
Both sell high purity and specialty metals in a variety of sizes and shapes. Good luck, WCB
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
At 10:04 AM -0500 10/4/00, "s_woodland-at-madasafish.com"-at-sparc5.microscopy.com wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Have you tried Carolina Biological? They have all kinds of pre-mounted slides for sale.
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
Anyone know of a easy, simple way to produce nice powder diffraction samples for illustrating the powder diffraction method for EM students? We use the International Centre for Diffraction Data For student identification of unknown samples. I have tried several methods that usually result in non-electron transparent samples (our TEM KV max is only 120). Any ideas?
_______________________________
Bill Carmichael Electron Microscopy Faculty
Madison Area Technical College 3550 Anderson St. Madison, WI 53704 608-243-4309
Macalester College (St. Paul, Minnesota) has a job opening which involves looking for someone with a microscopy background (as well as electronics skills). The job description is posted at: http://www.macalester.edu/~hr/jobs/job551.html
The description is quite brief, but Macalester has both a Zeiss EM109 TEM and a Zeiss DSM960A SEM. Maintenance and instruction for students/faculty on these machines will be one aspect of this position.
Yes, indeed! You can buy a sack of the stuff there that will keep you busy for all of eternity!
Also check with your neighbors where you live if they have pools and one of them might spare a handful!
Ed Sharpe archivist for SMECC
{ { Subj: Re: Ask a Microscopist - Diatomes Date: 10/4/00 3:20:58 PM US Mountain Standard Time From: nwwhite-at-mcdermott.com (White, Woody N.) To: woodland-at-madasafish.com ('woodland-at-madasafish.com'), Microscopy-at-sparc5.microscopy.com ('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 have not tried this source, but have heard:
You might try a swimming pool supply store. Look for Diatomaceous Earth filter media. As to preparation... Techniques will depend on the type of microscopy employed. If using a high vacuum electron microscope, an electrically conductive film will need to be applied to the diatoms which have been (typically) sprinkled onto double faced tape. The tape will not have to be conductive (if it is - so much the better) if the whole mount is conductively coated.
Woody White McDermott Technology, Inc. http://www.mtiresearch.com/
Take an oxide powder, grind it up with mortar and pestle and take a carbon coated grid and swipe it across the fines. You can also use two glass slides to grind the powder up.
You can evaporate a number of materials onto a carbon coated grid or onto a cleaved NaCl sample and then float that off on water onto a grid.
You could smoke a Mo wire in air to provide crystals if you leave it in the smoke long enough, you will probably have enough to cover the grid. Heat the wire across the terminals of an evaporator or heat with a torch to generate the white smoke. You could also burn some Mg and get MgO crystals.
If you don't have sufficient particles in a single diffraction pattern to give you rings, take multiple exposures from different areas on the same piece of film.
Incidentally, the MoO3 samples will provide you with a rotation calibration 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)
} -----Original Message----- } From: William Carmichael [mailto:wcarmichael-at-madison.tec.wi.us] } Sent: Wednesday, October 04, 2000 5:31 PM } To: microscopy-at-sparc5.microscopy.com } Subject: powder TEM diffraction sample 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 } -------------------------------------------------------------- } ---------. } } } Anyone know of a easy, simple way to produce nice powder } diffraction samples for illustrating the powder diffraction } method for EM students? We use the International Centre for } Diffraction Data For student identification of unknown } samples. I have tried several methods that usually result in } non-electron transparent samples (our TEM KV max is only } 120). Any ideas? } } } _______________________________ } } Bill Carmichael } Electron Microscopy Faculty } } Madison Area Technical College } 3550 Anderson St. } Madison, WI 53704 } 608-243-4309 } } wcarmichael-at-madison.tec.wi.us } http://electron-microscopy.madison.tec.wi.us } } }
What is the difference between diatoms and radiolaria (Actinopods)? They "look" the same to me.
gary g.
At 12:17 PM 10/4/00, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} --------------------------------------------------------------------------- } } Email: s_woodland-at-madasafish.com } Name: Simon Woodland } School: Kaskenmoor School } } Question: Where can I get some diatoms to put under our microscope? How } should they be prepared and mounted. } } ---------------------------------------------------------------------------
Simon -
You can go to a big home supply store and ask for "diatomaceous earth"; it's used for a lot of filtering applications. You'll get a big sack, so you can use it up by ordering the Lawrence Hall of Science/GEMS manual "River Cutters" (see the Prtoject MICRO URL below), which uses the same materiel to make a model stream bed. Or you can order a prepared slide from a variety of suppliers, Or you can contact Joe Neilly {joe.p.neilly-at-abbott.com} , keeper of Project MICRO's sandpile, and ask for samples of foraminefera sand or the very exotic "star sand". Preparation? Minimal. "Microscopic Explorations", MSA's GEMS manual, will tell you how to make filecard and tape slides that will work well.
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'm trying to solve a high resolution noise problem with a XL-30 Phillips FESEM. The room that it is in was recently checked and was ok, accept for some acoustic noise. To fix the noise I put up 3 inch sound proofing foam and quieter ducting for the air condition. The noise showed up in an image by a streak about 5 jaggies. After putting up the foam the noise was reduced, but it is still there. The noise is the same for 4mm working distance or a 10mm working distance.
There's a *lot* of difference. About that between a Redwood and an Elephant. Or more. And as distinctive.
Diatoms are about as good a plant as can be found among the protistans. They look like more-or-less fancy silicate pillboxes with lots of little holes.
Radiolaria are amoeboid beasts with a complicated silicate internal "skeleton". They tend to be nested, holey spheres, usually with a more-or-less complex profusion of spikes, needles, and the like.
Crude descriptions, but brief. The best way to see the difference is to get a copy of Ernst Haeckel's book published by Dover -- "Arts Forms in Nature". Beautiful drawings of many different organisms, including Radiolaria and Diatoms.
Phil
} What is the difference between diatoms and radiolaria (Actinopods)? } They "look" the same to me. } } gary g. } -- *** Be famous! Send a Tech Tip or article to Microscopy Today! *** Philip Oshel Technical Editor, Microscopy Today P.O. Box 620068 Middleton, WI 53562-0068 Voice: days (608) 263-4162, evening (608) 833-2885 Fax (608) 836-1969 (please make sure my name is on any fax)
Address for UPS, FedEx, or other couriers: Dept. of Animal Health and Biomedical Sciences University of Wisconsin 1656 Linden Drive Madison, WI 53706-1581 fax: (608) 262-7420 (dept. fax)
What certified (NIST traceable?) TEM mag. calibration samples are available for 20K to over 100K range? Diffraction grating replica sample will not provide sufficient accuracy- too few grating periods "fit" in the image at higher magnifications.
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax
Try Wards Natural History Establishment in upstate New York. The exact address escapes me.
Sam Purdy TechnicalCenter National Steel Corp Trenton MI
} ---------- } From: "s_woodland-at-madasafish.com"-at-sparc5.microscopy.com } Sent: 4, October 2000, 11:04 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Ask-A-Microscopist: Diatoms } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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: s_woodland-at-madasafish.com } Name: Simon Woodland } School: Kaskenmoor School } } Question: Where can I get some diatoms to put under our microscope? How } should they be prepared and mounted. } } -------------------------------------------------------------------------- } - } } }
We have the problems with our JEM-4000EX: the ion pump SIP-1 is not OK! The possible causes are: 1/- leak in vacuum system (after bake out of the SIP); 2/-some contamination into SIP (this SIP was go OK more 40,000 hour); 3/- ??? Does anyone know how to make testing of this situation?
How do to demount the pump, what order of actions?
I am not currently part of the list, so please mail to me directly.
-- Best regards, Anton Gutakovskii mailto:gut-at-thermo.isp.nsc.ru
Yes, mounting the Coolpix 990 to any photomicroscope is absolutely simple via the usual C-mount connection. For the coolpix you have to buy at NIKON the microscope adapter ( here in germany this adapter is around 1000.- DM or approx. 450.- US $; adapter is not listed in Nikon catalogue; contact the microscope section of Nikon) fitting on one side to the thread at the front of the lens and on the other side to your microscope-specific C-mount adapter ( c-mount adapter for Zeiss: buy the more expensive one which has the possibility of correction of the focus plane). The fantastic thing is that you can focus not only via the LCD- screen of the camera ( possible, but not so easy in my opinion) but via an external TV-Monitor in live modus, yielding really smashing results with a resolution satisfying publication needs. With my Coolpix sometimes the camera "hangs up" when the Auto- power off mode is activated (de-activating it helps). Does anybody have this problem as well ?? ( Contact me off-line). What is lacking for the Coolpix 990 at the moment is a cable release, but the Nikon people here in germany tell me it will be available at the end of the year (probably very expensive).
All in all, the Coolpix is the first digital camera under 5000 US-$ I have used or tested which is fully satisfying me in my daily lab routine (photomicroscope (even fluorescence), binocular, makro-photographs) especially due to its simple and clear operation, the possibility to adjust everything (speed, aperture, focus, etc.) manually and its excellent results.
Peter Heimann
************************************************** please reply to: peter.heimann-at-biologie.uni-bielefeld.de ("E-MAIL")
Dr. Peter Heimann Developmental Biology & Molecular Pathology; W7-107 University of Bielefeld D 33501 Bielefeld / Germany phone: xx49(0)521-106-5628 / 5627 FAX : xx49(0)521-106-5654 WEB-Site: http://www.uni-bielefeld.de/biologie/Entwicklungsbiologie WEB-Site: http://www.uni-bielefeld.de/SFB549 ******************************************************
Wednesday 15 November 2000, Moredun Institute, Edinburgh.
The annual symposium is supported by all area groups in Scotland. The programme for this meeting is comprised of the following three invited talks together with seven contributed talks on a wide range of microscopy with general appeal and a poster display. As in recent years, a comprehensive trade exhibition by leading companies will be present. The Trade Exhibition, Poster Display and Catering will all be together in one central area of this excellent venue.
15/11/2000 Programme
Multi-photon microscopy. Professor Alison Gurney, Dept.of Physiology and Pharmacology, University of Strathclyde.
Ultrastructure eggshell quality assessment: a way forward for captive breeding programmes. Avril Edmond, Dept. Veterinary Anatomy, Glasgow.
Non-invasive imaging of plant tissues using the confocal laser scanning microscope (CLSM) Dr. Karl Oparka, Unit of Cell Biology, SCRI, Dundee.
Functional dynamics of plasmodesmata in sink/source transition leaves. Alison Roberts, Unit of Cell Biology, SCRI, Dundee
COSMIC - a new interdisciplinary facility for advanced spectroscopy, micromanipulation and imaging. Professor Wilson Poon, Dept. of Physics and Astronomy, University of Edinburgh.
Imaging intracellular location of psychoactive drugs using fluorescence microscopy. Richard Horobin, IBLS, Glasgow.
Antigen retrieval allows improved visualisation of V-ATPase immuno-reactivity. Susan Lindsay, Caledonian University, Glasgow.
Techniques in Immunocytochemistry. Peter Jackson, Research Histology Unit, Dept. of Histopathology and Molecular Pathology, Leeds General Infirmary. Talk sponsored by DACO Ltd.
See web site for abstracts from previous meetings
http://www.abdn.ac.uk/emunit/smg99.htm
Kevin Mackenzie Electron Microscope unit Dept Zoology University of Aberdeen Tillydrone avenue Aberdeen AB24 2TZ ----------------- Tel 01224 272847 Fax 01224 272396 email k.s.mackenzie-at-abdn.ac.uk
I hope this isn't seen as an overt advert on the list server, I just want to let people know that this new book is available and hope it may be of general interest. Rest assured that I and my fellow co-authors get very little of the sales proceeds!
A.Patrick Gunning IFR Norwich
ATOMIC FORCE MICROSCOPY FOR BIOLOGISTS
by V J Morris, A R Kirby & A P Gunning
Atomic force microscopy (AFM) is part of a range of emerging microscopic methods for biologists which offer the magnification range of both the light and electron microscope, but allow imaging under the 'natural' conditions usually associated with the light microscope. To biologists AFM offers the prospect of high resolution images of biological material, images of molecules and their interactions even under physiological conditions, and the study of molecular processes in living systems. This book provides a realistic appreciation of the advantages and limitations of the technique and the present and future potential for improving the understanding of biological systems.
Contents: Apparatus Basic Principles Macromolecules Interfacial Systems Ordered Macromolecules Cells, Tissue and Biominerals Other Probe Microscopes
Readership: Undergraduates, postgraduates and researchers in biophysics. 352pp Pub. date: Dec 1999 Imperial College Press ISBN 1-86094-199-0 US$51 / £32
I think one can use the "self timer" function of the Coolpix 990 to expose an image without touching the camera body at the time of the exposure.
I can think of no engineering reason why the cable release would need to be expensive when it becomes available. I'll bet any electronics tinkerer could make one for $30.
I'm quite happy with my Cool-Pix 990. If only I could remember 10% of its function modes and how to access them.
High-purity Germanium detectors provide *better* resolution than Si(Li) detectors, and also have lower low-keV background. The higher atomic number of Ge means they have greater x-ray stopping power and therefore higher detection efficiency for the K lines of the heavier elements, and they have therefore been thought of as primarily useful at higher beam accelerating voltages for heavy element analysis. However, this may only be one of their advantages. The better resolution and lower background they offer at low keV suggests they would also be good, and perhaps preferable to Si for light element detection and peak discrimination which could mean more options for low-kV excitation in Field emission SEMs.
What are the flaws in this argument? It would be interesting to hear comments on the above from anyone who has direct knowledge and experience of both types of detectors.
BTW besides PGT, both Gresham and Oxford sell Ge detectors. I am not aware of any others Chris
} From: "NVo-at-IGC.com"-at-sparc5.microscopy.com To: Microscopy-at-sparc5.microscopy.com
Dear Listers:
I am trying to find an enlarger and digital image system to print the conventional and HREM negatives. What kind of enlarger and digital image system are better? manufacturer? prices?
} I'm looking to replace my two year old Polaroid Digital Microscope camera } (SCSI interface with color or black and white, 1600x1200 pixels). The } chip has failed and I don't want to buy another Polaroid camera. I do } primarily black and white photography from a Zeiss optical microscope for } materials engineering applications. I need for the camera to save in tiff } format without changing the dimensions of the objects. I don't need low } light applications. } } Any suggestions? } } Thanks, } } Robin Griffin }
We have found a simple method for preparing drug particles for electron diffraction. We suspend particles in a solvent by sonication for 2-3 seconds and then let the suspension settle for 10-20 seconds. The largest, heavy particles will settle first leaving the thin, light ones in suspension. Then take a drop (20 ul) from the top of the solution, place it on a carbon filmed grid and allow it to dry. The result is a distribution of particles laying flat accross the grid. You need a solvent in which the particles are not soluble. We typically use hexane.
Joe Neilly Abbott Laboratories D-45M, AP31 200 Abbott Park Rd. Abbott Park, IL 60064-6202 voice: (847)-938-5024 fax: (847)-938-5027 e-mail: joe.neilly-at-abbott.com
I have two questions on TEM of DNA and protein complex: 1. which method is the best , or most suitable ,to study such samples? Direct spray to the mica sheet and subsequent metal rotary shadowing, or cytochrome c spreading with metal coating examined by TEM, or atomic force microscopy? 2. Is there any computer software which is designed specifically for analyze these type of images?
This is a bit off the topic of microscopy, but most of us on this server do work in facilities that have XRD units. If anyone is selling, or knows of someone who is selling their XRD, please contact me. I am also curious if anyone has had any experience with the new desk tops like Rigaku has. I plan on demoing one in the future. At $50k-$60k, they are very cost efficient compared to standard XRD units, but I am wary of their performance as an analytical instrument. Lou Solebello
Dear members, I am a PhD student and I use a subscriber to find an answer to a problem I have. I am trying to find a method to fix and process whole fish eggs in order to cut sections for immunohistochemistry. Any suggestions?
Ioannis Vatsos Institute of Aquaculture University of Stirling Scotland, UK
i've installed a 35 and have some issues with a saw-tooth distortion in the scan. it stays constant regardless of KV, but the number of saw-tooths (teeth?) in a frame increases with slower scan rates. i suspected vibration so i mounted the roughing pumps on a big foam block.....no change. i also cycled off the chiller unit with no change, so its probably not mechanical vibrations, unless there is another vibration causing component i'm overlooking.
the distortion is also visible in the data field alphanumerics (they shift and shimmy a little) with the beam turned off, so i don't suspect a HV instability problem.
can i logically deduce that there is an electrical field leakage that is messing up the scan? or a ground loop? what about cable placement? or a problem with the 100V variac (its within 3' of the column).
right now the column is a bit dirty and needs cleaning....i've never seen, for example, dirty apertures cause this type of distortion though.
any suggestions would be appreciated!
brian ---------------------------------------------- Brian McIntyre Electron Microscopy Lab- River Campus Univ of Rochester Rochester, NY 14627 716-275-3058/4875
There currently is no NIST-traceable standard for high-magnification TEM on the market. The next best thing is the MAG*I*CAL TEM calibration sample, which is internally calibrated against the lattice spacing of silicon, a fundamental constant of nature. This statement has been used successfully in the past in fulfillment of the requirements for certification and traceability. The MAG*I*CAL sample allows magnification calibrations across the entire magnification range of TEMs, as well as the camera constant calibration and the image/diffraction pattern rotation calibration.
The sample is marketed by South Bay Technology, TEL: 800-728-2233, 1120 Via Callejon, San Clemente, CA 92673 USA, e-mail: henriks-at-southbaytech.com It is also marketed through various microscopy supply houses, (SPI, EMS, Pelco, etc.) if you have a favorite.
Your request again raises the concern that the microscopy community needs a high-magnification TEM calibration sample that is NIST-certified or NIST-traceable. NIST is currently considering investigating the need for a high-magnification TEM calibration reference, but will not proceed unless there is some interest shown from the user community. It would be helpful to the community in general if you could write a note or send an email to Ms. Nancy Trahey, stating the problem that you have encountered, and reiterating the need for such a NIST-certified or NIST-traceable standard. Her co-ordinates are listed below.
Ms. Nancy M. Trahey Standard Reference Materials Program (232) Engineering Mechanics (202), Room 113 NIST 100 Bureau Drive, Stop 2320 Gaithersburg, MD 20899-2320 Tel: (301) 975-2021 email: nancy.trahey-at-nist.gov
Disclaimer: I 'invented' the MAG*I*CAL TEM calibration sample and retain all bragging rights; it is listed in the Guinness Book of Records as "The World's Smallest Ruler".
John P. McCaffrey, Ph.D. National Research Council of Canada M-50, Montreal Rd. Ottawa, Ontario K1A 0R6 CANADA
-----Original Message----- } From: Vitaly Feingold [mailto:vitalylazar-at-worldnet.att.net] Sent: Wednesday, October 04, 2000 11:54 PM To: .
Dear listers,
What certified (NIST traceable?) TEM mag. calibration samples are available for 20K to over 100K range? Diffraction grating replica sample will not provide sufficient accuracy- too few grating periods "fit" in the image at higher magnifications.
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax
} Any comments on which would make a better detector, Ge or } Si ? only PGT sells Ge detectors and I'm aware of } the decrease in resolution with Ge detector. } Could someone lists or details the benefits of having a Ge } detector as oppose to a Si one.
We are using a "Gem" detector from Oxford Instruments with absoloutely no problems ... 115eV resolution, fast count rates, and very little energy shift with varying time constants. Another benefit is there are no escape peaks for x-ray lines 0~10keV. One drawback may be if you intend to allow the detector to come up to ambient temperatures at times. Si detectors seem to accommodate this, whereas it was suggested at the time of purchase we absolutely not allow this .. but I believe that was simply because the "Gem" was for the most part untested in this respect. I have no idea of this detector's ambient T survivability, we have always kept it at LN2 T.
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 am attaching CP dried salmon skins (approx. 2-3 mm thick) to stubs with Electrodag 502 for secondary electron imaging. The adhesive is a graphite paint in MEK base (a substitute for TV Tube Koat). After ethanol dehydration and CP drying in liquid CO2, the samples are relatively flat or slightly curved. However, once attached to the stub the skins develop a noticeable curl which leaves a gap between the stub surface and sample. In some cases I can see the "trail" in the adhesive left by the skin as it was curling! Filling the gap with more Electrodag is a waste of time because it evaporates and shrinks back away from the sample (thankfully it doesn't seem to produce more curling).
Could anyone suggest a better conductive adhesive? Also, is there any difference between aqueous (eg. Aquadag colloidal graphite) and MEK-based media as to its effect on biological samples?
As an SEM novice any advice you could share with me is greatly appreciated. Thank you very much,
Carla Aiwohi Western Fisheries Research Center Seattle, WA ph: (206) 526-6282 ext.242 fax: (206) 526-6654
Anyone know of a easy, simple way to produce nice powder diffraction samples for illustrating the powder diffraction method for EM students? We use the International Centre for Diffraction Data For student identification of unknown samples. I have tried several methods that usually result in non-electron transparent samples (our TEM KV max is only 120). Any ideas?
Dear Bill, The simplest way I know is to evaporate a layer of Au or Al onto a formvar-covered grid. By spreading the beam, you can get continuous rings, and by using a small selected area you can get discrete spots at various positions around the rings, so you can demonstrate directly that the rings consist of the summed intensities from many crystalites of differing orientations. Good luck. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
Ben, Our Zeiss DSM982 FESEM is also sensitive to acoustic noise. "The quieter the room the more noises we hear." There is no simple answer, just keep trying different things. I was told one lab installed a switch to turn the SEM PC cooling fans off for short periods of time during high resolution work. That is a risky fix if you are over 50 years old.
We strategically padded all connections to the column and redirected the room air flow but have not installed acoustic foam on the walls. Evenings are quieter than days. At 300kx we can detect a backhoe 600 yards away.
Good Luck Jim
} Date: Wed, 4 Oct 2000 19:52:12 -0700 (PDT) } From: Ben Craft {bcraft-at-uci.edu} } X-Sender: bcraft-at-taurus.oac.uci.edu } To: Microscopy-at-sparc5.microscopy.com } Subject: High Resolution noise } MIME-Version: 1.0 } } } } I'm trying to solve a high resolution noise problem with a XL-30 } Phillips FESEM. The room that it is in was recently checked and was ok, } accept for some acoustic noise. To fix the noise I put up 3 inch sound } proofing foam and quieter ducting for the air condition. The noise } showed up in an image by a streak about 5 jaggies. After } putting up the foam the noise was reduced, but it is still there. The } noise is the same for 4mm working distance or a 10mm working distance. } } Any one had a similar problem or have any advice? } } Thanks, } Ben } } } } } ####### } #####\_O -Ben Craft- } ####/\/} } #### /" } ### \ }
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
You could prepare your own. I find that the diatoms in diatomaceous earth are usually broken and like to prepare my own. You will need a plankton net, some potassium dichromate, and 30% peroxide. Once the diatoms are collected (plankton net tows from shoreline) you can clean them using the 30% peroxide and potassium dichromate. Place the diatoms in a large beaker (and I mean large here) then pour in 15 to 20 mls of peroxide followed by a large pinch of the potasium dichromate. The resultant reaction oxidizes organics leaving the glass frustules of the diatoms unharmed. It is better to be cautious with the amounts that you use as this is an exothermic reaction, sometimes violently so. Not that you'll have an explosion, but I've had to clean up boil overs on a number of occasions. Once the reaction subsides and the solution turns a golden yellow color it is just a matter of washing the diatoms. There are a couple of ways to do this. You could centrifuge the diatoms, then throw out the liquid, add water, centrifuge, throw out the liquid, etc. or you could fill the large beaker with water and let the diatoms settle to the bottom (1-2hrs), carefully pour off the fluid, and add more water, let the diatoms settle, etc. The diatoms will appear as a milky white residue in the bottom of your container (you may have to concentrate the material to really see this). If you need any more info please feel free to contact me.
} --------------------------------------------------------------------------- } } Email: s_woodland-at-madasafish.com } Name: Simon Woodland } School: Kaskenmoor School } } Question: Where can I get some diatoms to put under our microscope? How } should they be prepared and mounted. } } ---------------------------------------------------------------------------
-- ================================================================== 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 ==================================================================
Besides Nikon's own release (EU-1 wired remote control) for the CoolPix990, you can purchase a third-party release cable from the CKCpower.com website.
Jim
} } Hi Peter, } } I think one can use the "self timer" function of the Coolpix 990 to } expose an image without touching the camera body at the time of the } exposure. } } I can think of no engineering reason why the cable release would need to } be expensive when it becomes available. I'll bet any electronics } tinkerer could make one for $30. } } I'm quite happy with my Cool-Pix 990. If only I could remember 10% of } its function modes and how to access them. } } Bart Cannon } Cannon Microprobe } Seattle } }
We have a Ge detector and get considerably better resolution with it than we get with an older SiLi detector. The Ge detector has an ultrathin window, the Si detector has a Be window. They are both used for x-ray detection in the 0-20 KeV range.
Kent Ross
----------------------------------------- Kent Ross Research Associate Texas Center for Superconductivity University of Houston 77204-5932 DKROSS-at-uh.edu 713-743-8284 ------------------------------------------
} mounting the Coolpix 990 to any photomicroscope is absolutely } simple via the usual C-mount connection. For the coolpix you have } to buy at NIKON the microscope adapter ... } The fantastic thing is that you can focus not only via the LCD- } screen of the camera ( possible, but not so easy in my opinion) but } via an external TV-Monitor in live modus, yielding really smashing } results with a resolution satisfying publication needs. } With my Coolpix sometimes the camera "hangs up" when the Auto- } power off mode is activated (de-activating it helps). Does anybody } have this problem as well ?? ...
The AC adapter should allow for the video output to remain on ... leastwise, that is how my CP800 works.
} What is lacking for the Coolpix 990 at the moment is a cable } release, ... } } All in all, the Coolpix is the first digital camera under } 5000 US-$ ...
The 990 is indeed feature rich, and the cable release will be needed for microscopy (... altho you should be able to acquire with a steady camera if you take a picture with the time-delay feature ...).
I have realized just lately, altho the CP950 and 990 are the feature-rich and most desireable, the CP800 will work (altho somewhat awkwardly) ... providing the same NTSC/PAL out and lens threads. If you find a lesser expensive 1x C-mount adapter (as available at Optem International {www.optemintl.com} ), then you have a digital camera capable of 1600by1200 for approximately US$800. You should figure an additional $100 for AC adapter ($50) and several NiMH batteries and charger (~$50).
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/
Have you considered or looked into the Sony 390P color video camera?? It gives you a live image with an RGB output, C-mount and 1.3 million pixels (effective resolution) and sells for just under $ 3,000.
If you would like any more information, please let me know.
Sincerely,
Gary
Gary Liechty Manager, Technical Products
Allied High Tech Products, Inc. 2376 E. Pacifica Place Rancho Dominguez, CA 90220
310-635-2466 800-675-1118 US and Canada 310-762-6808 Fax
Products for Metallurgical Sample Preparation
www.alliedhightech.com ----- Original Message ----- } From: {"rgriffin-at-eng.uab.edu"-at-sparc5.microscopy.com} To: {microscopy-at-sparc5.microscopy.com} Sent: Thursday, October 05, 2000 5:54 AM
Simon,
I suspect that diatomaceous earth will have mostly fragments in it. If you let me know your address, I'll try to find an old diatom SEM preparation to send you and I'll also ask some folks here for a fresh sample of whole diatoms that you can prepare for viewing yourself.
Gary,
Being a microscopist, not a microbiologist/paleontologist/sedimentologist (though I've imaged my share of microplankton for them) I can only give a brief overview of the differences between diatoms and radiolaria, but here it is. Rads are floating, single celled marine protozoa that live inside ornate opaline tests (external shells) that look like little Christmas tree ornaments or something you might play badminton with. Their shells have lots of holes, spikes and spines and can be spherical, amorphously latticed, rather bulbous with long spikes from one end, sub-triangular, etc. The cell body inside sends out numerous fine cytoplasmic strands that radiate outward to form a sticky halo that captures food. Diatoms are single-celled plants that live wherever there's moisture - salt water, fresh water, soil, even on whale skin. They can be planktonic (free-floating) or benthic (attached to a substrate) or live on mud or rocks. Some live in the dark, but most need light to photosynthesize. Their siliceous tests are made of two valves that fit together like a pill box and come in two basic shapes, round and pennate. (The pennate ones can look like peanuts or canoes.) Diatom tests are also quite ornamental, with holes, radial designs, or long, sometimes feathery, projections, and sometimes they link together to form long chains. There's also a great variety in size among species.
Dee
*************************************************************** Dee Breger Mgr. SEM/EDX Facility Lamont-Doherty Earth Observatory 61 Route 9W Palisades, NY 10964 USA T: 914/365-8640 F: 914/365-8155
http://www.ldeo.columbia.edu/micro http://www.discovery.com/area/science/micro/micro1.html http://www.lsc.org/antarctica/front.html Journeys in Microspace (Columbia University Press, 1995)
You might also try contacting Bunton Instruments in the US:
{http://www.buntgrp.com/}
They sell microscope adaptors for the CoolPix. The adaptors are not listed on their web site, yet, but call or e-mail them for information. I know they are taking orders for adaptors now and I was told that they would be shipping "around the end of October."
I have no connection with Bunton Instruments other than as a customer.
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
You don't mention at what magnifications you see the sawtooth. However, if you see it in the alphanumerics, I can't see that it can be anything other than a problem in your display unit - a 60Hz interference in the line scan drive (but not the scan generator) or something like that. If that is the case, the magnification wouldn't make any difference.
Good luck,
Tony Garratt-Reed
} hi all- } } i've installed a 35 and have some issues with a saw-tooth distortion in the } scan. it stays constant regardless of KV, but the number of saw-tooths } (teeth?) in a frame increases with slower scan rates. i suspected } vibration so i mounted the roughing pumps on a big foam block.....no } change. i also cycled off the chiller unit with no change, so its probably } not mechanical vibrations, unless there is another vibration causing } component i'm overlooking. } } the distortion is also visible in the data field alphanumerics (they shift } and shimmy a little) with the beam turned off, so i don't suspect a HV } instability problem. } } can i logically deduce that there is an electrical field leakage that is } messing up the scan? or a ground loop? what about cable placement? or a } problem with the 100V variac (its within 3' of the column). } } right now the column is a bit dirty and needs cleaning....i've never seen, } for example, dirty apertures cause this type of distortion though. } } any suggestions would be appreciated! } } brian } ---------------------------------------------- } Brian McIntyre } Electron Microscopy Lab- River Campus } Univ of Rochester } Rochester, NY 14627 } 716-275-3058/4875 }
** Anthony J. Garratt-Reed ** MIT Room # 13-1027 ** 77 Massachusetts Avenue ** Cambridge, MA 02139-4307 ** USA ** ** Phone: (+) 1-617-253-4622 ** Fax: (+) 1-617-258-6478 **
We currently have 4 Ge EDX detectors and 1 Si(Li) detector, all from Oxford Instruments. The Ge detectors do have a lower background than the Si(Li) and the resolution is better. The Ge detectors range from 109 ev to 115 ev whereas the Si(Li) is around 132 ev. We routinely use all of them for low-kV work. The Ge detectors do a better job because of the resolution and the lower background. I do SEM of semiconductors, so 30kV is as high as I can go. I can't comment on the Ge detector's performance at higher than that, but they are my favorite detector in the 0.5 kV to 30kV range. N.B.: I have no financial interest in Oxford Instruments; I'm just a satisfied customer.
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 } -----------------------------------------------------------------------. } } High-purity Germanium detectors provide *better* resolution than } Si(Li) detectors, and also have lower low-keV background. The } higher atomic number of Ge means they have greater x-ray } stopping power and therefore higher detection efficiency for the K } lines of the heavier elements, and they have therefore been thought } of as primarily useful at higher beam accelerating voltages for } heavy element analysis. However, this may only be one of their } advantages. The better resolution and lower background they offer } at low keV suggests they would also be good, and perhaps } preferable to Si for light element detection and peak discrimination } which could mean more options for low-kV excitation in Field } emission SEMs. } } What are the flaws in this argument? } It would be interesting to hear comments on the above from anyone } who has direct knowledge and experience of both types of } detectors. } } BTW besides PGT, both Gresham and Oxford sell Ge detectors. I } am not aware of any others } Chris } } } From: "NVo-at-IGC.com"-at-sparc5.microscopy.com } To: Microscopy-at-sparc5.microscopy.com } Subject: Ge vs Si detector } Date sent: Wed, 4 Oct 2000 13:15:58 -0400 } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 Listners: } } } } Any comments on which would make a better detector, Ge or Si ? only PGT } } sells Ge detectors and I'm aware of the decrease in resolution with Ge } } detector. Could someone lists or details the benefits of having a Ge } } detector as oppose to a Si one. } } } } Sincerely } } } } N.V.Vo } } } } } } } } N.V.Vo, Ph.D. } } Intermagnetics General Corporation } } 450 Duane Avenue } } Schenectady, New York 12304 } } Phone: 518 346 1414 x 3107 } } Fax: 518 346 6080 } } E-mail: nvo-at-igc.com } } www.igc.com } } } } } } ========================================== } Dr. Chris Jeffree } University of Edinburgh } BIOSEM - Biological Sciences Electron Microscope Facility } Institute of Cell and Molecular Biology } Daniel Rutherford Building } King's Buildings, Mayfield Road } EDINBURGH, EH9 3JH, Scotland, UK } Tel. #44 (0) 131 650 5345 } FAX. #44 (0) 131 650 6563 } Mobile 07710 585 401 } email c.jeffree-at-ed.ac.uk } =========================================
Our 35C has several times displayed that saw-tooth interference in the past. Each time it was attributed to some ground loop problem. One place to look is in the ribbon aperture. Make sure that the insulators between the aperture strip and holder are properly set or 'not missing'.
I agree that the distortion is not from HV instability which usually displays some darkness or brightness irregularity.
Good luck! _ / \ Sam O. Mancuso /\ /\ Special Metals Corporation (__n__) New Hartford, NY
Brian McIntyre {mcintyre-at-optics.roch To: Microscopy-at-sparc5.microscopy.com ester.edu} cc: Subject: saw-tooth scan distortion: jeol35 10/05/2000 10:04 AM
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
hi all-
i've installed a 35 and have some issues with a saw-tooth distortion in the scan. it stays constant regardless of KV, but the number of saw-tooths (teeth?) in a frame increases with slower scan rates. i suspected vibration so i mounted the roughing pumps on a big foam block.....no change. i also cycled off the chiller unit with no change, so its probably not mechanical vibrations, unless there is another vibration causing component i'm overlooking.
the distortion is also visible in the data field alphanumerics (they shift and shimmy a little) with the beam turned off, so i don't suspect a HV instability problem.
can i logically deduce that there is an electrical field leakage that is messing up the scan? or a ground loop? what about cable placement? or a problem with the 100V variac (its within 3' of the column).
right now the column is a bit dirty and needs cleaning....i've never seen, for example, dirty apertures cause this type of distortion though.
any suggestions would be appreciated!
brian ---------------------------------------------- Brian McIntyre Electron Microscopy Lab- River Campus Univ of Rochester Rochester, NY 14627 716-275-3058/4875
-----Original Message----- } From: "s_woodland-at-madasafish.com"-at-sparc5.microscopy.com [mailto:"s_woodland-at-madasafish.com"-at-sparc5.microscopy.com] Sent: Wednesday, October 04, 2000 8:05 AM To: Microscopy-at-sparc5.microscopy.com
} } I am attaching CP dried salmon skins (approx. 2-3 mm thick) to stubs with } Electrodag 502 for secondary electron imaging. The adhesive is a graphite } paint in MEK base (a substitute for TV Tube Koat). After ethanol } dehydration and CP drying in liquid CO2, the samples are relatively flat or } slightly curved. However, once attached to the stub the skins develop a } noticeable curl which leaves a gap between the stub surface and sample. In } some cases I can see the "trail" in the adhesive left by the skin as it was } curling! Filling the gap with more Electrodag is a waste of time because } it evaporates and shrinks back away from the sample (thankfully it doesn't } seem to produce more curling). } } Could anyone suggest a better conductive adhesive? Also, is there any } difference between aqueous (eg. Aquadag colloidal graphite) and MEK-based } media as to its effect on biological samples? }
I have found the conductive double-sided adhesive stuff available from several EM suppliers in both disc and tape form to be pretty good. But maybe you can't press your samples down onto it.
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
} I was told one lab installed a switch to turn the } SEM PC cooling fans off for short periods of time during high resolution } work. That is a risky fix if you are over 50 years old.
??????????????????????????????
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
} } } } I was told one lab installed a switch to turn the } } SEM PC cooling fans off for short periods of time during high resolution } } work. That is a risky fix if you are over 50 years old. } } ??????????????????????????????
Clarification: I just turned 50. I if I attempted to turned these fans off manually for "a few minutes", chances are the PC processor would shut down on overheat before I remembered to turn them back on. It's an 'out of memory' problem. Jim
} } 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 }
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
EG&G ORTEC used to manufacture Germanium EDS detectors too. I do not know whether they still do it since they had become part of Perkin-Elmer. They have, however, a service group which does service Germanium detectors.
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax
This message is made of 100% recycled electrons. -----Original Message----- } From: Chris Jeffree {cjeffree-at-srv0.bio.ed.ac.uk} To: "NVo-at-IGC.com"-at-sparc5.microscopy.com {"NVo-at-IGC.com"-at-sparc5.microscopy.com} Cc: microscopy-at-sparc5.microscopy.com {microscopy-at-sparc5.microscopy.com}
Information in your posting strongly suggests ground loop(s) and possibly EM (electromagnetic) fields in the room too. It is possible for local EM interference to affect signal cables without affecting the SEM column much. Did you try EM shielding with sheets of soft Iron? If that makes no difference (it probably wouldn't, since HT set makes no difference for the distortion in your case), than some work needs to be done with shielding/grounding of the various units/components of the SEM. Let us say here, one who has the experience of eliminating 60Hz noise in audio circuits can do the job. Another possibility- did you check DC power supply voltages in your SEM for 60Hz noise? It must be filtered out well.
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax
This message is made of 100% recycled electrons. -----Original Message----- } From: Brian McIntyre {mcintyre-at-optics.rochester.edu} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
your SEM interference problem sounds almost the same as ours that I not long ago put on the listserver - the sawtooths in the scans. Sorry I haven't got an answer to the problem, but I'd be very interested if you manage to solve it! We are going to do a trial with mu-metal in the near future shielding the column of our SEM (Jeol 5410-LV) so I'll let you know how it goes. We suspect electromagnetic field interference.
Good luck,
Mark
******************************************** Mark West, Unidad de Microscopia Electronica, (Electron Microscopy Unit) Instituto de Fisiologia Celular, Universidad Nacional Autonoma de Mexico, 04510 Mexico D.F.
Dear Robin, For your application, you may find the Pixera camera to be a cost-effective solution. At the same resolutionoas your current camera, it costs about $1100. At 07:54 AM 10/5/00 -0500, you wrote:
} } I'm looking to replace my two year old Polaroid Digital Microscope camera } } (SCSI interface with color or black and white, 1600x1200 pixels). The } } chip has failed and I don't want to buy another Polaroid camera. I do } } primarily black and white photography from a Zeiss optical microscope for } } materials engineering applications. I need for the camera to save in tiff } } format without changing the dimensions of the objects. I don't need low } } light applications. } } } } Any suggestions? } } } } Thanks, } } } } Robin Griffin } } 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 Brian, You may have ripple on another power supply, such as lens or scan coils. I always try to put the variac as far from the column as possible. Check for something loose on the column or stage: I've had that effect when the gun HV supply cable was not tied down. Make sure there is a good vibration break (heavy weight) in the foreline hose. The service engineer at ours turned off the rotary pump for a few seconds, to see if it changed. Another source of vibration was a belt-drive pump in another room, but on the same floor beam. A dirty column won't cause this. It may be building vibration. At 10:04 AM 10/5/00 -0400, you wrote: } } hi all- } } i've installed a 35 and have some issues with a saw-tooth distortion in the } scan. it stays constant regardless of KV, but the number of saw-tooths } (teeth?) in a frame increases with slower scan rates. i suspected } vibration so i mounted the roughing pumps on a big foam block.....no } change. i also cycled off the chiller unit with no change, so its probably } not mechanical vibrations, unless there is another vibration causing } component i'm overlooking. } } the distortion is also visible in the data field alphanumerics (they shift } and shimmy a little) with the beam turned off, so i don't suspect a HV } instability problem. } } can i logically deduce that there is an electrical field leakage that is } messing up the scan? or a ground loop? what about cable placement? or a } problem with the 100V variac (its within 3' of the column). } } right now the column is a bit dirty and needs cleaning....i've never seen, } for example, dirty apertures cause this type of distortion though. } } any suggestions would be appreciated! } } brian Good luck!
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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what you see could be 60-Hz Noise. It follows the characteristics you are describing (more 'teeth' with lower scan rate). Can you do a back-of-the-envelope calculation of how many 'teeth' you get per second (check exposure settings and line times). If it's around 60, you should try to minimize the influence of 60 Hz Noise (ground loops, check all grounding contacts, 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: Brian McIntyre [mailto:mcintyre-at-optics.rochester.edu] Sent: Thursday, October 05, 2000 8:04 AM To: Microscopy-at-sparc5.microscopy.com
hi all-
i've installed a 35 and have some issues with a saw-tooth distortion in the scan. it stays constant regardless of KV, but the number of saw-tooths (teeth?) in a frame increases with slower scan rates. i suspected vibration so i mounted the roughing pumps on a big foam block.....no change. i also cycled off the chiller unit with no change, so its probably not mechanical vibrations, unless there is another vibration causing component i'm overlooking.
the distortion is also visible in the data field alphanumerics (they shift and shimmy a little) with the beam turned off, so i don't suspect a HV instability problem.
can i logically deduce that there is an electrical field leakage that is messing up the scan? or a ground loop? what about cable placement? or a problem with the 100V variac (its within 3' of the column).
right now the column is a bit dirty and needs cleaning....i've never seen, for example, dirty apertures cause this type of distortion though.
any suggestions would be appreciated!
brian ---------------------------------------------- Brian McIntyre Electron Microscopy Lab- River Campus Univ of Rochester Rochester, NY 14627 716-275-3058/4875
It is with great pleasure that I am announcing our 2001 FE-SEM and Image Analysis Courses
When: May 21-25, 2001
Guest Speaker: Dr. David C. Joy Professor, Tennessee University Marin Lagace, Researcher, Hydro-Quebec Research Center
Where: IREQ (Hydro-Quebec Research Centre), Varennes(Montreal), QC., Canada
Topics Covered: FE-SEM Components: gun, lenses, apertures, vacuum technology... Care & feeding of the FEG The secrets of successful FE-Microscopy Vacuum hygiene, cleaning and storing samples Unwanted Beam Interactions What is needed for High Resolution SEM Image Content Getting the most from your SEM How to assess resolution Low Voltage Work - High Resolution at low energy
LVSEM and charging FESEM and Microanalysis Low Energy EDS Work plus so much more
This 3-day workshop will be devided in Lecture and Practical Sessions
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What about using Superglue? I routinely use it when I need a good adhesion of samples. You really do not need a conductive adhesive with a sample thickness of about 3 mm - it's better just to draw a line with carbon adhesive from the edge of a sample to a stud prior conductive coating.
Vladimir Dusevich
-----Original Message----- } From: "carla_aiwohi-at-usgs.gov"-at-sparc5.microscopy.com To: Microscopy-at-sparc5.microscopy.com Sent: 10/5/00 8:46 AM
Hello all,
I am attaching CP dried salmon skins (approx. 2-3 mm thick) to stubs with Electrodag 502 for secondary electron imaging. The adhesive is a graphite paint in MEK base (a substitute for TV Tube Koat). After ethanol dehydration and CP drying in liquid CO2, the samples are relatively flat or slightly curved. However, once attached to the stub the skins develop a noticeable curl which leaves a p between the stub surface and sample. In some cases I can see the "trail" in the adhesive left by the skin as it was curling! Filling the gap with more Electrodag is a waste of time because it evaporates and shrinks back away from the sample (thankfully it doesn't seem to produce more curling).
Could anyone suggest a better conductive adhesive? Also, is there any difference between aqueous (eg. Aquadag colloidal graphite) and MEK-based media as to its effect on biological samples?
As an SEM novice any advice you could share with me is greatly appreciated. Thank you very much,
Carla Aiwohi Western Fisheries Research Center Seattle, WA ph: (206) 526-6282 ext.242 fax: (206) 526-6654
} i've installed a 35 and have some issues with a saw-tooth distortion in } the scan.
My 35C had an aperture heater which used to go into oscillation and produce the kind of problem you describe. It has long since been disconnected. It doesn't seem likely in your case because you see the problem with the beam off, but I thought it was worth mentioning just in case.
Sincerely yours, Andy Buechele
Andrew C. Buechele, Ph.D. Research Professor Vitreous State Laboratory The Catholic University of America 409 Hannan Hall Washington, D.C. 20064 (202) 319-4995 FAX: (202) 319-4469
I need good quality carbon coated grids for viewing purified plant virus particles with TEM. I have had little success with "store bought" coated grids. So unless someone could recommend a good cc grid source, I will need to buy a carbon coater. Should I purchase a carbon coater using flash evaporation or the more expensive carbon turbo evaporator? In addition, could anyone recommend an economically good microtome for thin sectioning plastic embedded specimens?
Thank You,
Nancy L. Robertson, Research Plant Pathologist USDA/ARS National Arctic Plant Germplasm Resources Unit 533 Fireweed Ave. Palmer, Alaska 99645 phone: 907-746-9465 fax:907-746-2677 e-mail: nancy_robertson-at-dnr.state.ak.us
Another way to pick up noise and voltage is to wires wrapped around each other. It is east to test just watch the screen and poke the wires with a wood stick. If there is any problem in the wiring it will show up on the screen. You only have to move them gently.
A wooden stick and light tapping can find a lot of problems. If you are working in the high voltage section make sure it is a long dry stick. It is great for finding intermittent problems. Just start tapping until the problem appears or goes away and then narrow in on the problem.
When you check the frequency if it comes up 120 Hz florescent lights become the number 1 suspect. You could be picking up electrical noise from them or you have something photo sensitive somewhere. Any silicone junction is sensitive to light. The quick test is to turn off the lights. If they are the problem it should at least get weaker.
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
----- Original Message ----- } From: "Mike Bode" {mb-at-Soft-Imaging.com} To: "'Microscopy-at-MSA.Microscopy.Com'" {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, October 05, 2000 6:53 PM
Thank you all for your replies on this topic. Several people seem to like Ge detectors for low keV work, and the replies have mostly been very positive about their resolution and performance. Some authors strongly advocate the use of Ge detectors for low kV, low probe current work with light element samples (See, for example, Block- van Hoek & Pinxter (1993) Chapter 1 in X-ray microanalysis in biology edited by Sigee et al, Cambridge University Press). Their opinion (and apparently yours) stands in contrast with the advice given by all of the XRMA vendors I am speaking to, whose opinion is that Si(Li) is the way to go (granted some of them don't sell Ge, so they would obviously have to advocate Si). So my question is "what is the Achilles heel of Ge detectors?". I am aware for example that their potentially very favourable characteristics at low kV are spoiled by the practical difficulty of manufacturing them without a large dead layer. Is this a fatal flaw in practice for analysis of elements with z {11? Are they less robust, less tolerant of thermal cycling? If anyone can help me get to the bottom of this I would be most grateful.
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
I am grinding a TEM specimen in a Multipol polishing machine, but a grinding plate is covered with some stains of grayish color whose origin I cannot identify. I tried to apply ethanol, atheton, and methanol to them, but these proved in vain.
Please suggest any cleaning liquid to wipe the grinding plate clean.
Alex
Aleksandr Tikhonovsky, Ph.D. tikhonov-at-mpi-halle.mpg.de tel 49 345 5582 929 Max-Planck-Institut für Mikrostrukturphysik Weinberg 2 D-06120 Halle Germany
You've had a lot of replies, but your symptoms are clear. In most cases, this interference is caused by an electrical device mounted too close to the SEM CRT - An EDS monitor, vacuum gauge controller, etc. If not, you probably have a bad filter or bypass capacitor in a power supply that the CRT uses. Since the effect is also seen in the alpha-numerics, it is directly affecting the view CRT itself. Keeping the area around the CRT clear of other instruments and checking for ripple in the power supplies to the CRT should solve the problem. The effects of external instruments near the view CRT are generally very local and movement of just a few inches can make a huge difference.
It is not mechanical vibrations and the CRTs are not that suseptible to electro-magnetic interference from distant sources.
If the alpha-numerics were not affected, I would suggest checking the relative difference in the noise when using a low and high working distance. If there is a noticably greater noise when at a larger working distance, then electro-magnetic fields affecting the beam would be suspect. The column of an SEM is actually well shielded from EM effects, as the lenses are encased in large ferro-magnetic structures to provide the focussing of the lens effects to the pole pieces. If lower working distances are used (sample closer to the final pole piece) then the effects of EM are reduced since they are primarily induced through the sample chamber and affect the beam from the final pole piece to the sample surface.
However, as I've said, if the alpha-numeric display is also affected, then more than likely the CRT beam is being affected by something close by or by a power supply to the CRT. The high voltage to the CRT (around 10KV) is not a likely suspect as the record CRT is much more suseptable to HV problems and in the JSM-35, as in most SEMs, is fed from the same supply. You would have seen some obvious focus problems in your micrographs if the high voltage was a problem. As the view CRT is an integral part of the electronics console, ground loop problems are unlikely.
The record CRT and view CRT are located close together in this instrument and as I recall they use the same power supplies. The record CRT is probably recording a problem as well, but it may be greatly reduced. The record cycle uses a greatly reduced scan speed, so these variations in the view CRT are going to show up as reduced edge sharpness in your micrographs or not at all if each record scan line is triggered by a zero crossing on the 60Hz input (I don't recall if the JSM-35 does this).
Allen R. Sampson, Owner Advanced Research Systems, St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
-----Original Message----- } From: Brian McIntyre [SMTP:mcintyre-at-optics.rochester.edu] Sent: Thursday, October 05, 2000 9:04 AM To: Microscopy-at-sparc5.microscopy.com
hi all-
i've installed a 35 and have some issues with a saw-tooth distortion in the scan. it stays constant regardless of KV, but the number of saw-tooths (teeth?) in a frame increases with slower scan rates. i suspected vibration so i mounted the roughing pumps on a big foam block.....no change. i also cycled off the chiller unit with no change, so its probably not mechanical vibrations, unless there is another vibration causing component i'm overlooking.
the distortion is also visible in the data field alphanumerics (they shift and shimmy a little) with the beam turned off, so i don't suspect a HV instability problem.
can i logically deduce that there is an electrical field leakage that is messing up the scan? or a ground loop? what about cable placement? or a problem with the 100V variac (its within 3' of the column).
right now the column is a bit dirty and needs cleaning....i've never seen, for example, dirty apertures cause this type of distortion though.
any suggestions would be appreciated!
brian ---------------------------------------------- Brian McIntyre Electron Microscopy Lab- River Campus Univ of Rochester Rochester, NY 14627 716-275-3058/4875
Have an EDS system attached? The EDS dewars, being generally supported by a cantilever structure, can be quite effective at coupling acoustic vibrations into an SEM. The large area and thin metal construction of the dewar as well as the large momentum arm of the cantilever provide for an amplification of acoustics into the instrument. You might also try to provide some form of isolated support and damping of the EDS system.
The size, shape and construction of the room can also have a considerable effect. Certain configurations can amplify acoustic and construction material mediated effects. Placement of the SEM within a particular room can make a big difference. Particularly bad is the placement in an alcove or bay of a room. Walls close to the sides of an instrument produce complex acoustic patterns. Each wall surface represents a resonator that can vibrate with unexpected sources. An instrument I had placed in this situation in a clean room of a major alphabet soup named manufacturer had incredible sensitivity to door closings and even footsteps in the adjoining airlock and preparation areas.
Your mention of 5 jaggies begs the question - at what scan rate were these viewed? What is the time span for each full scan? The SEM is an incredibly sensitive seismometer. I've seen the effects of waves crashing on the shores of an island as well as the vibrations from an M1-A1 Abrams tank at 40 miles per hour on a nearby proving ground. It can often be difficult to isolate the acoustic effects from the mechanically induced effects (those coupled through solid materials). Most SEMs provide good isolation from high frequency mechanical coupling, but low frequencies are problematic.
Vibrational problems, unlike electro-magnetic problems, affect the beam along it's entire path to some degree. In the column, most of these effects are canceled by the fact that a movement that causes the beam to be deflected outside of it's normal path will be countered by the increased magnetic effect of the lenses as the beam goes off-axis. The greatest effect though, is in the path from the final pole piece to the sample surface. Your determination of the effect being the same from 4mm to 10mm would probably not be true if you measured the difference from 4mm to 25mm.
Allen R. Sampson, Owner Advanced Research Systems, St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
-----Original Message----- } From: Ben Craft [SMTP:bcraft-at-uci.edu] Sent: Wednesday, October 04, 2000 9:52 PM To: Microscopy-at-sparc5.microscopy.com
I'm trying to solve a high resolution noise problem with a XL-30 Phillips FESEM. The room that it is in was recently checked and was ok, accept for some acoustic noise. To fix the noise I put up 3 inch sound proofing foam and quieter ducting for the air condition. The noise showed up in an image by a streak about 5 jaggies. After putting up the foam the noise was reduced, but it is still there. The noise is the same for 4mm working distance or a 10mm working distance.
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 all- } } i've installed a 35 and have some issues with a saw-tooth distortion in the } scan. it stays constant regardless of KV, but the number of saw-tooths } (teeth?) in a frame increases with slower scan rates. i suspected } vibration so i mounted the roughing pumps on a big foam block.....no } change. i also cycled off the chiller unit with no change, so its probably } not mechanical vibrations, unless there is another vibration causing } component i'm overlooking. } } the distortion is also visible in the data field alphanumerics (they shift } and shimmy a little) with the beam turned off, so i don't suspect a HV } instability problem. } } can i logically deduce that there is an electrical field leakage that is } messing up the scan? or a ground loop? what about cable placement? or a } problem with the 100V variac (its within 3' of the column). } } right now the column is a bit dirty and needs cleaning....i've never seen, } for example, dirty apertures cause this type of distortion though. } } any suggestions would be appreciated! } } brian } ---------------------------------------------- } Brian McIntyre } Electron Microscopy Lab- River Campus } Univ of Rochester } Rochester, NY 14627 } 716-275-3058/4875 Brian, You didn't say whether or not the sawtooth shows up on micrographs. If the problem were 60 Hz, they would not be apparent because the record scan speeds are sync'd to the line frequency. If you DO see them on micrographs, start by looking at your low voltage supplies, the CRT HV supply and the scan generator signals.
The problem has nothing to do with your column, since the characters show the problem. CRT HV instabilities should show up more the further from the center of the CRT you get. You should also be getting both light and dark lines as the vertical, as well as the horizontal, will be affected.
If you don't see the above symptoms, look at both scan signals going to the CRT. You may see noise on the horizontal signal, but if the sawtooth is showing up in the image, also, you probably won't see the noise on the horizontal signal to the column, because they aren't syncronized. If they both had the same noise, the image would look fine even though the alphanumerics shimmied.
Because of the alphanumerics, there is some processing of the CRT scan signals after the scan generator that does not happen to the scan signal sent to the mag control/column. A good place to look.
If the problem only occurs on 1 CRT, look at its drivers.
Ken Converse owner Quality Images third party SEM service Delta, PA
If you have not tried our carbon coated grids, we believe we make pretty good ones. If you have tried ours, please let me know what you didn't like and we can correct it since we make them up fresh for each order, and can make them thinner, thicker or whatever you wish. In a worst case scenario, we make what we consider the best evaporator in the business.
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--
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Microscope Supplies Since 1955
Nancy_Robertson wrote: }
} } Hi, } } I need good quality carbon coated grids for viewing purified plant virus } particles with TEM. I have had little success with "store bought" coated } grids. So unless someone could recommend a good cc grid source, I will } need to buy a carbon coater. Should I purchase a carbon coater using } flash evaporation or the more expensive carbon turbo evaporator? } In addition, could anyone recommend an economically good microtome for } thin sectioning plastic embedded specimens? } } Thank You, } } Nancy L. Robertson, Research Plant Pathologist } USDA/ARS } National Arctic Plant Germplasm Resources Unit } 533 Fireweed Ave. } Palmer, Alaska 99645 } phone: 907-746-9465 } fax:907-746-2677 } e-mail: nancy_robertson-at-dnr.state.ak.us
------- Forwarded message follows ------- } From: Charles Keith {chkeith-at-cb.uga.edu} To: farmer-at-emlab.cb.uga.edu Date sent: Thu, 5 Oct 2000 09:47:00 -0400 (Eastern Daylight Time) Priority: NORMAL
Mark -
Could you post a request to your microscopy listserv for the index of refraction of Dulbecco-Vogt PBS? I have done so to sci.techniques.microscopy
---------------------- Charles Keith Associate Professor, Department of Cellular Biology (http://www.uga.edu/~cellbio) University of Georgia chkeith-at-cb.uga.edu
John Shields EM Lab 151 Barrow Hall UGA campus 2-4080 fax: 2-4271 jshields-at-cb.uga.edu
Martin Microscope Company of Easley, SC, USA makes an adapter for the Coolpix 990 to fit the eyepiece tube of any microscope (both 23mm and 30mm diameter tubes) for $300. They can also get you an adapter for C-mount. Their phone number is 864-242-3424 or 864-859-2688
} } } Gary Radice {gradice-at-richmond.edu} 10/05/00 01:47PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
You might also try contacting Bunton Instruments in the US:
{http://www.buntgrp.com/}
They sell microscope adaptors for the CoolPix. The adaptors are not listed on their web site, yet, but call or e-mail them for information. I know they are taking orders for adaptors now and I was told that they would be shipping "around the end of October."
I have no connection with Bunton Instruments other than as a customer.
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 thought the only drawback was cost. How much of a cost differential are you being quoted?
I thought all manufacturers had to pick through crystals to find ones suitable for EDS detectors in regard to dead layer, impurity, etc. If good Ge chips are harder to fine, it would mean more expensive detectors.
We haven't tried thermally cycling our detector. I have heard that voltage applied to Si(Li) detectors while warm will lead to a loss of Li, ruining the detector. I don't know what effect it would have on a Ge detector. However, my manager is reluctant to find out if there would be a problem so we just keep our system cool.
Warren
At 09:10 AM 10/6/2000 +0100, you wrote:
} Thank you all for your replies on this topic. Several people seem to } like Ge detectors for low keV work, and the replies have mostly been } very positive about their resolution and performance. Some authors } strongly advocate the use of Ge detectors for low kV, low probe } current work with light element samples (See, for example, Block- } van Hoek & Pinxter (1993) Chapter 1 in X-ray microanalysis in } biology edited by Sigee et al, Cambridge University Press). Their } opinion (and apparently yours) stands in contrast with the advice } given by all of the XRMA vendors I am speaking to, whose opinion is } that Si(Li) is the way to go (granted some of them don't sell Ge, so } they would obviously have to advocate Si). So my question is "what } is the Achilles heel of Ge detectors?". I am aware for example that } their potentially very favourable characteristics at low kV are spoiled } by the practical difficulty of manufacturing them without a large dead } layer. Is this a fatal flaw in practice for analysis of elements with z } {11? Are they less robust, less tolerant of thermal cycling? If } anyone can help me get to the bottom of this I would be most } grateful. } } Chris
---------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
We have a new Nikon adaptor from Image Systems that allows us to attach our CoolPix 950 to our Zeiss Axioscope 2 via the C-mount or through the eye tube if we wanted to do it that way. Image Systems also sells something they call a cable release adaptor which is a "fancy piece" that hooks onto the camera over the shutter button so that a cable release can be screwed in there. This way we can take a picture without putting pressure on the adaptor set-up or programming the camera to do it. Image Systems in Columbia Maryland can be reached at 410-995-0748.
Just a recent customer.
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
Nancy - Try Ladd Research (www.laddresearch.com) for high quality grids. I've been using their C-coated grids for several years now and have found that nearly every grid has been perfect. Their grids are somewhat more expensive than those from other manufacturers, but well worth the price if you need good quality.
I haven't tried purchasing grids from every manufacturer so there may be other companies that also produce fine C-coated grids.
Dave Joswiak Dept. of Astronomy Univ. of Washington Seattle, WA 98195 (206)543-7702 joswiak-at-astro.washington.edu
On Thu, 5 Oct 2000, Nancy_Robertson wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Hi, } } I need good quality carbon coated grids for viewing purified plant virus } particles with TEM. I have had little success with "store bought" coated } grids. So unless someone could recommend a good cc grid source, I will } need to buy a carbon coater. Should I purchase a carbon coater using } flash evaporation or the more expensive carbon turbo evaporator? } In addition, could anyone recommend an economically good microtome for } thin sectioning plastic embedded specimens? } } Thank You, } } Nancy L. Robertson, Research Plant Pathologist } USDA/ARS } National Arctic Plant Germplasm Resources Unit } 533 Fireweed Ave. } Palmer, Alaska 99645 } phone: 907-746-9465 } fax:907-746-2677 } e-mail: nancy_robertson-at-dnr.state.ak.us } }
Dear Carla, The double-sided, conductive tabs that are sold in most EM supply catalogues are excellent and provide a good, strong hold. I use DAG 154 from Acheson Colloids, which is colloidal carbon in iso-propanol and ethanol, as a conductive paint to connect the coated surface to the stub. At 06:46 AM 10/5/00 -0700, you wrote: } } Hello all, } } I am attaching CP dried salmon skins (approx. 2-3 mm thick) to stubs with } Electrodag 502 for secondary electron imaging. The adhesive is a graphite } paint in MEK base (a substitute for TV Tube Koat). After ethanol } dehydration and CP drying in liquid CO2, the samples are relatively flat or } slightly curved. However, once attached to the stub the skins develop a } noticeable curl which leaves a gap between the stub surface and sample. In } some cases I can see the "trail" in the adhesive left by the skin as it was } curling! Filling the gap with more Electrodag is a waste of time because } it evaporates and shrinks back away from the sample (thankfully it doesn't } seem to produce more curling). } } Could anyone suggest a better conductive adhesive? Also, is there any } difference between aqueous (eg. Aquadag colloidal graphite) and MEK-based } media as to its effect on biological samples? } } As an SEM novice any advice you could share with me is greatly appreciated. } Thank you very much, } } Carla Aiwohi } Western Fisheries Research Center } Seattle, WA } ph: (206) 526-6282 ext.242 } fax: (206) 526-6654
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'm looking to replace my two year old Polaroid Digital } Microscope camera ...
If your camera-microscope interface is via 1x c-mount, then the Nikon CP990 and the 1x c-mount adapter is a very good choice ... or convert your microscope tube to c-mount (1x).
A point should be made ... the CP cameras acquire images to an internal memory card ("Compact Flash") ... than cannot acquire direct to a computer. The transfer is instead accomplished at some other time ... either a direct connection, camera to a serial or USB port, or (better/faster) by removing the memory card and using a "Compact Flash" card reader. This might be a hassle for a workstation type microscope, but if freedom of location is important, then probably works better.
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/
Another drawback is the need to keep them at 77 K all the time (see below).
I have heard that voltage applied to Si(Li) detectors while warm will lead to a loss of Li, ruining the detector.
Not actual loss of Li. To produce a Si(Li) detector, Li atoms are diffused into a Si crystal, which is then subjected to a bias voltage and heated. Li moves toward the cathode in such a way as to compensate for excess electron acceptors, and this results in a large fraction of the detector volume being depleted of carriers and, thus, having a very low dark current. When warmed to room temperature in the presence of bias voltage, Li is redistributed, and the detector no longer has the necessary large depletion region. It is possible to get the same effect with ultra pure Si or Ge, but such detectors--called "intrinsic"--are very expensive due to the stringent limit for impurities. It is now known that in the absence of bias voltage Si(Li) detectors can be at room temperature for a while without damage (although 15 years ago that was not thought to be the case), but Ge(Li) detectors must be kept at 77 K even when no voltage is applied. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
Chris I also have an Oxford intrinsic Ge detector on my SEM and enjoy all the advantages described by previous correspondents. However, as the max voltage on my SEM is 30kV the heaviest element for which I can get K line info (which is preferable to L-lines for quantification) with decent S/N ratio is around Sn (Z = 50). This of course depends on the amount of tin present in your sample.
If you have a Ge detector fitted to a TEM operating at 200 kV, then, in principle, you should be able to detect K lines for all elements. As a result, installation of an intrinsic Ge detector on a TEM may also be a good thing.
Alan Fox
Alan Fox Center for Materials Science and Engineering Naval Postgraduate School Monterey CA 93943
My company is interested in selling a fairly new SEM and we are having a difficult time finding an appropriate place to list it. Can anyone give some suggestions. Also is there any kind of depreciation formula for estimating an appropriate valuation.
Thanks
Raj
********************************************** 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 ********************************************** "Innovative Tools to Explore the Microworld"
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EM-listers: I frequently get asked for commercial sources for devices that plunge freeze TEM grids into a cryogen such as liquid ethane. We have our own in-house developed device so I have never kept up with what is on the market. Available devices also seem to come and go so some of my old sources no longer are current. Could I get vendors that have these devices to respond to me off the list. I would appreciate a reference to a web page if one exists and whatever ordering information is available. I will be posting this information on our lab web page and if there is enough interest on this list I can also post the responses.
Thanks Norm Olson
******************************************* Norm Olson Senior Research Electron Microscopist Department of Biological Sciences Lilly Hall of Life Sciences Purdue University West Lafayette, IN 47907
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What other liquids can be used for floating off thin sections other than water? I have a sample that swells in water. It swells slowly in alcohol. Thanks in advance.
I have been asked about the mechanism of Lugol's Iodine as a fixative and I am not sure how it works. Is it a coagulent or anitcoagulent fixative? Does it cross-bind proteins like formaldehyde or formalin? It is used often for fixing dinoflagellate samples and it seems to stabilize the structure of the organisms fairly well without alcohol. Can anyone explain to me the mechanism?
Charles T. Plybon Florida Marine Research Institute Research Staff Aquatic Health Group Tel: (727) 896-8626 e-mail: charles.plybon-at-fwc.state.fl.us
Does any one out there etch L. R. White sections? With sodium-meta-periodate, or something else? Does it really make a difference? Tim
Timothy G. Schneider Director of Electron Microscopy Department of Pathology Room 229 Jefferson Hall Thomas Jefferson University 1020 Locust St. Philadelphia Pa. 19107 215-503-4798 work 610-613-8170 cellular timothy.schneider-at-mail.tju.edu
Fabian-Baber Communications is a multimedia educational production company. We are currently producing a ten-part video series on the human body to be used in middle-school classrooms. We are looking for motion microscopy of human cells, such as blood cells coursing through arteries, cells during mitosis, etc. If you are interested in contributing to this project, or know of a good resource for these images, please contact , Somayyah Siddiqi, at sam-at-fabian-baber.com or 610 623 7812. Thanks so much!
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 would appreciate the benefit of your experience and knowledge in answering several questions for me as we prepare to purchase our CPD for use in teaching undergraduate biology courses and student/faculty research.
What is your recommendation re the following:
having / not having a viewing port: [I've heard a few recommendations for having a viewing port in a CPD [all accompanied by warnings re the inherent danger]. The only CPD I've used [years ago] had a port which I liked, especially for teaching purposes. I also been asked "Why would you want one?"
chamber size and orientation: vertical or horizontal:
type[s] of specimen holders:
electrically heated vs. hot water heated:
manufacturers and models that you use or have used recently:
costs of fluids and materials used in / with your CPD for a typical run:
compared to other models you might be familiar with: Specifically, is it too much more expensive to run a Polaron jumbo chamber vs. their regular size chamber? [again, for biological material].
What would you look for in your next CPD?
Thanks for your input and time -- they're greatly appreciated.
Lee Hadden Department of Biology Wingate University Wingate, NC 28174 hadden-at-wingate.edu 704.233.8238.
If you are trying to "antigen retrieve" on LR White, there is a nifty paper - Stirling and Graff, 1995, Journal of Histochem. and Cytochem., 43:115-123. "Antigen Unmasking for Immunoelectron Microscopy: Labeling is Improved by Treating with Sodium Ethoxide or Sodium Metaperiodate, then Heating on Retrieval Medium"
Tamara Howard CSHL
On Fri, 6 Oct 2000, Timothy Schneider 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 } -----------------------------------------------------------------------. } } } List severs: } } Does any one out there etch L. R. White sections? With } sodium-meta-periodate, or something else? Does it really make a difference? } Tim } } Timothy G. Schneider } Director of Electron Microscopy } Department of Pathology } Room 229 Jefferson Hall } Thomas Jefferson University } 1020 Locust St. } Philadelphia Pa. 19107 } 215-503-4798 work } 610-613-8170 cellular } timothy.schneider-at-mail.tju.edu } } }
Nancy Robertson wrote: =============================================================== } I need good quality carbon coated grids for viewing purified plant virus } particles with TEM. I have had little success with "store bought" coated grids. } So unless someone could recommend a good cc grid source, I will need to buy a } carbon coater. Should I purchase a carbon coater using flash evaporation or the } more expensive carbon turbo evaporator? In addition, could anyone recommend an } economically good microtome for thin sectioning plastic embedded specimens ? ================================================================ SPI Supplies has produced, in house, with our own (in-house) experienced staff, carbon coated grids, including holey and lacey types, for many years. I hope your less-than-expected results from purchased grids were not our grids, because we really are set up to make them however one does want to make them, mainly because we have our own TEM facilities at our side to inspect what we have made. In any case, many laboratories worldwide depend on the SPI carbon coated grids and we are not aware of any problems that anyone has encountered with them (lots not passing inspection are rejected and not sent to customers). Some want them thicker, some want them thinner, some want them with large holes, some smaller holes, but we can make them however someone wants them to be!
With regard to your question: "Should I purchase a carbon coater using flash evaporation", I will assume that you are asking about the "carbon coaters" that are operated with a rotary vane mechanical pump, usually in tandem with a sputter coater. We are not aware of anyone that has ever gotten acceptable carbon coatings that could be used as support films by TEM using such an equipment approach. Only a diffusion pumped vacuum system, or better, seems to be capable of producing the kind of carbon coatings that would meet that standard. A number of firms offer vacuum evaporators, and we have found the vacuum evaporators made by VG Microtech and also, Denton Vacuum to be especially good for this kind of applications. Quite possibly, others are equally good, but our experience has been with these two particular manufacturers. Note: We do not have any interest, financial or otherwise, in either Denton Vacuum or VG Microtech.
We had the good fortune of learning about a paper soon to be published comparing carbon grids from different commercial sources. The author is P. J. F. Harris, Dept. of Chemistry, U. Reading, Whiteknights, Reading RG6 6AD UK. The title is: Carbonaceous Contaminants on Carbon Support Films for Transmission Electron Microscopy, Carbon 1 (2000).
If Dr. Harris is correct, then one possible solution, since they are made using a different approach (but were not studied by Dr. Harris), might be the Quantifoil grids (my opinion), see URL http://www.2spi.com/catalog/grids/quantifoil-carbon-films.html
They are produced using a different kind of process and therefore would be expected to not have present some of the ambiguous features found on the carbon films studied in the paper mentioned above.
I don't know if this paper has been published yet or not, but I could send to any one requesting it, the entire preprint of the paper via pdf file. Such requests should be made off-line. Give me a day or two to respond.
Disclaimer: SPI Supplies has manufactured since 1975 custom made carbon support films for TEM applications. Grids made by SPI were one of those studied in the above referenced article. However, it is entirely possible that given some of the new demands being made on carbon coated grids generally, especially for high temperature stability, that one should consider as an alternative, the SPI Silicon Nitride Membrane Grids, as described on URL http://www.2spi.com/catalog/instruments/silicon-nitride.html
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 ######################## ============================================
Allen R. Sampson wrote: } } Have an EDS system attached? The EDS dewars, being generally supported by } a cantilever structure, can be quite effective at coupling acoustic } vibrations into an SEM. The large area and thin metal construction of the }
EDS dewars: In regard to EDS dewars and acoustic vibrations, if there is enough ice or dirt in the dewar to nucleate bubbles in the LN2 then you might get sufficient vibration as the LN2 boils off. You can often hear this simply by putting your ear against the dewar, especially while the LN2 is at a relatively low level in the dewar. Perhaps in really vibration sensitive systems such as a FESEM this could be another potential problem? The sound of the boiling can range from a continuos fizz to an occasional intense "bump" similar to the effect that you can get just as water begins to boil in a pot on a stove.
Stage motors: We had a bad problem with stage motors once. If you have a motorised stage then any AC ripple from bad power supply filtering remaining on top of the DC holding current can transmit enough vibration via the motor coils to cause problems at high magnification. Sometimes you can "feel" these kinds of vibrations in the motor bodies. Since this holding current is present even when the motors are not actually moving the stage, you have to actually shut off the power to the stage motors to determine if this is the cause.
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/
The October meeting of NESM (New England Society for Microscopy) will be held on October 17th at FEI Company in Peabody, MA.
Pre-registration deadline is Friday, October 13th. Please contact Mary McCann, NESM Treasurer at (617)484-7865 or by e-mail: mccanns-at-tiac.net. Registration is $5.00 for members and $20.00 for non-members (this fee includes a current-year membership in NESM).
The meeting will begin with a tour of FEI Company at 5:30pm. Also at this time, Dr. Ronald Mueller of Bristol Myers Squibb will speak on the utility of SEM within the Pharmaceutical Industry. A buffet dinner will follow at 6:00pm.
The scientific presentations will begin at 7:00pm. The speakers are: Dr. Ben Fabry of Harvard School of Public Health, Boston, MA-"Magnetic Twisting Cytometry with Sub-Micron Optical Detection"
Dr. Dan Friend of Brigham and Women's Hospital, Boston, MA-"Thin Section: Freeze Fracture Correlates-A Visit to the Good Old Days"
Barbara Foster-President, Microscopy, Marketing and Education, Springfield, MA- "Bridging the Microscopy-Spectroscopy Chasm"
Thanks to all who sent suggestions and options for connecting the CP990 to Zeiss Axioskop. There are several options. After examining each one, here is a good approach to using the CP990 on an Axioskop.
The first accessory is the attachment of the camera to the 'scope. This is done using an Axioskop trinoc photo port adapter to C-mount. Then, a CP990 thread ring to C-mount adapter is installed. These two adapters then mate the CP990 to the photo port of the Axioskop. Optmem makes all of the pieces needed to make the mating between 'scope and camera. The mate from photo port to C-mount can be done two ways.
Diagnostic Instruments makes a 38mm ISO mount (PN DZA, $275) which also accepts items for Leitz. This same ISO mount onto the Zeiss will accept a PA-41A adapter for Nikon F mount cameras. Optem makes a coupler DC10NN ($80) which mates the 38mm ISO port to C-mount. Then, their 25-70-14-03 ($320) is the CP990 to C-mount adapter. This completes the adaption for a total cost of $275 + $80 + $320 = $675.
Optem's alternate method is the DC-10AM ($150) which directly mounts in the trinoc photo port and provides a C-mount. Then, this mates with the same 25-70-14-03 CP990/C-mount adapter. Total cost of this approach is $150 + $320 = $470.
Zeiss also makes an Axioskop photo port C-mount adapter. This part number is 45 29 95. They can be found used from between $125-$250. Then, the Optem 25-70-14-13 CP990 adapter is used to complete the mate.
When using the CP990, or any other camera for that matter, vibration is a key issue. In this respect the CP990 has an advantage since it does not have a mechanical shutter as do other CCD cameras. But one still needs to reduce vibration when actually taking a pix. This is done via a remote cable release. There are several aftermarket units which perform this function. However, they are rather awkward and large. The best approach is the Nikon EU-1 wired remote ($195 list price). This is a small hand held puck which plugs into the USB port on the CP990. Pressing the large button down half way is like doing so on the CP990 shutter release button. Pressing all the way down takes the pix. The puck also indicates on an LCD how many frames are left.
When doing 'scope work, I recommend the Nikon EH-31 AC adaptor ($50 list price). This avoids drain on the camera batteries.
For storage, the CP990 uses CF type 1 media. The IBM microdrive will not work in this camera. Since the camera stores images in JPEG format, the file size of each image will vary according to how the image compresses. So far, the largest file I have taken is just over 1MB. I am using 128MB CF media. So I can reasonably expect to get about 100 frames per CF. I have two of these CF media. When one is full, it is pulled and replaced with an empty one. The full one is emptied using a Microtech USB CF reader ($38).
Finally, image focusing has always been a problem for me. The CP990 changes all of that. Using the CP990 real time NTSC video output jack, one can watch the image on a separate TV monitor (b/w or color) and adjust for perfect focus. Then take the pix. If you want the camera's LCD unit to stay on indefinitely while shooting, use the setup menu and disable the power off feature.
That's about it for the CP990. Next is the Fuji Finepix S1 Pro. One camp says it will work on a microscope. Another camp says it won't. The camera only does automatic modes when using a lens. When the lens is removed, the camera becomes dumb. What is will actually do remains to be seen. The folks at ElectroImage say that it does work.
For some time, Lugol's Iodine has been used not only to stain certain cells but also to fix and preserve samples of dinoflagellates and other zooplankton. I am curious as to what the mechanism is for stabilizing and fixing with Lugol's. Coagulent or anitcoagulent, does it cross-bind proteins preserving cellular integrity? Can anyone help?
Charles T. Plybon Florida Marine Research Institute Research Staff Aquatic Health Group Tel: (727) 896-8626 e-mail: charles.plybon-at-fwc.state.fl.us
Scott, Glycerol at different concentration (87% as a starter). I use it for PVP and PVA blocks. Marek.
At 15:35 2000-10-06 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Marek Malecki, M.D., Ph.D. Director and Principal Investigator
Molecular Imaging Laboratories University of California, San Diego
} For storage, the CP990 uses CF type 1 media. The IBM } microdrive will not work in this camera. Since the } camera stores images in JPEG format, the file size of } each image will vary according to how the image } compresses. So far, the largest file I have taken is } just over 1MB.
First ... thanx for compiling this info. However and regarding the statement above ... are you sure the CP990 does not provide for saving TIFFs? My CP800 will save a single high res TIFF to a 8Mb memory card. Presumably one could save ~8-10 CP990 TIFFs to a 128Mb CF card(?) Get back to us on this one ... I'm afraid "JPEG only" would dissuade those who believe in "non-lossy" TIFFs only.
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/
Hi All, Sorry to bother you with such a strange request. Does anyone have contact information for SeaLite Sciences (associated with bioluminescent markers)? All I have is a web site, and after several days of trying to get through, I've just about given up. Thanks for your help once again, Kristen
At 07:16 AM 10/9/00, you wrote: } Gary writes ... } } } For storage, the CP990 uses CF type 1 media. The IBM } } microdrive will not work in this camera. Since the } } camera stores images in JPEG format, the file size of } } each image will vary according to how the image } } compresses. So far, the largest file I have taken is } } just over 1MB. } } First ... thanx for compiling this info. However and regarding the } statement above ... are you sure the CP990 does not provide for saving } TIFFs? My CP800 will save a single high res TIFF to a 8Mb memory } card. Presumably one could save ~8-10 CP990 TIFFs to a 128Mb CF } card(?) Get back to us on this one ... I'm afraid "JPEG only" would } dissuade those who believe in "non-lossy" TIFFs only.
Yes, it does have a TIFF mode. But this is only available via the Manual mode. I figured that most folks wanted quick and easy access to image capture. If they want/need TIFF, they will have to attack the manual mode.
Larry: We would be very interested in hearing of what you come up with once you have a solution. Mike O'Keefe
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 } -----------------------------------------------------------------------. } } Alan: } } Just this morning we were discussing the same problem with our Gatan } CCD cameras on our HD-2000, one of which is retractable and used for } TEM imaging, and the other of which is at the end of the GIF for } energy-filtered imaging. These cameras are connected in series for } the water flow, and have very tiny water lines which can easily clog, } even with recirculated water. There is no provision by the } manufacturer to provide for a safety cut-off of the Peltier cooler in } the event of loss of water flow. We have recently experienced a } burn-out of the TEM camera due to a clog in the line, which resulted } in 2 weeks of downtime for repair (nicely done for free by Gatan), } but still no response to queries about some sort of protection. So } we have decided to find an appropriate water flow sensor, and to } install it in an appropriate place in the water line, and to connect } it to the camera controllers to provide a power cut-off when the } water flow diminishes. I'll post the details of the solution when it } is completed. } } Larry } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 } } } } Have any other microscopists had problems with Peltier coolers failing on } } water cooled CCD cameras? } } } } In my facility I have three such cameras all plumbed into the microscope } } water system and connected, by the camera manufacturer, into wall sockets. } } On one of the three cameras I have had the Peltier cooler fail four times } } in two years. After the latest failure the manufacturer asked if the } } camera had been run without water. I told him not intentionally but when } } we have power cuts and the power is restored, the camera comes back on but } } the microscope (and water) does not. The camera can therefore run for some } } time without water cooling. Power failures are not uncommon at this } } university! } } } } Many e-mails to the manufacturer have resulted in no response to my } } questions about reliability of the Peltier coolers. The manufacturer is } } treating the failure as an out of warranty issue but I believe there has } } been something wrong with this camera since it was installed. } } } } Any experiences of listservers with Peltier cooler failures would be } } appreciated. Please e-mail me directly. } } } } Regards } } } } Alan W Nicholls, PhD } } Electron Microscopy Service Director } } Research Resources Center - East (M/C 337) } } Room 100 Science and Engineering South Building } } The University of Illinois at Chicago } } 845 West Taylor St } } Chicago, IL 60607-7058 } } } } Tel: 312 996 1227 } } Fax: 312 996 8091 } } Office: Room 110 } } } } Web site www.rrc.uic.edu } } } } } } Alan W Nicholls, PhD } } Electron Microscopy Service Director } } Research Resources Center - East (M/C 337) } } Room 100 Science and Engineering South Building } } The University of Illinois at Chicago } } 845 West Taylor St } } Chicago, IL 60607-7058 } } } } Tel: 312 996 1227 } } Fax: 312 996 8091 } } Office: Room 110 } } } } Web site www.rrc.uic.edu } } 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
The 2000 Fall Meeting of the Texas Society for Microscopy will be held on October 26, 27, and 28 at the DoubleTree Hotel in Dallas, TX. The hotel is located in Campbell Centre at 8350 N. Central Expressway, Dallas, TX 75206. The phone number for the hotel is 800-222-TREE. A special workshop will be held on Thursday, Oct. 26, entitled "The SEM Today". It will cover the latest information concerning basic SEM, LVSEM, ESEM and detectors.
For more information and/or to register, please contact Pamela Neill, Program Chair TSM Alcon Research LTD 6201 South Freeway Forth Worth, TX 76134-2099 pamela.neill-at-alconlabs.com
We at Ladd may be able to help you. Please let us know what type of work you want to do with it ((target, size of sample, type of sample prep, etc) and I'll let you know what we have available.
Best Regards,
John Arnott
LADD RESEARCH 131 Dorset Lane Williston, VT 05495 USA
web site http://www.laddresearch.com
TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail sales-at-laddresearch.com
Microscope Supplies Since 1955
John andrew wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------.} } I need to purchase a sputter coater for under 5K. Can } this group recomend what would be a good purchase. } } Thank you } } __________________________________________________ } Do You Yahoo!? } Yahoo! Photos - 35mm Quality Prints, Now Get 15 Free! } http://photos.yahoo.com/
We appreciate the comments Dr. Joswaik and others concerning the quality of Ladd coated grids. Since there are still a number of researchers who coat their own grids, we are pleased to share some of the things we have learned over the past 45 years.
1. Clean the evaporator at least once a day. 2. Change the diffusion pump oil at least every two weeks. 3. Inspect and clean each individual grid prior to coating. 4. Out gas the garbon prior to evaporation. 5. Use fresh chemicals with each batch of grids to be coated.
Hopefully some of this will be of assistance. A recent posting mentioned that a diffusion pump system should be used for carbon coating. We agree 100% with that. In fact we designed the Ladd vacuum system with coated grids in mind by providing for "quick changing" the oil and speedy clean-up.
John Arnott --
LADD RESEARCH 131 Dorset Lane Williston, VT 05495 USA
web site http://www.laddresearch.com
TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail sales-at-laddresearch.com
Dear listers, We received several VERY large pieces of horse tendon, half of which was immersed in ethanol and the rest in formalin. The vet didn't know which would be best , so that's how it arrived, via air from New Mexico. We cut it into 1 x 3 mm pieces and processed it routinely, ie, glut, OsO4, ethanol dehydration and PO. We infiltrated in Spurr's and PO for 24 hrs, straight Spurr's for 2 hours and then embedded. What was left just fell out of the blocks . Needless to say, we have a lot of tissue left (still in EtOH and fomalin) and I was wondering if it's worth trying to salvage and if anyone has any suggestions as to the best way to handle this. I did ask the client to please contact me before sending any more tissue but for now they would like us to try and get something from this. Any ideas would really be appreciated. Thank you,
Mary Gail Engle Electron Microscopy & Imaging Facility University of Kentucky
We are considering purchase of a system for quantitative analysis of bacterial and fungal biovolume in soil samples. Could anyone provide info on equipment and preferred method (FITC labeling?). As always, I appreciate your help, Alice.
Alice Dohnalkova Environmental Microbiology Battelle, PNNL MS P7-50 Richland, WA 99352 tel. (509) 372-0692 fax (509) 376-1321
before i go through the effort of separating the ground wire from the rest of the supply system just a few more tidbits to throw out to the list:
1- the distortion in the scan seems to be about 60Hz (more cycles at slower scan rates). in reduced area raster (about 4 frames/sec??) the count is about 15. in "TV" (whatever rate that is on a 35) i get about 2 or 3 cycles.
2- the amplitude of the distortion increases a bit with decreasing KV (i.e. its worse at 10KV than at 30KV...but not much worse). and it seems to be worse at higher magnifications (at 10KX it just about covers the screen left to right; at 500X it is just a couple of bumps).
3- i tightened all the ground lugs....no change.
4- i tightened all the console connectors...no change.
5- i moved the variac farther away from the console...no change.
6- perhaps unrelated.....the conformation of the "spot" realized when the condenser lens current is decreased looks funny. i'm used to a realtively uniform circular cross section. this one looks like a central uniform area with dendritic "arms" radiating outwards...kinda like a spider web pattern. does this mean anything to anyone?? (the final apertures are pretty clean, and it looks funny with any of the three apertures)
thanks for all your previous comments...i'm sure to be closing in on fixing this with your help.
brian ---------------------------------------------- Brian McIntyre Electron Microscopy Lab- River Campus Univ of Rochester Rochester, NY 14627 716-275-3058/4875
Hi Scott. 'Bonne chance', as we say in Canada. A true water substitute (ie in both knife edge wetability and surface tension) is one of those Holy Grail things in ultramicrotomy. You could experiment with liquids like alcohol mixtures, ethylene glycol, etc, but my suggestion is to save yourself a lot of time and simply dry section. It's done more often than you think; water-soluble minerals, reactive metals like Mg, to name a few. Place a grid on the lower part of the diamond knife, section, then tease the section onto the grid with your hair tool. An important point is to then wet the section down onto the grid with something inert (perhaps an alcohol), then dry under a lamp, so that it doesn't fall off the grid on insertion into the TEM. If there is a danger of a somewhat brittle material breaking apart upon sectioning, use a coated grid and some of the segments should be retained.
Do let the Listserver know if someone out there suggests something that works as well as water. You'll have a lot of friends at the next MSA meeting!
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: Walck, Scott D. [SMTP:walck-at-ppg.com] Sent: Friday, October 06, 2000 3:35 PM To: 'Microscopy-at-MSA.Microscopy.Com' Subject: alternate fluid for ultramicrotomy
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What other liquids can be used for floating off thin sections other than water? I have a sample that swells in water. It swells slowly in alcohol. Thanks in advance.
Just wanted to say thank you for the many replies to my TEM/SEM problem with bacterial culture protocol. Problem solved, thanks again. Kristine C. Fambrough NMSU
Many years ago, I was trying lipid retention embedding for TEM of mycobacterium. GACH seemed to work but sectioning into water was a nightmare. It attracted water from the trough during the ultramicrotome's return stroke which then caused the block to swell. Freon worked better, but the sample from DuPont was quite volatile and evaporated quickly. The workaround was to section into water, but to coat the sides of the block with the thinnest possible film of silicone grease. The grease provided sufficient hydrophobicity to prevent water from jumping onto the face, and if I worked quickly the sections could be placed onto the grids. Needless to say, I only used a glass knife for this. It was a short project, so column contamination from volatiles in the grease wasn't a problem. Maybe a sample of the grease could be put under a high vacuum beforehand to remove volatile components.
Regards, Glen
"Malis, Tom" 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 Scott. 'Bonne chance', as we say in Canada. A true water substitute (ie } in both knife edge wetability and surface tension) is one of those Holy } Grail things in ultramicrotomy. You could experiment with liquids like } alcohol mixtures, ethylene glycol, etc, but my suggestion is to save } yourself a lot of time and simply dry section. It's done more often than } you think; water-soluble minerals, reactive metals like Mg, to name a few. } Place a grid on the lower part of the diamond knife, section, then tease the } section onto the grid with your hair tool. An important point is to then } wet the section down onto the grid with something inert (perhaps an } alcohol), then dry under a lamp, so that it doesn't fall off the grid on } insertion into the TEM. If there is a danger of a somewhat brittle material } breaking apart upon sectioning, use a coated grid and some of the segments } should be retained. } } Do let the Listserver know if someone out there suggests something that } works as well as water. You'll have a lot of friends at the next MSA } meeting! } } 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: Walck, Scott D. [SMTP:walck-at-ppg.com] } Sent: Friday, October 06, 2000 3:35 PM } To: 'Microscopy-at-MSA.Microscopy.Com' } Subject: alternate fluid for ultramicrotomy } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } What other liquids can be used for floating off thin sections other } than } water? I have a sample that swells in water. It swells slowly in } alcohol. } Thanks in advance. } } -Scott Walck }
--
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. ************************************************************************
Hello Guys I am a novice user of a SEM (LEO 1450 to be precise) equipment which was just installed in our central science lab. Is there a way to clean stubs for reuse or are stubs disposable once used. ( This may sound naive) soji Mr. O. O. ILORI DEPARTMENT OF ELECTRONIC/ELECTRICAL ENGINEERING OBAFEMI AWOLOWO UNIVERSITY, ILE-IFE, OSUN STATE NIGERIA. email: sojilori-at-oauife.edu.ng
Hi All, I'm tring to find users of JEOL JSM-840 (or similar) microscopes in the Atlanta area. In addition, I need to find contract SEM analysis labs here that might be able to run some fast turnaround samples for me (mostly cross sections). I know several of you contacted me recently after I posted for a microscopist, but I no longer have the information. Could you please send mail again to sbuckingham-at-excellatron.com . Many Thanks, Steve
Steve Buckingham Excellatron Solid State LLC 1460 Roswell Street, Suite J Smyrna, GA 30080 phone 770 438 2201 fax 617 812 5920
Dear Brian, The profile of the spot you describe sounds like an under-saturated filament. At 08:14 AM 10/10/00 -0400, you wrote:
} hi again- } } before i go through the effort of separating the ground wire from the rest } of the supply system just a few more tidbits to throw out to the list: } } 1- the distortion in the scan seems to be about 60Hz (more cycles at } slower scan rates). in reduced area raster (about 4 frames/sec??) the } count is about 15. in "TV" (whatever rate that is on a 35) i get about 2 } or 3 cycles. } } 2- the amplitude of the distortion increases a bit with decreasing KV } (i.e. its worse at 10KV than at 30KV...but not much worse). and it seems } to be worse at higher magnifications (at 10KX it just about covers the } screen left to right; at 500X it is just a couple of bumps). } } 3- i tightened all the ground lugs....no change. } } 4- i tightened all the console connectors...no change. } } 5- i moved the variac farther away from the console...no change. } } 6- perhaps unrelated.....the conformation of the "spot" realized when the } condenser lens current is decreased looks funny. i'm used to a realtively } uniform circular cross section. this one looks like a central uniform area } with dendritic "arms" radiating outwards...kinda like a spider web pattern. } does this mean anything to anyone?? (the final apertures are pretty } clean, and it looks funny with any of the three apertures) } } thanks for all your previous comments...i'm sure to be closing in on fixing } this with your help. } } brian } ---------------------------------------------- } Brian McIntyre
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
} ... } } 1- the distortion in the scan seems to be about 60Hz
This might imply you have a ground loop problem. Possibly one of the other devices (computer, EDX, ...) is providing a ground connection independent of your primary ground(?)
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/
)( / _ Jonathan Lipkin () \ _ Assistant Professor of Multimedia Design )( / _ Ramapo College of New Jersey () \ _ http://arts.ramapo.edu/lipkin )( / _ 201/ 684/ 7056/
Dear listserv:
Dee Breger was kind enough to forward this message for me. I'm writing a book on digital photography, to be published by Harry N. Abrams next year. The book will include a section on applied digital imaging, and I am particularly interested in scientific imaging. I am hoping that some of you folks might be able to provide some information.
For the purposes of the book, I am defining digital imaging as that which describes a scene using discrete digital information in a raster grid, or bitmap. So, I am particularly interested in scientific imaging applications which gather information in this way.
I would be interested to hear if there are other scientific imaging systems (DEE: besides SEMs) which collect information in this manner. For instance, it sounds as though atomic force microscopes work in this way: as they drag a stylus across a surface, the position is recorded (x and y), along with the height (z). This is later used to re-create the image in a raster grid (the fact that the AFM does not use light makes it another interesting foil to 'traditional' digital photography). How then, are colors in the final output determined from the data collected? I would also, of course, love to see any images you might have to show.
Thanks in advance, Jonathan Lipkin (jlipkin-at-ramapo.edu)
DEE: I'll forward to Jonathan any replies that are publically posted to the listserver. He also needs other detailed technical information on digital image formation which can be far more comprehensively clarified and amplified by some of the experts among my fellow listers than by me, though I've already given him the broad picture for analog scopes like my dear old Cambridge 250. His SEM impressions, which I'll pass along verbatim, include (1) "SEMs collect information, used to fill a raster grid, through the use of a PMT", (2) the signal is sent directly to the crt in analog form -I always had thought that the pmt sent out digital info, but it sounds like it just sends out analog data (a waveform? though it couldn't be a waveform if the raster grid is discrete in two dimensions)" and (3) "as the secondary electrons come off the sample, they hit a detector which generates a digital number (redundant) which is then placed in a pixel of a bitmap."
Many thanks to any who can help!
*************************************************************** Dee Breger Mgr. SEM/EDX Facility Lamont-Doherty Earth Observatory 61 Route 9W Palisades, NY 10964 USA T: 914/365-8640 F: 914/365-8155
http://www.ldeo.columbia.edu/micro http://www.discovery.com/area/science/micro/micro1.html http://www.lsc.org/antarctica/front.html Journeys in Microspace (Columbia University Press, 1995)
Does anyone in the Chicago area have an ultrasonic disc cutter we can use temporarily? Alternatively, does anyone have one they are not using and would be willing to sell or donate?
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
If you aren't able to find an ultrasonic cutter locally to use, please let me know. Perhaps I can arrange to have one of our SoniCut 380 systems sent from our applications lab for short term use or maybe we could arrange to cut some samples for you here. Please let me know if I can be of help.
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 "L. D. Marks" } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Does anyone in the Chicago area have an ultrasonic disc cutter we can use temporarily? Alternatively, does anyone have one they are not using and would be willing to sell or donate?
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
Well we've got a new vacuum evaporator and it has been so long since I've had one I've forgotten how to treat the bell jar to reduce the effort it takes to keep it clean. However, I do recall that there were little tricks to cleaning and treating the glass and other surfaces. Anyone care to shed a little light my darkened memory of these procedures?
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
Another fault this weekend, this time the subscription list became corrupted. This mean't a significant number of you (~ 1500 ) "disappeared" for a few days and hence did not receive mail. Iunfortunatley did not notice since the mail did go through to the rest of the subscribers on the list. In any event , I have restored the list to it's Oct 6th backup copy.
To those of you that unsubscribed between Oct 6th and Oct 10th. I apologize and ask you to unsubscribe again. I, unfortunately, do not keep records of those of you that have left.
I am trying to get the internal(cytoplasmic) side of a prepared erythrocyte ghost's plasma membrane attached to coverslip, so that the external side is free in water solution. Does anyone have an experience with this or suggest the reagent that preferably interacts with inner leaflet of plasma membrane or cytoskeletal protein ? How should I test the external side is free ?
Any suggestions on this problem would be greatly welcomed. Thank you
Nguyen
Lab. of Appl. Microbiology Dep. of Biochemistry Faculty of Agriculture Tohoku University Sendai, Japan.
The easiest way to keep your bell jar clean is to wipe a thin film of detergent over the surface of the clean glass and pump down to dry. After each coating run simply rinse off the glass, the detergent comes away and takes the coating with it. Recoat with detergent and dry.
Train ALL users in this technique, insist they clean the glass if they "FORGOT" to do it after their last run, and you will always have a clear, clean bell jar. Regards JVN Rick Harris 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 } -----------------------------------------------------------------------. } } Well we've got a new vacuum evaporator and it has been so long since I've } had one I've forgotten how to treat the bell jar to reduce the effort it } takes to keep it clean. However, I do recall that there were little tricks } to cleaning and treating the glass and other surfaces. Anyone care to shed } a little light my darkened memory of these procedures? } } 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
-- **************************************************** 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'm getting ready to purchase a new sputter coater for our FESEM. I'd like to hear how lucky others were with using a "regular" sputter coater for high resolution SEM work. Currently, there is a noise problem with our SEM, so the resolution is about 3-4nm. I've successfully imaged 5nm ZrO2 close-packed particles, and 4nm pores in glass using a thin coating of Au/Pd applied with an old Polaron. One thing I'm worried about is = once the noise is fixed and I can resolve the published 1.5nm resolution will I be able to resolve the grains in the coating?
Rick A. Harris wrote: =============================================================== Well we've got a new vacuum evaporator and it has been so long since I've had one I've forgotten how to treat the bell jar to reduce the effort it takes to keep it clean. However, I do recall that there were little tricks to cleaning and treating the glass and other surfaces. Anyone care to shed a little light my darkened memory of these procedures? =============================================================== One product that will do this is called Bell Bright™. A companion product for metal surfaces is called Metal Bright™. Both are described in detail on the SPI website at URL http://www.2spi.com/catalog/supp/supp3.html
The concept is to leave a thin water soluble polymer behind on the clean glass (or metal) surface, and then when it is time to remove the evaporated deposited, a lint free cotton wiper and water alone will wipe away most of the material (as the originally deposited polymer film dissolves away).
Disclaimer: SPI Supplies "invented" this product in the late 1970's and has offered it ever since. We would have a vested interest in seeing its use increase.
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
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Toc toc. Mi scusi, sono Emma Bonino e la prego di partecipare al gioco online che stiamo organizzando perche' anche cittadini come lei (che non votino affatto radicale o che lo facciano, che non votino o non intendano piu' votare) possano partecipare alle nostre decisioni, essere presenti o rappresentati nei nostri organi dirigenti.
In vista delle elezioni politiche occorre tentare di allearsi con il Polo o con l'Ulivo? O combattere da soli? O organizzare il boicottaggio di elezioni non democratiche? Quali Riforme istituzionali, politiche, del lavoro, dell'impresa: "americane" o "tedesche", o nessuna? Quali politiche sulle liberta' individuali e sulla scienza, oltre che sulle droghe: proibizioniste o antiproibizioniste, come gia' sul divorzio e sull'aborto? E i partiti devono aprirsi a tutti i cittadini attraverso Internet, o restare "chiusi" come finora?
Dal 27 novembre al 3 dicembre si votera' online per eleggere 25 nuovi membri del Comitato Radicale. Se, come vivamente spero, lei intende partecipare a questo gioco, lo preannunci e registri cliccando qui: http://www.radicali.it/p_register/
Mi scusi ancora. A presto.
Emma Bonino
PS Questa email e' stata inviata nel rispetto della legge sulla privacy; se desidera maggiori informazioni o se intende cancellarsi, puo' cliccare qui: http://www.radicali.it/p_privacy_com3/
Toc toc. Mi scusi, sono Emma Bonino e la prego di partecipare al gioco online che stiamo organizzando perche' anche cittadini come lei (che non votino affatto radicale o che lo facciano, che non votino o non intendano piu' votare) possano partecipare alle nostre decisioni, essere presenti o rappresentati nei nostri organi dirigenti.
In vista delle elezioni politiche occorre tentare di allearsi con il Polo o con l'Ulivo? O combattere da soli? O organizzare il boicottaggio di elezioni non democratiche? Quali Riforme istituzionali, politiche, del lavoro, dell'impresa: "americane" o "tedesche", o nessuna? Quali politiche sulle liberta' individuali e sulla scienza, oltre che sulle droghe: proibizioniste o antiproibizioniste, come gia' sul divorzio e sull'aborto? E i partiti devono aprirsi a tutti i cittadini attraverso Internet, o restare "chiusi" come finora?
Dal 27 novembre al 3 dicembre si votera' online per eleggere 25 nuovi membri del Comitato Radicale. Se, come vivamente spero, lei intende partecipare a questo gioco, lo preannunci e registri cliccando qui: http://www.radicali.it/p_register/
Mi scusi ancora. A presto.
Emma Bonino
PS Questa email e' stata inviata nel rispetto della legge sulla privacy; se desidera maggiori informazioni o se intende cancellarsi, puo' cliccare qui: http://www.radicali.it/p_privacy_com3/
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MULTI-LEVEL MARKETING (MLM) has finally gained respectability. It is being taught in the Harvard Business School, and both Stanford Research and the Wall Street Journal have stated that between 50% and 65% of all goods and services will be sold through multi-level methods by the mid to late 1990's. This is a Multi-Billion Dollar industry and of the 500,000 millionaires in the U.S., 20% (100,000) made their fortune in the last several years in MLM. Moreover, statistics show 45 people become millionaires everyday through Multi-Level Marketing.
You may have heard this story before, but over the summer Donald Trump made an appearance on the David Letterman show. Dave asked him what he would do if he lost everything and had to start over from scratch. Without hesitating, Trump said he would find a good network marketing company and get to work. The audience started to hoot and boo him. He looked out at the audience and dead-panned his response: "That's why I'm sitting up here and you are all sitting out there!"
The enclosed information is something I almost let slip through my fingers. Fortunately, sometime later I re-read everything and gave somethought and study to it. My name is Johnathon Rourke. Two years ago, the corporation I worked at for the past twelve years down-sized and my position was eliminated. After unproductive job interviews, I decided to open my own business. Over the past year, I incurred many unforeseen financial problems. I owed my family, friends and creditors over $35,000. The economy was taking a toll on my business and I just couldn't seem to make ends meet. I had to refinance and borrow against my home to support my family and struggling business. AT THAT MOMENT something significant happened in my life and I am writing to share the experience in hopes that this will change your life FOREVER FINANCIALLY!!!
In mid December, I received this program via e-mail. Six month's prior to receiving this program I had been sending away for information on various business opportunities. All of the programs I received, in my opinion, were not cost effective. They were either too difficult for me to comprehend or the initial investment was too much for me to risk to see if they would work or not. One claimed that I would make a million dollars in one year...it didn't tell me I'd have to write a book to make it!
But like I was saying, in December of 1997 I received this program. I didn't send for it, or ask for it, they just got my name off a mailing list.THANK GOODNESS FOR THAT!!! After reading it several times, to make sure I was reading it correctly, I couldn't believe my eyes. Here was a MONEY MAKING PHENOMENON. I could invest as much as I wanted to start, without putting me further into debt. After I got a pencil and paper and figured it out, I would at least get my money back. But like most of you I was still a little skeptical and a little worried about the legal aspects of it all. So I checked it out with the U.S. Post Office (1-800-725-2161 24-hrs) and they confirmed that it is indeed legal! After determining the program was LEGAL and NOT A CHAIN LETTER, I decided "WHY NOT."
Initially I sent out 10,000 e-mails. It cost me about $15 for my time on-line. The great thing about e-mail is that I don't need any money for printing to send out the program, and because all of my orders are fulfilled via e-mail, my only expense is my time. I am telling you like it is I hope it doesn't turn you off, but I promised myself that I would not "rip-off" anyone, no matter how much money it made me.
In less than one week, I was starting to receive orders for REPORT #1 By January 13, I had received 26 orders for REPORT #1. Your goal is to "RECEIVE at least 20 ORDERS FOR REPORT #1 WITHIN 2 WEEKS. IF YOU DON'T, SEND OUT MORE PROGRAMS UNTIL YOU DO!" My first step in making $50,000 in 90 days was done. By January 30, I had received 196 orders for REPORT #2. Your goal is to "RECEIVE AT LEAST 100+ ORDERS FOR REPORT #2 WITHIN 2 WEEKS. IF NOT, SEND OUT MORE PROGRAMS UNTIL YOU DO. ONCE YOU HAVE 100 ORDERS, THE REST IS EASY, RELAX, YOU WILL MAKE YOUR $50,000 GOAL." Well, I had 196 orders for REPORT #2, 96 more than I needed. So I sat back and relaxed. By March 1, of my e-mailing of 10,000, I received $58,000 with more coming in every day.
I paid off ALL my debts and bought a much needed new car. Please take time to read the attached program, IT WILL CHANGE YOUR LIFE FOREVER!! ! Remember, it won't work if you don't try it. This program does work , but you must follow it EXACTLY! Especially the rules of not trying to place your name in a different place. It won't work and you'll lose out on a lot of money! In order for this program to work, you must meet your goal of 20+ orders for REPORT #1, and 100+ orders for REPORT #2 and you will make $50,000 or more in 90 days. I AM LIVING PROOF THAT IT WORKS!!!
If you choose not to participate in this program, I am sorry. It really is a great opportunity with little cost or risk to you. If you choose to participate, follow the program and you will be on your way to financial security. If you are a fellow business owner and are in financial trouble like I was, or you want to start your own business, consider this a sign. I DID!
Sincerely, Johnathon Rourke
A PERSONAL NOTE FROM THE ORIGINATOR OF THIS PROGRAM:
By the time you have read the enclosed program and reports, you should have concluded that such a program, and one that is legal, could not have been created by an amateur.
Let me tell you a little about myself. I had a profitable business for 10 years. Then in 1979 my business began falling off. I was doing the same things that were previously successful for me, but it wasn't working. Finally, I figured it out. It wasn't me, it was the economy. Inflation and recession had replaced the stable economy that had been with us since 1945.I don't have to tell you what happened to the unemployment rate... because many of you know from first hand experience. There were more failures and bankruptcies than ever before.
The middle class was vanishing. Those who knew what they were doing invested wisely and moved up. Those who did not, including those who never had anything to save or invest, were moving down into the ranks of the poor. As the saying goes, "THE RICH GET RICHER AND THE POOR GET POORER." The traditional methods of making money will never allow you to "move up" or "get rich", inflation will see to that.
You have just received information that can give you financial freedom for the rest of your life, with "NO RISK" and "JUST A LITTLE BIT OF EFFORT." You can make more money in the next few months than you have ever imagined. I should also point out that I will not see a penny of this money, nor anyone else who has provided a testimonial for this program. I have already made over 4 MILLION DOLLARS!I have retired from the program after sending thousands and thousands of programs.
Follow the program EXACTLY AS INSTRUCTED. Do not change it in any way . It works exceedingly well as it is now. Remember to e-mail a copy of this exciting report to everyone you can think of. One of the people you send this to may send out 50,000...and your name will be on everyone of them!
Remember though, the more you send out the more potential customers you will reach.
So my friend, I have given you the ideas, information, materials and opportunity to become financially independent. IT IS UP TO YOU NOW!
"THINK ABOUT IT"
Before you delete this program from your mailbox, as I almost did, take a little time to read it and REALLY THINK ABOUT IT. Get a pencil and figure out what could happen when YOU participate. Figure out the worst possible response and no matter how you calculate it, you will still make a lot of money! You will definitely get back what you invested. Any doubts you have will vanish when your first orders come in. IT WORKS!
Jody Jacobs, Richmond, VA
HERE'S HOW THIS AMAZING PROGRAM WILL MAKE YOU THOUSANDS OF DOLLAR$
INSTRUCTIONS:
This method of raising capital REALLY WORKS 100% EVERY TIME. I am sure that you could use up to $50,000 or more in the next 90 days. Before you say "BULL... ", please read this program carefully.
This is not a chain letter, but a perfectly legal money making opportunity. Basically, this is what you do: As with all multi-level businesses, we build our business by recruiting new partners and selling our products. Every state in the USA allows you to recruit new multi-level business partners, and we offer a product for EVERY dollar sent. YOUR ORDERS COME BY MAIL AND ARE FILLED BY E-MAIL, so you are not involved in personal selling. You do it privately in your own home, store or office. This is the GREATEST Multi-Level Mail Order Marketing anywhere.
This is what you MUST do:
1. Order all 4 reports shown on the list below (you can't sell them if youdon't order them). -- For each report, send $5.00 CASH, the NAME & NUMBER OF THE REPORT YOU ARE ORDERING, YOUR E-MAIL ADDRESS, and YOUR NAME & RETURN ADDRESS (in case of a problem) to the person whose name appears on the list next to the report. MAKE SURE YOUR RETURN ADDRESS IS ON YOUR ENVELOPE IN CASE OF ANY MAIL PROBLEMS! -- When you place your order, make sure you order each of the four reports. You will need all four reports so that you can save them on your computer and resell them. -- Within a few days you will receive, via e-mail, each of the four reports. Save them on your computer so they will be accessible for you to send to the 1,000's of people who will order them from you.
2. IMPORTANT DO NOT alter the names of the people who are listed next to each report, or their sequence on the list, in any way other than is instructed below in steps "a" through "f" or you will lose out on the majority of your profits. Once you understand the way this works, you'll also see how it doesn't work if you change it. Remember, this method has been tested,and if you alter it, it will not work. a. Look below for the listing of available reports. b. After you've ordered the four reports, take this advertisement and remove the name and address under REPORT #4. This person has made it through the cycle and is no doubt counting their $50,000! c. Move the name and address under REPORT #3 down to REPORT #4. d. Move the name and address under REPORT #2 down to REPORT #3. e. Move the name and address under REPORT #1 down to REPORT #2. f. Insert your name/address in the REPORT #1 position.
Please make sure you COPY ALL INFORMATION, every name and address, ACCURATELY!
3. Take this entire letter, including the modified list of names, and save it to your computer. Make NO changes to the instruction portion of this letter.
Your cost to participate in this is practically nothing (surely you can afford $20). You obviously already have an Internet connection and e-mail is FREE!
There are two primary methods of building your downline:
METHOD #1: SENDING BULK E-MAIL
Let's say that you decide to start small, just to see how it goes, and we'll assume you and all those involved send out only 2,000 programs each. Let's also assume that the mailing receives a 0.5% response. Using a good list the response could be much better. Also, many people will send out hundreds of thousands of programs instead of 2,000. But continuing with this example, you send out only 2,000 programs. With a 0.5% response, that is only 10 orders for REPORT #1. Those 10 people respond by sending out 2,000 programs each for a total of 20,000. Out of those 0.5%, 100 people respond and order REPORT #2. Those 100 mail out 2,000 programs each for a total of 200,000. The 0.5% response to that is 1,000 orders for REPORT #3. Those 1,000 send out 2,000 programs each for a 2,000,000 total. The 0.5% response to that is 10,000 orders for REPORT #4. That's 10,000 $5 bills for you. CASH!!! Your total income in this example is $50 + $500 + $5,000 + $50,000 for a total of $55,550!!! REMEMBER FRIEND, THIS IS ASSUMING 1,990 OUT OF THE 2,000 PEOPLE YOU MAIL TO WILL DO ABSOLUTELY NOTHING AND TRASH THIS PROGRAM! DARE TO THINK FOR A MOMENT WHAT WOULD HAPPEN IF EVERYONE, OR HALF SENT OUT 100,000 PROGRAMS INSTEAD OF 2,000. Believe me, many people will do justthat, and more! By the way, your cost to participate in this is practically nothing. You obviously already have an Internet connection and e-mail is FREE!!! REPORT #2 will show you the best methods for bulk e-mailing, tell you where to obtain free bulk e-mail software and where to obtain e-mail lists.
METHOD #2 - PLACING FREE ADS ON THE INTERNET
Advertising on the internet is very, very inexpensive, and there are HUNDREDS of FREE places to advertise. Let's say you decide to start small just to see how well it works. Assume your goal is to get ONLY 10 people to participate on your first level. (Placing a lot of FREE ads on the Internet will EASILY get a larger response.) Also assume that everyone else in YOUR ORGANIZATION gets ONLY 10 downline members. Follow this example to achieve the STAGGERING results below:
1st level--your 10 members with $5.......................................$50 2nd level--10 members from those 10 ($5 x 100)..................$500 3rd level--10 members from those 100 ($5 x 1,000)...........$5,000 4th level--10 members from those 1,000 ($5 x 10,000).....$50,000 THIS TOTALS ----------} $55,550
Remember friends, this assumes that the people who participate only recruit 10 people each. Think for a moment what would happen if they got 20 people to participate! Most people get 100's of participants! THINK ABOUT IT! For every $5.00 you receive, all you must do is e-mail them the report they ordered. THAT'S IT! ALWAYS PROVIDE SAME-DAY SERVICE ON ALL ORDERS! This will guarantee that the e-mail THEY send out with YOUR name and address on it will be prompt because they can't advertise until they receive the report!
AVAILABLE REPORTS
*** Order Each REPORT by NUMBER and NAME *** Notes: -- ALWAYS SEND $5 CASH (U.S. CURRENCY) FOR EACH REPORT. CHECKS NOT ACCEPTED. -- ALWAYS SEND YOUR ORDER VIA FIRST CLASS MAIL. -- Make sure the cash is concealed by wrapping it in at least two sheets of paper. On one of those sheets of paper, include: (a) the number & name of the report you are ordering, (b) your e-mail address, and (c) your name & postal address.
PLACE YOUR ORDER FOR THESE REPORTS NOW:
REPORT #1 "The Insider's Guide to Advertising for Free on the Internet"
ORDER REPORT #1 FROM:
D. Baggett 316 gh lowery rd Chatsworth, Ga. 30705
REPORT #2 "The Insider's Guide to Sending Bulk E-mail on the Internet"
ORDER REPORT #2 FROM:
A. Picciotti 3202 The Highlands Drive Sugarland, Tx 77478
REPORT #3 "The Secrets of Multilevel Marketing on the Internet"
ORDER REPORT #3 FROM:
L. Daehlin 3208 Wellington Ct. Suite P Raliegh, N C 27615
REPORT #4 "How to Become a Millionaire Utilizing the Power of Multilevel Marketing and the Internet"
ORDER REPORT #4 FROM:
P. Drucker 1517 A Rolling Glen Rd Boothwyn, Pa. 19061
About 50,000 new people get online every month!
******* TIPS FOR SUCCESS ******* -- TREAT THIS AS YOUR BUSINESS! Be prompt, professional, and follow the directions accurately. -- Send for the four reports IMMEDIATELY so you will have them when the orders start coming in because: When you receive a $5 order, you MUST send out the requested product/report. -- ALWAYS PROVIDE SAME-DAY SERVICE ON THE ORDERS YOU RECEIVE. -- Be patient and persistent with this program. If you follow the instructions exactly, your results WILL BE SUCCESSFUL! -- ABOVE ALL, HAVE FAITH IN YOURSELF AND KNOW YOU WILL SUCCEED!
******* YOUR SUCCESS GUIDELINES ******* Follow these guidelines to guarantee your success:
If you don't receive 20 orders for REPORT #1 within two weeks, continue
advertising or sending e-mails until you do. Then, a couple of weeks later you should receive at least 100 orders for REPORT#2. If you don 't, continue advertising or sending e-mails until you do. Once you have received 100 or more orders for REPORT #2, YOU CAN RELAX, because the system is already working for you, and the cash will continue to roll in!
THIS IS IMPORTANT TO REMEMBER: Every time your name is moved down on the list, you are placed in front of a DIFFERENT report. You can KEEP TRACK of your PROGRESS by watching which report people are ordering from you. If you want to generate more income, send another batch of e-mails or continue placing ads and start the whole process again! There is no limit to the income you will generate from this business!
Before you make your decision as to whether or not you participate in this program. Please answer one question. DO YOU WANT TO CHANGE YOUR LIFE? If the answer is yes, please look at the following facts about this program:
1. You are selling a product which does not Cost anything to PRODUCE, SHIP OR ADVERTISE. 2. All of your customers pay you in CASH! 3. E-mail is without question the most powerful method of distributing information on earth. This program combines the distribution power of e-mail together with the revenue generating power of multi-level marketing. 4. Your only expense--other than your initial $20 investment--is your time! 5. Virtually all of the income you generate from this program is PURE PROFIT! 6. This program will change your LIFE FOREVER.
ACT NOW!Take your first step toward achieving financial independence. Orderthe reports and follow the program outlined above--SUCCESSwill be yourreward.
Thank you for your time and consideration.
PLEASE NOTE: If you need help with starting a business, registering a business name, learning how income tax is handled, etc., contact your localoffice of the Small Business Administration (a Federal Agency) 1-800-827-5722 for free help and answers to questions. Also, the InternalRevenue Service offers free help via telephone and free seminars aboutbusiness tax requirements. Your earnings are highly dependant on youractivities and advertising. The information contained on this site and in the report constitutes no guarantees stated nor implied. In the event that it is determined that this site or report constitutes a guarantee of any kind, that guarantee is now void. The earnings amounts listed on this site and in the report are estimates only. If you have any questions of the legality of this program, contact the Office of Associate Director for Marketing Practices, Federal Trade Commission, Bureau of Consumer Protection in Washington, DC.
///////////////////////////////////////////////////////////////// Remove at qmkx-at-netscape.net /////////////////////////////////////////////////////////////////
by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id GAA07967 for dist-Microscopy; Wed, 11 Oct 2000 06:27:21 -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 GAA07964 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Wed, 11 Oct 2000 06:26:50 -0500 (CDT) Received: from servizi-2.padova.com (elezioniradicali-2.padova.com [212.131.155.43]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id GAA07957 for {Microscopy-at-sparc5.microscopy.com} ; Wed, 11 Oct 2000 06:26:37 -0500 (CDT) Received: from servizi.padova.com (unverified [212.131.155.42]) by servizi-2.padova.com (Rockliffe SMTPRA 4.2.2) with ESMTP id {B0000005729-at-servizi-2.padova.com} for {mlistc3_emma.bonino-at-elezioniradicali-2.padova.com} ; Wed, 11 Oct 2000 11:17:42 +0200 Received: from padova08 (unverified [212.131.155.44]) by servizi.padova.com (Rockliffe SMTPRA 4.2.2) with SMTP id {B0000036121-at-servizi.padova.com} for {mlistc3_emma.bonino-at-elezioniradicali-2.padova.com} ; Wed, 11 Oct 2000 11:22:01 +0200 Message-ID: {001401c03365$51216370$2c9b83d4-at-pol.it} Reply-To: "Emma Bonino" {emma.bonino-at-elezioniradicali.padova.com}
Toc toc. Mi scusi, sono Emma Bonino e la prego di partecipare al gioco online che stiamo organizzando perche' anche cittadini come lei (che non votino affatto radicale o che lo facciano, che non votino o non intendano piu' votare) possano partecipare alle nostre decisioni, essere presenti o rappresentati nei nostri organi dirigenti.
In vista delle elezioni politiche occorre tentare di allearsi con il Polo o con l'Ulivo? O combattere da soli? O organizzare il boicottaggio di elezioni non democratiche? Quali Riforme istituzionali, politiche, del lavoro, dell'impresa: "americane" o "tedesche", o nessuna? Quali politiche sulle liberta' individuali e sulla scienza, oltre che sulle droghe: proibizioniste o antiproibizioniste, come gia' sul divorzio e sull'aborto? E i partiti devono aprirsi a tutti i cittadini attraverso Internet, o restare "chiusi" come finora?
Dal 27 novembre al 3 dicembre si votera' online per eleggere 25 nuovi membri del Comitato Radicale. Se, come vivamente spero, lei intende partecipare a questo gioco, lo preannunci e registri cliccando qui: http://www.radicali.it/p_register/
Mi scusi ancora. A presto.
Emma Bonino
PS Questa email e' stata inviata nel rispetto della legge sulla privacy; se desidera maggiori informazioni o se intende cancellarsi, puo' cliccare qui: http://www.radicali.it/p_privacy_com3/
It all depends: how busy you are, how expensive staff time in your country is and how costly the place is you buy your stubs from. Obviously some specimens should be stored in a desiccator for an eternity and other can be discarded immediately. There is nothing magical about cleaning stubs. Solvents are messy. Best to place on some newspaper a very coarse file or a sheet of coarse (50 grit) sandpaper and then rub the old stub over that file of sandpaper until specimen and glue are removed.
Disclaimer: ProSciTech sells stubs, but we are happy if people recycle and this leaves some money for useful purchases. 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 ABN: 99 724 136 560 www.proscitech.com
On Wednesday, October 11, 2000 3:09 AM, O. O. Ilori [SMTP:sojilori-at-oauife.edu.ng] wrote: } } Hello Guys } I am a novice user of a SEM (LEO 1450 to be precise) equipment which was } just installed in our central science lab. } Is there a way to clean stubs for reuse or are stubs disposable once used. } ( This may sound naive) } soji } Mr. O. O. ILORI } DEPARTMENT OF ELECTRONIC/ELECTRICAL ENGINEERING } OBAFEMI AWOLOWO UNIVERSITY, } ILE-IFE, OSUN STATE } NIGERIA. } email: sojilori-at-oauife.edu.ng } }
Dear Listers, The following are responses to my inquiry about automated critical point dryers. There was one other which I will post as well. Thanks to all who responded. Rosemary
I find automated CPDs easier only in the sense that they make training new users easier. But they also can encourage not paying sufficient attention to the drying instrument. I like the Polaron, in particular it's large chamber. Let's see ... the only other advantage to an automated system is if you have lots of samples to do in lots of runs. Otherwise, I've never seen enough of an advantage to justify the extra cost.
But! having said that, a CPD with autobleed is an excellent choice. This is a good safety feature, and can help protect specimens.
Phil
There are safety considerations, that is one of the several reasons why there is such a preference for the E3000 or E3100. BTW another good thing about the Polaron design and larger chamber is the less likelihood of turbulence, or putting it another way, you can do your exchanges at a faster rate because you can use larger flow rates before turbulence does set in.
Chuck (Garber)
We recently purchased a Tousimis 795 semi-automatic critical point dryer for our multi-user facilty. This unit replaces an old Denton "bomb". We considered ease of use for our customers to be an important consideration. Our unit has preset purge, bleed, vent and heating features which make it easier to use. We don't have to lug a large beaker of hot water over to the CPD to heat the chamber. The flow rates are factory preset but easily adjustable. Having these features, I believe, allows us to produce more consistent, reproducible results. The standard chamber size is rather small but a chamber extension is available and easy to install. We have found that when trying to critical point dry very large specimens, the chamber size can be a problem (most of our work is Drosophila flies or mouse embryos).
Although very few people do their own critical point drying here, our goal is to make the facility as user-friendly as possible, and this seemed to be a step in that direction.
Happy hunting.
Tom Januszewski Senior Electron Microscopist Molecular and Cellular Imaging Facility
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
i'd like to purchase a backscattered detector for an old Hitachi SEM. I am looking for vendors of such equipment, as Robinson detector, KE Si detector and so on. Could you help me with some addresses?
Thanks,
Peter
-- Dr NAGY Peter Chemical Research Center Institute of Chemistry of the Hungarian Academy of Sciences 1025 Budapest Pusztaszeri ut 59-67. Phone: 36-1-325-7548 Fax: 36-1-325-7509 E-mail: nagyp-at-chemres.hu
Dear Soji We usually treat the standard aluminium LEO pin stubs as disposable. Since we have a number of users each with a different specimen type and mounting procedure it becomes difficult to identify a single effective cleaning method, and staff time is more costly than the stubs.
If you are a novice user you may not yet realise that LEO pin stubs are obtainable at a fraction of LEO's price from most microscopy supply houses.
http://www.kaker.com/mvd/list.html is a list of microscopy vendors which you may find useful. Use this to look up web sites of some of the following companies: Agar Scientific, TAAB Laboratories, SPI, Electron Microscopy Science, etc. etc.
However, cleaning stubs can be cost-effective if you routinely produce many tens or hundreds of specimens in exactly the same way. If you have to do this it pays to find adhesives and conductive paints that redissolve readily in the same solvent, so things like epoxies are out. Choose your solvents carefully, because some like ethanol attack aluminium if it is immersed for long periods.
Another approach which avoids the need to clean stubs is to mount up your specimens on thin squares or discs of metal (e.g. thin aluminium sheet) which are then tacked to the stub using e.g. graphite dag. After use the metal supports can be cracked off and replaced with new ones. This way the stub can be re-used more often before it needs to be properly cleaned up. Good luck Chris
} On Wednesday, October 11, 2000 3:09 AM, O. O. Ilori } [SMTP:sojilori-at-oauife.edu.ng] wrote: } } } } Hello Guys } } I am a novice user of a SEM (LEO 1450 to be precise) equipment which was } } just installed in our central science lab. } } Is there a way to clean stubs for reuse or are stubs disposable once used. } } ( This may sound naive) } } soji } } Mr. O. O. ILORI } } DEPARTMENT OF ELECTRONIC/ELECTRICAL ENGINEERING } } OBAFEMI AWOLOWO UNIVERSITY, } } ILE-IFE, OSUN STATE } } NIGERIA. } } email: sojilori-at-oauife.edu.ng } } } } } }
========================================== Dr. Chris Jeffree University of Edinburgh BIOSEM - Biological Sciences Electron Microscope Facility Institute of Cell and Molecular Biology Daniel Rutherford Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JH, Scotland, UK Tel. #44 (0) 131 650 5345 FAX. #44 (0) 131 650 6563 Mobile 07710 585 401 email c.jeffree-at-ed.ac.uk =========================================
We are considering a purchase of a system for quantitative analysis of bacterial and fungal biovolume in soil samples. Could anyone provide info on preferred method (FITC labeling?) and equipment. As always, I appreciate your help, Alice.
Alice Dohnalkova Environmental Microbiology Battelle, PNNL MS P7-50 Richland, WA 99352 tel. (509) 372-0692 fax (509) 376-1321
Ref: Diode detector type... I have used GW Electronics. First on an ETEC SEM, and now on a Hitachi S-3500. I started with a model (many years ago) which is now discontinued and have upgraded to the current design. Both worked well for me. Check: http://www.gwelectronics.com/
---------------------------------------- Woody White McDermott Technology, Inc. http://www.mtiresearch.com/
Peter Try looking at this site. http://www.kaker.com/mvd/list.html It contains links to dozens of microscopy vendor sites Chris
Date sent: Wed, 11 Oct 2000 08:02:17 -0500 To: Microscopy-at-sparc5.microscopy.com } From: Nagy Peter {nagyp-at-chemres.hu}
The last time this subject came up, someone suggested wiping the inside of the jar with Polaroid print coaters. They seem to provide a water soluble layer that easily comes off. We used to have dozens of extras lying around and now had a use for them. However, now that we have largely gone to digital imaging we will have to watch our stock.
At 04:49 PM 10/10/2000 -0700, you wrote:
} Well we've got a new vacuum evaporator and it has been so long since I've } had one I've forgotten how to treat the bell jar to reduce the effort it } takes to keep it clean. However, I do recall that there were little } tricks to cleaning and treating the glass and other surfaces. Anyone care } to shed a little light my darkened memory of these procedures?
---------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
We agree with John Nailon that if a user forgets to prepare the evaporator prior to a run he should have to clean it. Since we have to clean our evaporators several times a day because of our substrate production, we do the following.
1. Place Victawet in a medium basket. 2. Evaporate the Victawet at 10(-5). This coats not only the bell jar, but all the other surfaces under the bell jar as well. 3. Clean the system with windex (or water) and wipe with a lint free cloth. 4. Re-coat with victawet prior to the next run.
John Arnott
Disclaimer: In additon to using, Ladd has been selling most of the above mentioned products, including Victawet, Lint-free cloth, Substrates, Evaporators and Baskets since 1955. --
LADD RESEARCH 131 Dorset Lane Williston, VT 05495 USA
web site http://www.laddresearch.com
TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail sales-at-laddresearch.com
Quality Since 1955
John Nailon 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 easiest way to keep your bell jar clean is to wipe a thin film of } detergent over the surface of the clean glass and pump down to dry. } After each coating run simply rinse off the glass, the detergent comes } away and takes the coating with it. Recoat with detergent and dry. } } Train ALL users in this technique, insist they clean the glass if they } "FORGOT" to do it after their last run, and you will always have a } clear, clean bell jar. } Regards } JVN } Rick Harris 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 } } -----------------------------------------------------------------------.} } } } Well we've got a new vacuum evaporator and it has been so long since I've } } had one I've forgotten how to treat the bell jar to reduce the effort it } } takes to keep it clean. However, I do recall that there were little tricks } } to cleaning and treating the glass and other surfaces. Anyone care to shed } } a little light my darkened memory of these procedures? } } } } 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 } } -- } **************************************************** } 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 need to have the phosphor screens recoated on an EM 400T we have on campus. I have been told that Grant Scientific does a good job. I have not been able to find a way to contact them. Can anyone help with a phone number or info.? I would also welcome any other suggestions. Thanks.
Joel McClintock EM Specialist Univ. of Kentucky Lexington, Ky 859-257-1242
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To minimize the area which needs cleaning, I obtained a pyrex glass cylinder large enough to fit vertically over my evaporating supports, etc. The ends of it were coated with a vacuum rated epoxy to prevent chipping. Before using, it is coated with a commercial release agent made for this "easy cleaning" purpose. The bottom sits on a sheet of disposable aluminum foil and the top is also covered by aluminum foil. To facilitate pumping, a 1/4" thick scrap of ceramic placed under the bottom edge of the "containment cylinder" aids pumping at a reasonable rate and provides a path for one of the power leads for my evaporating basket, insulating the lead from a metal support
platform. After use, the foil can be scrapped and only a 6" diameter cylinder requires cleaning!
Bernie Kestel Materials Science Division Argonne National Laboratory 9700 South Cass Avenue Argonne, Il., 60439
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Dear Rick, I used to use Victawet and it was the best, but it is not available, so I use a bar of hand soap. After you clean the bell jar or, if it is new, after you wet it, rub the dry bar of soap all over the inside and rub with a paper towel to spread it evenly and make it transparent. Good luck. At 04:49 PM 10/10/00 -0700, you wrote: } Well we've got a new vacuum evaporator and it has been so long since I've } had one I've forgotten how to treat the bell jar to reduce the effort it } takes to keep it clean. However, I do recall that there were little tricks } to cleaning and treating the glass and other surfaces. Anyone care to shed } a little light my darkened memory of these procedures? } } TIA } } Rick A. Harris, Director 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 Mr. Olori, The stubs used for SEM are reused for years, but the grids for a TEM are disposed of. I clean my SEM stubs by rubbing on a paper towel wet with either lab alcohol (ethanol) or acetone, depending what was on them. If I am using the carbon double-side sticky tabs, I soak the stubs in acetone for an hour, then rub on a paper towel to remove the tab. Congratulations on your new SEM. At 06:09 PM 10/10/00 +0100, you wrote: } } Hello Guys } I am a novice user of a SEM (LEO 1450 to be precise) equipment which was } just installed in our central science lab. } Is there a way to clean stubs for reuse or are stubs disposable once used. } ( This may sound naive) } soji } Mr. O. O. ILORI } DEPARTMENT OF ELECTRONIC/ELECTRICAL ENGINEERING } OBAFEMI AWOLOWO UNIVERSITY, } ILE-IFE, OSUN STATE } NIGERIA. } email: sojilori-at-oauife.edu.ng
Good 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
Peter Nagy wrote: ========================================================== i'd like to purchase a backscattered detector for an old Hitachi SEM. I am looking for vendors of such equipment, as Robinson detector, KE Si detector and so on. Could you help me with some addresses? =========================================================== Try Microscopy Vendor's Data Base http://207.137.96.185/mvd/products/back.html
They have them all, I think.
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 suppose that John Russ and others have stated the matter concisely and eloquently in their books and articles on the subject, but the concepts are not that difficult that I will take a stab at it.
A digital image is nothing more than a two-dimensional array of numbers where the numbers represent some characteristic of the image. Various image sources vary in the way that the arrays are assembled and what the numbers represent.
In an scanning electron microscope or a scanning probe microscope (which I suppose you could say that an SEM is), a single detector element is used to provide the signal at all points in the image. The X and Y coordinates of the scan are used to determine the array or pixel index, and the signal is digitized at each point.
Many systems utilize "active" control where the beam is directed to each coordinate in succession under computer control. A digital grid is predetermined within the computer and the coordinates are fed out through appropriate hardware to convert the digital coordinates to the necessary analog voltage to scan the probe to the desired point. A microscope signal is measured at that point and digitized for storage in the image array. The scan speed and resolution are under direct control of a computer.
Other systems are termed "passive" since the existing analog microscope scan system is used to control the probe scan as normal. The digitizing system monitors the x and y coordinates and records the signal level when the beam is in positions corresponding to the pixels in the image array.
There are a variety of signals available from electron microscopes. Detectors for secondary electrons and backscattered electrons operate as analog devices. (One might argue that since they are counting a finite number of electrons, then they could be setup as digital devices. However, the count rate is high enough that digital detectors are probably not feasible, and even if they were there remains a question of when to start counting.) A variety of detectors and devices are used (including PMTs - photo multiplier tubes) to measure the signal, but they are primarily, if not all, analog devices. An analog to digital convertor is used to convert the signal to digital for the image array. Some scopes may digitize the signal early on so that no analog signal is readily available, but it still exists somewhere inside the instrument.
X-ray signals in an electron microscope are probably treated as digital from the outset. Since the x-ray photons are detected as pulses from the x-ray detector, they are treated as discrete events and counted. Since counting is involved, x-ray signals are only used with active control systems (to the best of my knowledge). A counter is reset each time the probe is moved to a new location.
There is a whole other class of source for digital images. In this class there is a separate detector for each element of the array. The entire image is present at once as opposed to being built up by a scanned probe. The detectors operate in parallel as they build up and digitize the signal simultaneously. At some point the signal is read and the detectors are reset. CCD cameras are the most common device in this category. They find application in light microscopes and transmission electron microscopes as well as in consumer electronics such as video cameras and still cameras.
In these systems, the images are digital from the start. The readout is in terms of an electron count. It usually gets scaled to another digital range (0-255, 0-4095, etc) so that the original digital count may no longer be significant.
The size of the image array is determined by the size of the CCD array. Smaller image sizes can be obtained by grouping the signals from adjacent elements in the CCD array. Larger images are sometimes manufactured by interpolating the signals of neighboring elements, but that is not introducing new information.
I hope this helps. Feel free to ask for further clarification.
Warren S.
At 06:40 PM 10/10/2000 -0400, you wrote: } Dear listserv: } } Dee Breger was kind enough to forward this message for me. I'm writing a book } on digital photography, to be published by Harry N. Abrams next year. The book } will include a section on applied digital imaging, and I am particularly } interested in scientific imaging. I am hoping that some of you folks might } be able } to provide some information. } } For the purposes of the book, I am defining digital imaging as that which } describes a scene using discrete digital information in a raster grid, or } bitmap. So, I am particularly interested in scientific imaging applications } which gather information in this way. } } I would be interested to hear if there are other scientific imaging systems } (DEE: besides SEMs) which collect information in this manner. For instance, } it sounds as though atomic force microscopes work in this way: as they } drag a stylus across a surface, the position is recorded (x and y), along } with the height (z). This } is later used to re-create the image in a raster grid (the fact that the AFM } does not use light makes it another interesting foil to 'traditional' digital } photography). How then, are colors in the final output determined from the } data collected? I would also, of course, love to see any images you might have } to show. } } Thanks in advance, } Jonathan Lipkin (jlipkin-at-ramapo.edu) } } DEE: I'll forward to Jonathan any replies that are publically posted to the } listserver. He also needs other detailed technical information on digital } image formation which can be far more comprehensively clarified and } amplified by some of the experts among my fellow listers than by me, though } I've already given him the broad picture for analog scopes like my dear old } Cambridge 250. His SEM impressions, which I'll pass along verbatim, } include (1) "SEMs collect information, used to fill a raster grid, through } the use of a PMT", (2) the signal is sent directly to the crt in analog } form -I always had thought that the pmt sent out digital info, but it } sounds like it just sends out analog data (a waveform? though it couldn't } be a waveform if the raster grid is discrete in two dimensions)" and (3) } "as the secondary electrons come off the sample, they hit a detector which } generates a digital number (redundant) which is then placed in a pixel of a } bitmap." } } Many thanks to any who can help!
---------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
Grant does very fine work. As a mater of fact they are the source used by a few OEMs. Their address is 1385 Rock Island Road Gilbert South Carolina 29054 Phone: 803/892-2841
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 REPAIR -----Original Message----- } From: Joel McClintock {jmcclin-at-pop.uky.edu} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
We slice off the specimen with a razor blade and use the blade to scrape the stub fairly clean. As we use colloidal graphite, which is conductive, I do not mind some residual glue as it will get covered by glue from the next use. This glue can be removed, if required, by sonicating in butyl acetate.
Dave
On Wed, 11 Oct 2000 21:09:07 +1000 Jim at ProSciTech {jim-at-proscitech.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 } -----------------------------------------------------------------------. } } } It all depends: how busy you are, how expensive staff time in your country is } and how costly the place is you buy your stubs from. Obviously some specimens } should be stored in a desiccator for an eternity and other can be discarded } immediately. } There is nothing magical about cleaning stubs. Solvents are messy. Best to } place on some newspaper a very coarse file or a sheet of coarse (50 grit) } sandpaper and then rub the old stub over that file of sandpaper until specimen } and glue are removed. } } Disclaimer: ProSciTech sells stubs, but we are happy if people recycle and this } leaves some money for useful purchases. } 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 } ABN: 99 724 136 560 www.proscitech.com } } On Wednesday, October 11, 2000 3:09 AM, O. O. Ilori } [SMTP:sojilori-at-oauife.edu.ng] wrote: } } } } Hello Guys } } I am a novice user of a SEM (LEO 1450 to be precise) equipment which was } } just installed in our central science lab. } } Is there a way to clean stubs for reuse or are stubs disposable once used. } } ( This may sound naive) } } soji } } Mr. O. O. ILORI } } DEPARTMENT OF ELECTRONIC/ELECTRICAL ENGINEERING } } OBAFEMI AWOLOWO UNIVERSITY, } } ILE-IFE, OSUN STATE } } NIGERIA. } } email: sojilori-at-oauife.edu.ng } } } } } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
I need to have the phosphor screens recoated on an EM 400T we have on campus. I have been told that Grant Scientific does a good job. I have not been able to find a way to contact them. Can anyone help with a phone number or info.? I would also welcome any other suggestions. Thanks. Dear Joel, (803)892-2841. I just tried it, and it works. I also think that Grant did a good job for us. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
Hi List, This is a question/situation a colleague of mine wanted to post-her email for replies not on the list is rhealy-at-iastate.edu. Of all people out there, I know you all can help! Thanks so much! Tracey Pepper
We are working with Cochliobolus inoculated maize leaf tissue using jet-propane fixation, and freeze substitution using acetone (for immuno resins we switch to ultra pure ETOH prior to infiltration), and have attempted using several immuno-type resins such as: Lowicryl HM20 (with poly. at -20 under UV), LR White with accel. at 4deg., and Quetol for general morphology. Our ultimate goal is to visualize extra-cellular matrix of fungus within leaf tissues using immunocytochem. So far, fixation is great (general morph.) However, the immuno resins are not infiltrating adequately (under -30 deg.for Lowicryl and 4 deg. for LR White), lots of separation, and shattering. The longest we have infiltrated has been 1-2 hr. each step for LR White. The Lowicryl had an overnight in pure followed by 1/2 day for each of five total changes. Any and all suggestions would be greatly appreciated! Regards, Rosanne Healy
Tracey Pepper Supervisor Bessey Microscopy Facility Iowa State University ph: 515-294-3872 fax: 515.294.1337
Ben Craft wrote: ============================================================= I'm getting ready to purchase a new sputter coater for our FESEM. I'd like to hear how lucky others were with using a "regular" sputter coater for high resolution SEM work. Currently, there is a noise problem with our SEM, so the resolution is about 3-4nm. I've successfully imaged 5nm ZrO2 close-packed particles, and 4nm pores in glass using a thin coating of Au/Pd applied with an old Polaron. One thing I'm worried about is = once the noise is fixed and I can resolve the published 1.5nm resolution will I be able to resolve the grains in the coating? ============================================================= One system to consider is the Osmium Plasma Coater OPC-60 made by Nippon Laser and Electronics, Nagoya, Japan. It is distributed by SPI Supplies. You can find extensive information and examples on URL http://www.2spi.com/catalog/osmi-coat.html
The end result is an amorphous coating so there is no grain size with which one has to contend. The source of the osmium is an ampoule of osmium tetroxide. This is not just a simple matter of sputtering from an osmium metal cathode. And since the coating is an inert precious group metal, its shelf life is quite long, if not infinite. Safety concerns are address on the webpage mentioned above.
Contact me off-line should you want us to do a demo coating for you.
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 just bought a Pelco CPD2 with the hopes of replacing our old Polaron E3000, and so far, we're rather unimpressed just trying to get it through an initial setup. We were wondering if any other users could tell us how many runs they get out of an average CO2 cylinder? The cooling process with the CO2 seems somewhat inefficient at this point and we're using up a lot just trying to cool the chamber. Also any recommendations on the soak vs. the rinse methods of infiltrating the samples would be appreciated. I can only assume the operations get a little easier with practise, but right now we're wondering whether it's worth the investment in the long run. The Polaron has been very reliable and has served us well, but we're worried that its age is compromising its safety.
Our microscopy facility recently merged with the scientific photography and graphics production unit on campus. We need to quickly evaluate the potential value of a large format inkjet printer for producing posters for scientific meetings.
We welcome comments, suggestions and especially personal experiences using inkjet technology for poster production. Is it worth the expense, do researchers use it, etc... We are looking at the Epson 9000 printer in combination with the Fujifilm digital 9000 Photo Graphix RIP. Good idea? How about software for poster production?
Also, vendors are welcome to contact me and to submit quotations. We need all of this information by Friday (Oct 13). Sorry for the rush but I just now got the word that funds may be available. Whew....
Many thanks....
John B.
-- #################################################################### John J. Bozzola, Ph.D., Director IMAGE (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
Victawet is a stock item for Ladd. We sell and use it on a daily basis. Please let me know off-line if you want a quote.
Thanks,
JD Arnott
--
LADD RESEARCH 131 Dorset Lane Williston, VT 05495 USA
web site http://www.laddresearch.com
TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail sales-at-laddresearch.com
Quality Since 1955 Mary Mager 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 Rick, } I used to use Victawet and it was the best, but it is not available, so I } use a bar of hand soap. After you clean the bell jar or, if it is new, after } you wet it, rub the dry bar of soap all over the inside and rub with a paper } towel to spread it evenly and make it transparent. Good luck. } At 04:49 PM 10/10/00 -0700, you wrote: } } Well we've got a new vacuum evaporator and it has been so long since I've } } had one I've forgotten how to treat the bell jar to reduce the effort it } } takes to keep it clean. However, I do recall that there were little tricks } } to cleaning and treating the glass and other surfaces. Anyone care to shed } } a little light my darkened memory of these procedures? } } } } TIA } } } } Rick A. Harris, Director } 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
GRANT SCIENTIFIC CORP 1385 ROCK ISLAND ROAD GILBERT , SC 29054 803-892-2841 PHONE 803-892-2441 FAX
} Listers, } } I need to have the phosphor screens recoated on an EM 400T we have on } campus. I have been told that Grant Scientific does a good job. I have not } been able to find a way to contact them. Can anyone help with a phone number } or info.? I would also welcome any other suggestions. Thanks. } } Joel McClintock } EM Specialist } Univ. of Kentucky } Lexington, Ky } 859-257-1242
We are moving out our Philips EM400T TEM. It has suffered from vacuum logic and lens logic problems -- problems that currently mean that it is unusable. It has a STEM unit, a Quantum EDX detector, Kevex 7000 system, and a variety of holders, including tilt-rotate. Anyone interested in some or all of this equipment should get in touch ASAP, please. Thanks, Brian Robertson
*********************************************************** Assoc. Prof. Brian W. Robertson Department of Mechanical Engineering and Center for Materials Research and Analysis University of Nebraska-Lincoln, N124 WSEC, 17th & Vine Sts., Lincoln, NE 68588-0656, USA ** Office 402 472 8308; FAX 472 1465; Labs 472 8762 and 472 5498 **
I have a Reichert Jung Ultracut S/N382797, Type701701, acquired from Upjohn Japan. The unit has no stereoscope or power unit (lost in packaging or shipping). Any person interested in the unit to use or for parts, please don't hesitate to contact me.
Pauline Yu wrote: =========================================================== ........................ The Polaron has been very reliable and has served us well, but we're worried that its age is compromising its safety. ========================================================== Unless the unit has been dropped or otherwise abused, it is difficult for me to see how its "age" could have anything to do with its safety. But if this was your concern, there is a process, and we could help you arrange it, by which the chamber is returned to the UK, and retested in the same laboratory where it was tested originally, and essentially recertified. The testing is done at a pressure that is far beyond where a rupture disc would blow, so you could then use it with confidence for many years to come. The cost of the service is far less than the cost of a new system.
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 experimented with several different cleaning/mounting procedures and the one that is the most time effective for me is the following: I apply a conductive silicone release agent to my stubs before I apply carbon double stick tape to them, then I use a conductive Al or Cu tape ( I cut it in half length-wise so it doesn't take up so much space ) to insure conductivity, works for my samples which are mostly dry powders, industrial hygiene dusts, inks and papers. Then when I want to clean the stubs I just lift the tape off with a knife ( not to sharp ) and rinse and rub any residue off with paper towels using a 50/50 % toluene / acetone mix, then ethanol in a ultrasonic cleaner if I need them really clean.
Terry Ellis Hallmark Cards Inc. tellis2-at-hallmark.com USA.
Listers, I meet some problem how to electrical-polish 3mm pure copper disk. Would you plese tell me the good solution and the good condition of electrical-polishing copper on tenupol-5 machine. Thanks
Mr.Zhongwen Yao EPFL- CRPP Villingen, PSI Switzerland
You may also want to consider the option of Chromium coating which will give grain siz of less than 0.5NM Chromium grain size with thin film deposition typically 5NM.
We manufacture and sell the K575 Sputter Coater unit. It has the advantage of automated cycle coating Turbomolecular drag pumping for High Vaccum.
Please take a look at our web site for a detailed technical brief and the instrument specification.
HTTP://www.emitech.co.uk
Kind regards
Gareth } -----Original Message----- } From: Garber, Charles A. [SMTP:cgarber-at-2spi.com] } Sent: Wednesday, October 11, 2000 7:40 PM } To: MICROSCOPY BB } Subject: Coater for use with FESEM } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 ] -- } } Ben Craft wrote: } ============================================================= } I'm getting ready to purchase a new sputter coater for our FESEM. I'd } like to hear how lucky others were with using a "regular" sputter } coater } for high resolution SEM work. Currently, there is a noise problem } with our } SEM, so the resolution is about 3-4nm. I've successfully imaged 5nm } ZrO2 } close-packed particles, and 4nm pores in glass using a thin coating of } Au/Pd } applied with an old Polaron. One thing I'm worried about is = once the } noise } is fixed and I can resolve the published 1.5nm resolution will I be } able to } resolve the grains in the coating? } ============================================================= } One system to consider is the Osmium Plasma Coater OPC-60 made by } Nippon } Laser and Electronics, Nagoya, Japan. It is distributed by SPI } Supplies. } You can find extensive information and examples on URL } http://www.2spi.com/catalog/osmi-coat.html } } The end result is an amorphous coating so there is no grain size with } which } one has to contend. The source of the osmium is an ampoule of osmium } tetroxide. This is not just a simple matter of sputtering from an } osmium } metal cathode. And since the coating is an inert precious group } metal, its } shelf life is quite long, if not infinite. Safety concerns are } address on } the webpage mentioned above. } } Contact me off-line should you want us to do a demo coating for you. } } 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 looking for help concerning the following problem: Someone in our institute is trying to deposit a 10nm Au-film on a Ge single crystal by thermal evaporation from a tungsten boat (...rate: 0.5A/s). The main point to me is, if FESEM is an appropriate method to determine if we have really a Au-film or only gold islands ...
I do appreciate any comments concering both points (FESEM-tricks, because 99% of our samples are uncoated insulators) and the depostition technique/parameters for very thin gold films ...
I am looking for help concerning the following problem: Someone in our institute is trying to deposit a 10nm Au-film on a Ge single crystal by thermal evaporation from a tungsten boat (...rate: 0.5A/s). The main point to me is, if FESEM is an appropriate method to determine if we have really a Au-film or only gold islands ...
I do appreciate any comments concering both points (FESEM-tricks, because 99% of our samples are uncoated insulators) and the depostition technique/parameters for very thin gold films ...
Greetings, As seen below, Gareth Robinson mentions chromium as an option for fine coats to use in high res SEM.
A word of caution: Chromium is tricky to work with. You cannot use chromium sputter coating the same simply way you are used to sputtering with, say, gold or platinum. You have to keep the atmosphere in the coater squeaky clean, and even cleaner, and you must absolutely minimize the exposure of your coated sample to oxygen.
I expect that chromium coats can be superb, but you cannot expect to simply put a chromium target in your sputter coater and have decent results. We found this out the hard way.
As ever, Tobias
} } } } Ben, } } You may also want to consider the option of Chromium coating which will } give grain siz of less than 0.5NM Chromium grain size with thin film } deposition typically 5NM. } } We manufacture and sell the K575 Sputter Coater unit. It has the } advantage of automated cycle coating Turbomolecular drag pumping for } High Vaccum. } } Please take a look at our web site for a detailed technical brief and } the instrument specification. } } HTTP://www.emitech.co.uk } } Kind regards } } Gareth Gareth.Robinson-at-emitech.co.uk
At 5:38 PM -0500 10/11/00, Brian W Robertson wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Brian,
I am interested in the holders.
Regards, Lucille Giannuzzi
******************************************************************* 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 ********************************************************************
In upgrading our light microscopes, we are buying some new lenses for our confocal scope and a brightfield scope. The salesforce is regaling us with tales of improved resolution, clarity, brightness etc that comes with spending double or triple the price for a standard oil immersion or dry lens. Could someone please clarify, either on-line or off, what the advantages and disadvantages of each of the following ( my texts on light microscopy don't deal with them). Are they worth the extra dollars?
1) a water immersion lens for looking at cells suspended in buffer and covered with a coverslip (water is used rather than oil between coverslip and lens) These lenses typically have a lower numerical aperture than an oil immersion lens.
2) a water dipping lens, used to examine cells in culture directly, without a coverslip. I would expect the absence of refractive indice changes (media/coverslip/air or oil) would explain their improved brightness.
As you are so close by to our San Clemente, CA lab, I would suggest that you come by and bring some samples to try on our IBS/e Ion Beam Sputter Deposition and Etching System.
The IBS/e, a thin film deposition and etching system, is designed to improve high resolution electron microscopy imaging by depositing ultra-thin, fine grain metal and carbon films on specimens. Even low voltage imaging is improved.
We can deposit many different metals and carbon or show you examples of contrast enhancement on various types of specimens from our library of micrographs. I will send to you by separate mail some AFM images comparing evaporated coatings, sputtered coatings and ion beam deposited films.
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 Ben Craft } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I'm getting ready to purchase a new sputter coater for our FESEM. I'd like to hear how lucky others were with using a "regular" sputter coater for high resolution SEM work. Currently, there is a noise problem with our SEM, so the resolution is about 3-4nm. I've successfully imaged 5nm ZrO2 close-packed particles, and 4nm pores in glass using a thin coating of Au/Pd applied with an old Polaron. One thing I'm worried about is = once the noise is fixed and I can resolve the published 1.5nm resolution will I be able to resolve the grains in the coating?
The 2000 Fall Meeting of the Texas Society for Microscopy will be held
on October 26, 27, and 28 at the DoubleTree Hotel in Dallas, TX. The hotel is located in Campbell Centre at 8350 N. Central Expressway, Dallas, TX 75206. The phone number for the hotel is 800-222-TREE. A special workshop will be held on Thursday, Oct. 26, entitled "The SEM Today". It will cover the latest information concerning basic SEM, LVSEM, ESEM and detectors. Dr. David Joy, University of Tennessee, will be speaking on Low Voltage SEM and Variable Pressure SEM. YOU MUST BE REGISTERED FOR THE THURSDAY SEMINAR SERIES. It is free for members and new member applicants. If you plan to attend and have not registered, call or email Pam Neill at 817-568-6497 or pamela.neill-at-alconlabs.com
TSM PROGRAM SUMMARY OF DAILY ACTIVITIES
Thursday October 26, 2000 8:30-9:00 Registration at TI 9:00-5:00 Workshop coordinated by Kevin Cronyn, Hitachi and hosted by TI 12:30-7:00 Exhibitor setup and Poster setup DoubleTree 5:30-7:00 TSM Executive council Meeting, DoubleTree 7:00-9:00 Evening Social, DoubleTree 9:30-11:00 TSM Hospitality room DoubleTree
Friday October 27, 2000 8:00-8:30 Continental Breakfast in vendor's room 8:00-11:00 Registration 8:30-8:45 Introduction and announcements. 8:45-10:00 Platform presentations 10:00-10:15 Break in exhibitors’ room. 10:15-11:30 Guest speaker Russ Pinazotto "Electron Microscopy of Semiconductor Nanoparticles" 11:45-1:45 Lunch and business meeting with vendors at DoubleTree Hotel Campbell Center 1:45-3:00 Guest speaker Dr. Charles Mims “Use of High Pressure Freezing for Biological TEM". He will also include some information on plunge freezing. 3:00-4:30 Break in exhibitors’ room. Poster authors should be at their posts 5:30-7:00 Social in the hospitality suite. Dinner on your own. There will be a trip planned to the West End via DART for dinner and entertainment.
Saturday October 28, 2000 8:00-9:00 Registration 8:00-9:00 Continental Breakfast 8:00-9:15 Poster session continued 9:30-10:45 Platform presentations 10:45 adjourn
For more information and/or to register, please contact Pamela Neill, Program Chair TSM Alcon Research LTD 6201 South Freeway Forth Worth, TX 76134-2099 pamela.neill-at-alconlabs.com 817-568-6497 -- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-598-1291 (pager) KFAB Physical Analysis Lab--SEM/FIB/FA Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
a) 33% nitric acid, 67% methanol cooled to -25C and at 1.5 V
b) 45% phosphoric acid, 55% water at { 10C (no voltage given)
c) 10% orthophosphoric, 90% water at 0C and at 8 V.
I had success with the first solution in a Tenupol more years ago than I care to contemplate!
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: Zhongwen Yao [SMTP:zong-wen.yao-at-psi.ch] Sent: Thursday, October 12, 2000 9:02 AM To: Microscopy-at-sparc5.microscopy.com Subject: copper eclectrical polishing
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Listers, I meet some problem how to electrical-polish 3mm pure copper disk. Would you plese tell me the good solution and the good condition of electrical-polishing copper on tenupol-5 machine. Thanks
Mr.Zhongwen Yao EPFL- CRPP Villingen, PSI Switzerland
} Netters } } In upgrading our light microscopes, we are buying some new lenses for our } confocal scope and a brightfield scope. The salesforce is regaling us with } tales of improved resolution, clarity, brightness etc that comes with } spending double or triple the price for a standard oil immersion or dry } lens. Could someone please clarify, either on-line or off, what the } advantages and disadvantages of each of the following ( my texts on light } microscopy don't deal with them). Are they worth the extra dollars? } } 1) a water immersion lens for looking at cells suspended in buffer and } covered with a coverslip (water is used rather than oil between coverslip } and lens) These lenses typically have a lower numerical aperture than an } oil immersion lens. } } 2) a water dipping lens, used to examine cells in culture directly, without } a coverslip. I would expect the absence of refractive indice changes } (media/coverslip/air or oil) would explain their improved brightness. } } Thanks in advance for your input } } Steve Barlow } SDSU EM Facility
So they are saying that all of their old lenses are junk? Ask for a demo using your material at your site. Then you can see for yourself
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 **********************************************
Responding to the message of {Pine.GSO.4.10.10010111120030.19238-100000-at-aggie.pw.usda.gov} from "Pauline C. Yu" {splene-at-pw.usda.gov} : } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------.
} Hi listers } } We just bought a Pelco CPD2 with the hopes of replacing our old Polaron } E3000, and so far, we're rather unimpressed just trying to get it through } an initial setup. } We were wondering if any other users could tell us how many runs they get } out of an average CO2 cylinder? The cooling process with the CO2 seems } somewhat inefficient at this point and we're using up a lot just trying to } cool the chamber.
Pauline,
You should get very many runs out of an average cylinder. I'm not sure how the Pelco is designed, but with my Ladd Company critical point dryer, also cooled by CO2 gas flow-through, the "trick" is to set up a balance between the CO2 input valve and the exhaust or drain valve such that you get a BIG pressure drop from the CO2 tank pressure which is about 1000 psi - down to about 100-200 psi in the chamber as the gas flows through. You shouldn't need to open the input valve very much to achieve this effect. The cooling of the gas, hence the chamber, depends on a big pressure drop, the ol' Joule-Thomposon effect, just like in a refrigerator. When I first got my Ladd unit I had the same problem because I had too high a pressure in the chamber during blow through of the gas.
I hope this helps. It should only take just a few minutes to cool the chamber from room temp to about 5 C, for example, depending on size and mass of the chamber on your unit
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 looked in our archives and have found 2 solutions that Bernie Kestel from Argonne National Lab has developed for copper which may be of use. This work was done with our Model 550 single vertical jet electropolisher so references to jet height would be unique to our system. The other parameters may need to be adjusted slightly. Please be careful in using any of these chemicals and especially when adjusting the recipes. I owuld suggest that you contact Bernie Kestel directly prior to using these. You can reach him by email at kestel-at-anl.gov.
Recipe #2 150 ml HNO3 350 ml methanol 40 ml butyl cellosolve
Temp: -20 degrees C Jet height: 3.9mm Pump setting: 2.5 Volts: 40 Current: 75mA NOTES: excellent polish. Lower temperature result in an uneven foil surface.
I hope this information helps. If I can be of any additional assistance, please feel free to contact me.
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 Zhongwen Yao } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Listers, I meet some problem how to electrical-polish 3mm pure copper disk. Would you plese tell me the good solution and the good condition of electrical-polishing copper on tenupol-5 machine. Thanks
Mr.Zhongwen Yao EPFL- CRPP Villingen, PSI Switzerland
The K575 uses rotary and turbomolecular drag pumping to achive the high vaccum required.
The instrument has an automated shutter assembly to allow sputter target cleaning (Remove the oxide layer from the chromium target material) prior to sample sputtering.
We ahve had some very good application results witht he coater, please drop me a line for further info.
Best regards
Gareth } -----Original Message----- } From: Tobias Baskin [SMTP:BaskinT-at-missouri.edu] } Sent: Thursday, October 12, 2000 3:00 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: RE: Coater for use with FESEM } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society 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, } As seen below, Gareth Robinson mentions chromium as an option } for fine coats to use in high res SEM. } } A word of caution: Chromium is tricky to work with. You } cannot use chromium sputter coating the same simply way you are used } to sputtering with, say, gold or platinum. You have to keep the } atmosphere in the coater squeaky clean, and even cleaner, and you } must absolutely minimize the exposure of your coated sample to oxygen. } } I expect that chromium coats can be superb, but you cannot } expect to simply put a chromium target in your sputter coater and } have decent results. We found this out the hard way. } } As ever, } Tobias } } } } } } } } } Ben, } } } } You may also want to consider the option of Chromium coating which } will } } give grain siz of less than 0.5NM Chromium grain size with thin film } } deposition typically 5NM. } } } } We manufacture and sell the K575 Sputter Coater unit. It has the } } advantage of automated cycle coating Turbomolecular drag pumping for } } High Vaccum. } } } } Please take a look at our web site for a detailed technical brief and } } the instrument specification. } } } } HTTP://www.emitech.co.uk } } } } Kind regards } } } } Gareth Gareth.Robinson-at-emitech.co.uk } } -- } _ ____ __ ____ Tobias I. Baskin } / \ / / \ / \ \ 109 Tucker Hall } / / / / \ \ \ Biological } Sciences } /_ / __ /__ \ \ \__ University of } Missouri } / / / \ \ \ Columbia, } MO USA } / / / \ \ \ 65211-7400 } / / ___ / \ \__/ \ ____ voice: } 573-882-0173 } fax: 573-882-0123
Hi All, I have a JSM 840 that I need to relocate soon. The new location is very near a railroad and interstate! Does anyone have information on the optimum thickness of a concrete slab to prevent vibrations? Thanks,
Steve Buckingham Excellatron Solid State LLC 1460 Roswell Street, Suite J Smyrna, GA 30080 phone 770 438 2201 fax 770 438 2118
Hi E-beam buddies, After our last excimer laser was shut down, I had the idea of cleaning our Pt apertures with our plasma etcher using dry air. Optically they appear cleaner but in the TEM they are worse than before they went into the plasma. The aperture ID edge appears to be populated with hemispherical features that are prone to charging. I considered back streaming from the fomblin filled mech. pump but there is no residual smell of oil or sign of it anywhere about the chamber. Does anyone have any idea what is going on & perhaps how these apertures might be recovered? Some are cheap some are not.
Hi All, My goal is to localize active oxygen species/peroxidase activity in roots, and am wondering if anyone out there has any advice. I have several protocols, some using very different ways of utilizing DAB, and some using cerium chloride. They range from using whole mount tissue to processing tissue for TEM (and actually nothing in between). I am currently trying one of the DAB protocols, but would greatly appreciate hearing from anyone who has some insight. Thanks again for your help, Kristen Kristen A. Lennon Department of Plant Pathology 351 Bessey Hall Iowa State University Ames, IA 50011 kalen-at-iastate.edu
Funny thing is, even some really old lenses (old brass ones)! When it comes to low power still work pretty good!
I think many of the newer faster better sometimes what we will notice is very slight, but sales forces tend to extol the fact we need the latest even when perhaps we don't. i think anything produced today will only be just a little bit if at all better than the stuff form the 80's
Now on electron microscopes things are a bunch diff. with age especially on that old rust bucket of the edax computer We have on the AMR 1000... I suppose I should gather some spares or just consider replacing it with a PC when we finally get it set up and operational.
Ed Sharpe archivist for SMECC
{ { Subj: Re: light microscope lenses Date: 10/12/00 12:21:14 PM US Mountain Standard Time From: mcauliff-at-UMDNJ.EDU (Geoff McAuliffe) To: sbarlow-at-sunstroke.sdsu.edu (Steve Barlow) CC: CONFOCAL-at-LISTSERV.ACSU.BUFFALO.EDU, microscopy-at-sparc5.microscopy.com
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Steve Barlow wrote:
} Netters } } In upgrading our light microscopes, we are buying some new lenses for our } confocal scope and a brightfield scope. The salesforce is regaling us with } tales of improved resolution, clarity, brightness etc that comes with } spending double or triple the price for a standard oil immersion or dry } lens. Could someone please clarify, either on-line or off, what the } advantages and disadvantages of each of the following ( my texts on light } microscopy don't deal with them). Are they worth the extra dollars? } } 1) a water immersion lens for looking at cells suspended in buffer and } covered with a coverslip (water is used rather than oil between coverslip } and lens) These lenses typically have a lower numerical aperture than an } oil immersion lens. } } 2) a water dipping lens, used to examine cells in culture directly, without } a coverslip. I would expect the absence of refractive indice changes } (media/coverslip/air or oil) would explain their improved brightness. } } Thanks in advance for your input } } Steve Barlow } SDSU EM Facility
So they are saying that all of their old lenses are junk? Ask for a demo using your material at your site. Then you can see for yourself
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 **********************************************
Dear Mr. Yao, The Struers Tenupol instructions I have recommend a solution for Cu polishing consisting of: 250 ml phosphoric acid 500 ml distilled water 250 ml ethanol 2 ml Vogel's Sparbeize 5 g urea "to be mixed successively" and used at 15 degrees C, 0.3 A current, 15 V, flowrate 4 for 1 minute. I'm afraid I have no idea what "Vogel's Sparbeize" is. Another solution I have used for the window technique is 33% (v/v) nitric acid and 67% methanol at room temperature ( {25 degrees C), 8 V. You must remove the sample quickly to prevent oxidation and wash in methanol. This is a very flammable and reactive mixture used as jet fuel, so keep it away from heat, sparks and flame. 50 ml At 08:02 AM 10/12/00 -0500, you wrote: } } Listers, I meet some problem how to electrical-polish 3mm pure copper } disk. Would you plese tell me the good solution and the good condition } of electrical-polishing copper on tenupol-5 machine. } Thanks } } Mr.Zhongwen Yao
Good 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
Dear Gunnar, I would recommend masking a part of the Ge by putting a bit of Al foil over it before coating. You should be able to see the interface between the coated and uncoated surface clearly in the FESEM due to the atomic weight difference, and then zoom up to high magnification to see if there is any structure in the Au film. I know in a Au sputtered film this could be seen easily, but I haven't looked at a thermal evaporated film. It will show up best on a polished Ge wafer. You can use higher kV (15-20) and high resolution conditions in the FESEM, because the sample is conductive. At 03:58 PM 10/12/00 +0200, you wrote: } Hi, } } I am looking for help concerning the following problem: Someone in our } institute is trying to deposit a 10nm Au-film on a Ge single crystal by } thermal evaporation from a tungsten boat (...rate: 0.5A/s). } The main point to me is, if FESEM is an appropriate method to determine if } we } have really a Au-film or only gold islands ... } } I do appreciate any comments concering both points (FESEM-tricks, } because 99% of our samples are uncoated insulators) and the depostition } technique/parameters for very thin gold films ... } } ThanX very much in advance! } } } } Dipl.Ing.(FH) G.Glasser
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
MSA Listers, This is a long shot, but if anyone of you has information on the pin output of a DT2255 NuBus frame grabber and could forward to me off-list, I would be very grateful. DT does not have this info on their web page and seems to be selling support; I don't want to know how much they charge for pin output info on obsolete boards! Thanks.
-- Gregory Mulhollan, Ph.D. Extreme Devices Inc. 101 West 6th Street Suite 200 Austin, TX 78701 (512)479-7740 x2231 voice (512)479-7750 fax mulhollan-at-extremedevices.com
I would appreciate any advice/recommendations on TEM analysis of lipsomes, particularly vesicle size. Thanks Theresa
Theresa A. Fassel Core EM Unit, Mail Box MB-32 The Scripps Research Institute 10,550 N. Torrey Pines Rd. LaJolla, CA 92037-1027 tel: (858)-784-8182 fax: (858)-784-8193 tfassel-at-scripps.edu
Gatan Inc. has an immediate opening for a person to handle GIF and PEELS Product management. Gatan has a complete facility equiped with a new FEI Tecnai 20, a JEOL 2010/FasTEM and a Philips CM12.
The Individual should have either GIF or PEELS experience, a strong TEM background in Biological or Materials science, good computer skills, a willingness to travel and the vision and drive to help determine the future of these products.
The position is based in Pleasanton, CA and carries a salary comensurate with experience as well as a bonus plan and Corporate success share plan.
Please contact Ian Cotton, Director of Marketing at 925-224-7343 or email your resume to icotton-at-gatan.com.
********************************* Ian Cotton Marketing Director, Gatan, Inc. 5933 Coronado Lane Pleasanton, CA 94588 USA Phone 925-224-7343 Fax 925-463-0204 E-mail icotton-at-gatan.com Surf the Web to www.gatan.com *********************************
Gatan Inc. has an immediate opening for a person to manage our newly equiped demonstration facility. We have recently installed a new FEI Tecnai 20 and a JEOL 2010/FasTEM TEM. A full suite of Gatan analytical systems ( GIF and ENFINA PEELS ) and imaging systems are installed on these two microscopes.
The Individual should have either GIF or PEELS experience, a strong TEM background in Biological or Materials science, good computer skills, and excellent interpersonal skills. You should be experienced in the management of TEM instrumentation and have good organisational skills.
This is a challenging position for an individual who wants to work with state of the art equipment and contribute to the future development of our products.
The position is based in Pleasanton, CA and carries a salary comensurate with experience as well as a bonus plan and Corporate success share plan.
Please contact Ian Cotton, Director of Marketing at 925-224-7343 or email your resume to icotton-at-gatan.com.
********************************* Ian Cotton Marketing Director, Gatan, Inc. 5933 Coronado Lane Pleasanton, CA 94588 USA Phone 925-224-7343 Fax 925-463-0204 E-mail icotton-at-gatan.com Surf the Web to www.gatan.com *********************************
A colleague of mine is interested in purchasing a tissue processor. She has limited access to email so please reply directly to me and let me know what processor you use and would recommend. I'll pass the word along.
Thank you,
Ruth
*************************************** Ruth Yamawaki Department of Comparative Medicine Stanford University Stanford, CA 94305 ***************************************
*Date sent: Thu, 12 Oct 2000 08:02:18 -0500 *To: Microscopy-at-sparc5.microscopy.com *From: Zhongwen Yao {zong-wen.yao-at-psi.ch} *Subject: copper eclectrical polishing
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It seems that the best way, if you have enough money, is to buy proper electrolyte from the Struers. Based on our experiance, copper is difficult for TEM preparation. We use to say that even weather has an influance on electropolishing conditions. Good luck, Witold Zielinski
%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%% Witold Zielinski, Ph.D. Warsaw University of Technology Department of Materials Science and Engineering 02-507 Warszawa, Woloska 141 POLAND
Patton, David 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 slice off the specimen with a razor blade and use the } blade to scrape the stub fairly clean. As we use colloidal } graphite, which is conductive, I do not mind some residual } glue as it will get covered by glue from the next use. } This glue can be removed, if required, by sonicating in } butyl acetate. } } Dave } } On Wed, 11 Oct 2000 21:09:07 +1000 Jim at ProSciTech } {jim-at-proscitech.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 } } -----------------------------------------------------------------------. } } } } } } It all depends: how busy you are, how expensive staff time in your country is } } and how costly the place is you buy your stubs from. Obviously some specimens } } should be stored in a desiccator for an eternity and other can be discarded } } immediately. } } There is nothing magical about cleaning stubs. Solvents are messy. Best to } } place on some newspaper a very coarse file or a sheet of coarse (50 grit) } } sandpaper and then rub the old stub over that file of sandpaper until specimen } } and glue are removed. } } } } Disclaimer: ProSciTech sells stubs, but we are happy if people recycle and this } } leaves some money for useful purchases. } } 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 } } ABN: 99 724 136 560 www.proscitech.com } } } } On Wednesday, October 11, 2000 3:09 AM, O. O. Ilori } } [SMTP:sojilori-at-oauife.edu.ng] wrote: } } } } } } Hello Guys } } } I am a novice user of a SEM (LEO 1450 to be precise) equipment which was } } } just installed in our central science lab. } } } Is there a way to clean stubs for reuse or are stubs disposable once used. } } } ( This may sound naive) } } } soji } } } Mr. O. O. ILORI } } } DEPARTMENT OF ELECTRONIC/ELECTRICAL ENGINEERING } } } OBAFEMI AWOLOWO UNIVERSITY, } } } ILE-IFE, OSUN STATE } } } NIGERIA. } } } email: sojilori-at-oauife.edu.ng } } } } } } } } } } } } ---------------------------------------- } Patton, David } Email: David.Patton-at-uwe.ac.uk } "University of the West of England" If you have access to a glass bead blaster, it does a great job as long as you weren't looking for a mirror finish.
Ken Converse owner Quality Images third party SEM service Delta, PA
Bruce Brinson 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 E-beam buddies, } After our last excimer laser was shut down, I had the idea of } cleaning our Pt apertures with our plasma etcher using dry air. } Optically they appear cleaner but in the TEM they are worse than before } they went into the plasma. The aperture ID edge appears to be populated } with hemispherical features that are prone to charging. } I considered back streaming from the fomblin filled mech. pump but } there is no residual smell of oil or sign of it anywhere about the } chamber. Does anyone have any idea what is going on & perhaps how these } apertures might be recovered? Some are cheap some are not. } } thanks, } Bruce Brinson } Rice U. Bruce, I've been cleaning apertures of all types for many years with 1 micron diamond paste and a cut-knap polishing cloth, although any lint-free cloth will work if the apertures are flat and fairly thin. Heavily counter-sunk apertures like Siemens 2mm really benefit from the cut-knap cloths (metallography supplies).
Finish by cleaning ultrsonically in Joy and hot water, blow dry immediatly.
As to what is going on in your plasma system, I can't help you there.
Ken Converse Quality Images third party SEM service Delta, PA
thanks for all your help in diagnosing my 35 problem. turns that the aperture heater was the culprit. i'll make note of this for "next time"!
b- ---------------------------------------------- Brian McIntyre Electron Microscopy Lab- River Campus Univ of Rochester Rochester, NY 14627 716-275-3058/4875
MSA Listers: Is John Russ offering training course on image analysis this year? When? Where? TIA.
Joseph Fu National Institute of Standards & Technology 100 Bureau drive Stop 8212 Gaithersburg, MD. 20899-8212 Tel: 301-975-3495 Fax: 301-869-0822 Email: jofu-at-nist.gov
I've had some experience with water immersion lenses. Their main advantage is higher numerical aperture than dry lenses at equivalent magnification, so better theoretical resolution. They don't offer quite as good resolution as oil immersion lenses, but they tend to have longer working distances, I believe, and don't have the messy clean-up of oil immersion lenses. So, yes, they do have some distinct advantages.
However, as others have pointed out, you may already be able to see everything you need to see with your current lenses. And nothing beats a test-drive. -- 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
In a message dated 10/13/00 9:01:02 AM, jofu-at-nist.gov writes:
} Is John Russ offering training course on image analysis this year? When? } Where? TIA.
Thanks for the chance to plug the courses. There will be three sessions in 2001. Dates are May 9-11 and October 30-Nov. 1, in Raleigh (at N. C. State Univ) and June 6-8 in Denmark (at the Danish Technological Institute). Full info on the course contents, registration info, etc., (and a downloadable copy of the brochure for the 2000 courses, which is correct except that it does not have the year 2001 dates) is available on-line at http://members.aol.com/IPCourse/
It is also possible (and cost effective for groups of more than about 6-7 people) to arrange for an on-site course. Contact Cindy Allen in the NCSU Continuing Education department at 919 515 8171 for details.
I don't think there is an "optimum" thickness for the concrete slab. If vibration from the railroad (or any other source) is a concern most isolate the main slab by sawing & removing the concrete under the optics console. A new concrete slab is laid with "pea sized agregate" & felt on the sides. This lets the column stand free from the rest of the slab.
Regards,
Earl Weltmer
"Buckingham, Steve" 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 have a JSM 840 that I need to relocate soon. The new location is } very near a railroad and interstate! Does anyone have information on the } optimum thickness of a concrete slab to prevent vibrations? } Thanks, } } Steve Buckingham } Excellatron Solid State LLC } 1460 Roswell Street, Suite J } Smyrna, GA 30080 } phone 770 438 2201 } fax 770 438 2118
Many years ago we regularly used platinum crucibles in many analytical chemistry procedures. According to my analytical chemistry book, platinum can be cleaned by fusion with potassium or sodium bisulfite. I later used this procedure with good success to clean the platinum apertures for an RCA EML electron microscope. Get a small ceramic crucible, put a bit of the bisulfite in it, heat it until the bisulfite melts, then drop the apertures into the melt. After a few minutes allow the melt to solidify, dissolve the bisulfite with hot water, and your apertures should be as bright and shiny as new.
If this doesnn't work, try heating them in a mixture of equal parts of concentrated hydrofluoric acid and concentrated hydrochloric acid - using appropriate safety precautions, of course!
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
To whom it may concern, I am a materials engineering student at the Univ of Cincinnati (Ohio) currently co-oping at a local aerospace company. They're wanting some microprobe analysis done on some parts and have put me in charge of that. I have a descent understanding of X-Ray diffraction but would like some information on the specifics of a microprobe so that I know exactly what is going on when I'm sitting watching the analysis take place. Is there any electronic information you can send or any sites you can refer me to that will give in depth information on the workings of a microprobe? Please respond to keithj-at-meyertool.com.
Based on your information we believe there may be burrs on the ID edge of your holes. We could open up the holes a bit to provide burr-free apertures, or we would allow you a trade in since as the only US Electron Microscope aperture manufacturer they probably originated with us anyway. If an aperture has a burr-free edge, a flamer should do for cleaning Pt and if they are Moly you can clean them in an evaporator. Even if your aperture is imported we would allow a trade in on a new disc or strip. Let me know if I can be of further help,
John Arnott --
LADD RESEARCH 131 Dorset Lane Williston, VT 05495 USA
web site http://www.laddresearch.com
TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail sales-at-laddresearch.com
Quality Since 1955
Bruce Brinson wrote: }
} Hi E-beam buddies, } After our last excimer laser was shut down, I had the idea of } cleaning our Pt apertures with our plasma etcher using dry air. } Optically they appear cleaner but in the TEM they are worse than before } they went into the plasma. The aperture ID edge appears to be populated } with hemispherical features that are prone to charging. } I considered back streaming from the fomblin filled mech. pump but } there is no residual smell of oil or sign of it anywhere about the } chamber. Does anyone have any idea what is going on & perhaps how these } apertures might be recovered? Some are cheap some are not. } } thanks, } Bruce Brinson } Rice U.
Any feedback on the SEM images of the needles? Should I go ahead and finish the other 4 that way?
At only 25X mag, seems like LM would give better images, in color, for example, and the steel surfaces seem to show up better. The SEM images look a bit gray, as no metal coating and low kV imaging. Metal coating would show them up better, but I suppose that would not be allowed under the experimental constraints.
What do you think?
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
Hi Bruce, } } I have never tried the plasma etcher but I suspect it would remove organics } effectively, just as a flame does. After several cleanings in the flame, we } are usually left with deposits similar to what you describe. My old Philips } 300 manual suggests these are usually silica-- this makes sense for our } samples, maybe for yours as well? Same manual advises "24 hrs. immersion in } 10% solution of HF in water". I usually save the ones I can't clean until I } have several, then turn them over to a colleague who works with HF regularly.
} Most come back as clean as new. The hemispherical features suggest to me that } our contaminant will melt, but not vaporize in the flame, consistent with } silica. You would have to look into the power of your etcher. Naturally, } these techniques are for solid Pt apertures, and HF is not for the } accident-prone. Good luck! By the way, our much newer Philips manual (XL30 } ESEM-FEG) gives 4 methods of heating but no mention of HF. } } Matt }
Matthew J. Lynn, Ph.D. Center for Advanced Microscopy University of Miami (305)284-4736 mlynn-at-miami.edu
The optimum thickness of concrete is whatever thickness of concrete it takes to stop the train. Any other answer is incorrect.
David C
} } ----- Original Message ----- } From: Earl Weltmer {earlw-at-pacbell.net} } To: Buckingham, Steve {sbuckingham-at-excellatron.com} } Cc: 'Microscopy-at-MSA.Microscopy.Com' {Microscopy-at-sparc5.microscopy.com} } Sent: Friday, October 13, 2000 3:49 PM } Subject: Re: Concrete slab for 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 Steve, } } } } I don't think there is an "optimum" thickness for the concrete slab. } } If vibration from the railroad (or any other source) is a concern most } isolate } } the main slab by sawing & removing the concrete under the optics console. } A } } new concrete slab is laid with "pea sized agregate" & felt on the sides. } This } } lets the column stand free from the rest of the slab. } } } } Regards, } } } } Earl Weltmer } } } } "Buckingham, Steve" 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 have a JSM 840 that I need to relocate soon. The new location } is } } } very near a railroad and interstate! Does anyone have information on the } } } optimum thickness of a concrete slab to prevent vibrations? } } } Thanks, } } } } } } Steve Buckingham } } } Excellatron Solid State LLC } } } 1460 Roswell Street, Suite J } } } Smyrna, GA 30080 } } } phone 770 438 2201 } } } fax 770 438 2118 } } } } } } }
The optimum thickness of concrete is whatever thickness of concrete it takes to stop the train. Any other answer is incorrect.
David C
} } ----- Original Message ----- } From: Earl Weltmer {earlw-at-pacbell.net} } To: Buckingham, Steve {sbuckingham-at-excellatron.com} } Cc: 'Microscopy-at-MSA.Microscopy.Com' {Microscopy-at-sparc5.microscopy.com} } Sent: Friday, October 13, 2000 3:49 PM } Subject: Re: Concrete slab for 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 Steve, } } } } I don't think there is an "optimum" thickness for the concrete slab. } } If vibration from the railroad (or any other source) is a concern most } isolate } } the main slab by sawing & removing the concrete under the optics console. } A } } new concrete slab is laid with "pea sized agregate" & felt on the sides. } This } } lets the column stand free from the rest of the slab. } } } } Regards, } } } } Earl Weltmer } } } } "Buckingham, Steve" 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 have a JSM 840 that I need to relocate soon. The new location } is } } } very near a railroad and interstate! Does anyone have information on the } } } optimum thickness of a concrete slab to prevent vibrations? } } } Thanks, } } } } } } Steve Buckingham } } } Excellatron Solid State LLC } } } 1460 Roswell Street, Suite J } } } Smyrna, GA 30080 } } } phone 770 438 2201 } } } fax 770 438 2118 } } } } } } }
It is not the thickness, but rather the area of the slab. It is better be minimal. Also, the soil under the slab- rock is bad, sand is good. If you have any choice on the matter- construction is not started yet, etc.- than have the slab on the sand cushion, make it small, just the size of the SEM footprint, and separate from the rest of the building. It is possible, in some cases, to cut the existing concrete slab in order to separate it from the rest of the structure. (Consult the construction expert first!!) Then, fill the cuts with soft silicone, rubber, etc. in order to prevent hard objects from getting in the cuts, as they will make an acoustic contact, which you are trying to avoid.
Also, many options from many sources are available regarding antivibration supports and mounts, depending on your situation and requirements. Some are expensive, some are not. Contact me off list if you need further help.
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax
This message is made of 100% recycled electrons. -----Original Message----- } From: Buckingham, Steve {sbuckingham-at-excellatron.com} To: 'Microscopy-at-MSA.Microscopy.Com' {Microscopy-at-sparc5.microscopy.com}
Have you considered the speed, size, & direction of the train? In addition, the type of concrete & it's reinforcement material (ribar) also needs to be considered. Also whether you want the train to transmit vibration, collide with the SEM or land on top of the SEM needs to be calculated.
Any other answer is incorrect.
Regards,
Earl
David Cockayne wrote:
} Earl } } The optimum thickness of concrete is whatever thickness of concrete it } takes to stop the train. Any other answer is incorrect. } } David C } } } } } ----- Original Message ----- } } From: Earl Weltmer {earlw-at-pacbell.net} } } To: Buckingham, Steve {sbuckingham-at-excellatron.com} } } Cc: 'Microscopy-at-MSA.Microscopy.Com' {Microscopy-at-sparc5.microscopy.com} } } Sent: Friday, October 13, 2000 3:49 PM } } Subject: Re: Concrete slab for 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 Steve, } } } } } } I don't think there is an "optimum" thickness for the concrete slab. } } } If vibration from the railroad (or any other source) is a concern most } } isolate } } } the main slab by sawing & removing the concrete under the optics } console. } } A } } } new concrete slab is laid with "pea sized agregate" & felt on the sides. } } This } } } lets the column stand free from the rest of the slab. } } } } } } Regards, } } } } } } Earl Weltmer } } } } } } "Buckingham, Steve" 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 have a JSM 840 that I need to relocate soon. The new } location } } is } } } } very near a railroad and interstate! Does anyone have information on } the } } } } optimum thickness of a concrete slab to prevent vibrations? } } } } Thanks, } } } } } } } } Steve Buckingham } } } } Excellatron Solid State LLC } } } } 1460 Roswell Street, Suite J } } } } Smyrna, GA 30080 } } } } phone 770 438 2201 } } } } fax 770 438 2118 } } } } } } } } } } }
NYSEM Presidential Symposium 2000: "Cell Polarization: Mechanisms at the Membrane"
The New York Society for Experimental Microscopy will hold its annual Presidential Symposium on 19 October from 9:00 AM to 5:30 PM at Columbia University (Hammer Health Sciences Building, Room 401*). This year's Symposium is entitled: "Cell Polarization: Mechanisms at the Membrane". The Symposium will focus on recent breakthroughs in understanding the molecular mechanisms involved in the generation of cell polarity. Symposium participants will highlight recent advances in a number of model systems in which microscopic approaches can be combined with genetic and molecular biological approaches to understand cell polarity. One emerging theme that will be explored during the Symposium is the degree to which basic molecular pathways for cell polarization have been conserved despite the diversity of structures that result in the different systems. Registration is free, but preregistration is required for lunch (contact Dr. Philip Leopold, Cornell Univ. Medical School, email: pleopold-at-mail.med.cornell.edu; phone 212-746-2255)
* The Hammer Health Sciences Building is one block west of Broadway on the corner of Fort Washington and 168th Street. There is a subway stop at Broadway and 168th Street.
SCHEDULE - NYSEM PRESIDENTIAL SYMPOSIUM 2000 "Cell Polarization: Mechanism at the Membrane" 19 October 2000 Columbia University Hammer Health Sciences Building, Room 401
9:15 - 9:55 John Cooper (Department of Cell Biology & Physiology, Washington University, St. Louis): "Interactions of Microtubules with the Cortex During Mitosis in Budding Yeast"
9:55 - 10:35 Kerry Bloom (Department of Biology, University of North Carolina - Chapel Hill "How Microtubules Polarize Nuclear Movements in Yeast"
10:35 - 10:55 Coffee Break
10:55 - 11:35 Fred Chang (Department of Microbiology, Columbia University, New York) "Spatial control and cytoskeletal dynamics in fission yeast"
11:35 - 12:15 Carole Parent (National Cancer Institute, NIH, Bethesda) "Visualizing Signaling Events in Chemotaxing Eukaryotic Cells"
2:00 - 2:40 Gregg Gundersen (Department of Anatomy & Cell Biology, Columbia University, New York) "Rho GTPase Pathways Integrate the Microtubule and Actin Cytoskeletons During Cell Migration"
2:40 - 3:20 Clare Waterman-Storer (Department of Cell Biology, Scripps Research Institute, La Jolla) "Microtubule and Actin Interactions in Cell Polarization and Motility"
3:20 - 3:40 Coffee Break
3:40 - 4:20 Enrique Rodriguez-Boulan (Dyson Vision Research Institute, Cornell University, New York) "Role of Actin Filaments and Microtubules in Polarized Delivery of Plasma Membrane Proteins"
4:20 - 4:30 Gregg Gundersen - Closing Remarks
4:30 - 5:30 Reception - River View Lounge
**************************************************************************** Frank Macaluso tel: 718-430-3547 Analytical Imaging Facility fax: 718-430-8996 Albert Einstein College of Medicine e-mail: macaluso-at-aecom.yu.edu 1300 Morris Park Avenue http://www.aecom.yu.edu/aif/ Bronx, NY 10461 ****************************************************************************
---------------------- Forwarded by Paula M Churt/TMG/CSC on 10/13/2000 10:59 AM ---------------------------
Diane A Ciaburri/GDDS-at-GDDS 10/13/2000 09:55 AM
To: Paula M Churt/TMG/CSC-at-CSC cc:
Hello
Has anyone observed biological material-antibody-dynabead complex in EM? Our user has great results with Dynabeads M-450 Epoxy Product # 140.01 in his assay, but we would like to visualize the virus attached to the dynabead. The manufacturer suggests the iron core may leak on sectioning causing contamination. Any ideas? Thanks
Theresa
Theresa A. Fassel Core EM Unit, Mail Box MB-32 The Scripps Research Institute 10,550 N. Torrey Pines Rd. LaJolla, CA 92037-1027 tel: (858)-784-8182 fax: (858)-784-8193 tfassel-at-scripps.edu
The optimum thickness of concrete is whatever thickness of concrete it takes to stop the train. Any other answer is incorrect.
David C
----- Original Message ----- } From: Earl Weltmer {earlw-at-pacbell.net} To: Buckingham, Steve {sbuckingham-at-excellatron.com} Cc: 'Microscopy-at-MSA.Microscopy.Com' {Microscopy-at-sparc5.microscopy.com} Sent: Friday, October 13, 2000 3:49 PM
I run in the ETCH mode with Argon. Set power level to 5%. Put in the dummy Aluminum target. Hit PROCESS and keep the vacuum at 60mTorr-40mTorr. Try for about 30 seconds and then look at the standard. Repeat until clean. The lower the vacuum level, the less aggressive is the cleaning action. The plasma will strike when the vacuum reaches 20mTorr in the ETCH mode; 40mTorr in the PLATE mode. When done, remove the Al target and put back in the sputter coater target.
I coat (PLATE) at 80mTorr, 5nM/minute. This gives a nice, smooth and even coating with Au/Pd or Pt.
I'm using a Hummer VII. BTW, if you have trouble with the plasma not striking for PLATE when vacuum reaches 40mTorr, I have a simple fix that makes it virtually never fail to fire.
gary g.
At 07:59 AM 10/13/00, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } 10/13/00 Subject: Cleaning of SEM Mag } Standard } 09:36 } AM } } } } } } } } } Hi all, } } I've got an NIST 484e SEM Mag Standard that has years of carbon build-up } ("burn marks"). I want to clean it with my plasma etcher but I'm not sure } about the parameters to use. Does anyone have experience doing this? } } I have an Anatech Hummer plasma coater. Do I use the etch or plasma mode? } Argon or air for atmosphere? What vacuum and voltage, and for how long? } } Thanks! } } Diane Ciaburri } } dciaburri-at-gdds.com } (413)494-2847 } General Dynamics } 100 Plastics Ave. } Pittsfield Ma 01201
David is right. Our testing has shown that saw cutting the floor to isolate the column from the rest of the building has no affect other than to cost a lot. The vibration is transmitted through the ground, not the bldg.
Craig Franklin Vibration Engineering Consultants www.vibeng.com
----- Original Message ----- } From: David Cockayne {david.cockayne-at-materials.oxford.ac.uk} To: {Microscopy-at-sparc5.microscopy.com} Sent: Friday, October 13, 2000 5:16 PM
I don't know that the beads "leak", but they do not present a smooth appearance in sections, either internally or - sometimes - at the surface. the internal appearance can be discounted, but I always process some control ("clean") beads in parallel. Sometimes there is a little fluffy stuff (OK - flocculent material) on the surface of the blank beads, but it could never (in my experience) be confused with viruses.
One other thing I would suggest is to have your researcher try to get as much material aspossible on small beads. I once examined some IP samples that were not very efficiently done on large beads - between the large surface area of the beads and the low density of actual sample on the beads, a *lot* of time was wasted just looking for the IP material. A real aspirin project :)
I section Dynabeads on the not-so-great part of my diamond knife; I haven't noticed new scratches from it, but I suspect that the material is potentially dulling and scratchy.
Good luck!
Tamara Howard CSHL
On Fri, 13 Oct 2000, Theresa Fassel 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 } } Has anyone observed biological material-antibody-dynabead complex in EM? } Our user has great results with Dynabeads M-450 Epoxy Product # 140.01 in } his assay, but we would like to visualize the virus attached to the } dynabead. The manufacturer suggests the iron core may leak on sectioning } causing contamination. Any ideas? } Thanks } } Theresa } } } Theresa A. Fassel } Core EM Unit, Mail Box MB-32 } The Scripps Research Institute } 10,550 N. Torrey Pines Rd. } LaJolla, CA 92037-1027 } tel: (858)-784-8182 } fax: (858)-784-8193 } tfassel-at-scripps.edu } }
Hi Ruth; I would highly recommend the Ted Pella Microwave oven series 3450. I have used this oven for over 3 years now and have processed over 500 specimens (human and animal) without a single mishap to the tissue. In fact you may find the results better then you would get by conventional processing. You can contact Rick Giberson with Ted Pella's toll free number and he can give you more specific information and a price to work with. You can contact me at rla-at-mindspring.com as well.
Ron Austin (Research Associate) LSU Medical Ct. at Shreveport Dept. of Pathology Shreveport, LA 318-675-4557
The following is a request from a producer of the PBS TV show NOVA, which came in to the MSA Business office.
Can anyone help? Please correspond directly to the requester.
} } } } } }
Hi Folks --
I'm a producer with the science TV series NOVA. I'm working on a show about evolution and I'm desperately looking for a way to adapt a zeiss axiophot microscope to an Aaton 16mm film camera. So far I've struck out on finding anyone who has gear to accomplish this feat.
Any suggestiongs?
-- Chris Schmidt Powderhouse Productions 259 Elm Street Somerville MA 02144 617-629-2200 work 617-776-7905 fax 781-820-0727 mobile 617-403-8699 pager 781-646-0928 home Chris-at-powderhouse.net
Increasing the mass under the microscope works. An isolated room-size slab of 1m thickness can reduce vertical vibration by a factor of four and horizontal vibration by 10 to 12 times [1]. This foundation was a significant factor in allowing us the obtain a TEM resolution better than one Angstrom [2].
-Mike O'Keefe
1. "Design and implementation of a site for a one-Ångstrom TEM", John H. Turner, Michael A. O'Keefe and Robert Mueller, in 55th Ann. Proc. MSA, Cleveland, Ohio (1997) 1177-1178. 2. "The NCEM One-Ångstrom Microscope Project Reaches 0.89Å Resolution", Michael A. O'Keefe and Y.C. Wang in 58th Ann. Proc. MSA, Philadelphia, Pennsylvania (2000).
----- Original Message ----- } From: "Craig Franklin" {franklin-at-vibeng.com}
Craig, David, Earl, Steve,
Increasing the mass under the microscope works. An isolated room-size slab of 1m thickness can reduce vertical vibration by a factor of four and horizontal vibration by 10 to 12 times [1]. This foundation was a significant factor in allowing us the obtain a TEM resolution better than one Angstrom [2].
-Mike O'Keefe
1. "Design and implementation of a site for a one-Ångstrom TEM", John H. Turner, Michael A. O'Keefe and Robert Mueller, in 55th Ann. Proc. MSA, Cleveland, Ohio (1997) 1177-1178. 2. "The NCEM One-Ångstrom Microscope Project Reaches 0.89Å Resolution", Michael A. O'Keefe and Y.C. Wang in 58th Ann. Proc. MSA, Philadelphia, Pennsylvania (2000).
----- Original Message ----- } From: "Craig Franklin" {franklin-at-vibeng.com}
Craig, David, Earl, Steve,
Increasing the mass under the microscope works. An isolated room-size slab of 1m thickness can reduce vertical vibration by a factor of four and horizontal vibration by 10 to 12 times [1]. This foundation was a significant factor in allowing us the obtain a TEM resolution of better than one Angstrom [2].
-Mike O'Keefe
1. "Design and implementation of a site for a one-Ångstrom TEM", John H. Turner, Michael A. O'Keefe and Robert Mueller, in 55th Ann. Proc. MSA, Cleveland, Ohio (1997) 1177-1178. 2. "The NCEM One-Ångstrom Microscope Project Reaches 0.89Å Resolution", Michael A. O'Keefe and Y.C. Wang in 58th Ann. Proc. MSA, Philadelphia, Pennsylvania (2000).
----- Original Message ----- } From: "Craig Franklin" {franklin-at-vibeng.com}
Craig, David, Earl, Steve,
Increasing the mass under the microscope works. An isolated room-size slab of 1m thickness can reduce vertical vibration by a factor of four and horizontal vibration by 10 to 12 times [1]. Such a foundation was a significant factor in allowing us to obtain a TEM resolution better than one Angstrom [2].
-Mike O'Keefe
1. "Design and implementation of a site for a one-Ångstrom TEM", John H. Turner, Michael A. O'Keefe and Robert Mueller, in 55th Ann. Proc. MSA, Cleveland, Ohio (1997) 1177-1178. 2. "The NCEM One-Ångstrom Microscope Project Reaches 0.89Å Resolution", Michael A. O'Keefe and Y.C. Wang in 58th Ann. Proc. MSA, Philadelphia, Pennsylvania (2000).
----- Original Message ----- } From: "Craig Franklin" {franklin-at-vibeng.com}
Craig, David, Earl, Steve,
Increasing the mass under the microscope works. An isolated room-size slab of 1m thickness can reduce vertical vibration by a factor of four and horizontal vibration by 10 to 12 times [1]. Such a foundation was a significant factor in allowing us to obtain a TEM resolution better than one Angstrom [2].
-Mike O'Keefe
1. "Design and implementation of a site for a one-Ångstrom TEM", John H. Turner, Michael A. O'Keefe and Robert Mueller, in 55th Ann. Proc. MSA, Cleveland, Ohio (1997) 1177-1178. 2. "The NCEM One-Ångstrom Microscope Project Reaches 0.89Å Resolution", Michael A. O'Keefe and Y.C. Wang in 58th Ann. Proc. MSA, Philadelphia, Pennsylvania (2000).
----- Original Message ----- } From: "Craig Franklin" {franklin-at-vibeng.com}
} I have a descent understanding of X-Ray diffraction but would like some } information on the specifics of a microprobe so that I know exactly what is } going on when I'm sitting watching the analysis take place. Is there any } electronic information you can send or any sites you can refer me to that } will give in depth information on the workings of a microprobe?
Keith
There is a wonderful book "Handbook of Silicate Analysis", by P J Potts, that has great chapters on many modern instrumental techniques, including XRF and EPMA. Although written with a geological slant, Potts combines lucidity with solid info, and it should be interesting to you also.
cheers
rtch
ps I hope your XRD is located downstairs.
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
Thanks for the heads-up. I was working from home this morning (Sunday) and finally noticed and stopped it. I hoped it had only gone out once or twice, but I had received five copies this afternoon when I checked the listserver. :-(
Cheers, Mike
----- Original Message ----- } From: "Ritchie Sims" {r.sims-at-auckland.ac.nz}
To Mike, Craig, Earl and others,
Many years ago there was an article in JEOL News detailing the design of floors for electron microscopes at the John Innes Institute in the UK, and more recently I saw similarly detailed plans for a new facility in Australia (in Brisbane, I think). We have incorporated here aspects of the John Innes Institute plans, and do not have a vibration problem, but as we do not have trains that run anywhere nearby, unfortunately I have no idea whether or not these floors would be effective in reducing the vibration being discussed!
Rob
On 15 Oct 00, at 19:03, MAOKeefe-at-lbl.gov 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 } ---------------------------------------------------------------------- } -. } } } Craig, David, Earl, Steve, } } Increasing the mass under the microscope works. An isolated room-size } slab of 1m thickness can reduce vertical vibration by a factor of four } and horizontal vibration by 10 to 12 times [1]. This foundation was a } significant factor in allowing us the obtain a TEM resolution better } than one Angstrom [2]. } } -Mike O'Keefe } } 1. "Design and implementation of a site for a one-Ångstrom TEM", John } H. Turner, Michael A. O'Keefe and Robert Mueller, in 55th Ann. Proc. } MSA, Cleveland, Ohio (1997) 1177-1178. 2. "The NCEM One-Ångstrom } Microscope Project Reaches 0.89Å Resolution", Michael A. O'Keefe and } Y.C. Wang in 58th Ann. Proc. MSA, Philadelphia, Pennsylvania (2000). } } ----- Original Message ----- } } From: "Craig Franklin" {franklin-at-vibeng.com} } Date: Saturday, October 14, 2000 8:11 am } Subject: Re: Concrete slab for SEM } } } ------------------------------------------------------------------- } } ----- The Microscopy ListServer -- Sponsor: The Microscopy Society } } of America To Subscribe/Unsubscribe -- Send Email to } } ListServer-at-MSA.Microscopy.ComOn-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html-------- } } ---------------------------------------------------------------. } } } } } } Steve, } } } } David is right. } } Our testing has shown that saw cutting the floor to isolate the } } column from the rest of the building has no affect other than to } } cost a lot. The vibration is transmitted through the ground, not the } } bldg. } } } } Craig Franklin } } Vibration Engineering Consultants } } www.vibeng.com } } } } } } ----- Original Message ----- } } } From: David Cockayne {david.cockayne-at-materials.oxford.ac.uk} } } To: {Microscopy-at-sparc5.microscopy.com} } } Sent: Friday, October 13, 2000 5:16 PM } } Subject: Re: Concrete slab for 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} ------ } } -----------------------------------------------------------------. } } } } } } } } } Earl } } } } } } The optimum thickness of concrete is whatever thickness of } } concrete it } } } takes to stop the train. Any other answer is incorrect. } } } } } } } } } David C } } } } } } } } } ----- Original Message ----- } } } } From: Earl Weltmer {earlw-at-pacbell.net} } } } To: Buckingham, Steve {sbuckingham-at-excellatron.com} } } } Cc: 'Microscopy-at-MSA.Microscopy.Com' } } {Microscopy-at-sparc5.microscopy.com} } Sent: Friday, October 13, 2000 } } 3:49 PM } } } Subject: Re: Concrete slab for 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 Steve, } } } } } } } } I don't think there is an "optimum" thickness for the concrete } } slab.} } If vibration from the railroad (or any other source) is a } } concern most } } } isolate } } } } the main slab by sawing & removing the concrete under the optics } } console. } } } A } } } } new concrete slab is laid with "pea sized agregate" & felt on } } the sides. } } } This } } } } lets the column stand free from the rest of the slab. } } } } } } } } Regards, } } } } } } } } Earl Weltmer } } } } } } } } "Buckingham, Steve" 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 have a JSM 840 that I need to relocate soon. The new } } location } } } is } } } } } very near a railroad and interstate! Does anyone have } } information on } } the } } } } } optimum thickness of a concrete slab to prevent vibrations? } } } } } Thanks, } } } } } } } } } } Steve Buckingham } } } } } Excellatron Solid State LLC } } } } } 1460 Roswell Street, Suite J } } } } } Smyrna, GA 30080 } } } } } phone 770 438 2201 } } } } } fax 770 438 2118 } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } }
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 **
From root Mon Oct 16 02:48:57 2000 Return-Path: {Microscopy-request-at-sparc5.microscopy.com} Received: (from daemon-at-localhost) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id CAA19424 for dist-Microscopy; Mon, 16 Oct 2000 02:31:45 -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 CAA19421 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Mon, 16 Oct 2000 02:31:14 -0500 (CDT) Received: from oxmail.ox.ac.uk (oxmail2.ox.ac.uk [163.1.2.1]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id CAA19414 for {Microscopy-at-sparc5.microscopy.com} ; Mon, 16 Oct 2000 02:31:03 -0500 (CDT) Received: from ermine.ox.ac.uk ([163.1.2.13]) by oxmail.ox.ac.uk with esmtp (Exim 3.12 #1) id 13l4j0-0003Lg-00 for Microscopy-at-sparc5.microscopy.com; Mon, 16 Oct 2000 08:30:22 +0100 Received: from cockayne.materials.ox.ac.uk ([163.1.65.199] helo=cockayne) by ermine.ox.ac.uk with smtp (Exim 3.13 #1) id 13l4j0-00073J-00 for Microscopy-at-sparc5.microscopy.com; Mon, 16 Oct 2000 08:30:22 +0100 Message-ID: {001701c03742$383d8920$c74101a3-at-materials.ox.ac.uk}
Dear Mike
I am sure that increasing the mass under the microscope works, as you say. But the question was what is the "optimum thickness to stop vibrations". There is only one "optimum", and I believe my answer was it.
David ----- Original Message ----- } From: {"MAOKeefe-at-lbl.gov"-at-sparc5.microscopy.com} To: Craig Franklin {franklin-at-vibeng.com} Cc: MSA List Server {Microscopy-at-sparc5.microscopy.com} ; DavidCockayne {david.cockayne-at-materials.oxford.ac.uk} Sent: Sunday, October 15, 2000 8:03 PM
Craig, David, Earl, Steve,
Increasing the mass under the microscope works. An isolated room-size slab of 1m thickness can reduce vertical vibration by a factor of four and horizontal vibration by 10 to 12 times [1]. This foundation was a significant factor in allowing us the obtain a TEM resolution better than one Angstrom [2].
-Mike O'Keefe
1. "Design and implementation of a site for a one-Ångstrom TEM", John H. Turner, Michael A. O'Keefe and Robert Mueller, in 55th Ann. Proc. MSA, Cleveland, Ohio (1997) 1177-1178. 2. "The NCEM One-Ångstrom Microscope Project Reaches 0.89Å Resolution", Michael A. O'Keefe and Y.C. Wang in 58th Ann. Proc. MSA, Philadelphia, Pennsylvania (2000).
----- Original Message ----- } From: "Craig Franklin" {franklin-at-vibeng.com}
Colleagues...
Can anyone recommend a introductory text for this person. Remember Ask-A-Microscopist is a forum that is used by NON-MICROSCOPISTS looking for help so keep try to your technical details at the right level.
Below is the result of your feedback form. It was submitted by (sfield-at-island.net) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Sunday, October 15, 2000 at 23:18:56 ---------------------------------------------------------------------------
Email: sfield-at-island.net Name: Shane Field
School: Hard Knocks
State: BC
Zip: V0N 3E0
Question: I recently came into possession of an old Bushnell microscope (the kind that is kept in some high school science labs for 2 or 3 generations). On the top of the scope it says--CENCO 506-L Bushnell Triple Tested 667157. (This may have been the 57 Chev 283 of scopes found in biology classrooms all over North America) Anyway, I took Physics and Chemistry in high school and never did learn how to operate a simple microscope. Imagine my chagrin at not being able to figure out how it works --I am able to view objects at low power but not high power. Would you be able to reccomend a good (but simple) Coles Notes kind of booklet that will give me the basics of lighting,focussing/eye piece selection etc? etc.? Thank you-TY TY TY!
At 4:03 PM -0500 10/13/00, Theresa Fassel wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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
Greetings, Years ago, I made 16 mm movies from a microscope as follows. Hooked up an ordinary video camera to the scope and displayed the image on a high quality monitor. In front of the monitor was a wooden square box that telescoped and was black on the inside. At the end of that was a 16 mm movie camera, focussed on the monitor. One frame of the move was exposed in a 1/4 sec exposure so we were running much slower than the tv raster rate. We were actually doing timelapse studies like this and got very nice images for playback and analysis. This was done in the lab of Ray Keller at UC Berkeley who would have much more of the details to hand if you want to follow this route up.
Could this help?? Tobias Baskin
} The following is a request from a producer of the PBS TV show NOVA, which } came in to the MSA Business office. } } Can anyone help? Please correspond directly to the requester. } } } } } } } } } } Hi Folks -- } } I'm a producer with the science TV series NOVA. I'm working on a show } about evolution and I'm desperately looking for a way to adapt a zeiss } axiophot microscope to an Aaton 16mm film camera. So far I've struck } out on finding anyone who has gear to accomplish this feat. } } Any suggestiongs? } } -- } Chris Schmidt } Powderhouse Productions } 259 Elm Street } Somerville MA 02144 } 617-629-2200 work } 617-776-7905 fax } 781-820-0727 mobile } 617-403-8699 pager } 781-646-0928 home } Chris-at-powderhouse.net
Another poster related his success at using an intermediate monitor for getting the image onto 16 mm film. I might take that a step further.
I don't know film production techniques, but was of the impression that much of the editing has gone digital. If that be the case, are there not utilities to record high resolution digital movies from microscope cameras. I thought Sony had a rather decent digital video camera that John McKenzie was touting a year or two ago. Even better cameras should be available now.
Please remember the final use and display of the sequence. I know that film can provide greater detail than digital for still photographs, but I doubt that this sequence would be enlarged to the degree that resolution would be an issue. Then there is the question of final display. If I viewed the final result on my plain, old TV, most any camera system would provide more resolution than I could appreciate. If it was viewed on an HDTV, more detail would be visible, but still not as much as a good video camera should be able to provide. Maybe an IMAX presentation would require all the resolution a camera could provide and then some. But by then, I would guess the resolution of the microscope would be more of a limitation than the resolution of the camera.
In short, I would suggest collecting and processing the sequence in digital format until the final stage and then transferring it to 16-mm if need be.
Warren Straszheim
At 01:04 PM 10/15/2000 -0400, you wrote:
} The following is a request from a producer of the PBS TV show NOVA, which } came in to the MSA Business office. } } Can anyone help? Please correspond directly to the requester. } } } } } } } } Hi Folks -- } } I'm a producer with the science TV series NOVA. I'm working on a show } about evolution and I'm desperately looking for a way to adapt a zeiss } axiophot microscope to an Aaton 16mm film camera. So far I've struck } out on finding anyone who has gear to accomplish this feat. } } Any suggestiongs? } } -- } Chris Schmidt } Powderhouse Productions } 259 Elm Street } Somerville MA 02144 } 617-629-2200 work } 617-776-7905 fax } 781-820-0727 mobile } 617-403-8699 pager } 781-646-0928 home
---------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
After attempting a test roll of Polapan CT 35mm instant film, I discovered what "outdated" means in the Polaroid context - the negative failed to strip off the celluloid and the strip looked unusable.
Later, after finding that there are no known dealers of this film in my area (SE PA) I retrieved the film from the waste basket & washed it in warn water, whereupon, to my utter amazement, usable positive images appeared out of the murk.
I am now waiting for the film strip to dry, after dipping it in PhotoFlo as I am used to doing with Type 55 P/N negatives.
Is there a more conservative way to strip the negative off the film ? My method seems rather crude ...
Best regards, George Langford amenex-at-amenex.com
} Craig, David, Earl, Steve, } } Increasing the mass under the microscope works. An isolated room-size } slab of 1m thickness can reduce vertical vibration by a factor of four } and horizontal vibration by 10 to 12 times [1]. This foundation was a } significant factor in allowing us the obtain a TEM resolution better } than one Angstrom [2]. } } -Mike O'Keefe } } 1. "Design and implementation of a site for a one-Ångstrom TEM", John } H. Turner, Michael A. O'Keefe and Robert Mueller, in 55th Ann. Proc. } MSA, Cleveland, Ohio (1997) 1177-1178. 2. "The NCEM One-Ångstrom } Microscope Project Reaches 0.89Å Resolution", Michael A. O'Keefe and } Y.C. Wang in 58th Ann. Proc. MSA, Philadelphia, Pennsylvania (2000). } The concrete slab need to sit on dry sand to minimize coupling to the soil/rock below it. A layer of gravel with drain tile laid in it covered by a 2 or 3 foot layer of sand would be a starting point. If the drain tile won't drain by gravity a sump will be needed to pump out any water. A water barrier need to be placed below the gravel. The truly paranoid would put down 6 inches of tamped sand, 4 inches of benotnite clay, a layer of heavy plastic enough sand to keep the installation of gravel from puncturing the plastic. The benonite serves as a self sealing moisture barrier. It is a clay that swells when it gets wet. It is not particularly expensive if you buy it from a oil field mud company. They buy it in train car loads and sell it in truck loads or sacks.
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
} Question: I recently came into possession of an old Bushnell microscope } (the kind that is kept in some high school science labs for 2 or 3 } generations). On the top of the scope it says--CENCO 506-L Bushnell Triple } Tested 667157. (This may have been the 57 Chev 283 of scopes found in } biology classrooms all over North America) Anyway, I took Physics and } Chemistry in high school and never did learn how to operate a simple } microscope. Imagine my chagrin at not being able to figure out how it works } --I am able to view objects at low power but not high power. Would you be } able to reccomend a good (but simple) Coles Notes kind of booklet that will } give me the basics of lighting,focussing/eye piece selection etc? etc.? } Thank you-TY TY TY! } Shane -
I hope that you'll be willing to go a step beyond "booklet". There's a very good basic book; here's the review from the Project MICRO bibliography (URL below):
Nachtigall, W. 1995 Exploring With the Microscope 160 pp. hardbound. 6.5x9.5". $19.95. ISBN 0-8069-0866-1 Sterling Publishing Co., NY. Although this book is intended for adult amateur microscopists, it is well written and will provide teachers and classroom volunteers with much useful information on "serious" light microscopy; it's unequalled as a basic reference for beginners. Almost half of the book is devoted to simple preparation methods for biological specimens and descriptions (with good illustrations) of commonly encountered organisms. Adult. RECOMMENDED
If that's beyond your butget, shop around the MICRO bibliography. You may find enough basic info in the website hotlink list.
Caroline
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
Hi George, Well, George if you have talked to Rick and have been to a workshop I don't know if I have any solutions for you, however, you said you were mostly processing yeast and Drosphila Fly eyes. In my earlier day in EM I work for a very short time on plant tissue and I found that I had to use a vacuum device of some kind to get the resin to penetrate the tissue. Pella has a Vacuum device you can use in the model of oven you have (which by the way is the same as mine). I found that nerve tissue take a little bit more fixation (3% Glut. in 0.1M Cacodylate buffer pH 7.3) in the oven then other tissue does. The nerve I mostly work with is sural nerve and I find that a temp setting of 37C with an irritation time of 40sec x3. Cooling the tissue down, in wet ice, to about 7-8C before and between each irritation period works for me. What sort of fixative do you use with these tissues?
I have a vacuum system for that microwave here at Shreveport which is only a 3 or 4 hour trip from Dallas. Maybe you could work sometime in to come over, bring your specimens with you.
Ron Austin (Research Associate) LSU Medical Ct. Dept. of Pathology 318-675-4557 Shreveport, LA e-mail: rla-at-mindspring.com
Could you please help us - please send replies to my email address:
c.hayward-at-kingston.ac.uk
Hi, We have a 30 year old Philips T301 transmission electron microscope, and the rubber roll membrane (original part number 5322 360 40071) in the pneumatic film lifting mechanism of the camera has split. In view of the age of the instrument it looks as if we would only be able to replace it if some nice person out there has a spare one that they might be prepared to part company with. Is there anybody out there who has a spare camera lift roll membrane please?
Thanks,
Bill Edwards.
Dr. C. Hayward-Costa School of Life Sciences Kingston University Penrhyn Road, Kingston upon Thames Surrey KT1 2EE, UK 44(0)208 547 2000 x 2240 Email: c.hayward-at-kingston.ac.uk
The principle of mounting the microscope on a massive slab is good, but I suspect that to isolate your microscope from the very low frequency, long-wavelength vibrations caused by rail traffic you will need a much softer and compliant isolation than can be provided by any depth of sand or compacted aggregate. I have recently built a garage / workshop within 50 metres of the London - Edinburgh track. The building is on a heavy reinforced concrete raft foundation. The slab is on 12" of sand over 3 feet of 2 to 3" aggregate and the substrate is several metres of heavy gravel material described by the geologists as raised beach. Nevertheless you can feel the vibration caused by passing trains through your feet.
My guess is that you need a fully floating slab, either mounted on very compressible pads of rubber or similar elastomer, or raised on an air cushion.
Vibration is a problem which many people worry about as demands for high resolution and traffic increase, and solutions to it are expensive. It strikes me that it would be a good move to do some modelling of the physics of the situation before committing money to it. Does anyone know how to go about this? Does software exist?
Chris
} From: "Gordon Couger" {gcouger-at-couger.com} To: {R.Cross-at-ru.ac.za} , {"MAOKeefe-at-lbl.gov"-at-sparc5.microscopy.com} Copies to: {Microscopy-at-sparc5.microscopy.com}
Food Structure & Functionality Symposium 2001 - An international symposium leading Food Structure & Functionality studies through the 21st century
Being held in conjunction with the 92nd AOCS Annual Meeting and Expo , May 13-16, 2001, Minneapolis Convention Center, Minneapolis, Minnesota, USA. For information, e-mail us at meetings-at-AOCS.org
The mandate of the Food Structure & Functionality Forum : *To promote global collaboration between Food and Agriculture professionals in Structure and Functionality disciplines by facilitating and providing a forum for exchange of knowledge, expertise and research findings*. The symposium has two themes: * new and novel approaches (including microscopy, rheology and spectroscopy) to the study of structure-function relationships in foods; * food system studies covering any part of the processing chain - from the raw material to the final product, and including trouble shooting. preliminary program May 13th - Short Courses (each short course will take a full day, they will be run in parallel) 1) Food Contaminants. Contact person - Mark Auty 2) Specific Labelling in Foods. Contact person - Marcel Paques
May 15th-17th inclusive - Technical sessions (6 sessions will run over 3 days)
May 14th Morning. Session 1. Dairy applications and fat based foods (Chairs: Mark Auty and Tony McKenna)
Afternoon Session 2. Food Safety (Chairs: Judy Arnold and Maria Brandl )
Evening -Round Table Discussion (RTD) -Social program for Division members and FS&F symposium participants
May 15th Morning Session 3. New Methods and Techniques for Food Structure and Functionality Analysis Chair: Kathy Groves
Afternoon Session 4. Agricultural Applications of Microscopy and Imaging (Chairs: Delilah Wood and Paula Allan-Wojtas)
Evening FS&F Division member meeting
May 16th Morning Session 5. Ingredients and Food Processing (Chairs: Diana Kittleson and Jim Charbonneau)
Afternoon 6. Colloidal and Interfacial Sciences (Chairs: Marcel Paques and David Pechak)
Poster Session Posters will be displayed during the meeting and presented during breaks between technical sessions
Contact information for the chairs is shown below, in alphabetical order:
Paula Allan-Wojtas Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Noval Scotia Canada B4N 1J5 Tel: (902) 679-5566 FAX: (902) 679-2311 email: allanwojtasp-at-em.agr.ca
Judy Arnold USDA -ARS - RRC 950 College Station Rd. P.O. Box 5677 Athens, GA 30604-5677 USA Tel: (706) 546-3515 FAX: (706) 546-3068 email: jarnold-at-ars.usda.gov
Mark Auty Dairy Products Research Centre TEAGASC Moorepark, Femoy, Co. Cork Ireland Tel: 011-353-25-42447 FAX: 011-353-25-42340 email: mauty-at-moorepark.teagasc.ie
Maria Brandl USDA - ARS - WWRC 800 Buchanan Street Albany, CA 94710 USA Tel: (510) 559-5885 FAX: (510) 559-5948 email:mbrandl-at-pw.usda.gov
James E. Charbonneau National Food Processors Association Food Chemistry and Packaging Department 1401 New York Ave, NW Washington, D.C. 20005 USA Tel: (202) 639-5972 FAX: (202) 639-5991 email: jcharbo-at-nfpa-food.org
Kathy Groves Leatherhead Food Research Association Randalls Road, leatherhead Surrey KT22 7RY England Tel: 44 0132 822330 FAX: 44 0132 386228 email: kgroves-at-lfra.co.uk
Tony McKenna New Zealand Dairy Institute Private Bag 11 029 Palmerston North, New Zealand Tel: 011 64 6 350 4649 FAX: 011 64 6 356 1476 email: tony.mckenna-at-nzdri.org.nz
Marcel Paques Wangeningen Centre for Food Sciences/Unilever Research Vlaardingen P.O. Box 20, 6710 BA Ede The Netherlands Tel: 011 31 318 659690 FAX: 011-31-318 650400 email: paques-at-nizo.nl
David Pechak Kraft Technology Centre 801 Waukegan Road Glenview, IL 60025 USA Tel: (847) 646-4808 FAX: (847) 646-3864 email: dpechak-at-kraft.com
Delilah Wood USDA - ARS - WWRC 800 Buchanan Street Albany, CA 94710 USA Tel: (510) 559-5653 FAX: (510) 559-5777 email: wood-at-pw.usda.gov
All this talk of huge amounts of concrete and agregate and sand. . .
What about isolation platforms? While it might be slightly more expensive - at least you won't be left with a huge construction bill if your concrete slab doesn't isolate enough vibration.
TMC makes specific floor platforms. We were looking at their tables when ordering a Confocal Mircosope. And from the catalog and price list you can get a platform that will easily isolate the column from nearly all vibration for around $5000 to $6000 dollars. It looks like they also make active vibration cancelling systems too.
Simple things like elastomer and rubber bases for concrete slabs have a fatigue life, which could be shorter than the remain life of the microscope.
I have no finacial interest in TMC: http://www.techmfg.com
I cannot see how you will isolate train vibrations without a system a little more complex than a concrete slab.
Geoff Williams
Electron Microscope Facility Supervisor Biology Department Brooks Hall Central Michigan University Mt. Pleasant, MI 48859
} } } "Buckingham, Steve" {sbuckingham-at-excellatron.com} 10/12/00 18:55 PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi All, I have a JSM 840 that I need to relocate soon. The new location is very near a railroad and interstate! Does anyone have information on the optimum thickness of a concrete slab to prevent vibrations? Thanks,
Steve Buckingham Excellatron Solid State LLC 1460 Roswell Street, Suite J Smyrna, GA 30080 phone 770 438 2201 fax 770 438 2118
Steve,
Have you tried something cheaper? When I was with The Johns Hopkins University School of Medicine we were remodeling the lab. A new darkroom sink for developing negatives was being put in right beside the TEM. The plumbers had to drill for water lines and a sink drain within 3-5 feet of the microscope. We lifted up each corner of the microscope and installed 60 or 80 durometer(I think it was 80), 4"x4"x1" rubber pads and could use the microscope without vibration. The lab was located just below street level. This might be a way to go before trying concrete slabs. The only way I've heard of using concrete slabs was to isolate the slab from the rest of the floor. A concrete slab could be very expensive. Most rubber supply houses can supply the pads. Hope you can work it out.
Phil Rutledge USDA/ARS 151 Dixon Drive Chestertown, MD 21620 voice: 410.778.4136 or 2120 fax: 410.778.4399 e-mail: rutledge-at-ars.usda.gov
Videotapes from M&M 2000 are now available for purchase. A new list of all tapes in the collection can be found at the MSA web site or directly at : http://www.biotech.ufl.edu/sems/newtape00.htm
Many of the older tapes are available for immediate shipment. Others, including the 2000 tapes, must be ordered from the video duplication service. This usually requires about two weeks. Please remember to order the tapes by number and title. Make checks payable to "MSA" in US funds only. Purchase orders can be accepted. Orders may be placed by phone, mail, fax or e-mail to the contact at the bottom of this message.
If you are a presenter in any of the tapes and feel that your tape should not be offered for sale in the future for whatever reason (obsolete information, bad hair day, etc.) please let me know.
Greg Erdos Videotape Wrangler Gregory W. Erdos, Ph.D. Assistant Director, Biotechnology Program PO Box 110580 University of Florida Gainesville, FL 32611
Can anyone answer this one?. Remember Ask-A-Microscopist is a forum that is used by NON-MICROSCOPISTS and K-12 Students looking for help so keep try to your technical details at the right level.
Nestor --------------------------------------------------- Below is the result of your feedback form. It was submitted by (crowj-at-mail.leon.k12.fl.us) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, October 17, 2000 at 11:33:21 ---------------------------------------------------------------------------
Email: crowj-at-mail.leon.k12.fl.us Name: students
School: The Academic Resource Center
State: Florida
Zip: 32304
Question: Students are working with bacteria and fungi and yeast. They want to know why yeast is killed at temperatures lower than body temperatures, yet bacteria can grow on human bodies with a much higher temperature (98.2 according to most sources is the average body temp).
Hello List - Have I waited long enough to receive my proceedings volumes from the Philadelphia M&M meeting? If so, whom should I contact regarding "re-shipment"? Robert J. Palmer Jr., Ph.D. Natl Inst Dental Craniofacial Res - Natl Insts Health Oral Infection and Immunity Branch Bldg 30, Room 308 30 Convent Drive Bethesda MD 20892 ph 301-594-0025 fax 301-402-0396
We have both technical and postdoctoral openings for transmission electron microscopists interested in studying biological macromolecules and cellular assemblies to high resolution using electron cryomicroscopy. The recent research emphasis of our laboratory has been on the molecular mechanisms underlying the assembly, organization, and dynamics of the cytoskeleton. Stable funding is available for these positions for 5 years. Salaries are negotiable.
The Department of Biological Sciences provides outstanding modern TEM facilities including Philips CM300-FEG, CM200-FEG, and EM420 electron cryo-microscopes dedicated to structural studies. Both the 300kV and 200kV field emission gun microscopes are outfitted with CCD cameras, 2k x 2k and 1k x 1k respectively, in addition to film cameras.
Purdue is home to over 50,000 students, faculty, and staff, and is located in West Lafayette, Indiana - approximately 1 hour northwest of Indianapolis and 2.25 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.
The ideal candidates will be enthusiastic, self-motivated individuals with strong communication skills who are committed to applying an integrated structure-function approach to the study of biological macromolecules. Interested individuals should contact Dr. McGough by e-mail or phone. Formal applications for the technical positions should be made via the Personnel Services department http://www.adpc.purdue.edu/Personnel/job-home.htm.
Purdue is an equal access/equal opportunity university.
Amy M. McGough, Ph.D. Assistant Professor, Department of Biological Sciences and Editorial Board Member, FEBS Letters Purdue University West Lafayette, IN 47907-1392 E-mail: amcgough-at-bilbo.bio.purdue.edu Office: 765-496-7501; Lab: 765-496-7716 Secretary: 765-494-4954; Fax: 765-496-1189 CM200F: 765-496-2746; CM300F: 765-496-7508 http://bilbo.bio.purdue.edu/~amcgough/ Biophysics & Cell Biology Symposium: http://bilbo.bio.purdue.edu/~rbernal/
Aside from all the concrete, sand and aggregate issues, the column on our Cameca microprobe rests on inflatable "feet" that helps reduce vibrations as well. This of course is in addition to an isolation slab that it sits on that was poured first.
-----Original Message----- } From: Buckingham, Steve [mailto:sbuckingham-at-excellatron.com] Sent: Thursday, October 12, 2000 10:59 AM To: 'Microscopy-at-MSA.Microscopy.Com'
Hi All, I have a JSM 840 that I need to relocate soon. The new location is very near a railroad and interstate! Does anyone have information on the optimum thickness of a concrete slab to prevent vibrations? Thanks,
Steve Buckingham Excellatron Solid State LLC 1460 Roswell Street, Suite J Smyrna, GA 30080 phone 770 438 2201 fax 770 438 2118
Maybe the rubber pads might help a bit, but is there actually going to be a problem? The 840 has really good spring suspension anyway, I can't see that rubber pads would add significant isolation to what's there already.
Might be instructive to ask if anyone's actually experienced vibration sensitivity with an 840.
rtch
} } } Hi All, } I have a JSM 840 that I need to relocate soon. The new location is } very near a railroad and interstate! Does anyone have information on } the optimum thickness of a concrete slab to prevent vibrations? } Thanks, } } Steve Buckingham } Excellatron Solid State LLC } 1460 Roswell Street, Suite J } Smyrna, GA 30080 } phone 770 438 2201 } fax 770 438 2118 } } Steve, } } Have you tried something cheaper? When I was with The Johns Hopkins } University School of Medicine we were remodeling the lab. A new } darkroom sink for developing negatives was being put in right beside } the TEM. The plumbers had to drill for water lines and a sink drain } within 3-5 feet of the microscope. We lifted up each corner of the } microscope and installed 60 or 80 durometer(I think it was 80), } 4"x4"x1" rubber pads and could use the microscope without vibration. } The lab was located just below street
Phil Rutledge USDA/ARS 151 } Dixon Drive Chestertown, MD 21620 voice: 410.778.4136 or 2120 fax: } 410.778.4399 e-mail: rutledge-at-ars.usda.gov } } } } 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 List Members, } } We are trying to section by microtome samples of solution-cast PEG. PEG } is dissolved in water and THF and allowed to rest until it becomes a } viscous solution by the evaporation of the solvent. It is then put into } molds and under vacuum to further rid the material of solvent. Finally, } the PEG becomes solid. } } THE PROBLEM: The PEG moldings can not be sectioned because they are too } brittle and crumble in the chuck. } } Does anyone have a protocol for preparing PEG samples that does not lead } to this end? } } Thank you very much, } } Jennifer Taylor
Thanks for your interest. There are around 500-600 different species of yeast, many of them unrelated genetically. Some of these, such as some species of Sporobolomyces have temperature requirements that are below normal body heat (37C). Others, such as Candida albicans, are pathogenic on humans, with their optimum temperature at normal body heat. The bakers' yeast, Saccharomyces cerevisiae, can survive 40C for short periods, but has its growth optimum at around 27C. There are specially selected strains (especially used in brewing) which have other growth optima.
Whilst I do not know the details, I believe that bacteria have similarly varied optimal growth temperatures (e.g. the iron bacterium Gallionella at 20C, the yoghurt bacterium Streptococcus thermophilus at over 45C). I hope this helps.
Cheers
Stephan Helfer PhD Mycologist, Royal Botanic Garden Edinburgh
} Email: crowj-at-mail.leon.k12.fl.us } Name: students } } School: The Academic Resource Center } } State: Florida } } Zip: 32304 } } Question: Students are working with bacteria and fungi and yeast. They } want to know why yeast is killed at temperatures lower than body } temperatures, yet bacteria can grow on human bodies with a much higher } temperature (98.2 according to most sources is the average body temp). } } --------------------------------------------------------------------------- } } } }
Cheers
Stephan 0131 248 2865 FAX 248 2901 http://www.rbge.org.uk
I called on Monday regarding the missing Proceedings, and found that there were 3 mailing lists, and they are now working on the third list, so it could be an additional 2 weeks before the final volumes are received. Hang in there...
Larry
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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
PEG comes in different molecular weights. You might try a lower molecular weight PEG whcih would be less crumbly.
Also, John Wolosewick developed a technique to section biological tissue embedded in PEG. When I worked in his lab, we used to attach the PEG embedded tissue to aluminum stubs with dental wax and put the stub in the chuck of the microtome. That would prevent the chuck of the microtome from crushing your sample.
Joe Neilly Abbott Laboratories D-45M, AP31 200 Abbott Park Rd. Abbott Park, IL 60064-6202 voice: (847)-938-5024 fax: (847)-938-5027 e-mail: joe.neilly-at-abbott.com
jtaylor-at-stevens-tech.edu on 10/17/2000 09:21:39 PM Please respond to jtaylor-at-stevens-tech.edu To: Microscopy-at-sparc5.Microscopy.Com cc:
Dear List Members, } } We are trying to section by microtome samples of solution-cast PEG. PEG } is dissolved in water and THF and allowed to rest until it becomes a } viscous solution by the evaporation of the solvent. It is then put into } molds and under vacuum to further rid the material of solvent. Finally, } the PEG becomes solid. } } THE PROBLEM: The PEG moldings can not be sectioned because they are too } brittle and crumble in the chuck. } } Does anyone have a protocol for preparing PEG samples that does not lead } to this end? } } Thank you very much, } } Jennifer Taylor
At 8:23 PM -0400 10/17/00, Jennifer E. Taylor wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
The problem may be the molcular weight of PEG that you are using. Years ago, I used PEG to embed heart muscle for TEM, and if memory serves, we used a mixture of MW 10,000 and 6,000 to obtain a "sectionable" consistancy. Our source of information at that time was work by John Wolosowick (this was in the early-to-mid 1980's). A similar mixture may work for you. I'lm sorry I can't be more speciic, but that was work done at another institution, for a PI who has since passed away, so I don't have immediate access to the records. Try looking up John's work (It was Wolosowick & Keith Porter, I believe).
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'm planning to BSA-gold uptake into human cell-culture for TEM. Cell lines are grown in DMEM-F12. With the aim to study the ultrastructure of the endocytic system and determine the pH of the various organelles. Many methods state that they grew the cells overnight in serum-free media before starting the uptake. Does anybody know if this alters the morphology and/or pH of the cell?
Jennifer Jackson University of Natal, Pietermaritzburg South Africa email: jacksonj-at-nu.ac.za.
When I did some PEG work, a piece of the PEG embedded sample was glued to a blank epoxy block and then sectioned. There are several papers out there on the technique.
Polyethylene Glycol (PEG) Embedding and Subsequent De-embedding as a Method for the Structural and Immunocytochemical Examination of Biological Specimens by Electron Microscopy. Histake Kondo. 1984. J. Electr. Micros. Tech., 1:227-241.
The application of polyethylene glycol (PEG) to electron microscopy. J.J. Wolosewick. 1980. J. Cell Biol., 86:675-681.
Greg
"Jennifer E. Taylor" 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, } } } } We are trying to section by microtome samples of solution-cast PEG. } PEG } } is dissolved in water and THF and allowed to rest until it becomes a } } viscous solution by the evaporation of the solvent. It is then put } into } } molds and under vacuum to further rid the material of solvent. } Finally, } } the PEG becomes solid. } } } } THE PROBLEM: The PEG moldings can not be sectioned because they are } too } } brittle and crumble in the chuck. } } } } Does anyone have a protocol for preparing PEG samples that does not } lead } } to this end? } } } } Thank you very much, } } } } Jennifer Taylor
-- ================================================================== 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 ==================================================================
On Oct 10 I posted a request for digital imaging info on behalf of graphics prof Jonathan Lipkin (jlipkin-at-ramapo.edu). I've seen a reply from Warren Straszheim (thanks, Warren!) but I've discovered that in that time period I didn't receive an unknown quantity of incoming email (including my own posting) and have no idea if anyone else responded also. So if any of you posted a reply to Jonathan and assumed I'd been able to forward it to him, I'd appreciate it if you'd send it to me again privately.
Many thanks, Dee
*************************************************************** Dee Breger Mgr. SEM/EDX Facility Lamont-Doherty Earth Observatory 61 Route 9W Palisades, NY 10964 USA T: 914/365-8640 F: 914/365-8155
http://www.ldeo.columbia.edu/micro http://www.discovery.com/area/science/micro/micro1.html http://www.lsc.org/antarctica/front.html Journeys in Microspace (Columbia University Press, 1995)
I have 3 print processors looking for homes - 2 free, one (new) for some fee.
Check out details at MSA's surplus equipment site: http://www.msa.microscopy.com/SurplusEquipment/SurplusListings.html
Ann Hein Lehman EM Facility Manager Trinity College Hartford, CT 06106 v. 860-297-4289 f. 860-297-2538 e. ann.lehman-at-trincoll.edu w. http://www.trincoll.edu/~alehman
Some time back Gatan made a ±60+ single hole, non-rotary specimen holder for the JEOL 200-CX. They are no longer in stock and I was hoping to find one, or the equivalent, gathering dust somewhere that I could buy and put to good use. Reply privately to {treese-at-marinebio.mbl.edu} . I would also be interested in any surplus single tilt holders for the JEOL that I could try to modify for ±60 tilt. Thanks!....Tom Reese "
Steve, The chief advantage to using water immersion lenses has to do with, as you surmised, the refractive indices in the optical system. Live cells are most happy in aqueous media, so therefore it is best to look at them in such an environment. Coverslip corrected water lenses have the advantage of eliminating oil from the system, and no-coverslip "dipping" lenses eliminate the gross errors that can occur when looking through the unpredictable surface of water (try to take a picture of a fish from a dock, then jump in the water and try an underwater photo).
An air-water interface produces problems similar to an air-glass interface. In the same way that an oil lens will optimize the image of a fixed prep, a water immersion lens will optimize observation of an aqueous prep. The resolution of your optical system is determined generally by the lowest refractive index. Therefore it is beneficial to eliminate air from the equation, but there is no advantage to using an oil lens with a water prep. An oil lens may have higher potential NA, say 1.4 at 60X rather than 1.2 for the water 60X, but the system NA is limited by the 1.33 refractive index of water, not the 1.515 refractive index of the oil. The highest potential NA of the water prep system is 1.33, just as an oil system is potentially 1.5, yet practical design concerns result in about a 1.2 limit for the water prep, 1.4 for the oil.
An advantage to using a coverslip corrected water lens, your case #1, is that the lens designers need only account for two refractive indicies when dealing with spherical aberration, and these two will always be in similar proportion. As these are live cells and somewhat unpredictable, we cannot always know how close they will be to the coverglass. Between the cell and the coverglass is water, and between the coverglass and lens is again water. It does not matter for the system whether the cell is very close to the coverslip or a little further away - there will just be a little more or less water on the objective side. Thus the optical path through water and glass is always the same. With oil immersion, the oil and water in the path will change depending on whether the cell is relatively near or far from the coverslip, as the objective moves closer or further from the coverglass. This can result in spherical aberration. A sensitive instrument like a confocal microscope is sensitive to these errors.
As for your case #2, non-coverslip corrected water lenses, or "dipping lenses", are crucial for live cell experiments where a number of devices may be manipulating or even connected to the cells. For one thing we have cells in a dish with a number of devices penetrating the water from above, giving an irregular surface. Additionally we have a sometimes significant amount of buffer above the cells, giving rise to working distance issues. A dipping lens typically has a narrow tapered end, teflon coated, and can be brought nicely into the buffer. The NA is typically a little lower than a coverslip objective but the needs for working distance (~2-3mm) and narrow profile outweigh the need for very high NA (0.9 vs. 1.2 for a 60X). A dry or oil lens simply will not perform in such conditions.
I hope this gives you a bit more background.
Regards, Bill Fester Olympus America Microscope Division - NY
} } Netters } } In upgrading our light microscopes, we are buying some new lenses for our } confocal scope and a brightfield scope. The salesforce is regaling us with } tales of improved resolution, clarity, brightness etc that comes with } spending double or triple the price for a standard oil immersion or dry } lens. Could someone please clarify, either on-line or off, what the } advantages and disadvantages of each of the following ( my texts on light } microscopy don't deal with them). Are they worth the extra dollars? } } 1) a water immersion lens for looking at cells suspended in buffer and } covered with a coverslip (water is used rather than oil between coverslip } and lens) These lenses typically have a lower numerical aperture than an } oil immersion lens. } } 2) a water dipping lens, used to examine cells in culture directly, without } a coverslip. I would expect the absence of refractive indice changes } (media/coverslip/air or oil) would explain their improved brightness. } } Thanks in advance for your input } } Steve Barlow } SDSU EM Facility } }
I am posting this request to help Paul, a graduate student here. If anyone can help him out, please respond to me.
Thanks, Jo Ann Moore Senior Biological Scientist Anatomy Department, College of Medicine University of South Florida Tampa FL
Hello folks -
I am in the process of refurbishing an old Aus-Jena microscope in our laboratory, and would like to be able to connect a video camera to the camera mount, but unfortunately it seems nearly impossible to find an appropriate J-mount adapter anywhere costing less than my left leg and right hand. I am hoping that one of the photomicroscopists around might have a spare one of these squirreled away in a drawer or cabinet somewhere that they wouldn't mind parting with. And if there's a disabled Aus-Jena scope around that could be available for parts, that'd be great as well.
Many thanks, Paul Jantzen Alzheimer's Research Laboratory
I have an Oxford/Link exL WDS/EDX (detector not included) system that I will gladly send to anyone for cost of shipping. The monitor and printer are still operational (or was the last time I could get the system to boot up), the rest does not work at all. I also have a monitor to a Tracor 2000 system is anyone needs it.
Phoebe Doss Manager Electron Microscope Lab Oklahoma State University
Good Morning, We moved the Instrument( JEOL 100CX II ) down the hall. Now the Diff. Pump Light comes on for the INITIAL WARM-UP, But after ~ 20min, the Diffusion Pump light goes OFF, and after another 10 min. the Scope shuts down. We checked & rechecked all the cables connections, didn't help. ANY IDEAS!!!!! Thanks Chuck
} We moved the Instrument( JEOL 100CX II ) down the hall. Now the Diff. Pump } Light comes on for the INITIAL WARM-UP, But after ~ 20min, the Diffusion } Pump light goes OFF, and after another 10 min. the Scope shuts down. } We checked & rechecked all the cables connections, didn't help. } ANY IDEAS!!!!!
Here's one - clean the penning gauge and see if it behaves.
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* ****************************************************************************
{ { Subj: HELP WITH JEOL 100CX II Date: 10/19/00 6:26:30 PM US Mountain Standard Time From: buiocchi-at-astro.ocis.temple.edu (Chuck Buiocchi) To: Microscopy-at-sparc5.microscopy.com
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Good Morning, We moved the Instrument( JEOL 100CX II ) down the hall. Now the Diff. Pump Light comes on for the INITIAL WARM-UP, But after ~ 20min, the Diffusion Pump light goes OFF, and after another 10 min. the Scope shuts down. We checked & rechecked all the cables connections, didn't help. ANY IDEAS!!!!! Thanks Chuck
I haven't worked with the 100 CX but I am familiar with the JEOL way of doing things.
The diffusion pump LED is a warning lamp (assuming the usual JEOL way) and probably indicates that the cooling water or diff pump power is off. Check to see if the diffusion pump is heating and that you have water flow. I suspect bad water flow is keeping the pump heater from being turned on. The other 10 minutes to shut down is just another JEOL safety circuit. If the vacuum system is stays in rough pumping mode, the scope turns off after 30 minutes.
Ron Vane XEI Scientific
-----Original Message----- } From: Chuck Buiocchi {buiocchi-at-astro.ocis.temple.edu} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
} We moved the Instrument( JEOL 100CX II ) down the hall. Now the Diff. } Pump Light comes on for the INITIAL WARM-UP, But after ~ 20min, the } Diffusion Pump light goes OFF, and after another 10 min. the Scope } shuts down. We checked & rechecked all the cables connections, didn't } help. ANY IDEAS!!!!!
Sounds to me as if a connection to one of the diff pump heaters has broken. The wires become brittle from the heat and a bump while you were moving it make have broken one of them. A more costly possibility is that one of the heaters may have expired.
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 Chuck, For the luck of details in your description- is the cooling water runs properly through all circuits? Both diff. pumps, electronics, lenses? Perhaps some line(s) (partially) clogged? Check 2 solenoid water valves on the water hoses on the floor behind the TEM. Are the wires broken? Those are easy to damage during the move. Feel sides of the diff. pumps and electronics return water line 15 minutes after start up. Are they getting hot? Flowmeter in the water line never hurts...
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax
This message is made of 100% recycled electrons. -----Original Message----- } From: Chuck Buiocchi {buiocchi-at-astro.ocis.temple.edu} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
Dear Chuck, This could be a cooling problem; first DP. overheats and trips off, then a part of electronics - lens coils or power supply (whatever is cooled in your scope) -this would shut down the scope completely.
Good luck! Dan
Dan Kaszubski Micro-Time Co. Olathe,KS
Good Morning, We moved the Instrument( JEOL 100CX II ) down the hall. Now the Diff. Pump Light comes on for the INITIAL WARM-UP, But after ~ 20min, the Diffusion Pump light goes OFF, and after another 10 min. the Scope shuts down. We checked & rechecked all the cables connections, didn't help. ANY IDEAS!!!!! Thanks Chuck
I had a similar problem recently (SEM) with a JSM 35C, it pumped for 20-30 minutes and then cut out. Hints: 1. make sure the water flow is ok 2. make sure the diff pumps are getting hot 3. make sure the RP belts are tight. My JSM problem was a loose RP belt on the pump which 'roughs' the column - the pre-vac. in the column wasn't good enough. A new belt cured it.
But I still have the same problem on a 200CX TEM and it's none of the above - awaiting help from the answers to your original question! Let me know of any good ones you might get off-line, please!
Keith Ryan Marine Biological Association Plymouth UK
{ { { Chuck Buiocchi {buiocchi-at-astro.ocis.temple.edu} 10/20 4:23a } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Good Morning, We moved the Instrument( JEOL 100CX II ) down the hall. Now the Diff. Pump Light comes on for the INITIAL WARM-UP, But after ~ 20min, the Diffusion Pump light goes OFF, and after another 10 min. the Scope shuts down. We checked & rechecked all the cables connections, didn't help. ANY IDEAS!!!!! Thanks Chuck
} } } Chuck Buiocchi {buiocchi-at-astro.ocis.temple.edu} 10/20/00 00:19 AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Good Morning, We moved the Instrument( JEOL 100CX II ) down the hall. Now the Diff. Pump Light comes on for the INITIAL WARM-UP, But after ~ 20min, the Diffusion Pump light goes OFF, and after another 10 min. the Scope shuts down. We checked & rechecked all the cables connections, didn't help. ANY IDEAS!!!!! Thanks Chuck
Chuck,
I used a 100 CX and when thsat happened to me, it was usually the top diff pump heater. They go bad after a while. In 13 years I've only replaced the bottom diff pump heater 1 time. I've never used a CXII so I don't know if it has 2 diff pumps. Hope this helps.
We had a similar problem recently, but it turned out to be my fault, rather than the scope's. When you said you had just moved the scope, it rang a bell...
I didn't move the scope, but I rearranged the position of its water chiller. A couple days later, the scope was shutting down and the chiller was laboring mightily with a water temp of over 100 F. I checked the water level and refrigerant level and internal water flow in the chiller tank and all seemed well. So I called the refrigeration guy.
After looking around a bit, the first thing he says is, "Did you move this recently?" I said yes, and he felt behind the chiller and found two hoses that had crimped when I moved it against the wall. Poor thing was hardly getting any outside cooling water flow.
The refrigeration guy was kind. He didn't make fun of me and he didn't charge me for the visit.
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: Chuck Buiocchi [mailto:buiocchi-at-astro.ocis.temple.edu] Sent: Thursday, October 19, 2000 5:46 PM To: Microscopy-at-sparc5.microscopy.com
Good Morning, We moved the Instrument( JEOL 100CX II ) down the hall. Now the Diff. Pump Light comes on for the INITIAL WARM-UP, But after ~ 20min, the Diffusion Pump light goes OFF, and after another 10 min. the Scope shuts down. We checked & rechecked all the cables connections, didn't help. ANY IDEAS!!!!! Thanks Chuck
Have you checked the compressed air pressure or the pressure switch. I don't have drawings but I assume that one is fitted.
Good luck, Ron
} } Chuck } } I had a similar problem recently (SEM) with a JSM 35C, it pumped for } 20-30 minutes and then cut out. } Hints: } 1. make sure the water flow is ok } 2. make sure the diff pumps are getting hot } 3. make sure the RP belts are tight. } My JSM problem was a loose RP belt on the pump which 'roughs' the } column - the pre-vac. in the column wasn't good enough. A new belt } cured it. } } But I still have the same problem on a 200CX TEM and it's none of the } above - awaiting help from the answers to your original question! Let } me know of any good ones you might get off-line, please! } } Keith Ryan } Marine Biological Association } Plymouth UK } } { { { Chuck Buiocchi {buiocchi-at-astro.ocis.temple.edu} 10/20 4:23a } } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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, } We moved the Instrument( JEOL 100CX II ) down the hall. Now the Diff. } Pump } Light comes on for the INITIAL WARM-UP, But after ~ 20min, the } Diffusion } Pump light goes OFF, and after another 10 min. the Scope shuts down. } We checked & rechecked all the cables connections, didn't help. } ANY IDEAS!!!!! } Thanks } Chuck } =========================================================================== 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. ============================================================================
please find enclosed a summary of the first circular of the 5th Multinational Congress on Electron Microscopy, that will be held in Lecce (ITALY), on September 20-25 2001.
An overseas participation is strongly desired, and we hope that many scientists from all over the world will come to Italy in September 2001.
The congress is organized by the following societies:
Austrian Society for Electron Microscopy Croatian Society for Electron Microscopy Czechoslovak Society for Electron Microscopy Hungarian Society for Microscopy Italian Society for Electron Microscopy Slovenian Society for Electron Microscopy
The purpose of the congress is to provide an interdisciplinary forum where scientists, both in the materials science and in the biomedical research can present the most recent results of their activity.
Some of the topics which will be covered are:
Architecture of the nucleus and nuclear import and export, art, biomaterials, biosensors, environmental microscopy, cell and cell components, cell death in biology and pathology, chemical analysis and mapping, microscopy in human health, microscopy of nanostructural materials, microscopy of structural and functional materials, energy-filtered electron microscopy, instrumental development, investigations of thin films for sensor technology, microbiology, microscopy of semiconducting materials, parassitology, plant cell biolgy, remote microscopy, strain analysis, structure biology, structural determination and refining, viruses, teleoperation and computer assisted operation, nanotechnology, single molecule microscopy and manipulation.
You can find further information and eventually download the brochures (soon available) and also preregister at the web site:
www.mcem5.unile.it
The Congress contacts are:
Laboratory of Comparative Anatomy and Cytology Dept. of Biology University of Lecce Via per Monteroni, 73100 Lecce, Italy Phone: +39 0832 320657 (855) Fax: +39 0832 320654
Institute for the study of new Materials for Electronics University of Lecce Via per Arnesano, 73100 Lecce, Italy Phone: +39 0832 322362 Fax: +39 0832 325299
The JEOL scopes have a heat sensor that ensures the DP is hot. They sometimes have two sensors, one at the bottom of the DP, the hottest point, and one at the top of the DP the coolest point. My guess is during your move the sensor(s) came out of the clamp that holds it against the DP. Look for a cylinder that has two wires coming out of it and find the metal band or clamp where it fits into. I hope this helps!
Bill
_______________________________
Bill Carmichael Electron Microscopy Faculty
Madison Area Technical College 3550 Anderson St. Madison, WI 53704 608-243-4309
This e-mail came in to the MSA Business Office. If you sell light microscopes you will want to respond directly to the writer. Please do not copy the listserver.
Ron
} } } } } }
To whom this may concern: I am Karim El Masry, Assistant Brand Manager on Haircare products in Procter & Gamble Egypt.
I was wondering if you could help me find video microscopes that magnify up to x200. I will be using the microscopes in one of my programs demonstrating the video microscopes capabilities.
Please advise with the company producing or supplying them. I will need at least a good 50 microscopes within the next two weeks.
Hope to hear from you very soon. Best Regards, Karim +202 394 7609
{elmasry.k-at-pg.com}
} } } } } } } }
Ron Anderson, IBM, Hopewell Jct., New York, USA. anderron-at-us.ibm.com
IBM Analytical Services; http://www.chips.ibm.com/services/asg
Dear List, Does anyone out there know where I can order parts for my LKB Ultrotome V? I need to order new chucks and can't find a dealer who deals with the LKB's. Are parts available anymore? A textbook listed Cambridge Instruments as a supplier. Anyone have a phone, e-mail or address? Thanks for your help, Jo Dee
-- Jo Dee Fish Coordinator of Electron Microscopy Cell Analysis Facility The Burnham Institute 10901 N. Torrey Pines Rd. La Jolla, CA 92037 (858)646-3100 ext. 3620
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The following is a brief note from a graduate student who is going to try in situ localization on plant tissue for light microscopy. Any help you can give her is appreciated. Send replies to me and I will forward to her. Thanks in advance for your help. Debby ------ I am planning in situ hybridization using ribo-probes labelled with DIG with maize and rice tissues (coleoptiles, young leaves, roots and root tips). I am afraid that some of the organs like the root tips are very small to be embedded in paraffin and still be able to get nice cross sections. Is there any other resin that I can try? Also the fixative that I planned to use consists of 50% methanol, 5% acetic acid, and 2% formaldehyde (40%). Do you have other suggestions for a fixative. I would appreciate any protocols which may help in this project. Claudia Elena Vergara
---- Debby Sherman Phone: 765-494-6666 Life Science Microscopy Center FAX: 765-494-5896 c/o Dept.of Botany & Plant Path. E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id SAA12359 for dist-Microscopy; Fri, 20 Oct 2000 18:47:40 -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 SAA12347 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Fri, 20 Oct 2000 18:47:09 -0500 (CDT) 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 SAA12340 for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 20 Oct 2000 18:46:58 -0500 (CDT) X-Sender: zaluzec-at-ultra5.microscopy.com Message-Id: {v03130300b61689b97429-at-[206.69.208.21]} Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii" X-Priority: 1 (Highest)
Good Morning Everyone, First let me say THANK YOU, to all who responsed to my earlier message, greatly appreciated!!! However some NEW additional info. The pump is HEATING up, but the Scope dies after it switches over to pump out the colum. Any Ideas??? Thanks again, Chuck
Listers, Anyone out there with a spare vac power supply & vac controller board for a JSM 35 SEM that they would be willing to sell? JEOL wasn't much help. Please reply off line, either by email or phone. Thanks in advance. Gary M. Easton Scanners Corporation 410-857-7633 x102
A colleague's student came and asked me how to prepare a ceramic powder for TEM observation. As I understand it, the powder is composed of alumina particles, average size 90 microns. The particles are very porous, with holes/pores ranging in size from around 1 to 30 microns. Their aim is to show the presence of internal pores. And yes they have done SEM on fractured particles, but they have been asked to do TEM as well.
Since these samples are rather different from what I generally work with, I'm hoping that someone can give us some advice on the specimen prep.
Many thanks in advance for any help you can give us.
Gill
Dr. Gillian M. Bond Department of Materials & Metallurgical Engineering New Mexico Tech Socorro, NM 87801
Leica may be able to help you Their US and UK addresses are copied below
The following web site has these and other microscopy vendor addresses, phone numbers, emails and links to their web pages
http://www.kaker.com/mvd/vendors.html Chris
Leica Instruments, Inc. 111 Deer Lake Road Deerfield, IL 60015 USA Tel: 800 248 0123, 708 405 0123 Fax: 708 405 8139 Leica UK Ltd Davy Avenue Knowlhill, Milton Keynes MK5 8LB UK Tel: +44 (0) 1908 24 62 46 Fax: +44 (0) 1908 60 99 92
Date sent: Fri, 20 Oct 2000 13:03:05 -0700 } From: Jo Dee Fish {jofish-at-burnham-inst.org} To: Microscopy-at-sparc5.microscopy.com
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Gillian M. Bond wrote: ================================================================ A colleague's student came and asked me how to prepare a ceramic powder for TEM observation. As I understand it, the powder is composed of alumina particles, average size 90 microns. The particles are very porous, with holes/pores ranging in size from around 1 to 30 microns. Their aim is to show the presence of internal pores. And yes they have done SEM on fractured particles, but they have been asked to do TEM as well.
Since these samples are rather different from what I generally work with, I'm hoping that someone can give us some advice on the specimen prep.
Many thanks in advance for any help you can give us. ================================================================== We have been preparing these types of samples in our own laboratory for a number of years for our commercial clients. We have found that such samples ,
1] Have to be vacuum embedded in order to make sure all the internal pores are infiltrated with the resin,
2] Diamond knife ultramicrotomy is always required, because glass knives just won't "cut" it, and
3] We prefer our own SPI-Pon™ 812 resin (although at least some of the other "Epon substitute" resins should work just as well), in part because you have the possibility of varying the hardness of the final resin through a considerable range, a feature sometimes important in terms of ending up with a block with the "right" properties (e.g. hardness).
4] If these 90 µm particles were formed in situ, and not as the result of grinding, then they should be sputter coated with (preferably) Pt but Au will do if Pt is not available, prior to embedding, so that once in the TEM, one would always be able to keep track of the original outside surfaces, since this is the way one can determine the presence of a skin (of different morphology).
Contrast can be a problem for resolving the pores of the smallest dimensions , so one would probably be wise to think about the use of "holey" support films. We prefer our own SPI Materials Science Diamond Knives for this type of work because these kinds of samples will immediately impart to the knife edge the kinds of fine striations one works so hard to get out in the production of a more expensive "life science" diamond knife. In other words , using a new life science knife on these kinds of samples (in our opinion) is quite wasteful economically.
Disclaimer: SPI Supplies offers the embedding resins, diamond knives and holey support films for doing this kind of work. We also provide this kind of laboratory service for those not set up or wanting to do their own sample preparation.
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 ######################## ============================================
} } } Good Morning Everyone, } First let me say THANK YOU, to all who responsed to my earlier } message, greatly appreciated!!! However some NEW additional info. } The pump is HEATING up, but the Scope dies after it switches over to } pump out the colum. Any Ideas??? Thanks again, Chuck } } }
Is that after starting to evacuate via the rotary pump, or later, when it switches over to the DP?
And does it switch off as soon as it starts the column evacuation, or after a while?
Maybe you've got such a massive air leak that the watchdog senses that there's not sufficient vacuum backing for the DP(s).
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
Hello folks, I am looking for a used microtome which will work well in sectioning resin (1-3 micron). A manual microtome would be fine, especially if is a good bargain. Thanks!
Tonino Traini CDT-MDT-DDS Electron Microscopy Center Ud'A University * Tel: (871) 3554143 * Fax: (735) 90514 * E-mail: ttrain-at-tin.it
} The pump is HEATING up, but the Scope dies after it switches } over to pump out the colum. Any Ideas???
There are two diff pumps - are they both heating?
If both are heating I think the most likely possibility, as mentioned by others here in response to your original enquiry, is overheating of the diff pumps because of a water cooling problem. On the other hand, if the instrument turns off at or very soon after it tries to switch over to high vacuum, and if the diff pumps are hot but not overheating, then it sounds as if the rough pumping by the rotary pump has been inadequate. Is there any pumping noise from the rotary pumps? It may be that one of the hoses was not properly re- connected after the move, or one of the hoses may be leaking. They perish after a while.
Good luck.
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 ** ------- End of forwarded message -------
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 **
Email: ed_bachmann-at-unc.edu Name: Ed Bachmann School: University of North Carolina at Chapel Hill
Question: I am looking for a high quality new or used stereoscopic dissecting scope for examining mosses. I would like as high a magnification as I can get consistent with very good resolution. Is there anyplace I can find unbiased reviews or comparisons of brands and models? Can you comment on the use of adaptor lenses that attach to the objective lens to boost magnification. I've been told they cause image degradation (loss of resolution?). Thank you.
We have begun using a Gatan Bio-Scan digital camera on our LEO EM910 and we like it very much. I would like to improve on this camera's capabilities by increasing its spatial resolution. One method I've heard of and that is used in some digital cameras is to acquire multiple images at sub-pixel spacing and then recombine these into one image. My microscope has the capability of shifting the image in any direction in sub-pixel units. Has anyone tried this, or does anyone know where I can get information on how this might be done?
_____________________________________________________________ C. Robert Bagnell Jr., Ph.D., Res.Assoc.Prof. Microscopy Services Laboratory Department of Pathology & Laboratory Medicine CB #7525 UNC-CH, Chapel Hill, N.C. 27599 ph 919-966-2413 fx 919-966-6718 http://www.pathology.med.unc.edu/path/microscopy/welcome.htm
I was talking to a friend of mine this morning about what software he should have to do digital imaging. Reflexively I said PhotoShop and was asked how he would use it. I told him the standard things - contrast enhancement, color matching across different media, annotation, resizing, etc. His response was that for the price was that all it did (a little naivety but a fair question) . You know, sometimes you get in a rut and it set me to wondering how other people are using PhotoShop? It's not like I'm selling the software but I'd like to be able to give a better answer before telling someone they should plunk down that much money. Aside from image analysis plug-ins (which I had forgotten about when we were talking) how else is PhotoShop being used - anything fancy to justify the cost?
We are trying to track down the location of "Cambridge Image Technology," manufacturers of cathodoluminescence stages for optical microscopes. Can anyone help? Thanks in advance,
Mati
Mati Raudsepp, Associate Professor (Hon.) Dept. of Earth & Ocean Sciences 6339 Stores Road The University of British Columbia Vancouver, BC V6T 1Z4
You should add the ability to do montaging with layers, selecting and adjusting contrast ranges, selecting edges. you also have the capability of using John Russ' Image Processing Toolkit or Provea Pro to do relatively inexpensive image 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: Bill Miller [mailto:microbill-at-mohawk.net] } Sent: Monday, October 23, 2000 5:25 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: PhotoShop usage } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: 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 } } } -------------------------------------------------------------- } ---------. } } } I was talking to a friend of mine this morning about what software he } should have to do digital imaging. Reflexively I said } PhotoShop and was } asked how he would use it. I told him the standard things - contrast } enhancement, color matching across different media, } annotation, resizing, } etc. His response was that for the price was that all it did } (a little } naivety but a fair question) . You know, sometimes you get } in a rut and it } set me to wondering how other people are using PhotoShop? } It's not like } I'm selling the software but I'd like to be able to give a } better answer } before telling someone they should plunk down that much } money. Aside from } image analysis plug-ins (which I had forgotten about when we } were talking) } how else is PhotoShop being used - anything fancy to justify the cost? } } Bill Miller } }
One of the reasons that Photoshop is in such widespread use may be Adobe's sales policy. Like Apple years ago, they sell to Academic Institutions at very greatly discounted prices, so that for us we aren't "plunking down that much money" - in fact, the cost to us of Photoshop is competitive with far less able program suites. As a result, all our graduates go off into their jobs and immediately buy Photoshop!
In reality, though, I would still consider Photoshop because it has everything I could possibly need (other, cheaper packages are almost always deficient in some way or another), because it will handle 16 bit images, because its user interface is not frilled up but is fully accessible, because of the extensive file format support, and so on, as well, of course, as the plug-ins. Although Photoshop has power that I will never ever use, I'm also surprised at how often I use some feature that I would never have considered as important, if I did a feature by feature price-performance evaluation of cheaper products.
Tony Garratt-Reed
At 10:24 AM 10/23/2000 +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: (+) 1-617-258-6478 **
This is a re-post; please excuse us if you've seen it before.
Also, please respond to Dr. Spector (e-mail given in post), not to the poster...although I will forward any messages I receive on to him. +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
Microscopy Core Technician -
Cold Spring Harbor Laboratory is seeking an experienced and responsible microscopy technician for the laboratory's core biological microscopy facility. The individual should have practical expertise in transmission electron microscopy, confocal and widefield fluorescence microscopy, digital imaging, and microinjection. The successful candidate will be involved in designing and carrying out experimental protocols for users, training individuals in the use of various microscopes, and aligning microscopes and keeping the facility operating at an efficient and high level of productivity. Interested individuals should send their resume, including a description of their expertise and the names and addresses of 3 references to: Dr. David L. Spector, Cold Spring Harbor Laboratory, One Bungtown Road, Cold Spring Harbor, New York 11724, email: spector-at-cshl.org
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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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One nice action we do with PhotoShop is an automated process that compresses a large tiff file or similar into an e-mailable jpeg. In the automated batch process, it can compress a whole folder of large files in a matter of a minute or two.
-----Original Message----- } From: Bill Miller [mailto:microbill-at-mohawk.net] Sent: Monday, October 23, 2000 2:25 AM To: Microscopy-at-sparc5.microscopy.com
I was talking to a friend of mine this morning about what software he should have to do digital imaging. Reflexively I said PhotoShop and was asked how he would use it. I told him the standard things - contrast enhancement, color matching across different media, annotation, resizing, etc. His response was that for the price was that all it did (a little naivety but a fair question) . You know, sometimes you get in a rut and it set me to wondering how other people are using PhotoShop? It's not like I'm selling the software but I'd like to be able to give a better answer before telling someone they should plunk down that much money. Aside from image analysis plug-ins (which I had forgotten about when we were talking) how else is PhotoShop being used - anything fancy to justify the cost?
} } We are trying to track down the location of "Cambridge Image } Technology," manufacturers of cathodoluminescence stages for } optical microscopes. Can anyone help? Thanks in advance, } } Mati
Don't know that one, but we've been happy with the Luminoscope we got from Premier American Technologies, PA, phone 814/867-8600.
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
Easy. Take multiple images with each having some amount of overlap. Then feed these images into Visual Stitcher Pro ($50) and voile, one single big image.
gary
At 07:12 AM 10/23/00, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi Listers: Since all of us are heavily involved with the use of the internet, I thought this message I just received from Prof. Galler, one of the true computer experts here at the Univ. of Michigan, and one of the poeople who has been deeply involved in the development of our computer system, might be of interest to you. It not only sheds light on some of the political aspects of the development of the internet, but also shows how risky it can be to deal with news media.
I know this has some political overtones (and perhaps undertones, too) However, I think it is also of importance for all of us who use the Internet to have accurate information about it's development from reliable sources. Wil Bigelow - - - - - - - - - - - - - - - - - - -
} X-Sender: galler-at-g.imap.itd.umich.edu (Unverified) } Mime-Version: 1.0 } Date: Sat, 21 Oct 2000 11:00:26 -0400 } To: coe-fac-staff-at-umich.edu } From: "Bernard A. Galler" {galler-at-umich.edu} } Subject: Setting the record straight ... } Status: } } Many of us recently saw the excellent statement by Vint Cerf and Bob Kahn } on Gore's "Inventing the Internet" claim (appended below). I asked for and } received permission from Vint to send it to our local paper, the Ann Arbor } News. When I sent it to them, I added a preface which said: "I am } submitting this important Letter to the Editor, with permission. It was } written by two people who are generally regarded in the computer industry } as the real inventors of the Internet." } } Unfortunately, they did publish "my letter" without my preface and without } the first half of the Cerf/Kahn statement, but with my name at the bottom. } Not only did they remove the validation of the statement, since there is no } longer any mention of the real authors, but they cast me in the role of } plagiarist. I have received several email messages and phone calls } congratulating me on my letter and my excellent writing! } } You may be sure that I will visit the Ann Arbor News when I return to Ann } Arbor on Monday. } Bernie } =================================================================== } Al Gore and the Internet } } By Robert Kahn and Vinton Cerf } Al Gore was the first political leader to recognize the importance of the } Internet and to promote and support its development. } } No one person or even small group of persons exclusively "invented" the } Internet. It is the result of many years of ongoing collaboration among } people in government and the university community. But as the two people } who designed the basic architecture and the core protocols that make the } Internet work, we would like to acknowledge VP Gore's contributions as a } Congressman, Senator and as Vice President. No other elected official, to } our knowledge, has made a greater contribution over a longer period of time. } } Last year the Vice President made a straightforward statement on his } role. He said: "During my service in the United States Congress I took the } initiative in creating the Internet." We don't think, as some people have } argued, that Gore intended to claim he "invented" the Internet. Moreover, } there is no question in our minds that while serving as Senator, Gore's } initiatives had a significant and beneficial effect on the still-evolving } Internet. The fact of the matter is that Gore was talking about and } promoting the Internet long before most people were listening. We feel it } is timely to offer our perspective. } } As far back as the 1970s Congressman Gore promoted the idea of high speed } telecommunications as an engine for both economic growth and the } improvement of our educational system. He was the first elected official } to grasp the potential of computer communications to have a broader impact } than just improving the conduct of science and scholarship. Though easily } forgotten, now, at the time this was an unproven and controversial } concept. Our work on the Internet started in 1973 and was based on even } earlier work that took place in the mid-late 1960s. But the Internet, as we } know it today, was not deployed until 1983. When the Internet was still in } the early stages of its deployment, Congressman Gore provided intellectual } leadership by helping create the vision of the potential benefits of high } speed computing and communication. As an example, he sponsored hearings on } how advanced technologies might be put to use in areas like coordinating } the response of government agencies to natural disasters and other crises. } } As a Senator in the 1980s Gore urged government agencies to consolidate } what at the time were several dozen different and unconnected networks into } an "Interagency Network." Working in a bi-partisan manner with officials } in Ronald Reagan and George Bush's administrations, Gore secured the } passage of the High Performance Computing and Communications Act in } 1991. This "Gore Act" supported the National Research and Education } Network (NREN) initiative that became one of the major vehicles for the } spread of the Internet beyond the field of computer science. } } As Vice President Gore promoted building the Internet both up and out, as } well as releasing the Internet from the control of the government agencies } that spawned it. He served as the major administration proponent for } continued investment in advanced computing and networking and private } sector initiatives such as Net Day. He was and is a strong proponent of } extending access to the network to schools and libraries. Today, } approximately 95% of our nation's schools are on the Internet. Gore } provided much-needed political support for the speedy privatization of the } Internet when the time arrived for it to become a commercially-driven } operation. } } There are many factors that have contributed to the Internet's rapid growth } since the later 1980s, not the least of which has been political support } for its privatization and continued support for research in advanced } networking technology. No one in public life has been more intellectually } engaged in helping to create the climate for a thriving Internet than the } Vice President. Gore has been a clear champion of this effort, both in the } councils of government and with the public at large. } } The Vice President deserves credit for his early recognition of the value } of high speed computing and communication and for his long-term and } consistent articulation of the potential value of the Internet to American } citizens and industry and, indeed, to the rest of the world. } } } Bernard A. Galler } E-mail: galler-at-umich.edu } Fax: 734-668-9998 }
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
Photoshop is used for all of the things mentioned, and that only scratches the surface of what is possible. In addition to the full blown graphics aspects, the current full versions of Photoshop (version 5.5 with 6.0 set to release any day now) include PDF support and Adobe ImageReady, an application for preparing images for web viewing. If you require basic image manipulation tools and Text capabilities, consider PhotoShop LE. Its $99 and has the most commonly used features of its big brother. A comparison of "LE" vs 6.0 is available at : http://www.adobe.com/products/photoshople/comparison.html
George Laing National Graphic Supply
Bill, It's not from the microscopist's point of view but there's an article in a magazine called Photo Electronic Imaging (PEI) that may help answer some of your questions about Photoshop. I really enjoy this magazine. The October issue gives lots of good info on Photoshop 6 (pages 16-17 show a 6.0 feature roundup). And, every month there's an article called Photoshop - Instant Expert by Ben Willmore. The web site is www.peimag.com.
Beth
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
NATIONAL INSTITUTES OF HEALTH Bethesda, Maryland Division of Bioengineering & Physical Science Supramolecular Structure & Function Lab.
Physical /life scientist (Biologist GS9/GS11/GS12) at BS or MS level with solid technical experience in thin-section transmission electron microscopy. Experience in advanced techniques in analytical electron microscopy and structural biology (cryosectioning, high-pressure freezing, and macromolecular preparation methods) is desirable, although on-the-job training is possible for an experienced and meticulous microscopist. PROOF OF U.S. CITIZENSHIP IS REQUIRED for this appointment.
For further information please contact:
Dr. Richard Leapman Division of Bioengineering & Physical Science Bldg. 13, Rm. 3N17 National Institutes of Health Bethesda, MD 20892 Tel: (301) 496-2599 FAX: (301) 496-6608 E-mail: leapman-at-helix.nih.gov
Applications (with curriculum vitae or federal employment form SF-171) should include the reference number ORS-00-0231, and should be post marked by November 7, 2000 and sent to:
Ms. Karen Harris National Institutes of Health ORS Human Resources Office 31 Center Drive Msc 2157 Bldg 31, Rm 4B41 Bethesda, MD 20892
Our health and safety officer wants to know if Uranyl Nitrate is used as an EM stain. Does anyone use Uranyl Nitrate? He did not tell me why he needs to know.
Thanks,
Ruth
*************************************** Ruth Yamawaki Department of Comparative Medicine Stanford University Stanford, CA 94305 ***************************************
At 12:05 PM 10/23/00, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Amazing and informative. I truly thought that the Internet was created by DARPA and BBN. I had no idea that Gore played such a pivotal role in its creation. When I was working with DARPA in the '70s, Gore's name never came up. I suppose it was because the lower level creators were just computer scientists and engineers.
Email: hma-at-tupphysiol1.bp.dal.ca Name: HuaLong Ma School: Dalhousie University
Question: I am reading a paper using BrdU staining the cell and observe the fluorescence micrograph and confocal micrograph, could you give some explainations about these techniques and what is special about them. By the way, what is " Camera lucida" ? thanks
I don't believe that one person may claim that he or she has invented/created the Internet. Internet is a result of join efforts of many people from many countries (for this reason it called INTERnet). As my knowledge, Internet was grew as a fast way to communicate between members of the scientific community at the beginning. Because of international nature of the scientific community, Internet was "INTER" from the very early steps. I have no idea how Al Gore's was involved in such spontaneous out of control process. Moreover, I do believe, that the beauty of the Internet is their "deregulation" and non-controlling by any government-related agency. This is my personal opinion.
Sergey
} Date: Mon, 23 Oct 2000 15:05:17 -0400 } From: Wil Bigelow {bigelow-at-engin.umich.edu} } Subject: Setting the record straight ... } 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
Wil, With all due respect, no more political mesages. Bob Ruscica
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 } -----------------------------------------------------------------------. } } Hi Listers: } Since all of us are heavily involved with the use of the internet, I } thought this message I just received from Prof. Galler, one of the true } computer experts here at the Univ. of Michigan, and one of the poeople who } has been deeply involved in the development of our computer system, might } be of interest to you. It not only sheds light on some of the political } aspects of the development of the internet, but also shows how risky it can } be to deal with news media. } } I know this has some political overtones (and perhaps undertones, too) } However, I think it is also of importance for all of us who use the } Internet to have accurate information about it's development from reliable } sources. } Wil Bigelow } - - - - - - - - - - - - - - - - - - - } } } X-Sender: galler-at-g.imap.itd.umich.edu (Unverified) } } Mime-Version: 1.0 } } Date: Sat, 21 Oct 2000 11:00:26 -0400 } } To: coe-fac-staff-at-umich.edu } } From: "Bernard A. Galler" {galler-at-umich.edu} } } Subject: Setting the record straight ... } } Status: } } } } Many of us recently saw the excellent statement by Vint Cerf and Bob Kahn } } on Gore's "Inventing the Internet" claim (appended below). I asked for and } } received permission from Vint to send it to our local paper, the Ann Arbor } } News. When I sent it to them, I added a preface which said: "I am } } submitting this important Letter to the Editor, with permission. It was } } written by two people who are generally regarded in the computer industry } } as the real inventors of the Internet." } } } } Unfortunately, they did publish "my letter" without my preface and without } } the first half of the Cerf/Kahn statement, but with my name at the bottom. } } Not only did they remove the validation of the statement, since there is no } } longer any mention of the real authors, but they cast me in the role of } } plagiarist. I have received several email messages and phone calls } } congratulating me on my letter and my excellent writing! } } } } You may be sure that I will visit the Ann Arbor News when I return to Ann } } Arbor on Monday. } } Bernie } } =================================================================== } } Al Gore and the Internet } } } } By Robert Kahn and Vinton Cerf } } Al Gore was the first political leader to recognize the importance of the } } Internet and to promote and support its development. } } } } No one person or even small group of persons exclusively "invented" the } } Internet. It is the result of many years of ongoing collaboration among } } people in government and the university community. But as the two people } } who designed the basic architecture and the core protocols that make the } } Internet work, we would like to acknowledge VP Gore's contributions as a } } Congressman, Senator and as Vice President. No other elected official, to } } our knowledge, has made a greater contribution over a longer period of time. } } } } Last year the Vice President made a straightforward statement on his } } role. He said: "During my service in the United States Congress I took the } } initiative in creating the Internet." We don't think, as some people have } } argued, that Gore intended to claim he "invented" the Internet. Moreover, } } there is no question in our minds that while serving as Senator, Gore's } } initiatives had a significant and beneficial effect on the still-evolving } } Internet. The fact of the matter is that Gore was talking about and } } promoting the Internet long before most people were listening. We feel it } } is timely to offer our perspective. } } } } As far back as the 1970s Congressman Gore promoted the idea of high speed } } telecommunications as an engine for both economic growth and the } } improvement of our educational system. He was the first elected official } } to grasp the potential of computer communications to have a broader impact } } than just improving the conduct of science and scholarship. Though easily } } forgotten, now, at the time this was an unproven and controversial } } concept. Our work on the Internet started in 1973 and was based on even } } earlier work that took place in the mid-late 1960s. But the Internet, as we } } know it today, was not deployed until 1983. When the Internet was still in } } the early stages of its deployment, Congressman Gore provided intellectual } } leadership by helping create the vision of the potential benefits of high } } speed computing and communication. As an example, he sponsored hearings on } } how advanced technologies might be put to use in areas like coordinating } } the response of government agencies to natural disasters and other crises. } } } } As a Senator in the 1980s Gore urged government agencies to consolidate } } what at the time were several dozen different and unconnected networks into } } an "Interagency Network." Working in a bi-partisan manner with officials } } in Ronald Reagan and George Bush's administrations, Gore secured the } } passage of the High Performance Computing and Communications Act in } } 1991. This "Gore Act" supported the National Research and Education } } Network (NREN) initiative that became one of the major vehicles for the } } spread of the Internet beyond the field of computer science. } } } } As Vice President Gore promoted building the Internet both up and out, as } } well as releasing the Internet from the control of the government agencies } } that spawned it. He served as the major administration proponent for } } continued investment in advanced computing and networking and private } } sector initiatives such as Net Day. He was and is a strong proponent of } } extending access to the network to schools and libraries. Today, } } approximately 95% of our nation's schools are on the Internet. Gore } } provided much-needed political support for the speedy privatization of the } } Internet when the time arrived for it to become a commercially-driven } } operation. } } } } There are many factors that have contributed to the Internet's rapid growth } } since the later 1980s, not the least of which has been political support } } for its privatization and continued support for research in advanced } } networking technology. No one in public life has been more intellectually } } engaged in helping to create the climate for a thriving Internet than the } } Vice President. Gore has been a clear champion of this effort, both in the } } councils of government and with the public at large. } } } } The Vice President deserves credit for his early recognition of the value } } of high speed computing and communication and for his long-term and } } consistent articulation of the potential value of the Internet to American } } citizens and industry and, indeed, to the rest of the world. } } } } } } Bernard A. Galler } } E-mail: galler-at-umich.edu } } Fax: 734-668-9998 } } } } 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 Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } We are trying to track down the location of "Cambridge Image } Technology," manufacturers of cathodoluminescence stages for } optical microscopes. Can anyone help? Thanks in advance, } } Mati
Here is a link to the web page for Cambridge Image Technology Ltd.
_____________________________ Dr Su Sajip Characterisation Development Engineer IQE (Europe) Ltd. United Kingdom Cypress Drive, St. Mellons Cardiff CF3 OEG * Wales, UK
Bill, I use PhotoShop for: Brightness/contrast/gamma/colour balance adjustments Re-size, rotate and annotate Select colour channels from RGB images Background correction Distortion of images to show make small thickness variations more visible Fiducial markers (very handy, I have a marker for each mag of my optical microscopes, SEM and TEM) Getting rid of blemishes/unwanted features (should I own up to this?) Making false colour SEM images Making movie sequences And probably a few more things I can't think of at the moment. I've used it to process } 10,000 images over the last 3 years.
I would only add that although it is expensive when bought by itself, it can often be obtained much more cheaply as part of a package for a scanner or camera.
Richard
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 was talking to a friend of mine this morning about what software he } should have to do digital imaging. Reflexively I said PhotoShop and was } asked how he would use it. I told him the standard things - contrast } enhancement, color matching across different media, annotation, resizing, } etc. His response was that for the price was that all it did (a little } naivety but a fair question) . You know, sometimes you get in a rut and it } set me to wondering how other people are using PhotoShop? It's not like } I'm selling the software but I'd like to be able to give a better answer } before telling someone they should plunk down that much money. Aside from } image analysis plug-ins (which I had forgotten about when we were talking) } how else is PhotoShop being used - anything fancy to justify the cost? } } Bill Miller }
e-mail richard.beanland-at-marconi.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."
this message is to advertise the 5th multinational congress on electron microscopy, that will be held in Lecce (Italy) from September 20 to September 25, 2001.
The Congress is organized by the microscopy societies of the following countries:
Austria Croatia Czech Rep Hungary Italy Slovak Rep Slovenia
The meeting is a forum for all scientists active in all fields of microscopy.
The topics covered will be:
Architecture of the nucleus and nuclear import and export, art, biomaterials, biosensors, environmental microscopy, cell and cell components, cell death in biology and pathology, chemical analysis and mapping, microscopy in human health, microscopy of nanostructural materials, microscopy of structural and functional materials, energy-filtered electron microscopy, instrumental development, investigations of thin films for sensor technology, microbiology, microscopy of semiconducting materials, parassitology, plant cell biolgy, remote microscopy, strain analysis, structure biology, structural determination and refining, viruses, teleoperation and computer assisted operation, nanotechnology, single molecule microscopy and manipulation
A large exhibition area will be available to manufacturers. Open labs and tutorials will be organized.
Information can be found at www.mcem5.unile.it, where brochures can be downloaded too, or directly writing to me (I am president of the congress (materials science section) together with Prof. L. Dini, (biomedical field)).
We espect and hope to have a large participation from all around the world. Hope to see you all in Lecce.
with all due respect to anyone, and keeping in mind that this listserver is mailny populated by Americans (so I belong to a minority, here, being Italian) I would like to espress my opinion on Robert Ruscica's message.
I strongly believe that internet is one of the few places where a human being can "still" express his own opinion on something.
Expressing an idea, reporting an opinion, is a way to express his own "freedom" and freedom is important to anyone.
Provided that anyone has the possibility of replying a message, of expressing a different opinion, of stating what he thinks too, I find that "discussing" in a positive way is always stimulating, and a high level auditorium, as this listserver is supposed to be, is a good place to discuss, as science and research are (unfortunately or fortunately) strictly related to politics.
} Email: hma-at-tupphysiol1.bp.dal.ca } Name: HuaLong Ma } School: Dalhousie University } } Question: I am reading a paper using BrdU staining the cell and observe the } fluorescence micrograph and confocal micrograph, could you give some } explainations about these techniques and what is special about them. By the } way, what is " Camera lucida" ? thanks }
HuaLong -
I can't help you much with your flourescence and confocal questions, but I do know what a Camera lucida is - it's a simple optical device which allows you to "project" and enlarge the image seen in a microscope onto a piece of paper, so that you can then draw it more accurately. It may be as simple as a tiltable mirror mounted on a pole with a base which also tilts, allowing the user to make whatever adjustments necessary to optimize the "projection". These devices have been around for a long time, (at least a hundred years, and probably much longer) and were often used in the original production of some of the fine, meticulous drawings one sees in antique texts in a wide variety of subjects. I wouldn't be surprised if some are still in use in anatomical and paleontological studies - sometimes a hand drawing is the best way to illustrate a certain feature (and since you need fingers to hold a pencil, such drawings may have been the first "digital" micrographs (sorry)). In my own former specialty, micropaleontology, Camera lucida's are still being used now and then, and they may even still be commercially available. I see you're at Dalhousie; if you ever want to have a look at one, I'd be pleased to show you ours (after wiping the dust off it); we're just across the bridge.
Frank Thomas MicroAnalysis Facility Geological Survey of Canada (Atlantic) Bedford Institute of Oceanography Dartmouth, Nova Scotia Canada B2Y 4A2
(902) 426-4635
From root Tue Oct 24 07:27:37 2000 Return-Path: {Microscopy-request-at-sparc5.microscopy.com} Received: (from daemon-at-localhost) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id HAA19848 for dist-Microscopy; Tue, 24 Oct 2000 07:15:41 -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 HAA19845 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Tue, 24 Oct 2000 07:15:10 -0500 (CDT) Received: from mercury.mis-hq.com (mercury.mis-hq.com [64.208.103.66]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id HAA19838 for {microscopy-at-sparc5.microscopy.com} ; Tue, 24 Oct 2000 07:14:59 -0500 (CDT) Received: from SETH (jupiter.mis-hq.com [64.208.103.70]) by mercury.mis-hq.com (8.9.3/8.9.3) with SMTP id HAA04815; Tue, 24 Oct 2000 07:13:50 -0500 Reply-To: {sethg-at-paxit.com}
Let me join on the bandwagon....unless we want to start posting "Bush for President" signs on this listserv, we ought to drop the clever, some might even say "subliminal", political agendas.
SethG
Wil, With all due respect, no more political mesages. Bob Ruscica
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 } -----------------------------------------------------------------------. } } Hi Listers: } Since all of us are heavily involved with the use of the internet, I } thought this message I just received from Prof. Galler, one of the true } computer experts here at the Univ. of Michigan, and one of the poeople who } has been deeply involved in the development of our computer system, might } be of interest to you. It not only sheds light on some of the political } aspects of the development of the internet, but also shows how risky it can } be to deal with news media. } } I know this has some political overtones (and perhaps undertones, too) } However, I think it is also of importance for all of us who use the } Internet to have accurate information about it's development from reliable } sources. } Wil Bigelow } - - - - - - - - - - - - - - - - - - - } } } X-Sender: galler-at-g.imap.itd.umich.edu (Unverified) } } Mime-Version: 1.0 } } Date: Sat, 21 Oct 2000 11:00:26 -0400 } } To: coe-fac-staff-at-umich.edu } } From: "Bernard A. Galler" {galler-at-umich.edu} } } Subject: Setting the record straight ... } } Status: } } } } Many of us recently saw the excellent statement by Vint Cerf and Bob Kahn } } on Gore's "Inventing the Internet" claim (appended below). I asked for and } } received permission from Vint to send it to our local paper, the Ann Arbor } } News. When I sent it to them, I added a preface which said: "I am } } submitting this important Letter to the Editor, with permission. It was } } written by two people who are generally regarded in the computer industry } } as the real inventors of the Internet." } } } } Unfortunately, they did publish "my letter" without my preface and without } } the first half of the Cerf/Kahn statement, but with my name at the bottom. } } Not only did they remove the validation of the statement, since there is no } } longer any mention of the real authors, but they cast me in the role of } } plagiarist. I have received several email messages and phone calls } } congratulating me on my letter and my excellent writing! } } } } You may be sure that I will visit the Ann Arbor News when I return to Ann } } Arbor on Monday. } } Bernie } } =================================================================== } } Al Gore and the Internet } } } } By Robert Kahn and Vinton Cerf } } Al Gore was the first political leader to recognize the importance of the } } Internet and to promote and support its development. } } } } No one person or even small group of persons exclusively "invented" the } } Internet. It is the result of many years of ongoing collaboration among } } people in government and the university community. But as the two people } } who designed the basic architecture and the core protocols that make the } } Internet work, we would like to acknowledge VP Gore's contributions as a } } Congressman, Senator and as Vice President. No other elected official, to } } our knowledge, has made a greater contribution over a longer period of time. } } } } Last year the Vice President made a straightforward statement on his } } role. He said: "During my service in the United States Congress I took the } } initiative in creating the Internet." We don't think, as some people have } } argued, that Gore intended to claim he "invented" the Internet. Moreover, } } there is no question in our minds that while serving as Senator, Gore's } } initiatives had a significant and beneficial effect on the still-evolving } } Internet. The fact of the matter is that Gore was talking about and } } promoting the Internet long before most people were listening. We feel it } } is timely to offer our perspective. } } } } As far back as the 1970s Congressman Gore promoted the idea of high speed } } telecommunications as an engine for both economic growth and the } } improvement of our educational system. He was the first elected official } } to grasp the potential of computer communications to have a broader impact } } than just improving the conduct of science and scholarship. Though easily } } forgotten, now, at the time this was an unproven and controversial } } concept. Our work on the Internet started in 1973 and was based on even } } earlier work that took place in the mid-late 1960s. But the Internet, as we } } know it today, was not deployed until 1983. When the Internet was still in } } the early stages of its deployment, Congressman Gore provided intellectual } } leadership by helping create the vision of the potential benefits of high } } speed computing and communication. As an example, he sponsored hearings on } } how advanced technologies might be put to use in areas like coordinating } } the response of government agencies to natural disasters and other crises. } } } } As a Senator in the 1980s Gore urged government agencies to consolidate } } what at the time were several dozen different and unconnected networks into } } an "Interagency Network." Working in a bi-partisan manner with officials } } in Ronald Reagan and George Bush's administrations, Gore secured the } } passage of the High Performance Computing and Communications Act in } } 1991. This "Gore Act" supported the National Research and Education } } Network (NREN) initiative that became one of the major vehicles for the } } spread of the Internet beyond the field of computer science. } } } } As Vice President Gore promoted building the Internet both up and out, as } } well as releasing the Internet from the control of the government agencies } } that spawned it. He served as the major administration proponent for } } continued investment in advanced computing and networking and private } } sector initiatives such as Net Day. He was and is a strong proponent of } } extending access to the network to schools and libraries. Today, } } approximately 95% of our nation's schools are on the Internet. Gore } } provided much-needed political support for the speedy privatization of the } } Internet when the time arrived for it to become a commercially-driven } } operation. } } } } There are many factors that have contributed to the Internet's rapid growth } } since the later 1980s, not the least of which has been political support } } for its privatization and continued support for research in advanced } } networking technology. No one in public life has been more intellectually } } engaged in helping to create the climate for a thriving Internet than the } } Vice President. Gore has been a clear champion of this effort, both in the } } councils of government and with the public at large. } } } } The Vice President deserves credit for his early recognition of the value } } of high speed computing and communication and for his long-term and } } consistent articulation of the potential value of the Internet to American } } citizens and industry and, indeed, to the rest of the world. } } } } } } Bernard A. Galler } } E-mail: galler-at-umich.edu } } Fax: 734-668-9998 } } } } 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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Yes, Wil's posting definitely has political overtones. So does the timing of the posting just 2 weeks before a presidential election. Are we now going to be inundated by apologists for every outrageous claim made by the Democrat candidate? I don't think I'd have time to read all email displaying torturous mental gymnastics justifying all the prevarications. Save it for the ballot box and keep it off the listserver.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi Listers: Since all of us are heavily involved with the use of the internet, I thought this message I just received from Prof. Galler, one of the true computer experts here at the Univ. of Michigan, and one of the poeople who has been deeply involved in the development of our computer system, might be of interest to you. It not only sheds light on some of the political aspects of the development of the internet, but also shows how risky it can be to deal with news media.
I know this has some political overtones (and perhaps undertones, too) However, I think it is also of importance for all of us who use the Internet to have accurate information about it's development from reliable sources. Wil Bigelow - - - - - - - - - - - - - - - - - - -
} X-Sender: galler-at-g.imap.itd.umich.edu (Unverified) } Mime-Version: 1.0 } Date: Sat, 21 Oct 2000 11:00:26 -0400 } To: coe-fac-staff-at-umich.edu } From: "Bernard A. Galler" {galler-at-umich.edu} } Subject: Setting the record straight ... } Status: } } Many of us recently saw the excellent statement by Vint Cerf and Bob Kahn } on Gore's "Inventing the Internet" claim (appended below). I asked for and } received permission from Vint to send it to our local paper, the Ann Arbor } News. When I sent it to them, I added a preface which said: "I am } submitting this important Letter to the Editor, with permission. It was } written by two people who are generally regarded in the computer industry } as the real inventors of the Internet." } } Unfortunately, they did publish "my letter" without my preface and without } the first half of the Cerf/Kahn statement, but with my name at the bottom. } Not only did they remove the validation of the statement, since there is no } longer any mention of the real authors, but they cast me in the role of } plagiarist. I have received several email messages and phone calls } congratulating me on my letter and my excellent writing! } } You may be sure that I will visit the Ann Arbor News when I return to Ann } Arbor on Monday. } Bernie } =================================================================== } Al Gore and the Internet } } By Robert Kahn and Vinton Cerf } Al Gore was the first political leader to recognize the importance of the } Internet and to promote and support its development. } } No one person or even small group of persons exclusively "invented" the } Internet. It is the result of many years of ongoing collaboration among } people in government and the university community. But as the two people } who designed the basic architecture and the core protocols that make the } Internet work, we would like to acknowledge VP Gore's contributions as a } Congressman, Senator and as Vice President. No other elected official, to } our knowledge, has made a greater contribution over a longer period of time. } } Last year the Vice President made a straightforward statement on his } role. He said: "During my service in the United States Congress I took the } initiative in creating the Internet." We don't think, as some people have } argued, that Gore intended to claim he "invented" the Internet. Moreover, } there is no question in our minds that while serving as Senator, Gore's } initiatives had a significant and beneficial effect on the still-evolving } Internet. The fact of the matter is that Gore was talking about and } promoting the Internet long before most people were listening. We feel it } is timely to offer our perspective. } } As far back as the 1970s Congressman Gore promoted the idea of high speed } telecommunications as an engine for both economic growth and the } improvement of our educational system. He was the first elected official } to grasp the potential of computer communications to have a broader impact } than just improving the conduct of science and scholarship. Though easily } forgotten, now, at the time this was an unproven and controversial } concept. Our work on the Internet started in 1973 and was based on even } earlier work that took place in the mid-late 1960s. But the Internet, as we } know it today, was not deployed until 1983. When the Internet was still in } the early stages of its deployment, Congressman Gore provided intellectual } leadership by helping create the vision of the potential benefits of high } speed computing and communication. As an example, he sponsored hearings on } how advanced technologies might be put to use in areas like coordinating } the response of government agencies to natural disasters and other crises. } } As a Senator in the 1980s Gore urged government agencies to consolidate } what at the time were several dozen different and unconnected networks into } an "Interagency Network." Working in a bi-partisan manner with officials } in Ronald Reagan and George Bush's administrations, Gore secured the } passage of the High Performance Computing and Communications Act in } 1991. This "Gore Act" supported the National Research and Education } Network (NREN) initiative that became one of the major vehicles for the } spread of the Internet beyond the field of computer science. } } As Vice President Gore promoted building the Internet both up and out, as } well as releasing the Internet from the control of the government agencies } that spawned it. He served as the major administration proponent for } continued investment in advanced computing and networking and private } sector initiatives such as Net Day. He was and is a strong proponent of } extending access to the network to schools and libraries. Today, } approximately 95% of our nation's schools are on the Internet. Gore } provided much-needed political support for the speedy privatization of the } Internet when the time arrived for it to become a commercially-driven } operation. } } There are many factors that have contributed to the Internet's rapid growth } since the later 1980s, not the least of which has been political support } for its privatization and continued support for research in advanced } networking technology. No one in public life has been more intellectually } engaged in helping to create the climate for a thriving Internet than the } Vice President. Gore has been a clear champion of this effort, both in the } councils of government and with the public at large. } } The Vice President deserves credit for his early recognition of the value } of high speed computing and communication and for his long-term and } consistent articulation of the potential value of the Internet to American } citizens and industry and, indeed, to the rest of the world. } } } Bernard A. Galler } E-mail: galler-at-umich.edu } Fax: 734-668-9998 }
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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Colleagues....
I've already replied privately to a number of you but before it start to grow let's nip this one in the bud.
Unless the topic is Microscopy and Microanalysis related please keep tangential commentary off-line. Comments about the Internet and what any politican's may have or have not said has no direct bearing on our agenda.
I met a problem in our EDS system, we have a TX 1255 Nortan detector and 4pai system (DTSA software). The problem is even the input counts is high to 2000 to 3000 and the dead time is below 30%, the output counts is less than 200, and when the input is less than 300 and dead time is less than 30%, there is no out put. I checked the discriminator adjustment in the pulse processor, it is almost there. How can I adjust the system?
Regards!
J.G. Wang
Jinguo Wang Materials Characterization Laboratory 194 MRI Building 5-9285
On Mon, 23 Oct 2000 15:52:51 -0700 Ruth Yamawaki {Ryamawaki-at-cmexchange.stanford.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 } -----------------------------------------------------------------------. } } Ruth,
Uranyl nitrate is used to make Reynolds lead citrate, a widely used TEM post stain for ultrathin tissue sections. In this formula, the lead nitrate is chelated by citrate. Hope this is of some use
} Our health and safety officer wants to know if Uranyl Nitrate is used } as an EM stain. Does anyone use Uranyl Nitrate? He did not tell me } why he needs to know. }
Full-time position available for an Electron Microscopy/Histology Technician to work on a NSF funded research project on biological structural colors. Responsibilities include specimen preparation, observation, and imaging using transmission electron microscopy (TEM) and light microscopy. Additional opportunties for active participation in research and publication. Required qualifications: Bachelor's degree or equivalent in an appropriate field; previous experience in microscopy or histology; previous experience in laboratory research. Preferred qualifications: previous experience in specimen preparation and sectioning for TEM; previous experience operating transmission eletron microscopes; previous experience in fixation, sectioning, and staining for light microscope histology; previous experience in computer data base management; graduate degree in an appropriate field. Salary: $25,000 per year (full-time) with benefits. Send a CV, a written statement describing previous experience and qualifications, and the names addresses, phone, and email addresses of three professional references to: Dr. Richard O. Prum, Natural History Museum, Dyche Hall, University of Kansas, Lawrence, KS 66045-2454; (785) 864-3897; prum-at-ukans.edu. Review of applications begins Nov. 1, 2000 and will continue until position is filled. EO/AA Employer
------------------------------------------------------------------------------ -- Richard O. Prum Associate Professor, Department of Ecology and Evolutionary Biology Curator, Division of Ornithology KU Natural History Museum University of Kansas Lawrence, KS 66045 Phone: (785)864-3897 Fax: (785)864-5335 Email: prum-at-ukans.edu ------------------------------------------------------------------------------ --
} ... I said PhotoShop, and was asked how he would use it. } I told him the standard things - contrast } enhancement, color matching across different media, } annotation, resizing, etc. ... } ... } how else is PhotoShop being used - anything fancy to } justify the cost?
We use PS5 here for annotation and presentation of spacial elemental mapping (elemental x-ray maps), and average atomic number maps (backscatter electron images). There isn't better software for presenting this type of data in a "perceptually" uniform way. For example, if you have a grayscale elemental map which represents SiO2 from 0% to 100%, with most softwares and typical monitors the change from 20% to 30% isn't perceived the same as 70% to 80%, if perceived at all. Opening and converting such an image into a Photoshop gamma=2.2 working space makes this gradation perceptually uniform, and much better for presentation. The caveat here is the "conversion" when you do this ... that is, PS makes the simple opening of the file such an innocent process, the user may not notice his/her data has been modified. What was once a pixel value of '128' is now '183'!! The data is now useless for quantitative analysis. Make it a practice to keep (possibly even write-protect) your original image files, and put your PS projects in a separate directory.
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/
} Date: Mon, 23 Oct 2000 15:52:51 -0700 } From: Ruth Yamawaki {Ryamawaki-at-cmexchange.stanford.edu} } To: "'Microscopy-at-sparc5.microscopy.com'" {Microscopy-at-sparc5.microscopy.com} } Subject: Uranyl Nitrate } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 health and safety officer wants to know if Uranyl Nitrate is used as an } EM stain. Does anyone use Uranyl Nitrate? He did not tell me why he needs } to know. } } Thanks, } } Ruth } } *************************************** } Ruth Yamawaki } Department of Comparative Medicine } Stanford University } Stanford, CA 94305 } *************************************** } Uranyl nitrate, like uranyl acetate and other uranium salts, is used as a stain in EM. The salt most frequently used is uranyl acetate because of its staining efficiency. Uranium has a heavy atomic nucleus (92) and is the heaviest metal used as a stain in EM. In tissue, it stabilizes nucleic acids and membranes when used prior to embedding. It is frequently used after sections are cut to "post stain" or further increase the contrast of membranes, etc., in sections. Uranyl salts, particularly acetate, are also used as negative stains for small things such as viruses and small cellular organelles in suspension. The stain darkens the background support and cracks and crevasses in the sample to add contrast. Uranyl salt solutions should be handled with care because they are radioactive. The alpha radiation is generally contained by glass (lead not necessary), but nonetheless, one should not leave dried pipets and droplets lying around to become airborne and possibly inhaled.
Hayat MA. Positive Staining for Electron Microscopy. 1975. Van Nostrand Reinhold, NY.
Hayat MA, Miller SE. 1990. Negative Staining. McGraw-Hill, NY.
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
I am currently trying to source an used heating holder, cooling holder and a power supply for use in an Hitachi H600 TEM. If you have either of these holders that are not in use and would like to sell them (or donate them to a University) please contact me to see if we can come to a mutually agreeable price.
Chas
Charlie Cooney Metallographer Dept. of Materials and Metallurgical Engineering Queen's University Nicol Hall Kingston, Ontario K7L 1N6
We are looking into purchasing a digital microscope camera for our Zeiss axioplan upright compound microscope. We are particularly interested in color cameras that can capture good fluorescent images in addition to brightfield images.
I would very much appreciate some suggestions regarding a good camera.
Thanks in advance.
Soumitra
************************************************************* Soumitra Ghoshroy Ph.D. Electron Microscopy Lab and Fluorescence Imaging Facility Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3600 Fax: 505-646-5665 e-mail:sghoshro-at-nmsu.edu http://confocal.nmsu.edu/eml
} } Dear Listservers, } } with all due respect to anyone, and keeping in mind that this listserver is } mailny populated by Americans (so I belong to a minority, here, being } Italian) I would like to espress my opinion on Robert Ruscica's message. } } I strongly believe that internet is one of the few places where a human } being can "still" express his own opinion on something. } } Expressing an idea, reporting an opinion, is a way to express his own } "freedom" and freedom is important to anyone.
Of course it is. Of course the internet is a wonderful thing. Of course we all want to discuss our political philosophies (vote for Bush).
However, the internet is also a *big* thing. There are lots and lots of people discussing lots and lots of different things here. Not everyone needs to discuss all topics in every conversation.
Nader is an idiot.
It is not a restriction on any inalienable liberties for a group of people to assemble and say "Here, today, we have chosen to talk about microscopy. If you want to talk about Gore, feel free to go to the political group. If you want to talk about woodworking, there is a fine woodworking discussion group over there. If you want to talk about Dungeons and Dragons, there is a great D&D group across the hall." That part and parcel of the right of association, and it is just as important as a right of speech.
Bush is wonderful.
It is further inappropriate for someone to come in and say "I don't care what everyone has agreed to! I want to talk about Gore's policiy towards D&D--playing woodworkers, and I don't care if it *is* a microscopy group. *I* care about Gore and woodworking and you microscopists had just better sit down and listen!"
Buchanan is a Nazi.
That is inappropriate, and it is not an abrogation of liberty for folk to say "Well, I'd rather you didn't."
Gore is dangerous.
On a practical note, that is *why* we have mailing lists and news groups. Do you suggest that we do away with all of them and just have one large mailing list and one large newsgroup called misc.stuff? Of course not.
Gore bad. Bush good.
In large part, we have managed to avoid direct moderation of most newsgroups and mailing lists by agreeing to abide by self-selected and agreed upon rules of behavior. Refusing to abide by these choices will not result in greater "freedom," but will instead result in the opposite -- the increasing direct moderation and censorship of newsgroups as a last-ditch attempt to control spam and intrusion.
M.A. Hayat mentions the use of uranyl nitrate on page 34 of his book Positive Staining (c. 1975) as an electron stain. He says that it is soluble in water, ethanol, acetone, and ether, but not in benzene, toluene and chloroform. The aqueous solution can hold up to 56% of anhydrous salt (w/v) at 25 C. Uranyl nitrate solutions are more stable than are uranyl acetate solutions but its less efficient as a stain than the acetate.
I have never used it. Ask your saftey officer why he needs to know.
Gib Ahlstrand
} Responding to the message } from Ruth Yamawaki {Ryamawaki-at-cmexchange.stanford.edu} : } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 health and safety officer wants to know if Uranyl Nitrate is used as an } EM stain. Does anyone use Uranyl Nitrate? He did not tell me why he needs } to know. } } Thanks, } } Ruth } } *************************************** } Ruth Yamawaki } Department of Comparative Medicine } Stanford University } Stanford, CA 94305 } ***************************************
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
According to the Polysciences Catalog Uranyl Nitrate is a negative stain. Also a photopolymerizing reagent for the polymerization of acrylamide gels at low pH. I personally have no experience with the stuff, but would imagine that it is similar to uranyl acetate which is a commonly used negative or positive stain. Tim
Timothy G. Schneider Director of Electron Microscopy Department of Pathology Room 229 Jefferson Hall Thomas Jefferson University 1020 Locust St. Philadelphia Pa. 19107 215-503-4798 work 610-613-8170 cellular timothy.schneider-at-mail.tju.edu
Hi All, I am looking for a good replacement for D19b developer (for nuclear emulsion plates, as used in our X-ray radiography and topography images, no longer available). We are using Suprol at the moment, but would like something as good as the old stuff. Anyone have any ideas?
e-mail richard.beanland-at-marconi.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."
Hi all: I am shopping around, looking for recommendation of glass suppliers for that could make a chamber for our CRC-100 magnetron sputter coater. The manufacturer wants alot for theirs and I am looking for alternative suppliers that this community might be aware of . Any suggestions would be greatly appreciated. Thanks in advance for your assistance.
Regards, Mike Coviello Lab Manager Unversity of Texas -at- Arlington
} -----Original Message----- } From: Berryhill, Bridgette } Sent: Tuesday, October 24, 2000 10:15 AM } To: Azizan, Azliyati; Barber, Janet; Chen, Lan; Gahagan, Jackie; } Govindraj, Prasanthi; Grimm, David; Hall,Laura; Hernandez, Dan; Howell, } Troy; Ingrassia, Angela; Kane, Brad; McKinnon,William; McPherson, } Charlene; Su, Phy-Huynh; Vallet, Noelle; Westling, Jennifer; 'David } Alexandrou'; 'Elyse Drossman'; 'Jane Dunlevy'; 'MaryBeth Colter'; 'Stacey } Kennedy' } Subject: FW: Easy way to help a good cause TOMORROW } } } From: Amy Wolfson } Sent: Monday, October 23, 2000 9:59 PM } To: Debby Greenberg; Marilyn Weisman; Michele Schointuch; Fran Silver; } Carol Bassett; Victoria Rennesund; Stephanie Shaw; Andrea Sands; Bridgette } Berryhill; Shari Hamilton; Sharon Bevis; Jane Birkhold; Darbi Bossman; } Jennifer Capouya; Cindy Daniels; Pat Fox; Bev Kinsey; Nancy Leeds; Wendy } McCoy; Ilene Beer; Sarah Shanks; Alina Simonds; Erin Stuart; Katya Bock; } Lowie Bock; Connie Zink } Cc: Mary DeLong } Subject: Fw: Easy way to help a good cause TOMORROW } } } } Subject: Easy way to help a good cause TODAY - Oct .24 } } } } } } } } } } } } Subject: FW: Breast Cancer & the NFL } } } } } } The NFL has chosen October 24, 2000 as the NFL's Breast Cancer } } } } } Awareness day. Every time someone logs onto www.NFL.com, the NFL } } will } } } } } donate $5 to the Susan G. Komen Breast Cancer Foundation. This } is } a } } } } simple } } } } } opportunity to learn about one of our fabulous National Series } } Sponsor } } } } of } } } } } the Komen Race for the Cure(R), as well as earn up to $50,000 for } the } } } } Komen } } } } } Foundation. Tell all your friends and co-workers on October 24 } to } } log } } } } on!! } } }
Photographic developers that are no longer available can often be made from scratch using just a few standard ingredients that are fairly readily available. It may be that Eastman Kodak would consent to give you the formula if asked, but if not, chances are pretty good that the formula is out there floating around somewhere in an old book, journal, or even in cyberspace.
I know that most people don't want to mess around with making their own developers, or don't feel secure using them, but if you're interested (and not in a huge hurry) I can check some sources to try and locate the formula. It is probably just a minor variant of regular Kodak D-19. I have a small collection of darkroom chemistry oriented literature, plus some good leads on where to ask.
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: Richard Beanland +44 1327 356363 [mailto:richard.beanland-at-marconi.com] Sent: Tuesday, October 24, 2000 12:38 PM To: Microscopy Listserver
Hi All, I am looking for a good replacement for D19b developer (for nuclear emulsion plates, as used in our X-ray radiography and topography images, no longer available). We are using Suprol at the moment, but would like something as good as the old stuff. Anyone have any ideas?
e-mail richard.beanland-at-marconi.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."
Does anyone know of software that will do a batch conversion of the .IM_ image files captured using Link ISIS v. 3 Autobeam, to a standard format (eg. pcx, jpg, etc)?
Fiona ____________________ Dr Fiona Graham EM Unit, University of Natal, Durban South Africa
**** Remember ICEM-15 in 2002, is in Durban, South Africa **** www.icem15.com
Ruth, 0.01% Uranyl Nitrate is used as a local catalyst in the polymerization of methacrylates.
Hayat MA. Principles and Techniques of Electron Microscopy. 1970. Van Nostrand Reinhold, NY.
Hope this helps! Jo Dee
Sara Miller 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 } -----------------------------------------------------------------------. } } On Mon, 23 Oct 2000, Ruth Yamawaki wrote: } } } Date: Mon, 23 Oct 2000 15:52:51 -0700 } } From: Ruth Yamawaki {Ryamawaki-at-cmexchange.stanford.edu} } } To: "'Microscopy-at-sparc5.microscopy.com'" } {Microscopy-at-sparc5.microscopy.com} } } Subject: Uranyl Nitrate } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 health and safety officer wants to know if Uranyl Nitrate is used as an } } EM stain. Does anyone use Uranyl Nitrate? He did not tell me why he needs } } to know. } } } } Thanks, } } } } Ruth } } } } *************************************** } } Ruth Yamawaki } } Department of Comparative Medicine } } Stanford University } } Stanford, CA 94305 } } *************************************** } } } Uranyl nitrate, like uranyl acetate and other uranium salts, is used as a } stain in EM. The salt most frequently used is uranyl acetate because of } its staining efficiency. Uranium has a heavy atomic nucleus (92) and is } the heaviest metal used as a stain in EM. In tissue, it stabilizes } nucleic acids and membranes when used prior to embedding. It is } frequently used after sections are cut to "post stain" or further } increase the contrast of membranes, etc., in sections. Uranyl salts, } particularly acetate, are also used as negative stains for small things } such as viruses and small cellular organelles in suspension. The stain } darkens the background support and cracks and crevasses in the sample to } add contrast. Uranyl salt solutions should be handled with care because } they are radioactive. The alpha radiation is generally contained by } glass (lead not necessary), but nonetheless, one should not leave dried } pipets and droplets lying around to become airborne and possibly inhaled. } } Hayat MA. Positive Staining for Electron Microscopy. 1975. Van Nostrand } Reinhold, NY. } } Hayat MA, Miller SE. 1990. Negative Staining. McGraw-Hill, NY. } } Sara E. Miller, Ph. D. } P. O. Box 3712 } Duke University Medical Center } Durham, NC 27710 } Ph: 919 684-3452 } FAX: 919 684-3265
-- Jo Dee Fish Coordinator of Electron Microscopy Cell Analysis Facility The Burnham Institute 10901 N. Torrey Pines Rd. La Jolla, CA 92037 (858)646-3100 ext. 3620
} -Hi All, } I am looking for a good replacement for D19b developer (for nuclear emulsion plates, as used in our X-ray radiography and topography images, no longer available). We are using Suprol at the moment, but would like something as good as the old stuff. Anyone have any ideas? } } Richard } } ============================================================== } Richard Beanland, } Structural Analysis Lab, } Caswell Technology, } Caswell, } Towcester, } Northants NN12 8EQ } } e-mail richard.beanland-at-marconi.com } Tel. +44 1327 356363 } Fax. +44 1327 356398 }
How about mixing your own from scratch? I will see if I can dig up the formula tomorrow (Wed).
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 **********************************************
Just for your information: A camera Lucida is also called a Drawing Tube and even sometimes Une Chambre claire. It does not project the image observed thru the microscope but allowed to overimposed the image seen thru the microscope and a piece of paper located on the side of the microscope and at the same time the pencil on this paper. The brightness if the paper as the be the same as the image observed to see both field at the same time. Than one just have to follow the contour of the object to obtain the right drawing. It exist drawing tubes for Compond microscopes as well as stereo`s microscopes My two cents information Norm
----- Original Message ----- } From: Frank Thomas {thomasf-at-agc.bio.ns.ca} To: {Microscopy-at-sparc5.microscopy.com} ; {hma-at-tupphysiol1.bp.dal.ca} Sent: Tuesday, October 24, 2000 07:28
Folks we have some pretty definitive formularies for photographic chemicals here at the museum. Contact us off list with a wish list of developer recipes if you have not found anything.
thanks Ed Sharpe archivist for SMECC
{ { Subj: RE: D19b developer Date: 10/24/00 1:43:45 PM US Mountain Standard Time From: TindallR-at-missouri.edu (Tindall, Randy D.) To: richard.beanland-at-marconi.com ('Richard Beanland +44 1327 356363') CC: microscopy-at-sparc5.microscopy.com ('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 -----------------------------------------------------------------------.
Richard,
Photographic developers that are no longer available can often be made from scratch using just a few standard ingredients that are fairly readily available. It may be that Eastman Kodak would consent to give you the formula if asked, but if not, chances are pretty good that the formula is out there floating around somewhere in an old book, journal, or even in cyberspace.
I know that most people don't want to mess around with making their own developers, or don't feel secure using them, but if you're interested (and not in a huge hurry) I can check some sources to try and locate the formula. It is probably just a minor variant of regular Kodak D-19. I have a small collection of darkroom chemistry oriented literature, plus some good leads on where to ask.
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/
} The caveat here is the "conversion" when you do this ... that is, PS } makes the simple opening of the file such an innocent process, the } user may not notice his/her data has been modified. What was once a } pixel value of '128' is now '183'!!
I don't want to put down Photoshop, especially since we are producing a competing product, but I think this remark points to one problem that many "photo" applications have -- they do not treat the intensity as scientific data. I may be mistaken, but I think Photoshop is conceptually a "beautifying" software to make photos "look nicer" -- and it does a great job at this, but it may also be lacking in certain areas. I'd be interested if Photoshop can deal with inverse length scales such as on diffraction patterns, or if it can calibrate the intensity in values such as non linear length scales or other values. In the case Michael mentions above, for example, one should be able to change the look-up table to optimize the display without changing the underlying data.
Don't misunderstand me, please. I think Photoshop is a great product for what it does, but for heavy duty data reduction and quantitative analysis another software may be a better choice (Sorry if this sounds like a "plug" for ours, there are many others available as well).
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: michael shaffer [mailto:epmalab-at-darkwing.uoregon.edu] Sent: Tuesday, October 24, 2000 8:32 AM To: Microscopy-at-sparc5.microscopy.com
Bill Miller writes ...
} ... I said PhotoShop, and was asked how he would use it. } I told him the standard things - contrast } enhancement, color matching across different media, } annotation, resizing, etc. ... } ... } how else is PhotoShop being used - anything fancy to } justify the cost?
We use PS5 here for annotation and presentation of spacial elemental mapping (elemental x-ray maps), and average atomic number maps (backscatter electron images). There isn't better software for presenting this type of data in a "perceptually" uniform way. For example, if you have a grayscale elemental map which represents SiO2 from 0% to 100%, with most softwares and typical monitors the change from 20% to 30% isn't perceived the same as 70% to 80%, if perceived at all. Opening and converting such an image into a Photoshop gamma=2.2 working space makes this gradation perceptually uniform, and much better for presentation. The caveat here is the "conversion" when you do this ... that is, PS makes the simple opening of the file such an innocent process, the user may not notice his/her data has been modified. What was once a pixel value of '128' is now '183'!! The data is now useless for quantitative analysis. Make it a practice to keep (possibly even write-protect) your original image files, and put your PS projects in a separate directory.
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/
At last a chance to use some ancient knowledge to help out.
For Kodak products, there was a convention that said anything with a number as a name was a public domain type formula. So if you go into some old Kodak books you will find the formulas for things like D-19, D-11, D-76 etc.
Products that used a name for a name, like Dektol, were proprietary formulas, not generally available.
I probably have the formulas for most of the standard type numbered products. Let me know if you need one.
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
Knock off the political mumbo-jumbo. It is surely inappropriate here.
Nestor can you filter this stuff? -----Original Message----- } From: William R. Oliver {oliver-at-cpt.afip.org} To: Massimo Catalano {massimo.catalano-at-ime.le.cnr.it} Cc: microscopy-at-sparc5.microscopy.com {microscopy-at-sparc5.microscopy.com}
I think De Vere 504 is a good one. Depending on your need and the type of paper you use, you can choose different enlarger heads such as Ilford Ilfospeed Multigrde enlarger head.
Regards,
Kun Li
Systems on Silicon Manufacturing Company Pte Ltd FA Section --
On Tue, 24 Oct 2000 09:03:27 Jinguo Wang wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Get your FREE Email at http://www.mailcityasia.com
I have read that some dilution of Kodak HC110 made a good substitute for D 19. A call to Kodak's help line 800 242 2424 should get an answer. HC110 keeps for ever and can be replenished.
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger
} From: "Geoff McAuliffe" {mcauliff-at-UMDNJ.EDU} } } -Hi All, } } I am looking for a good replacement for D19b developer (for nuclear emulsion plates, as used in our X-ray radiography and topography images, no longer available). We are using Suprol at the moment, but would like something as good as the old stuff. Anyone have any ideas? } } } } Richard } } } } ============================================================== } } Richard Beanland, } } Structural Analysis Lab, } } Caswell Technology, } } Caswell, } } Towcester, } } Northants NN12 8EQ } } } } e-mail richard.beanland-at-marconi.com } } Tel. +44 1327 356363 } } Fax. +44 1327 356398 } } } } How about mixing your own from scratch? I will see if I can dig up the formula tomorrow (Wed). } } 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 } ********************************************** } } }
Here is a formulae for Developer D19b I have in an old file.
Metol 2.2 Hydroquinone 8.8 Sodium sulphite(anhydrous) 72 Sodium carbonate(anhydrous) 48 Potassium bromide 4 Water to 1 L.
D-19b was generally regarded as a high contrast developer for X-ray and industrial photography. Used undiluted at 68F, the average time for tank development is five minutes. This is a well balanced developer of good keeping properties.
D19b formula (quoted as a "manufacturer's formula" in British Journal Photographic almanac for 1955)
metol 2.2g hydroquinone 8.8g sodium sulphite cryst. 144g sodium carbonate cryst. 130g potassium bromide 4g water to 1litre
Chris
Date sent: Tue, 24 Oct 2000 17:37:32 +0000 (GMT) } From: Richard Beanland +44 1327 356363 {richard.beanland-at-marconi.com}
If it's ordinary D-19 developer there are numerous sources for this formula published. There's also one for a replenisher called D-19R (replenisher). However, I've not seen a D-19b. I've used D-19 for TEM plates (Kodak SO-163 and 4489) for years and it's readily available in those familiar yellow packages, at least here in the states. The formula (from Kodak's publication J-1):
Water, about 50 degrees C 500ml
Kodak ELON Developing agent (aka Metol or p-methylaminophenol sulfate) 1.0 gram Kodak Sodium sulfite (anhydrous) 75.0 grams Kodak Hydroquinone 9.0 grams Kodak Sodium Carbonate (monohydrate) 30.0 grams Kodak Potassium Bromide (anhydrous) 5.0 grams Cold water to make 1 liter.
At a state-side price, for working solution, of about $5.00 for 3 gallons I buy those little yellow packages. In formulating related photographic solutions, I've found no difference in using equivalent bulk chemicals of different manufacture. As I recall the above formula is also in the Ilford manual (probably sans the Kodak trade names).
See what develops,
John
No financial interest in the Eastman-Kodak Company
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 } -----------------------------------------------------------------------. } } At last a chance to use some ancient knowledge to help out. } } For Kodak products, there was a convention that said anything with a number } as a name was a public domain type formula. So if you go into some old } Kodak books you will find the formulas for things like D-19, D-11, D-76 } etc. } } Products that used a name for a name, like Dektol, were proprietary } formulas, not generally available. } } I probably have the formulas for most of the standard type numbered } products. Let me know if you need one. } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu
My posting for the D-19 formula was wrong. My message was for D-11 which I've used for higher contrast work. Looks like the correct D-19b formula has been posted. It's different from regular D-19.
D.M. de Leeuw et al. Physica C 158 (1989) 391 prepared air and moisture sensitive samples for TEM by crushing them in a mortar under decanol and putting a droplet of the suspension onto a grid. Decanol has low evaporation rate and thus protects the sample. However, it evaporates inside microscope in air-lock but probably also in the column. I would like to know if the procedure is save for modern UHV microscopes and/or there are other procedures that could be safely used for similar air sensitive samples.
Leszek Kepinski
Institute of Low Temperature and Structure Research, Polish Academy of Sciences, P.O.Box 1410, 50-950 Wroclaw, Poland
} Knock off the political mumbo-jumbo. It is surely inappropriate here. } } Nestor can you filter this stuff?
Sheesh. Gals and guys, it was a *joke,* see. You know, there was this big flap about a certain politician inserting subliminal messages into things (even though he couldn't *pronounce* the word), and I was trying to leaven my reply to Dr. Catalano with a little humor. You know -- inserting inappropriate short statements sotto voce. Ever see Kevin Nealon's schtick on "Subliminal Man"?
As described by Dr. Gary Radford in "Subliminal Persuasion in the Mass Media" http://alpha.fdu.edu/~gradford/subliminal.html:
Kevin Nealon, a comic on Saturday Night Live, portrays a character called "Subliminal Man." Subliminal Man persuades people to do his bidding by uttering "hidden," but in fact wholly audible messages under his breath. In a commercial for Miller Lite, he says: "What they won't do in commercials these days [buy Miller Lite] - I mean, they resort to all sorts of tactics [brainwash]. Now take Miller Lite [six pack], it has a great pilsner taste [gonna love it]." At the end of the ad, Subliminal Man propositions a beautiful blonde: "Hi [your place], would you like to join me for a Lite [your treat]? It's on me." She coos back: "OK, but it's my treat."
And, you see, it *had* to be Bush, because he was the one in the news about it.
Sorry, I had made the obviously silly assumption that this was a pretty transparent schtick which merely attempted to sugar-coat the real message, which was that frank poltical discussions *were* inappropriate. You chose to ignore *that* message and instead only read the bullet insertions between the paragraphs.
I would make some correllations between stiffness, lack of sense of humor and likely candidate support, but I will leave that be for now.
Henceforth I promise never to use humor in this group and *only* reply in terms of self-righteous outrage and stern condemnation.
I hope this isn't an inappropriate question, but we have some quick decisions to make.
I've queried the list a couple times before about peoples' experiences with non-OEM equipment insurance contracts, but now I need to ask a more specific question. I would be very interested in hearing from people who have experience with CIC Corp and/or Kemper Cost Management, particularly with electron microscopes and ancillary equipment. Both positive and negative comments are very welcome.
Please (pretty please?) reply to me directly and not to the listserver, especially if you have negative comments. I think Nestor frowns on public company bashing and I don't blame him.
Thanks 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/
Hi, I am a graduate student and I need some help trying to figure out a quantitative way to evaluate my samples before running my experiments with them. I am using a technique we call nanosphere lithography by which we deposit polystyrene spheres onto a substrate. We want a single layer of hexagonally packed spheres. My spheres are 400nm across which means the spaces between them are just under 100nm. We get this packing in some areas and can see a nice, dark blue color. However, just as in crystal growth we also see grain boundries and other defects where we may have no spheres or multiple layers of spheres. My substrates are 12 - 15 mm square ITO coated glass. AFM is not a good solution because it takes entirely too long to look at an entire sample and even then, I only have my observations to say which area is the best. Someone mentioned stereology to me but I am unsure of what that technique entails and what information I can gain from it. I am looking for something that will be able to look at the entire sample and eventually give me some sort of number such as the % that is perfect or even the % that is defective. We have an 80X objective but that isn't enough resolution to see the tiny spheres in enough detail. However, I have access to some other facilities on campus but need to know what to look for. Does anyone have any suggestions? Thanks for your time and any comments are appreciated.
I have been very happy with my Durst Point Source enlarger with a varicontrol illuminator. However before you invest over 10K in such a device, you really should investigate the possibilities of a CCD camera upgrade for your scope. I was at the Microscopy and Microanalysis 2000 meeting this past summer in Philadelphia, and based on the many different vendors of such devices, it is obvious that the days of film are not unlimited. While a CCD camera and its related computer needs are expensive up front, the product they produce is very manuscript and grant application form friendly. For labs with space restrictions, a computer work station certainly takes up less square footage of floor space compared to a dark room and does not have to expose the operator and the environment to potentially hazardous chemistry that is associated with conventional dark room operations. The down side of CCD imaging is the upfront cost and not quite as high resolution compared to film. Tim
Timothy G. Schneider Director of Electron Microscopy Department of Pathology Room 229 Jefferson Hall Thomas Jefferson University 1020 Locust St. Philadelphia Pa. 19107 215-503-4798 work 610-613-8170 cellular timothy.schneider-at-mail.tju.edu
Speaking of which I have a self-righteous stage from a condemned stem free to a good home. Unfortunately it's contaminated with subliminable materials, but could be cleaned with a solution of vacuous politicians in HF. Replies by bush telegraph only please Chris } } Henceforth I promise never to use humor in this group and *only* } reply in terms of self-righteous outrage and stern condemnation. } } } billo } }
========================================== Dr. Chris Jeffree University of Edinburgh BIOSEM - Biological Sciences Electron Microscope Facility Institute of Cell and Molecular Biology Daniel Rutherford Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JH, Scotland, UK Tel. #44 (0) 131 650 5345 FAX. #44 (0) 131 650 6563 Mobile 07710 585 401 email c.jeffree-at-ed.ac.uk =========================================
We are presently purchasing several student polarizing microscopes, plus one which can be used with a digital camera for classroom presentation. We are initially considering the Coolpix 950/990 with its appropriate adapter on a 1X C-mount. During a demo yesterday, we were quite impressed with a Coolpix's NTSC output ... color as good as we've seen with more expensive cameras. However, we were a bit disappointed when we tried to grab a digital image. The camera would vignette much more than the video display would indicate (there was actually no vignetting with the NTSC out) ... although the color is quite accurate. But before I point you at examples, let me first explain we tried this demo with my personal CP800, believing we should get performance very similar to the CP950. The vignetting problem may be due to a feature setting available on the 950 I am not aware of. Point your browser at the URLs below ... the 1st is full image, so you can inspect the detail ... the 2nd will show you the vignetting.
As a starter, you need to get your hands on John Russ' Image Analysis Handbook to find out what stereology or Quantitative Microscopy can do for you. There are other books that you can find on the subject, but they are cited in his book.
I would suspect that you might be able to process your using an FFT to filter the periodic structure so that the grain boundaries are enhanced and then further enhance the grain boundaries so that you can quantify them. There are a couple of standard quantitative measurements that you can make at this point to characterize your microstructure. You can do the measurements by hand, but the computer is much easier. John Russ has a series of Plug-ins for doing these measurements and image enhancements in Photoshop and NIH image that are a companion to his book.
There are also a number of other commercial products out there that might be able to help you with this, but they would be more expensive. NIH Image is an alternative software package that is free and John's plug-ins also work there.
-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: Anjeanette Ormonde [mailto:ormonde-at-chem.nwu.edu] } Sent: Wednesday, October 25, 2000 10:27 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Suggestions needed/stereoscopy? } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.ht} ml } } } -------------------------------------------------------------- } ---------. } } } Hi, } I am a graduate student and I need some help trying to } figure out } a quantitative way to evaluate my samples before running my } experiments } with them. I am using a technique we call nanosphere } lithography by which } we deposit polystyrene spheres onto a substrate. We want a } single layer } of hexagonally packed spheres. My spheres are 400nm across } which means } the spaces between them are just under 100nm. We get this } packing in some } areas and can see a nice, dark blue color. However, just as } in crystal } growth we also see grain boundries and other defects where we } may have no } spheres or multiple layers of spheres. My substrates are 12 - 15 mm } square ITO coated glass. AFM is not a good solution because it takes } entirely too long to look at an entire sample and even then, } I only have } my observations to say which area is the best. Someone mentioned } stereology to me but I am unsure of what that technique } entails and what } information I can gain from it. I am looking for something } that will be } able to look at the entire sample and eventually give me some sort of } number such as the % that is perfect or even the % that is } defective. We } have an 80X objective but that isn't enough resolution to see the tiny } spheres in enough detail. However, I have access to some } other facilities } on campus but need to know what to look for. Does anyone have any } suggestions? Thanks for your time and any comments are appreciated. } } Angie Ormonde } } }
Does anybody know a good way to clean a dirty Faraday cage? (I know, I shouldn't have let it get dirty in the first place....:-() I'm thinking sonification in soapy water with lots of rinsing afterwards, but I wonder if there are any drawbacks to this. I'm hesitant to use solvents because I don't know what "glue" was used to attach the little aperture on top, and it was way too expensive to screw up that way. I tried a puff of air from one of those little rubber squeeze-bulb thingies (whatever the hell they're called) but that just didn't do the trick. I don't really know what the contamination is but it's partially transparent in a 20KeV beam. Could be anything but metal, I guess.....
F.C. Thomas MicroAnalysis Facility Bedford Institute of Oceanography Dartmouth, Nova Scotia Canada Geological Survey of Canada (Atlantic) Bedford Institute of Oceanography Dartmouth, Nova Scotia Canada B2Y 4A2
The enlargers made by Durst, Beseler and Omega have always been considered among the best. A condenser system for the light source and a point light source will give you the sharpest image. I have had several Durst enlargers because of their nice point light source. It helps to buy the best lens available. Schneider lenses have a good reputation (Apo Rodagon etc). The enlarger usually must be one designed for use with up to 4" X 5" negatives in order to acommodate the EM negatives. In the past I purchased special negative carrierers from a small company on the east coast named Carlwen. I recommend getting assistance from your local professional photography dealer. Larry D. Ackerman Lily & Yuh Nung Jan Laboratories Howard Hughes Medical Institute UCSF, Box 0725, Rm U226 533 Parnassus Ave. San Francisco, CA 94143
A huge thankyou to the many of you who responded to my query about D19b. Several people gave me the recipe, and one kind person even had a can of the stuff they were willing to send to me! I am a happy man.
e-mail richard.beanland-at-marconi.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 assume that you mean oxygen sensitive samples when you say air sensitive.
In that event you might try crushing the specimen in a glove box/bag purged with a continual flow of dry nitrogen gas. You can mount the specimen on the TEM grid and then the grid in the TEM holder all within the glove bag. Exposure to oxygen and atmospheric water vapor would be limited to the brief interval between removing the holder from the bag and inserting it into the microscope column. In fact, I have seen people do all this in a nitrogen purged plastic bag that also encloses the TEM's airlock to avoid oxygen exposure during transfer to the airlock.
This may be worth a try if you are concerned about Decanol evaporation in the TEM column.
} D.M. de Leeuw et al. Physica C 158 (1989) 391 prepared air and moisture
} sensitive samples for TEM by crushing them in a mortar under decanol and } putting a droplet of the suspension onto a grid. Decanol has low evaporation } rate and thus protects the sample. However, it evaporates inside microscope } in air-lock but probably also in the column. I would like to know if the } procedure is save for modern UHV microscopes and/or there are other } procedures that could be safely used for similar air sensitive samples.
} } Leszek Kepinski
-- _____________________________________________________ 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
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I am a graduate student and I need some help trying to figure out a quantitative way to evaluate my samples before running my experiments with them. I am using a technique we call nanosphere lithography by which we deposit polystyrene spheres onto a substrate. We want a single layer of hexagonally packed spheres. My spheres are 400nm across which means the spaces between them are just under 100nm. We get this packing in some areas and can see a nice, dark blue color. However, just as in crystal growth we also see grain boundries and other defects where we may have no spheres or multiple layers of spheres. My substrates are 12 - 15 mm square ITO coated glass. AFM is not a good solution because it takes entirely too long to look at an entire sample and even then, I only have my observations to say which area is the best. Someone mentioned stereology to me but I am unsure of what that technique entails and what information I can gain from it. I am looking for something that will be able to look at the entire sample and eventually give me some sort of number such as the % that is perfect or even the % that is defective. We have an 80X objective but that isn't enough resolution to see the tiny spheres in enough detail. However, I have access to some other facilities on campus but need to know what to look for. Does anyone have any suggestions? Thanks for your time and any comments are appreciated.
Dear Angie, Here are a couple of suggestions. I don't know how practical they are, but they may suggest more practical possibilities. To get the average number of spheres per unit area, you could try to measure the fraction of incident light absorbed. Illuminate the specimen with a wavelength of light which is at the absorption max, and either collect all the light which is not absorbed (including scattered light), or measure the energy absorbed with a calorimeter. The latter does not require collecting the scattered light, but may have other problems, such as accounting for heat re-radiated. You would need to determine the absorption of one sphere by calculating it or by measurement of a dilute suspension of spheres in a medium which does not absorb light at the wavelength selected and which has the same index of refraction as the spheres. To get a measurement of the packing, you could try to measure the diffraction of a parallel beam of light incident on the specimen. This time, the spheres should not absorb at the wavelength selected. You could calculate the expected intensity and compare this with what is observed. The spread of the diffraction spots will give a measurement of the regularity of the crystal, and the appearance of two or more distinct hexagonal patterns will tell you that there is more than one patch being illuminated, and that the patches are about the same size as the illuminating spot--assuming that the spot is not on the boundary between two large patches (which is easily determined). The appearance of rings or circles of many spots will indicate that many small patches are being illuminated, and other possible patterns can be interpreted. As I said, there are no guarrantees that either of these measurements are useful, but good luck. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
Hello everyone, I was wondering if anyone had suggestions on a protocol for the fixation of either newt or squid eye to be viewed in the SEM. I will be viewing a crossection of the eye. Would it be best to fix after cutting the eye, or try to perform a frozen fracture after fixation and dehydration. Any suggestions will be appreciated. Thanks, Thomas
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Sorry I don't know the answer to your quaestion, but I just can't stop myself from asking:-
- How do you know it's dirty? - How would you have prevented it from getting dirty? - Does it matter, anyway?
cheers
rtch
} } Does anybody know a good way to clean a dirty Faraday cage? (I } know, I } shouldn't have let it get dirty in the first place....:-() I'm } thinking sonification in soapy water with lots of rinsing } afterwards, but I wonder if there are any drawbacks to this. I'm } hesitant to use solvents because I don't know what "glue" was used } to attach the little aperture on top, and it was way too expensive } to screw up that way. I tried a puff of air from one of those little } rubber squeeze-bulb thingies (whatever the hell they're called) but } that just didn't do the trick. } I don't really know what the contamination is but it's partially } transparent in a 20KeV beam. Could be anything but metal, I } guess.....
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
} To: John Heckman {heckman-at-pilot.msu.edu} } From: Gary Gaugler {gary-at-gaugler.com} } Subject: D-19 } } Kodak still makes D-19. Part numbers are: } } 1464593 1 gallon } 1946045 5 gallons } } gary g.
Have you tried SEM? The size should be easily resolvable in any SEM. You may have to coat your sample to avoid charging.
Then take your images and run some FFTs on them. That should give you information about packing and also faults in the packing. For a quantitative analysis you probably need to design some filters in Fourierspace, and do a back transform, then analyze those images.
There are a bunch of books out there that explain things in detail. John Russ' book comes to mind.
Good luck.
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: Anjeanette Ormonde [mailto:ormonde-at-chem.nwu.edu] Sent: Wednesday, October 25, 2000 8:27 AM To: Microscopy-at-sparc5.microscopy.com
Hi, I am a graduate student and I need some help trying to figure out a quantitative way to evaluate my samples before running my experiments with them. I am using a technique we call nanosphere lithography by which we deposit polystyrene spheres onto a substrate. We want a single layer of hexagonally packed spheres. My spheres are 400nm across which means the spaces between them are just under 100nm. We get this packing in some areas and can see a nice, dark blue color. However, just as in crystal growth we also see grain boundries and other defects where we may have no spheres or multiple layers of spheres. My substrates are 12 - 15 mm square ITO coated glass. AFM is not a good solution because it takes entirely too long to look at an entire sample and even then, I only have my observations to say which area is the best. Someone mentioned stereology to me but I am unsure of what that technique entails and what information I can gain from it. I am looking for something that will be able to look at the entire sample and eventually give me some sort of number such as the % that is perfect or even the % that is defective. We have an 80X objective but that isn't enough resolution to see the tiny spheres in enough detail. However, I have access to some other facilities on campus but need to know what to look for. Does anyone have any suggestions? Thanks for your time and any comments are appreciated.
I am not familiar with the CoolPix cameras and the NTSC out function. Do they have that available right off the camera? Is your video monitor overscanning? Many do so that you do not see the entire height or width of the image. If so, the vignetting would not appear too bad until you grabbed the image and saw it all. Some monitors include an underscan switch so you can see the whole image.
I suppose someone (Barbara Foster maybe) is going to weigh in on the subject of vignetting in general. It seems that there have been several discussions on this a few months back. I thought the basic problem was using a transfer lens of too low a power for the camera. I have the opposite problem with our Pixera (or any other 1/3" camera). We are using a 0.5x transfer lens. A 0.33x would be best to match the field of view and the picture, but they were quite a bit more than the 0.5x lenses.
Warren S.
At 07:43 AM 10/25/2000 -0700, you wrote:
} We are presently purchasing several student polarizing } microscopes, plus one which can be used with a digital camera for } classroom presentation. We are initially considering the Coolpix } 950/990 with its appropriate adapter on a 1X C-mount. } During a demo yesterday, we were quite impressed with a Coolpix's } NTSC output ... color as good as we've seen with more expensive } cameras. However, we were a bit disappointed when we tried to grab a } digital image. The camera would vignette much more than the video } display would indicate (there was actually no vignetting with the NTSC } out) ... although the color is quite accurate. But before I point you } at examples, let me first explain we tried this demo with my personal } CP800, believing we should get performance very similar to the CP950. } The vignetting problem may be due to a feature setting available on } the 950 I am not aware of. } Point your browser at the URLs below ... the 1st is full image, so } you can inspect the detail ... the 2nd will show you the vignetting. } } http://darkwing.uoregon.edu/~mshaf/dogsci/x-biot.jpg } http://darkwing.uoregon.edu/~mshaf/dogsci/x-biot-sm.jpg } } 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/
We just received our Fuji FinePix Pro S1 camera. Beautiful instrument!!
I have not had luck interfacing it the a light microscope since the camera stubornly insists on having an autoNikkor lens attached to it. The usual microscope adapters cause the camera to be uncommunicative with the computer.
I KNOW that it can be interfaced with a LM and computer because I saw it in action at M&M2000.
So, does anyone know how to do this?
Several people are holding their breaths on this one.....
John B. -- #################################################################### John J. Bozzola, Ph.D. Professor & Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
I just had a demo yesterday of the 990 on a Leica Orthoplan II. The vignetting you showed was not present on our demo. Perhaps the zoom was set wrong or the wrong type of relay lens was used. I did see a slight amt of vignetting but it was minor--nothing like what you showed.
We are definitely going to buy the 990 along with relay lens, wired remote control and AC adapter. We got a price of $1864.
I really liked the software too.
JB
-- #################################################################### John J. Bozzola, Ph.D. Professor & Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
I expect that Faraday cage is actually a F cup - since it has an aperture on top. The F cage attracts low energy electrons towards the scintillator and the cup is of course used to trap electrons and measure their current with a picoampmeter for analysis. We made our own when turning an SEM into a Probe about 20 years ago. Used were an aperture holder, with a bit of machined carbon rod at the end and this was capped with a 50um aperture. The carbon/aperture cup had to be insulated from the holder. A fine wire and the aperture were held in place with a bit of carbon dag. Worked well, though insertion was not nearly as convenient as currently used pneumatic F cups.
The relevance is that there is nothing "magical" about that device. The cup most only conduct through the wire and meter to ground, but the cup must be electrically conducting. I would not hesitate to remove the aperture, clean this properly and replace it. A couple of small spots of dag would hold it in place. Contamination could cause the insulating layer around the cup to conduct and this would make the cup's reading useless. If you can measure with a good meter any continuity between cup and holder itself, then you may need to replace the insulating layer, which I expect is a bit of plastic sleeving; you should be able to improvise something and that may be easier than cleaning that plastic. 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 ABN: 99 724 136 560 www.proscitech.com
On Thursday, October 26, 2000 2:26 AM, Frank Thomas [SMTP:thomasf-at-AGC.BIO.NS.CA] wrote: } } Listers - } } Does anybody know a good way to clean a dirty Faraday cage? (I know, I } shouldn't have let it get dirty in the first place....:-() I'm thinking } sonification in soapy water with lots of rinsing afterwards, but I wonder } if there are any drawbacks to this. I'm hesitant to use solvents because I } don't know what "glue" was used to attach the little aperture on top, and } it was way too expensive to screw up that way. I tried a puff of air from } one of those little rubber squeeze-bulb thingies (whatever the hell they're } called) but that just didn't do the trick. } I don't really know what the contamination is but it's partially } transparent in a 20KeV beam. Could be anything but metal, I guess..... } } F.C. Thomas } MicroAnalysis Facility } Bedford Institute of Oceanography } Dartmouth, Nova Scotia } Canada } Geological Survey of Canada (Atlantic) } Bedford Institute of Oceanography } Dartmouth, Nova Scotia } Canada B2Y 4A2
In a message dated 10/24/00 6:05:02 PM, mb-at-Soft-Imaging.com writes:
} I don't want to put down Photoshop, especially since we are producing a } competing product, but I think this remark points to one problem that many } "photo" applications have -- they do not treat the intensity as scientific } data. I may be mistaken, but I think Photoshop is conceptually a } "beautifying" software to make photos "look nicer" -- and it does a great } job at this, but it may also be lacking in certain areas.
Michael is partly right - Photoshop does not include native capabilities for calibration of intensity values for optical density, or the ability to directly measure hue in color images, or to calibrate spatial distances in microns, etc. It also has very limited image measurement capabilities. But it has many advantages (particularly ease of use, consistency on various computer platforms, a huge base of users so that few bugs survive, etc.) and all of the other features that other responders to the original question have pointed out, plus it is relatively inexpensive (particularly if you can take advantage of the academic price). Many microscopists already have it, as well. And the shortcomings are easily filled in using plug-ins. Those are the reasons that my son and I decided to use it as the platform for writing plug-ins to do scientific imaging. With the layers and actions capability in the most recent versions (5.0 and 6.0), and the Tool Kit or Fovea plug-in package (no Nestor, I won't make any further plug for the product - others have already mentioned it anyway), the capabilities for scientific quality image processing actually exceed those in many of the costly "commercial" packages.
I had a similar problem on my metallurgical microscope when I first installed a camera to it, at 25X and 50X
it would vignette but at higher magnifications it wouldn't . Kodak suggested a higher C-mount magnification and
that took care of it.
This info is from the Kodak site:
"3.What is a C-Mount and why does it need to be 1.0X? To accommodate the need for photo documentation, many microscopes feature an additional photo port or tube that delivers an optical path to a camera. It is the C-Mount that adapts the microscope to the KODAK MDS Universal Optical Adapter and MDS 290 digital camera. C-Mounts of less than 1.0X magnification may cause images to vignette (darkening around the edges of the image), and are therefore not recommended. "
I'm one of the breath holders, as I'm currently waiting for my microscope adapter to arrive. Got the camera and other goodies, but the adapter is on back order. As I read the book, it seems you can't use all the automatic bells and whistles unless you have a Nikon type D lens. Perhaps you need to switch to aperture priority or shutter priority mode? Anyone else have any ideas?
David Bell Scientist Electron Microscopy Lab Millipore Corporation 80 Ashby Road Bedford, MA 01730 (781) 533-2108
"John J. Bozzola" To: microscopy-at-sparc5.microscopy.com {bozzola-at-siu. cc: edu} Subject: Fuji S1 digital cams on LMs
10/25/00 06:28 PM
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We just received our Fuji FinePix Pro S1 camera. Beautiful instrument!!
I have not had luck interfacing it the a light microscope since the camera stubornly insists on having an autoNikkor lens attached to it. The usual microscope adapters cause the camera to be uncommunicative with the computer.
I KNOW that it can be interfaced with a LM and computer because I saw it in action at M&M2000.
So, does anyone know how to do this?
Several people are holding their breaths on this one.....
John B. -- #################################################################### John J. Bozzola, Ph.D. Professor & Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
At 07:43 AM 10/25/00 -0700, michael shaffer wrote: } However, we were a bit disappointed when we tried to grab a } digital image. The camera would vignette much more than the video } display would indicate (there was actually no vignetting with the NTSC } out) ... although the color is quite accurate.
In your second image, in the lower right corner for example, I can see what I'd call true vignette, the sharp edge of the lens tube. I'd guess you do see that small dark area in the NTSC-out view. The rest of the ring of darkness could be the drop-off in the light intensity of the microscope's output, and perhaps an automatic exposure only properly exposed the center of the view. Were you using manual or automatic exposure settings? Did you try both methods? Where was the zoom set?
I recently purchased an Olympus C-2500L. This is a true SLR digital camera, roughly $1,000. I had high hopes of using it in an afocal coupling manner with my Ortholux and a Dobsonian telescope. In other words, I just wanted to point the camera into the eyepiece and get a good view. There are also ~$150 adapters that will hold the camera in place in these situations, like http://www.photosolve.com/xtendascope.asp .
I was very disappointed to learn that I should've bought a less expensive camera non-SLR for this purpose - although I've heard the Coolpix 8xx/9xx are suitable and otherwise nice, too. Those with non-SLR lenses are much better at this sort of afocal coupling. With the C-2500L, the front lens element doesn't move when I zoom/tele. I can't avoid vignette-ing at all.
A 43 mm Plossl eyepiece might help on the telescope, and I haven't had time to figure out which sort of microscope eyepiece might help. I have other archived e-mails from a few experts that I'd be glad to forward to anyone interested.
Good morning fellow listers, we have a patron that we are helping find a set of RCA EMU3-F manuals and prints. any info leading to that would be great. also if I could have the info on the folks that redo the florcesent screen i will pass that along too ... thanks Ed Sharpe archivist for SMECC
We have a home-grown program that we wrote for the purpose of translating batches of files into 8-bit TIFF. We then use other utilities to convert those TIFF files to JPEG.
I use it only occasionally, and it may still have some bugs in it. But let me check on its status and I can probably get it to you with some instructions.
At 09:38 PM 10/24/2000 +0200, you wrote:
} Does anyone know of software that will do a batch conversion of the } .IM_ image files captured using Link ISIS v. 3 Autobeam, to a standard } format (eg. pcx, jpg, etc)? } } Fiona } ____________________ } Dr Fiona Graham } EM Unit, } University of Natal, Durban } South Africa
---------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
I just got off the phone with the Fuji representative (thank you Gary) who worked with me to solve the interface problem with the Fuji S1 camera. Turns out that the software had to be set up properly. Now the system works without a hitch. The images are FANTASTIC.
We had originally purchased the camera for copystand, macro photography but I suspect that the S1 will be spending most of its life on the light microscope from this point on. It is supposed to work even on fluorescent images -- so I am looking forward to checking that out too.
Thanks also to Matt Irwin from ElectroImage who called and also helped point me in the right direction.
This camera is really wonderful!!!!
JB -- #################################################################### John J. Bozzola, Ph.D. Professor & Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
I have a Zeiss EM 9S 2, vintage 1975, which I would like to keep running for as long as possible. Parts are very difficult to obtain. If any of you are considering discarding one of these instruments, please get in touch with me. I'd be happy to take it off your hands. Thanks.
Good morning everyone, I' m looking for specimen preparation to Low-Vacuum SEM observation. I work with a Jeol, JSM-5600LV and I' d like to see lactic microorganisms and microalgae with that SEM. Does anyone Know "SEMpore" which is a registered mark of Jeol A. B. used for some authors to observe Cryptosporidium using a LV SEM? I' m hoping that someone can give me some advice to specimen preparation. Regards, Patrícia. _____________________________________________ Patrícia João Reis Escola Superior de Biotecnologia Rua Dr. António Bernardino de Almeida 4200 - 072 Porto Portugal Telef. +351 225580044 Fax. +351 22 5090351 E-mail: pmreis-at-morango.esb.ucp.pt
I am planning to use reflectance microscopy on live cells and I am looking for bathing solutions that can increase the reflectance from the cell membrane without harming the cells. Any suggestions ? Are there such products out there ? Thanks,
Isaac Bankman Johns Hopkins University Applied Physics Laboratory
FWIW: When IXRF images are exported to TIFF, the code allows a max array of 1024. Larger images are reduced to that on export. Sometimes a high resolution / low mag image is useful to have. I have had limited success using photoshop to read the native files. I change the file type to .raw then open w/photoshop. PShop will try to "guess" the header size. Seems to work well with 4Kx4K arrays, but fails with 2K square. Since I'm after an image allowing me to digitally zoom in and roam around, this is not sooo bad.
Woody White McDermott Technology, Inc Lynchburg Research Center
Me: http://home.att.net/~woody.white
{ {SNIP} } } } We have a home-grown program that we wrote for the purpose of } translating } batches of files into 8-bit TIFF. We then use other utilities } to convert } those TIFF files to JPEG.
I have very good luck cleaning things that go into electron optical instruments using Tiles Soap Scum Remover. This is a mixture of high powered detergents combined with a organic solvent (diethylene glycol butyl ether). I just scrub parts with this thoroughly, or treat them with it ultrasonically, then rinse them thoroughly with hot running water, and finish by rinsing with reagent grade isopropyl alcohol and blowing off the residual alcohol with a duster. I've been able to remove pump oils, machining oils, and even silicone grease in this way, and have had no problems when parts so cleaned are later put in a high vacuum system.
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 thought I'd get back to you all and let you know the problem of vignetting was not a question of "zoom" (although you cannot zoom out and not see true vignetting because of the tube). The problem was apparently my CP800 and its aperture adjustment for proper exposure.
Thanx to those who suggested I turn off the "flash" which sets a different aperture priority, ... and fixing the focus at infinity. I can now take photos similar to what was posted previously and not see vignetting ... 'cept very slightly which suggests I ought to increase the zoom only slightly, but might be corrected properly with the larger CP990's CCD.
However, I still experience similar vignetting for very bright images .. the CP800 will still close down the aperture for compensation. However, this occurs (for example) if I take picture with crossed polars, and then want the same image without crossed polars ... but I could fix this with a neutral density filter.
I haven't yet had a chance to play with a CP950 or 990, but presumably these "better" cameras come with more features ... possibly a "aperture priority" mode which would lock-out the problem.
I cannot yet post images until I've had time to download to my home computer ... possibly next week.
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/
From root Fri Oct 27 18:58:46 2000 Return-Path: {Microscopy-request-at-sparc5.microscopy.com} Received: (from daemon-at-localhost) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id OAA10026 for dist-Microscopy; Fri, 27 Oct 2000 14:48:54 -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 OAA10022 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Fri, 27 Oct 2000 14:42:44 -0500 (CDT) Received: from srvr20.engin.umich.edu ([141.213.75.22]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id OAA10013 for {microscopy-at-sparc5.microscopy.com} ; Fri, 27 Oct 2000 14:42:12 -0500 (CDT) Received: from [141.212.131.74] (dow2146amac74.engin.umich.edu [141.212.131.74]) by srvr20.engin.umich.edu (8.9.3/8.9.1) with ESMTP id PAA03915 for {microscopy-at-msa.microscopy.com} ; Fri, 27 Oct 2000 15:38:18 -0400 (EDT) X-Sender: bigelow-at-srvr5.engin.umich.edu Message-Id: {v03007803b61f7b42c740-at-[141.212.131.74]} Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
I have very good luck cleaning things that go into electron optical instruments using Tiles Soap Scum Remover. This is a mixture of high powered detergents combined with a organic solvent (diethylene glycol butyl ether). I just scrub parts with this thoroughly, or treat them with it ultrasonically, then rinse them thoroughly with hot running water, and finish by rinsing with reagent grade isopropyl alcohol and blowing off the residual alcohol with a duster. I've been able to remove pump oils, machining oils, and even silicone grease in this way, and have had no problems when parts so cleaned are later put in a high vacuum system.
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 next meeting of the Minnesota Microscopy Society will be on Thursday, November 9, 2000
The theme of the meetng is "Everything you wanted to know about SPM but were afraid to ask"; A talk to be given by Chuck Mooney, SPM Assistant Product Manager, JEOL USA, Inc.
Abstract: The first scanning probe microscope (SPM), the scanning tunneling microscope, was introduced to the world in 1981 as a frightfully complex laboratory experiment where conductors could be probed at the atomic scale. Five years later, the Nobel Prize in Physics was awarded to its inventors, even as the basic technique was expanded to include probing insulators with the invention of the atomic force microscope (AFM). Since that time, many new methods have been added to the general field of scanning probe microscopy that allow for the probing of all manner of samples and forces. Recent breakthroughs have allowed the measurement of the strength of bonds between atomic species as well as the potential for 3-D sub-surface imaging and spectroscopy of atomic species near the sample surface. Many research laboratories have added scanning probe microscopy techniques to their analytical arsenals aided by the introduction of commercially available instruments. Unfortunately, the strengths and weaknesses of SPM remain a mystery to many who could benefit from a better understanding of both the techniques that are routine and those that are difficult, the relative levels of information that can be extracted from those techniques, and the types of samples that are suitable for most SPMs. In the time allowed, an introduction to the various SPM techniques will be given, applications examples shown, as well as providing insight into the future of probe microscopy.
Location: Medtronic, Inc., 6700 Shingle Creek Parkway, Brooklyn Center
Dinner: New King Buffet, 5927 John Martin Drive, Brooklyn Center
The New King Buffet is a full menu Chinese buffet. Cost is $10.00 per person.
We will first meet at the New King Buffet for dinner. After dinner, we will reconvene at Medtronic for the business and technical meetings.
For reservations contact Mike Coscio by Friday, November 3: (763-514-1331, or mike.coscio-at-medtronic.com).
__________________ Stuart McKernan stuartm-at-tc.umn.edu 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
I'd like more CPD advice again. I just had a very bad run on our new Pelco CPD2--stuff came out visibly wet! And the o ring on the door is already stretched out of shape. It's the last straw for my supervisor, we're sending it back. But now we would like to know what to get. Does anyone have a Tousimis Samdri-type drier? How is it? Any other *users* with recommendations who want to tell us why they like their CPDs? Mail from company reps, while not solicited, will be nonetheless noted.
Thanks for the help from those who wrote last time, and thanks in advance to those with responses.
From root Fri Oct 27 19:04:09 2000 Return-Path: {Microscopy-request-at-sparc5.microscopy.com} Received: (from daemon-at-localhost) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id OAA10063 for dist-Microscopy; Fri, 27 Oct 2000 14:57:44 -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 OAA10059 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Fri, 27 Oct 2000 14:51:16 -0500 (CDT) Received: from darkwing.uoregon.edu (darkwing.uoregon.edu [128.223.142.13]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id OAA10052 for {Microscopy-at-sparc5.microscopy.com} ; Fri, 27 Oct 2000 14:51:00 -0500 (CDT) Received: from mshaf (shaf.uoregon.edu [128.223.94.20]) by darkwing.uoregon.edu (8.10.1/8.10.1) with SMTP id e9RJYN907892 for {Microscopy-at-sparc5.microscopy.com} ; Fri, 27 Oct 2000 12:34:23 -0700 (PDT)
I thought I'd get back to you all and let you know the problem of vignetting was not a question of "zoom" (although you cannot zoom out and not see true vignetting because of the tube). The problem was apparently my CP800 and its aperture adjustment for proper exposure.
Thanx to those who suggested I turn off the "flash" which sets a different aperture priority, ... and fixing the focus at infinity. I can now take photos similar to what was posted previously and not see vignetting ... 'cept very slightly which suggests I ought to increase the zoom only slightly, but might be corrected properly with the larger CP990's CCD.
However, I still experience similar vignetting for very bright images .. the CP800 will still close down the aperture for compensation. However, this occurs (for example) if I take picture with crossed polars, and then want the same image without crossed polars ... but I could fix this with a neutral density filter.
I haven't yet had a chance to play with a CP950 or 990, but presumably these "better" cameras come with more features ... possibly a "aperture priority" mode which would lock-out the problem.
I cannot yet post images until I've had time to download to my home computer ... possibly next week.
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/
Al Coritz Sales & Service Manager RMC-Boeckeler Instruments 4650 S. Butterfield Dr. Tucson, AZ 85714 Voice: 520-745-0001 Cell: 520-465-3598 Fax: 520-745-0004 Email:Al-at-Boeckeler.com Website:RMCProducts.com
Before damning the Pelco, are you certain your methods are good? I do not know how experienced you are with CPD, so forgive me if you have thought of the following already. However, CPDs are very simple systems and most problems arise from operator errors, or failure to understand the basic principles
If the specimens are visibly wet then they are, presumably, wet either with the transition fluid or water - What fluid are you using? amyl acetate, acetone, ethanol, or something else? Can you smell it? Is the wetness of the specimen with the transition fluid or with water? If water, the specimens are not properly solvent-dehydrated. If transitional solvent, the sovent has not been removed by the CO2
The immersion in liquid CO2 should very quickly remove the transition fluid unless
1) the specimens were not fully immersed in liquid CO2, but only in gas - you must either use the dip-tube type of CO2 cylinder, or invert a non-dip type cylinder to obtain a supply of liquid CO2 to the CPD chamber. Remember that the dip-tube type must actually contain a liquid level - for some time after the available liquid is exhausted the cylinder will continue to supply gas, but you cannot do CPD with CO2 gas
2) insufficient liquid CO2 flushing took place, so some of the transition fluid remained in the chamber: flush for longer, or for short bursts with soak-time between
3) The specimens were very large, and insufficient CO2 rinsing time was allowed - see 2
4) The specimens were still wet with water: use more solvent-drying time with drier solvents
I think I am right in saying that the dehydration and transition fluid stages are where your problems should be sought, and that errors of method or hardware faults in or leading up to the actual critical point stage would not lead to wet specimens if the earlier procedures were correctly followed.
Chris
} Dear listers,
{color} {param} 0000,0000,0000 {/param} }
} I'd like more CPD advice again. I just had a very bad run on our new Pelco
} CPD2--stuff came out visibly wet! And the o ring on the door is
} already stretched out of shape. It's the last straw for my supervisor,
} we're sending it back. But now we would like to know what to get. Does
} anyone have a Tousimis Samdri-type drier? How is it? Any other *users*
} with recommendations who want to tell us why they like their CPDs? Mail
} from company reps, while not solicited, will be nonetheless noted.
}
} Thanks for the help from those who wrote last time, and thanks in advance
} to those with responses.
}
} Pauline Yu
} splene-at-pw.usda.gov
} Microscopist Technician
} USDA-ARS-WRRC
}
}
{nofill} ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 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
Pauline, I agree with Chris Jeffree on two important points: 1. use bone-dry liquid carbon dioxide with a dip tube 2. use several steps of 100% ethanol (I recommend that all using our older Polaron E3000 use an unopened pint bottle of 100% ethanol in the last two ethanol steps. This is esp. important during the summer months when it is humid here) and would add the following tips --when dehydrating plant samples esp. vegetative or reproductive meristems, leave samples overnight in the third 100% ethanol and change again the next day before transferring to the CPD --when dehydrating drosophila or other whole insects, add several overnights in 100% ethanol --after filling the pressure chamber with the liquid carbon dioxide, check to see a meniscus, an indication that you have liquid carbon dioxide (for safe viewing, it's recommended that one do this on our system by directing a flashlight into the chamber while viewing the chamber via a mirror placed opposite to the chamber window) -- if the temperature of your chamber is higher than 19C, the carbon dioxide will fill the chamber and the pressure gauge will read 800psi but what you may have is vapor --use a flushing step just after filling with liquid carbon dioxide, i.e., with the inlet open, open the drain valve and monitor the drain line (at first, you'll see liquid (ethanol), a slush (ethanol/CO2) and then white powder (dry ice) -- many of our users place samples in the polypropylene specimen carriers which hold the ethanol when capped and we find it advantageous to use this step each time so that our subsequent exchanges are replacing the ethanol within tissues or organisms --any problems we had with gaskets were solved by purchasing those supplied by Energy Beam Sciences. Rosemary
Try staying simple. We have used two Polaron E3000 CP Dryers for at least 25 years and they have been very reliable. Some gaskets and seals have had to be changed but other than that we have had no problems. They do not have the automated features that some of the others have but they produce good results.
I think they are sold in the US by SPI, and perhaps other vendors too.
Good luck.
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 **
Right now Stormfront Studios is adding to its staff and looking for people to create Next Gen Games for the X-BOX and PS2. We're looking for an Entry Programmer, AI Programmer, Sr. Programmer, Lead Programmer, Development Director, Sr. 3D Max Artist, 3D Animator and/or Art Director. Interested or know of anyone who may be interested? Please let me know if you can help us out - we need your talent!
Thanks, Marta Daglow EMAIL: mdaglow-at-earthlink.net WEB: www.stormfront.com San Rafael, CA 94903
Dear colleagues, I would like to carry out experiment of irradiating an object with electrons in presence of gas media (about 1 torr) and intend to use CamScan4 SEM for this purpose. This device doesn't contain low vacuum accessories, so I hope to make them by myself. Could you tell me which are the tipical holes sizes in the differential pumping apertures, the distance between them and expected pressure increasing in diff. pump entrance. Thanks in advance. Eugeny Jikharev, Physics & Technology Institute of Russian Academy of Sciences, Moscow.
I am looking for some TEM negative film holders for Philips/FEI microscopes. These are holders with the format 3,25 x 4’’ and have two components. The holder itself and the insertion (for loading film and imaging plates). Placing an order in Germany it takes 6 months to get them – quite a long time. Is anybody out there who knows an other source to get the mentioned holders faster or has someone any used material available ?
Ruediger Kuenzler -- DIBIS Digital Biomedical Imaging Systems AG Gewerbestraße 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
Our laboratory is looking for an assistant to help prepare grants and publications. Below is the text of an announcement written for our HR department. In the past this position has been held by recent college graduates preparing applications for medical or graduate school or by trained research personnel interested in re-entering the workforce. -Brandon Poe, Ph.D.
===
We are looking for someone to fill a full- or part-time, temporary job, helping our neuroscience research laboratory to prepare reports on our research for publications and grant applications. This will require using image analysis software (e.g. Adobe Photoshop) and some darkroom work. Learning specific skills on the job is acceptable, but some familiarity with computers and illustration techniques will be preferred. We also need help with literature searches (w/ Ovid & Medline) and running a computer-based bibliography program (i.e. Papyrus). For this work, a background in college-level biology would be preferred. Time permitting, participation in quantitative data analyses would be an option, and a familiarity with statistical methods would be desirable. This job is available immediately for a period of 6-9 months. It would be suitable for a part-time college science major, a college graduate preparing for professional school or for a return to the job market after interruption of a career in science.
Contact: D. Kent Morest, M.D. Professor, Department of Neuroscience Director, Center for Neurological Sciences The University of Connecticut Health Center 263 Farmington Avenue Farmington, CT 06030-3401 tel: 860-679-2645 fax: 860-679-8766 email: kent-at-neuron.uchc.edu web: http//www3.uchc.edu/~kent
-- Brandon H. Poe, Ph.D. Department of Neuroscience University of Connecticut Health Center Farmington, CT 06030 email: bpoe-at-ravenbrand.com
} Pauline, Here's my 2-cents.... } I just had a very bad run on our new Pelco CPD2--stuff came out visibly wet!
} } } } } } } It sounds as though you had not fully purged the transitional fluid } } } } } } } and/or you did not have a sufficient liquid CO2 level before you } } } } } } } heated.
And the o ring on the door is already stretched out of shape.
} } } } This sounds like you are getting the O-ring wet with your transitional } } } } fluid (ethanol?) when you are loading the chamber.
These problems will persist unless care is taken when loading samples and doing the "run". CPD is not something you can do well without paying close attention to each step no matter whose unit you use.
As a "veteran" of some of these same problems, with a different brand unit than yours, I speak from painful experience!
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 have put images up on my wwwsite for your examination. Again, the Coolpix 800 was mounted on a 1X C-mount with the Nikon CP adapter.
A full-size crossed polar image of biotite which exhibits only minor vignetting: http://darkwing.uoregon.edu/~mshaf/dogsci/biotite-x.jpg
... the same image reduced in size: http://darkwing.uoregon.edu/~mshaf/dogsci/biotite-x-s.jpg
... the same biotite image ... uncrossed polars (no analyzer) ... which exhibits the problem vignetting because the CP800 stopped down its aperture: http://darkwing.uoregon.edu/~mshaf/dogsci/biotite-nx-s.jpg
A interference figure (calcite): http://darkwing.uoregon.edu/~mshaf/dogsci/bertrand.jpg
I had previously implied the CP800 would be a very low cost alternative for putting a "pro-sumer" digital camera on a microscope ...~US$800. Apparently I was wrong .. only under some lighting conditions will it work correctly. You do need one of the CP9xx series and use their "aperture priority" mode in most cases of illumination.
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/
Pauline - A user had a similar problem on a recent run using our stalwart Tousimis Samdri unit that has been in use for at least 20 years. The problem was due to an operator error - the purge valve was accidentally left closed so that the CO2 flow was too little or non-existent. The next run was perfect.
Good luck. 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
Sorry to bother the list over this, but I received an email from Steve Poe, USDA, APHIS, responding to a question posted to the microscopy list. I tried to reply to the address given in his message:
Steve.R.Poe%aphisn1-at-hq.usda.gov
but my message was returned with a "user unknown" error.
If anyone has an email address for Steve other than the one above, please let me know.
Thanks, Ed
Ed Bachmann Odum Institute for Research in Social Science Manning Hall CB 3355 University of North Carolina at Chapel Hill Chapel Hill, NC 27599-3355 (919) 962-0512
Thanks for the replies to my question about thin films grown on PMMA. My original attempts to do cross sections in the PIPs have not yielded much success. Mainly because my samples are highly allergic to both heat and acetone so normal things like low temperature wax on the dimpler are out ... my next plan was to use the microtome.
However, my attempts at reviving an old (1986) unused (no one here knows how long ago) Sorvall Microtome have been for naught! I finally located (I think) all the blown fuses, burnt out light bulbs and loose wires, but I don't think the cutting arm is retracting after the down stroke. The block face seems to be rubbing against the back of the knife on the return (upward) stroke and dragging water and cut sections from the boat up with it.
Also, all my attempts at locating the Sorvall Company have not proven fruitful. I get all kinds of web addresses, all in German (not my native tongue) and they all take me to the same error message (Maybe this whole project is just not supposed to happen). So if any one can help with an good address or a service number for Sorvall, I would say prayers in your name to the electron gods!
Does anyone know if there is available for the JEOL 840/733 spectros a multilayer which covers O to P, same as the TAP?
And about how much they cost?
I'm having some difficulty getting any real info from Osmic.
Are there any other suppliers?
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
} Also, all my attempts at locating the Sorvall Company have not proven } fruitful. I get all kinds of web addresses, all in German (not my native } tongue) and they all take me to the same error message (Maybe this whole } project is just not supposed to happen). So if any one can help with an } good address or a service number for Sorvall, I would say prayers in your } name to the electron gods! } } Thanks in advance for all your help. } Dorrance.
*************** Hi Dorrance,
Sorvall was bought years ago by a company called RMC (Research Manufacturing Company), which has just become part of Boeckeler Instruments in Tucson, AZ.
Boeckeler Instruments, Inc. 4650 South Butterfield Drive Tucson, AZ 85714
Tel: (520) 745-0001 Fax: (520) 745-0004
email: info-at-boeckeler.com
I'm not sure that they can help you much, since RMC stopped supporting many of the older instruments (Parts are no longer available). Give them a call though, you never know....
Lee
(I have no financial interest in Sorvall, RMC or Boeckeler)
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 Colleagues: I have made great contacts through this listserver in the past and hope that I can again this time. I am in search of a postdoctoral fellow to continue a program of research using modern optical imaging methods to follow membrane trafficking in vascular endothelial cells. It is a very exciting project that cuts across the fields of cell biology, immunology, and pathology, and uses techniques from standard transmission EM to tracking GFP-labeled proteins. If you have the right background and are interested, or know of a colleague who fits these criteria, please get in touch with me. The following is the text of an ad that appeared in the August 10 issue of Nature and the September issue of Nature Cell Biology:
Postdoctoral position in Membrane Trafficking Postdoctoral position open to study membrane trafficking in endothelial cells. The ideal candidate would have experience studying regulated membrane trafficking at the cellular and molecular levels using biochemical, microscopic, and molecular biology approaches. Experience with modern imaging systems is critical. Experience with endothelial cells is a definite "plus", but not necessary. The candidate should demonstrate excellent communication skills in English, be able to work independently, as well as to interact with a lively group of productive investigators studying leukocyte-endothelial cell interactions. We are based at the Weill Medical College of Cornell University in the nicest and safest area of New York City. We interact extensively with investigators Weill as well as at The Rockefeller University and Memorial Sloan-Kettering Cancer Center, which are both just across the street. The position is fully-funded and available immediately.
Submit CV and names (and contact numbers) of three references to:
William A. Muller, MD, PhD Department of Pathology and Program in Immunology Box 69 Weill Medical College of Cornell University 1300 York Avenue New York, NY 10021 e-mail: wamuller-at-med.cornell.edu
Thank you very much for your help.
Sincerely, Bill Muller
William A. Muller, MD, PhD Associate Professor of Pathology Weill Medical College of Cornell University 1300 York Avenue New York, NY 10021
Why do you not try to section your thin film samples in a cryostat, we have manufactured and supplied many for this application with great success.
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: McLean, Dorrance [mailto:dmclea-at-sandia.gov] Sent: 30 October 2000 20:23 To: 'microscopy'
Dear All,
Thanks for the replies to my question about thin films grown on PMMA. My original attempts to do cross sections in the PIPs have not yielded much success. Mainly because my samples are highly allergic to both heat and acetone so normal things like low temperature wax on the dimpler are out ... my next plan was to use the microtome.
However, my attempts at reviving an old (1986) unused (no one here knows how long ago) Sorvall Microtome have been for naught! I finally located (I think) all the blown fuses, burnt out light bulbs and loose wires, but I don't think the cutting arm is retracting after the down stroke. The block face seems to be rubbing against the back of the knife on the return (upward) stroke and dragging water and cut sections from the boat up with it.
Also, all my attempts at locating the Sorvall Company have not proven fruitful. I get all kinds of web addresses, all in German (not my native tongue) and they all take me to the same error message (Maybe this whole project is just not supposed to happen). So if any one can help with an good address or a service number for Sorvall, I would say prayers in your name to the electron gods!
Dear Evgeny, 1. Could you please tell more precise what signals you are going to use and what kind of objects you are going to investigate? 2. It depends on your vac system (with or without IGP, number of pirani gauges) but you will have to change some thresholds in the VAC electronics. 3.If you intend to insert the aperture in the objective lens (typical size 600-800micron) and use a BSD as a detector you should change: READY threshold in a high vac measuring (gray box behind the column) PREVAC thresholds in the VAC PCB 4. Anyway I wouldn't recommend to do this way because you will get problems with vacuum system - it's very sensitive.
Good luck,
Dmitri
Evgeny Zhikharev 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 would like to carry out experiment of irradiating an object with } electrons in presence of gas media (about 1 torr) and intend to use } CamScan4 SEM for } this purpose. This device doesn't contain low vacuum accessories, so I } hope to make them by myself. Could you tell me } which are the tipical holes sizes in the differential pumping apertures, } the distance between them and expected pressure increasing in } diff. pump entrance. } Thanks in advance. } Eugeny Jikharev, } Physics & Technology Institute of Russian Academy of Sciences, } Moscow. } }
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 } -----------------------------------------------------------------------. } } Hi, Listers } } Does anyone know if there is available for the JEOL 840/733 spectros } a multilayer which covers O to P, same as the TAP? } } And about how much they cost? } } I'm having some difficulty getting any real info from Osmic. } } Are there any other suppliers? } } 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 Ritchie, Try Jim Nicholino at
JNicolin-at-xraydetectors.com
His regular crystals are terrific and I imagine if he doesn't supply multilayer crystals, he knows who does.
I am looking for references (current if possible) for thermal imaging using a thin insulating layer or other thermal imaging techniques that have a minimum of one micron spatial resolution. I am currently working with ridge lasers and would like to characterize the temperatures at the facet and along the ridge. Any help would be appreciated.
I am also looking for a beginner to advanced text on EBIC imaging/contrast mechanisms.
Yes, you can process an emulsion by freezing the sample, fracturing it and viewing it it by cryoSEM (Low Temperature SEM). The sample is kept frozen on a cold stage.
Dave
On Tue, 31 Oct 2000 08:31:14 +0100 (WAT) "O. O. Ilori" {sojilori-at-oauife.edu.ng} 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 } -----------------------------------------------------------------------. } } } Is it possible to view an emulsion under a SEM? By emulsion I mean } particles suspended in a matrix. } If possible, how? } } } Mr. O. O. ILORI } DEPARTMENT OF ELECTRONIC/ELECTRICAL ENGINEERING } OBAFEMI AWOLOWO UNIVERSITY, } ILE-IFE, OSUN STATE } NIGERIA. } } email: sojilori-at-oauife.edu.ng } } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
Long time no hear from me. Did you miss me? I am now on the East Coast in Washington, DC and am looking for someone to be a EM tech. with me. Here is the job announcement:
TEM position available at George Washington University. The Department of Pathology is hiring a TEM technician. Must have experience with embedding, thick and thin sectioning, scoping and darkroom work. Immunochemistry experience a plus. Bachelor's degree and 2 years experience required. Excellent tuition benefits available. If interested please check the GW human resources web site at: www.gwu.edu/cgi-bin/hrs/vacancies Position title= Electron Microscope Technologist 2 Or contact me at patpxs-at-gwumc.edu or 202-994-2930
This is a clinical laboratory and we work mostly on clinical samples, though we do have some research samples come through (mostly HIV and AIDS related). The lab is pretty low key, but we do have a high standards.
George Washington University is smack dab in the heart of downtown Washington, DC. Walk a few blocks and you're at the White House (my friend even got to pet Buddy the Prez's dog), the mall with all the monuments and all sorts of other cool stuff.
The tuition benefit basically means you can take courses at GW for close to free (pay a reg. fee and buy books). Spouses and kids are eligible for a more limited benefit. Salary commensurate with experience, yada, yada, yada.
So if you're interested, please contact me.
Paula Sicurello patpxs-at-gwumc.edu 202-994-2930 (phone) 202-994-2518 (fax)
I have semi-thin sections of a sample embedded in methacrylate resin containing alginate and flour among other things. Could anyone give me advice how to stain the alginate for light microscopy?
a word of caution - unless GREAT care is taken in the freezing process there will be significant production of artifactual structure.... i.e. just like freezing biological tissue. See any of the many freeze fracture/freeze etch TEM references.
Bill Miller
At 09:19 AM 10/31/00, Patton, David wrote: } Microscopy-at-sparc5.microscopy.com
I spoke or wrote too soon regarding a source for Sorvall microtome parts. The following company took over the Sorvall product line from Ventana/RMC:
Boeckeler Instruments, Inc. 4650 South Butterfield Dr. Tucson, AZ 85714 Ph: 520-745-0001
Again, I have no connection with Ventana or Boeckeler Instruments. I just want to fix our Sorvall microtomes.
Greg Lum EM Facility San Francisco State University Ph: 415/338-1345 Email: glum-at-sfsu.edu
Greg Lum Computer/Microscopy Consultant Electron Microscopy Lab Mgr Department of Biology College of Science & Engineering San Francisco State University Ph: 415/338-1339 Fax: 415/338-2295 EMail: glum-at-sfsu.edu
We are thinking of purchasing a Kodak EDAS 290 Electrophoresis system. Does anyone have any experience with this system ( pros & cons), or can recommend one that they are happy with.
We currently doing some EM work on rat cultured hippocampal cells. The purpose of this experiment is to visualize individual synaptic vesicles by transmission electron microscopy. The problem we had is that the resolution/contrast is not high enough to distinguish the synaptic vesicles.
The following is what we did, cells were cultured on coverslips, they were fixed in situ in 2% glutaraldehyde for 1 hr to 24 hrs, postfixed with 1% OsO4 for 1 hour, and en bloc stained with 2% aqueous uranyl acetate for 30 min. dehydrated in ethanol and embedded in Polybed812. 40-50 nm sections were stained by 5% uranyl acetate and lead citrate prior to EM observation.
I would appreciate it very much if someone would provide suggestions on section staining or cell processing.
Xinran
******************************************** Xinran Liu, M.D., Ph.D. Center for Basic Neuroscience The University of Texas Southwestern Medical Center at Dallas 6000 Harry Hines Blvd., NA4.214A Dallas, TX 75390-9111 Phone: (214) 648-1830 Fax: (214) 648-1801 E-mail: xinran.liu-at-utsouthwestern.edu
My amateur results with a microtome have been less than satisfactory. Embedding in paraffin has been tire some and not very successful. Sectioning raw or fix material has only worked on a very limited range of things.
I picked up an old A0 Microtome and a new knife that I believed used CO2 as a coolant. I can get CO2 but it is a hassle and I have to build a stage and delivery system for it since the stage is missing.
I am planning on building a simpler stage using copper on an insulating base and use propane or 1,1,1,2,tetrafloroethane. The tests I have done with tetrafloroethane look very promising. It reaches -62 f and is very easy to use. It is normally used to chill parts on circuit boards when looking for problems. It comes in a spray can with a plastic tube.
Modifying a propane torch is also an easy solution. I have worked with propane all my life and I am well aware of the dangers. Having to go outside to use it is one of the main reasons for using the tetrafloroethane.
My questions are: Will spraying the sample directly with the coolant cause more damage to the sample than freezing indirectly through the stage? If so can I just use a larger sample and trim away the damaged area? Are there any better readily available coolants?
Thanks Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
Perhaps the membranes are leaching out during the dehydration and infiltration. Try a rapid dehydration, such as, two minutes each in 50, 70, 95, and one change of 100% EtOH. Then infiltrate in 3:1 EtOH:Epon for ten minutes, 1:1 for 10 minutes, 1:3 for twenty minutes, full strength Epon for thirty minutes, change to fresh full strength Epon and polymerize. Glutaraldehyde fix for only one hour, maximum, and that is longer than actually needed for cell monolayers. Stick with your en bloc staining method prior to dehydration. Also, perhaps thicker sections, like 70 nm, would increase your contrast. Good luck!
Dotty Sorenson Microscopy and Image Analysis Laboratory Department of Cell and Developmental Biology University of Michigan Medical School Ann Arbor, Michigan (734)763-1170 dsoren-at-umich.edu
On Tue, 31 Oct 2000, Xinran Liu 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 currently doing some EM work on rat cultured hippocampal cells. } The purpose of this experiment is to visualize individual synaptic vesicles } by transmission electron microscopy. The problem we had is that the } resolution/contrast is not high enough to distinguish the synaptic vesicles. } } The following is what we did, cells were cultured on coverslips, they were } fixed in situ in 2% glutaraldehyde for 1 hr to 24 hrs, postfixed with 1% } OsO4 for 1 hour, and en bloc stained with 2% aqueous uranyl acetate for 30 } min. dehydrated in ethanol and embedded in Polybed812. 40-50 nm sections } were stained by 5% uranyl acetate and lead citrate prior to EM observation. } } I would appreciate it very much if someone would provide suggestions on } section staining or cell processing. } } Xinran } } ******************************************** } Xinran Liu, M.D., Ph.D. } Center for Basic Neuroscience } The University of Texas Southwestern Medical Center at Dallas } 6000 Harry Hines Blvd., NA4.214A } Dallas, TX 75390-9111 } Phone: (214) 648-1830 } Fax: (214) 648-1801 } E-mail: xinran.liu-at-utsouthwestern.edu } }
} We currently doing some EM work on rat cultured hippocampal cells. } The purpose of this experiment is to visualize individual synaptic vesicles } by transmission electron microscopy. The problem we had is that the } resolution/contrast is not high enough to distinguish the synaptic vesicles. } } The following is what we did, cells were cultured on coverslips, they were } fixed in situ in 2% glutaraldehyde for 1 hr to 24 hrs, postfixed with 1% } OsO4 for 1 hour, and en bloc stained with 2% aqueous uranyl acetate for 30 } min. dehydrated in ethanol and embedded in Polybed812. 40-50 nm sections } were stained by 5% uranyl acetate and lead citrate prior to EM observation. } } I would appreciate it very much if someone would provide suggestions on } section staining or cell processing. } } Xinran } } ******************************************** } Xinran Liu, M.D., Ph.D. } Center for Basic Neuroscience } The University of Texas Southwestern Medical Center at Dallas } 6000 Harry Hines Blvd., NA4.214A } Dallas, TX 75390-9111 } Phone: (214) 648-1830 } Fax: (214) 648-1801 } E-mail: xinran.liu-at-utsouthwestern.edu
Sounds like your sections are too thin for adequate (for your needs) contrast. Thinner sections are not automatically "better". Try cutting pale gold sections and see if that helps. ALSO, check the alignment and stigmation of your microscope. You may need a different size condernser aperature and/or a different size objective aperature. All of these can affect contrast and resolution.
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 **********************************************
From root Wed Nov 1 13:57:03 2000 Return-Path: {Microscopy-request-at-sparc5.microscopy.com} Received: (from daemon-at-localhost) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id NAA08322 for dist-Microscopy; Wed, 1 Nov 2000 13:34:55 -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 NAA08319 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Wed, 1 Nov 2000 13:34:24 -0600 (CST) Received: from rhea.king.ac.uk (rhea.king.ac.uk [141.241.2.9]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id NAA08312 for {Microscopy-at-sparc5.Microscopy.Com} ; Wed, 1 Nov 2000 13:34:13 -0600 (CST)
Dear Cameron,
Thank you very much for your reply (I had a week off and found it just now) - it was the only reply I have received.
The letter was written and signed by our Senior Technician and when we sent it, it bounced back, obviously because his name is not on the list. Therefore we added in the second posting that the responses should be forwarded to my address - I did not intend to deprive the microscopy community of the replies and am quite happy to post this again for all to see - especially your helpful reply.
Thank you very much - if that saves our aging TEM it would make me very happy.
Regards Claudia
} From: Cameron Hind {Cameron_Hind-at-baxter.com}
EMPLOYMENT OPPORTUNITY
HKL Technology, a global leader in electron diffraction systems for scanning electron microscopes, announces an opening in its North American headquarters in Burnt Hills, New York. Burnt Hills is located in Saratoga County, in New York’s Capital District, centrally located about 3 hours from New York City, Boston, and Montreal. HKL Technology is a small, high technology firm growing in a competitive market. The company is currently seeking an individual with advanced skills and experience in materials characterization, which must include scanning electron microscopy and electron or x-ray diffraction, to take responsibility for our applications lab. A masters or doctoral degree in physics, materials science, or geological science is required. Candidates should have positive experience in teaching others, as training new and existing users is a vital role for this position. A successful candidate will be gregarious and confident, and must enjoy traveling. If you are interested in this position, send e-mail to sutliff-at-hkltechnology.com providing your full name, current address, and the contact information of two references. Resumes should be sent by regular mail to the address below.
John A. Sutliff HKL Technology, Inc. P.O. Box 179 (801 Saratoga Road) Burnt Hills, NY 12027 sutliff-at-hkltechnology.com www.hkltechnology.com TEL:(518)384-0101 FAX:(518)384-0281
Anyone know about Semper & Sprynt Boards and can help? (Send replies to Richard Hunton {Hunton-at-Cardiff.ac.uk} , not me!)
Hi Folks:
I'm fairly new to this business so I hope that you'll forgive what might seem to be a 'dumb' question.
I am currently using a LEO / Cambridge S360 SEM with a 4QBSD. My problem is that when I use probe currents of greater than about 18nA I get what looks like a reflection of the four diode quadrants of the detector superimposed on the backscattered image.
This appears (pretty much) independant of the working distance and magnification. The SE image *does* have a dark spot on the picture where the centre of the 'reflected' quadrant ghost appears.
I *would* like to be able operate at much higher probe currents than this if possible. I am using 20 kV and I even changed the filament (you always hope, don't you?).
Any help would be most gratefully recieved.
Robert
Robert McDonald SEM & EPMA Laboratories Department of Geography - Division of Earth Sciences Gregory Building Lilybank Gardens Glasgow University Glasgow G12 8QQ Scotland
Tel 0141 330 5505 (SEM lab) or 0141 330 5442 (Microprobe Lab) fax 0141 330 4817
Specimen Preparation Engineer Northwestern University
The Electron Probe Instrumentation Center (EPIC) at Northwestern University has an immediate opening for a specimen preparation expert. EPIC is a part of the world renowned Materials Research Center (MRC) and the Department of Materials Science & Engineering at Northwestern. The EPIC facility serves over 120 users in all aspects of Scanning and Transmission Electron Microscopy. The role of the specimen preparation engineer is to assist users with their specimen preparation needs, including instruction in TEM and SEM sample preparation using IBT, FIB, PIPS, electropolishing, ultramicrotomy, cutting/grinding/polishing, vacuum evaporation etc. All microscopes in EPIC are under full service contract. Thus, the duties include training students/users, development of specialized techniques and applications, minor maintenance, record keeping and billing. A BS or technical degree in physical/biological sciences is required. The candidate must have hands-on experience in all aspects of specimen preparation as well as considerable familiarity with digital acquisition, processing and computer assisted techniques. All levels of experience will be considered. Compensation will be commensurate with experience and qualifications. Send cover letter, resume and three references to: Prof. Vinayak P. Dravid, Director EPIC Materials Science & Engineering Northwestern University, 3013A MLSB Evanston, IL 60208 E-mail: v-dravid-at-northwestern.edu Fax: (847) 491-7820 http://epic.ms.nwu.edu/epic/index.htm
Northwestern University is an Affirmative Action/Equal Opportunity Employer. Hiring is contingent upon eligibility to work in the United States.
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It is possible to use an osmium-potassium ferrocyanide fix to enhance membranes (see Karnovsky, 1971, Russell and Burguet, 1977). However, the downside is potential bleaching of ribosomes. I would be interested in hearing from others as to their experience with this type of tissue preparation. Debby
Debby Sherman Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: sherman-at-btny.purdue.edu 1057 Whistler Building West Lafayette, IN 47907-1057
On Wednesday, November 1, 2000, Geoff McAuliffe {mcauliff-at-UMDNJ.EDU} wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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Hi all, I need to find someone to work on a RMC 6000-XL ultramicrotome. The machine has lots of internal vibration on the up/retracted stroke but it is stable in the cutting stroke...it will cut a section then shake the dookey out of it so that it is impossible to form a ribbon. The machine is on an anti-vibration table. any help would be greatly appreciated. thanks much, Beth
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
Hello Xinran, I en bloc stain my cell cultures in 2% UA in 70% EtOH. I leave them in the 4 degree refrigerator overnight, at least 16 hours. The rest of my processing sounds comparable to yours. This seems to give good contrast. I haven't worked with hippocampal cell cultures, though, so don't now how it would work on them. Good luck, Jo Dee
Xinran Liu 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 currently doing some EM work on rat cultured hippocampal cells. } The purpose of this experiment is to visualize individual synaptic vesicles } by transmission electron microscopy. The problem we had is that the } resolution/contrast is not high enough to distinguish the synaptic vesicles. } } The following is what we did, cells were cultured on coverslips, they were } fixed in situ in 2% glutaraldehyde for 1 hr to 24 hrs, postfixed with 1% } OsO4 for 1 hour, and en bloc stained with 2% aqueous uranyl acetate for 30 } min. dehydrated in ethanol and embedded in Polybed812. 40-50 nm sections } were stained by 5% uranyl acetate and lead citrate prior to EM observation. } } I would appreciate it very much if someone would provide suggestions on } section staining or cell processing. } } Xinran } } ******************************************** } Xinran Liu, M.D., Ph.D. } Center for Basic Neuroscience } The University of Texas Southwestern Medical Center at Dallas } 6000 Harry Hines Blvd., NA4.214A } Dallas, TX 75390-9111 } Phone: (214) 648-1830 } Fax: (214) 648-1801 } E-mail: xinran.liu-at-utsouthwestern.edu
-- Jo Dee Fish Coordinator of Electron Microscopy Cell Analysis Facility The Burnham Institute 10901 N. Torrey Pines Rd. La Jolla, CA 92037 (858)646-3100 ext. 3620
I routinely prepare hippocampal neurons grown on glass coverslips for thin section EM using microwave fixation. I fix for 16 seconds (8 on-20 off-8 on) on ice with chilled fixative (2% glutaraldehyde in 0.1M cacodylate buffer). For secondary fixation, I fix 16 seconds on ice with chilled 1-2% osmium tetroxide in 0.1M cacodylate buffer containing 0.8% potassium ferricyanide. I let this sit for 5 mins before proceeding with 30 seconds of 5% en bloc uranyl acetate. Let the dish sit for 30 mins. Proceed with microwave dehydration (10 seconds each ethanol change, 3 times 100%) and infiltration (5 mins each). I cut 70 nm thin sections (pale gold/silver) and stain with 5% aqueous UA for 15-30 mins. followed by 3 mins. with lead citrate (made fresh each time). In regards to Xinran Liu's problem, there could be several factors. The fixation time is too long- 20 minutes of bench fixation is adequate. Are you putting sections on formvar? This could reduce contrast. Also, make thicker sections (60-70nm). Thirdly, check your uranyl acetate. I was using old UA that had expired and was useless. A new bottle made enormous difference. Freshly made stains help a lot, particularly for cell cultures that have low contrast. You should be able to see synaptic vesicles no problem. If these things don't help, the problem could be with the health of your cultures. Synaptic vesicles are the first to go in sick cultures. So make sure the cultures are healthy and save yourself a lot of time and effort! Good luck.
JoAnn Buchanan Molecular and Cellular Physiology Stanford University School of Medicine Stanford, CA 94305 650-723-5856
Dookey? I haven't come across that one yet up here in the Great White North. Just in case you missed it, RMC announced not too long ago that they had 're-emerged' from the hiatus caused by Ventana getting out of the EM products business. They are alive and well in Tucson and could likely point you in the direction of someone in your area. Contact info:
Al Coritz Sales & Service Manager RMC-Boeckeler Instruments 4650 S. Butterfield Dr. Tucson, AZ 85714 Voice: 520-745-0001 Cell: 520-465-3598 Fax: 520-745-0004 Email:Al-at-Boeckeler.com Website:RMCProducts.com
Hope you find someone suitable, eh? (That's Canuck-talk).
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: } beth-at-dogwood.botany.uga.edu[SMTP:beth-at-dogwood.botany.uga.edu] } Sent: Thursday, November 02, 2000 10:01 AM } To: microscopy-at-sparc5.microscopy.com } Subject: RMC microtome service } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 find someone to work on a RMC 6000-XL ultramicrotome. The } machine } has lots of internal vibration on the up/retracted stroke but it is stable } in the cutting stroke...it will cut a section then shake the dookey out of } it so that it is impossible to form a ribbon. The machine is on an } anti-vibration table. } any help would be greatly appreciated. } thanks much, } Beth } } ************************************** } Beth Richardson } EM Lab Coordinator } Botany Department } University of Georgia } Athens, GA 30602 } } Phone - (706) 542-1790 } FAX - (706) 542-1805 } Email - beth-at-dogwood.botany.uga.edu } } "Between the two evils, } I always pick the one I never tried before". Mae West (1893-1980) } ************************************** } } }
{html} {font face="Arial, Helvetica"} POSITION OPEN # 1 of 2 {br} {br} Postdoctoral Scholar/Research Associate {br} {br} Cryo-Electron Microscopy of Soft and Hybrid Nanostructures {br} {br} Northwestern University {br} Materials Science & Engineering, Evanston, IL {br} {br} A postdoctoral scholar/research associate position is immediately available at Northwestern University in the area of advanced Analytical Cryo-TEM of soft and hybrid nanostructured materials. {br} This project is being supervised by Prof. Vinayak P. Dravid, and concerns with analysis of nanoscale structure, assembly/organization and chemistry of advanced soft and hybrid nanostructures (such as patterned DNA/Lipids, nanoparticle-DNA assemblies and nanoparticle-Lipid complexes etc..) using Analytical TEM and Cryo-EM techniques. This is a part of extensive cross-disciplinary and collaborative activities in nanotechnology at Northwestern. As a result, the candidate would have ample opportunity to learn diverse aspects of nanotechnology, from soft nanolithography to dynamic measurements, while contributing to Cryo-TEM analysis of soft and hybrid nanostructures. {br} Northwestern’s electron probe instrumentation center (EPIC: {a href="http://epic.ms.nwu.edu/" eudora="autourl"} http://epic.ms.nwu.edu {/a} ) is well equipped with several modern SEMs, TEMs and extensive specimen preparation equipment ranging from focused ion beam (FIB) to cryo-transfer with LN2 and LHe stages. One of the cold FEG TEMs (Hitachi HF-2000) will soon be equipped with a Gatan Imaging Filter (GIF) which will be extensively utilized in 2-D mapping of element- and bonding specific spectral signatures in soft and hybrid nanostructures. {br} The position requires a PhD in physical/biological sciences/engineering. Considerable hands-on experience in cryo-preparation techniques (e.g. freeze fracture/drying, ultramicrotomy) for soft/hybrid specimen is required. Experience in analytical and cryo-TEM techniques and computation/simulations is necessary. Prior knowledge of imaging filter/spectral imaging is desirable but not mandatory. {br} The position is available immediately for at least two years with possibility for extension for additional years upon mutual agreement. {br} Salary and compensation would commensurate with experience, in the range: $ 30,000-$40,000 per year. {br} Please forward resume with three names of references to: {br} Prof. Vinayak P. Dravid {br} Materials Science & Engineering {br} Director, Electron Probe Instrumentation Center (EPIC) {br} 2225 N. Campus Drive, 1133 MLSF {br} Northwestern University, Evanston, IL 60208, USA {br} Ph.: (847) 467-1363, Fax: (847) 491-7820 {br} E-mail: v-dravid-at-northwestern.edu {br} URL: {/font} {a href="http://vpd.ms.nwu.edu/" eudora="autourl"} {font face="Arial, Helvetica" color="#0000FF"} {u} http://vpd.ms.nwu.edu {br} {br} {br} {/a} {/font} {/u} {font face="Arial, Helvetica"} POSITION OPEN # 2 of 2 {br} {br} Postdoctoral Scholar/Research Associate {br} {br} Analytical TEM and Electron Holography of Nanostructured Materials {br} Northwestern University, Evanston, IL, {u} USA {br} {br} {/u} A postdoctoral scholar/research associate position is immediately available at Northwestern University in the area of advanced Analytical TEM of nanostructured materials. {br} This project is being supervised by Prof. Vinayak P. Dravid, and concerns with analysis of nanoscale chemistry, structure and electrostatic fields at interfaces and junctions in advanced nanostructures (nanocrystals, nanotubes, thin film interfaces) using analytical TEM and holography techniques. This is a part of extensive cross-disciplinary and collaborative activities in nanotechnology at Northwestern. As a result, the candidate would have ample opportunity to learn diverse aspects of nanotechnology, from soft nanolithography to dynamic property measurements while contributing to TEM analysis of nanostructures. {br} Northwestern’s electron probe instrumentation center (EPIC: {a href="http://epic.ms.nwu.edu/" eudora="autourl"} http://epic.ms.nwu.edu {/a} ) is well equipped with several modern SEMs, TEMs and extensive specimen preparation equipment ranging including a focused ion beam (FIB) system. One of the cold FEG TEMs (Hitachi HF-2000) will soon be equipped with a Gatan Imaging Filter (GIF) which will be utilized in 2-D mapping of element- and bonding specific spectral signatures in nanostructures. {br} The position requires a PhD in physical sciences/engineering. Considerable hands-on experience in advanced TEM techniques such as HRTEM, EDS/EELS, CBED and computation/simulations is required. Experience in electron holography is desirable but not mandatory. {br} The position is available immediately for two years with possibility for extension upon mutual agreement. {br} Salary and compensation would commensurate with experience, in the range: $ 30,000-$40,000 per year. {br} Please forward resume with three names of references to: {br} Prof. Vinayak P. Dravid {br} Materials Science & Engineering {br} Director, Electron Probe Instrumentation Center (EPIC) {br} 2225 N. Campus Drive, 1133 MLSF {br} Northwestern University, Evanston, IL 60208, USA {br} Ph.: (847) 467-1363, Fax: (847) 491-7820 {br} E-mail: v-dravid-at-northwestern.edu {br} URL: {/font} {a href="http://vpd.ms.nwu.edu/" eudora="autourl"} {font face="Arial, Helvetica" color="#0000FF"} {u} http://vpd.ms.nwu.edu {br} {br} {br} {br} {/a} {/font} {/u} {br} {div} ******************************************************* {/div} {div} (Vinayak P. Dravid) {/div} {div} Materials Science & Engineering {/div} {div} Director, Electron Probe Instrumentation Center (EPIC) {/div} {div} 2225 N. Campus Drive, 1133 MLSF {/div} {div} Northwestern University, Evanston, IL 60208, USA {/div} {div} Ph.: (847) 467-1363, Fax: (847) 491-7820 {/div} {div} E-mail: v-dravid-at-northwestern.edu {/div} {br} {div} {a href="http://nuinfo.nwu.edu/materials/faculty/vpd.html" EUDORA=AUTOURL} http://nuinfo.nwu.edu/materials/faculty/vpd.html {/a} {/div} {div} or {/div} {div} {a href="http://vpd.ms.nwu.edu/" EUDORA=AUTOURL} http://vpd.ms.nwu.edu {/a} {/div} ******************************************************* {/html}
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We are in the preliminary throes of writing a grant proposal for a new SEM. This scope will have the following characteristics: EDX, cathodoluminescence, beam etching, BS, "conventional" SEM use, possibly variable pressure. Questions 1. What are the advantages/disadvantages of Schottky over cold FE. 2. I have personally dealt with conventional filament replacement (W), I gather that FE emitter replacement is much more involved than W filament replacement. What are practicalities of FE (thermal or cold) emitter replacement with respect to who performs the work and the cost of the emitter. 3. Is there any reason that variable pressure will preclude the use of the scope for beam etching. Thanks to all Bruce Bruce Cutler Director, Microscopy & Electronic Imaging Lab University of Kansas
by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id OAA01294 for dist-Microscopy; Thu, 2 Nov 2000 14:30:06 -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 OAA01284 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Thu, 2 Nov 2000 14:29:35 -0600 (CST) Received: from mail (madison.tec.wi.us [198.150.15.232]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id OAA01274 for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 2 Nov 2000 14:29:08 -0600 (CST) Message-Id: {200011022029.OAA01274-at-ultra5.microscopy.com} Received: from stu.madison.tec.wi.us (nimsstu1.madison.tec.wi.us [198.150.15.195]) by mail; Thu, 02 Nov 2000 14:27:37 -0600 Received: from dflatoff [198.150.245.87] by stu.madison.tec.wi.us with Novell Internet Messaging System Web Client; Thu, 02 Nov 2000 14:27:39
Greetings All, I would like to request guidance on protocols for preparing TEM cross section to view the interface between diamond like carbon (DLC) coating on a steel substrate which has been preimplanted to increase the adhesion of the coating. Any assistance with be greatly appreciated.
Greetings All, I would like to request guidance on protocols for preparing TEM cross section to view the interface between diamond like carbon (DLC) coating on a steel substrate which has been preimplanted to increase the adhesion of the coating. Any assistance with be greatly appreciated.
In the early to mid-90s, our library, due to budget constraints at the time, let our subscriptions to microscopy journals lapse. We are now in the position to resubscibe to a number of journals. Could I get advice on which journals microscopists feel are the most useful. This includes journals in both the biolgical and physical sciences, applied research electron microscopy, as well as techniques oriented periodicals. Thanks for your input.
Mark J. Grimson Electron Microscopy Facility Dept. of Biology, MS 3131 Texas Tech University Lubbock, Texas 79409
MICROSCOPE INTERFACE DESIGNER - Public Imaging Facility Fixed-term Position
The Exploratorium is a museum dedicated to the public understanding of science, art, and human perception. Founded in 1969 by Frank Oppenheimer, it has pioneered the role of museums as active teaching centers with original programming based on an interactive approach to learning. It serves as an interdisciplinary resource for schools, universities, scientists, and artists, as well as for the public.
SUMMARY The Microscope Interface Designer will provide technical expertise to a three-year project to develop a microscopic imaging facility for the public, including museum visitors, students, teachers and Internet users, to view a variety of living specimens via video and the Internet. The facility will utilize the highest quality optics and state-of-the-art microscopic techniques and provide a unique experience for the general public to interact with the technology and tools used by biomedical researchers. The project will require extensive inter-organizational and external collaborations with research facilities, equipment and software companies, teachers and curriculum developers, and media technology developers. The Microscope Interface Designer will work collaboratively with the Exploratorium's Life Sciences staff and report to the Life Sciences Area Manager.
ESSENTIAL FUNCTIONS · Design and develop approaches for presentation of living specimens to the public in an accessible and comprehensible manner · Work collaboratively with the Visitor Research & Evaluation Department to incorporate visitor feedback in the development process · Design and develop hardware and software interface systems to enable public to easily use laboratory grade microscopes · Work closely with technical experts at equipment and software companies to develop design and technical options that enhance the project's success · Design and oversee fabrication of all necessary technical hardware for visitor interfaces with microscopes · Work collaboratively with software developers to implement software designs · Maintain and operate microscopes during the development phase
MINIMUM QUALIFICATIONS · MS degree in biological sciences with a focus in microscopy, Ph.D. desirable · Minimum 3 years experience with microscopic imaging techniques · Demonstrated experience in working with biological specimens of various sorts, particularly living tissue preparations · Familiarity with a variety of tissue culturing techniques · Familiarity with device control and software interfaces for computers · Proven ability to solve technical design problems · Ability to work collaboratively as well as independently
HOW TO APPLY: This is a three-year, fixed-term, exempt, union position which pays $788.63/week. This position starts as soon as possible and ends on or before September 30, 2003. Please apply to: Dept. LS-3L Exploratorium, 3601 Lyon Street, San Francisco, CA 94123 Fax: (415) 561-0370 E-Mail: resume-at-exploratorium.edu (attachments not accepted) No phone calls, please The Exploratorium is committed to a diverse workforce
See our web site at www.exploratorium.edu
Kellin Defiel Human Resources Exploratorium San Francisco, CA 94123 kellind-at-exploratorium.edu (415) 561-0337
Does anyone out there have a 40 kV/LaB6 EHT power supply box (aka Wallace unit) for a Cambridge 250 Mark 2 that they're willing to donate, sell, or lend for diagnostics? Or even just the mains inverter board? My users are stacked in a holding pattern and we'd all be grateful for some help!
Many thanks, Dee
*************************************************************** Dee Breger Mgr. SEM/EDX Facility Lamont-Doherty Earth Observatory 61 Route 9W Palisades, NY 10964 USA T: 914/365-8640 F: 914/365-8155
http://www.ldeo.columbia.edu/micro http://www.discovery.com/area/science/micro/micro1.html http://www.lsc.org/antarctica/front.html Journeys in Microspace (Columbia University Press, 1995)
Can anyone answer this question . Remember Ask-A-Microscopist is a forum that is used by NON-MICROSCOPISTS looking for help so keep try to your technical details at the right level.
Nestor
================================================================ Email: adam.boyes-at-virgin.net Name: Adam Boyes School: Horndean Sixth Form
Question: When looking at plant cells containing chloroplasts through a microscope, why is it that they move around the edges of the cell clockwise and counterclockwise in adjacent cells?
What kind of steel? If you have a heat treated hard steel such as 440C or M50, you are in for a lot of trouble. When you thin these down, the internal stresses can bend the sample like crazy. You are best off doing a cross section using the slotted-D approach. If your sample is magnetic, an added benefit is that it minimizes the amount of magnetic material in the sample. Slot a rod that fits into a 3mm OD tube. I used 304SS tubing and rods from Small Parts Catalog. You must thin the sample to where they don't curl. Cut them down to fit in the slot and epoxy them in. Cut your samples so that the blade cuts perpendicular to the normal of the layers, i.e. along the direction of the slot. You should be able to dimple and ion mill normally. By normally, I mean low angle. The carbon resists ion milling. You might need to use a gas mixture. We had a little paper in the MRS Sample Prep IV, Vol 480, that showed a comparison of Ar, Ne, Ne/O2 or Ar/O2 to help with the carbon. Another option is to put water vapor into the ion mill while you are milling with a leak valve.
-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: Daniel L Flatoff [mailto:dflatoff-at-stu.madison.tec.wi.us] } Sent: Thursday, November 02, 2000 3:28 PM } To: Microscopy-at-sparc5.microscopy.com; Microscopy-at-sparc5.microscopy.com } Subject: TEM prep of DLC coating on steel } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: 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 } } } -------------------------------------------------------------- } ---------. } } } Greetings All, } I would like to request guidance on protocols for } preparing TEM cross section to view the interface between } diamond like carbon (DLC) coating on a steel substrate } which has been preimplanted to increase the adhesion of the } coating. Any assistance with be greatly appreciated. } } } } Daniel L. Flatoff } } }
haven't received a response from my last message so I'll send it again: does anyone know who supports and supplies parts for the Technics Micro Ion Mill? (ours was made 1976)
cheers Liz McKenzie
******************************************************* Geomicrobiology and Electron Microscopy Laboratory Room S9 Cramer Hall 1721 SW Broadway Portland State University Portland OR97201
It is possible that your car engine is driven by psycho-kinetic energy. But if it looks like a petrol engine and smells like a petrol engine, a sensible working hypothesis is that it is a petrol engine.
I would appreciate any procedures or references concerning preparation of solder joints for microelectronic applications. The solders that I am concerned with are high Pb, Pb-Sn-Ag and Au-Sn. Thanks.
The advantages/disadvantages of cold vs. Schottky FE guns lie at the extremes of performance. Cold FE will give you better ultimate image resolution (roughly 1.5 vs. 2.5 nm, depending on manufacturer, of course) and will give you noticably better images at low voltage (1kV or so). Schottky will give you much higher currents at larger spot sizes (for x-ray mapping or back-scatter diffraction pattern acquisition, for example) and much better medium-term (tens of minutes to a few hours) beam current stability, if that is important to you. It is also not necessary to flash the tip periodically in a Schottky gun. For most other SEM needs, either form will work wonderfully for you.
We have one of each, and we do not notice any difference in the maintenance needs between them. Don't worry about tip replacements. Our cold FE is nearly 6 years old and is still running with the original tip, as is our Schottky, which is over three years old. I don't think this is a function of vendor - it is my impression that microscopes from other vendors have similar records. The tip replacement is performed by the service engineers, and depending on your service contract, may be included in that. It takes about 3 days (I am told!) on either instrument.
It is much more important to be clear about your microscopy needs. You say in your posting "possibly variable pressure" ... there is a big difference between the ESEM of FEI/Philips and the variable pressure designs of other manufacturers, so you need to be clear about how important this factor is to you. What structures will you need to examine by "conventional" SEM, how important is high-speed x-ray mapping, etc. etc. etc. By asking these kinds of questions the right microscope will eventually become clear to you. Don't worry about the mechanics of the gun - they all work fine.
All in my humble opinion, of course!!
Tony Garratt-Reed.
At 02:19 PM 11/02/2000 -0600, 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: (+) 1-617-258-6478 **
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{Subject: Re: Freeze-plunge device availability {From: Dennis C. Winkler, DCWinkler-at-aol.com
{Last month you posted a query about the availability of freeze-plunge devices.
{I am interested in the responses you received. I didn't find them {posted on the listserver or on your web site. Would you mind sharing {them?
I have not had a chance to post them yet but here is the information. I received one response and I knew of one other vendor.
1) BAL-TEC Plunge Freezing System TFD 010 Contact Johnny Hagen at 603-622-5011 for the details. They have a single picture on their website www.bal-tec.com List price for the system is $8674
2) This summer at MSA I saw that Gatan had a system. Our sales person is Paul Miller, 724-779-2501 Their website is www.gatan.com but I saw no information on this system there the last time I looked. List price ~$20,000 (ball park figure)
The two systems do quite different things. The Bal-tec system is designed to keep the cryogens at the proper temperature and it has a pneumatic cylinder plunging arm. The Gatan system has an enclosed chamber around the sample to maintain humidity and you program in how long you want to do wicking before you plunge. You can do single sided or double sided wicking and it looks like it has a fair amount of flexibility for doing different types of plunge experiments.
As usual... the disclaimer is that I am not receiving kickbacks from either company on this.
Norm Olson
******************************************* Norm Olson Senior Research Electron Microscopist Department of Biological Sciences Lilly Hall of Life Sciences Purdue University West Lafayette, IN 47907
by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id JAA01667 for dist-Microscopy; Fri, 3 Nov 2000 09:09:03 -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 JAA01663 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Fri, 3 Nov 2000 09:08:33 -0600 (CST) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id JAA01656 for {microscopy-at-sparc5.microscopy.com} ; Fri, 3 Nov 2000 09:08:03 -0600 (CST) Received: from northrelay02.pok.ibm.com (northrelay02.pok.ibm.com [9.117.200.22]) by e4.ny.us.ibm.com (8.9.3/8.9.3) with ESMTP id KAA203496; Fri, 3 Nov 2000 10:06:38 -0500 Received: from d01ml065.pok.ibm.com (d01ml065.pok.ibm.com [9.117.250.65]) by northrelay02.pok.ibm.com (8.8.8m3/NCO v4.95) with ESMTP id KAA24862; Fri, 3 Nov 2000 10:06:15 -0500 Importance: Normal
We tripod polish with some modification to the procedure: You can't use water to polish the specimen, we use propylene glycol from a squeeze bottle when actually polishing the solder; The thin solder section is mechanically weak, we mount on a Mo grid with a small aperture size prior to second side polishing; Use low-angle ion milling to keep the solder from melting and to minimize element-selective milling in the ion mill if you have to ion mill the specimen. Use a cold stage in the TEM or restrict the beam heating.
Good luck,
Ron
Ron Anderson, IBM, Hopewell Jct., New York, USA. anderron-at-us.ibm.com
IBM Analytical Services; http://www.chips.ibm.com/services/asg
} } Hello all - } } I've written to the list in the past regarding embedding of bovine } oocytes and I wish to express thanks to all of the helpful suggestions that } I received. } } I'm still having a coupld of problems, though, so I'm again seeking } advice. I am fixing oocytes in gluteraldehyde and then embedding them in } agarose. I am using 1% low melting point agarose, creating a bubble into } which I place my oocytes and then sealing the bubble with 0.5% agarose. I } then trim the agarose into a 1 mm^2 chip and dehydrate through a series of } ethanol steps, including staining with eosin to help visualize the eggs. } Finally, I clear in xylenes and embed the chip in a paraffin block. } } My problem is twofold: 1) following sectioning and rehydration, } when I } look at the agarose sections, I can certainly see oocytes (they stain } specifically for an anti-ZP antibody)and they're consistent in placement } among consequent sections, but frankly they're horribly ugly. The zona is } "wrinkled", and the eggs look like they've undergone an osmotic trauma. 2) } The agarose sections seem to frequently fold or rip and I'm not quite sure } at what stage of the process that this is occuring - I feel when placing } the sections into the waterbath or retrieving them on slides (treated with } HistoPrep) they seem fine. Is it possible the folding is occuring during } the deparaffination/rehydration, maybe because the agarose sections don't } stick well to the slides? } } Any help in this matter is greatly appreciated! Thanks! } } --Carrie
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
Folks, heres a new thread.... We are curious about how well automated ICC/IHC machines work How easy are they to use? How flexible are the programs? How much time do they save? Can you program sophisticated multicolor protocols? (of course the inflammatory question) which is the best machine? Whats the reagent usage like? etc
Looking forward to hearing from you all Simon
--------------------------- Simon C. Watkins Ph.D. MRC Path Associate Professor, Cell Biology and Physiology Director: Center for Biologic Imaging BSTS 225 University of Pittsburgh Pittsburgh PA 15261 Tel:412-648-3051 Fax:412-648-8330 http://sbic6.sbic.pitt.edu -----------------------------------
In the past, cold FE sources gave the best resolution, however, Schottky (thermal) FE sources have improved significantly over the years. Since the "resolution" specifications provided by the SEM vendors cannot always be compared directly, it is best to use your own samples and observe the imaging capabilities of each SEM for yourself. Such demos are common before the sale of FE SEMs.
A difference between cold FE and Schottky FE sources that is not subjective is the stability of the beam current. Cold FE sources will typically have frequent spikes and/or jumps of the beam current on the order of +/-3% or more. Readings of the beam current taken just seconds apart may show changes on this order, and over 1 hour intervals changes of +/-10% or more are not unusual. In contrast, the FE sources are exceptionally stable with total noise and drift of less than 1%/hour. (I've measured ~1% drift over 12 hours in one case.)
If you are concerned about beam current noise and drift, it may be helpful to request a graph of beam current vs. time showing the typical performance of the SEM in question. (But be wary of chart recording traces that may have a slow time constant which will hide fast changes in the beam current!)
Note that for imaging, such noise/drift in the beam is not generally an issue, because the cold FE SEMs dynamically adjust the brightness displayed on the imaging screen to compensate for the changes in the actual beam current. In effect, the beam intensity on the specimen is still changing, but the displayed image does not show it.
If you need high beam currents, the Schottky source will typically give much more current than a cold FE source.
Regarding replacement of the FE source, that is generally done under the SEM service contract. The price I have heard for the FE source itself is ~$20k US, although I'm sure others can give more exact numbers. The SEM vendors themselves will be the best source of information regarding pricing.
Variable pressure SEMs will typically have a "high vacuum" mode for conventional viewing. However, a variable pressure SEM may have an extra aperture to support the differential pumping in the column that may limit the lowest magnification, even in non-VP mode.
I hope this helps.
Joe _________________________________________ Joe Nabity, Ph.D. JC Nabity Lithography Systems E-Beam Lithography using Commercial SEMs & STEMs PO Box 5354, Bozeman, MT 59717 USA Voice: (406) 587-0848 FAX: (406) 586-9514 E-mail: info-at-jcnabity.com Web: www.jcnabity.com
In a message dated 11/2/2000 7:04:37 PM Mountain Standard Time, bcutler-at-eureka.idl.ukans.edu writes:
} We are in the preliminary throes of writing a grant proposal for a new SEM. } This } scope will have the following characteristics: EDX, cathodoluminescence, } beam } etching, BS, "conventional" SEM use, possibly variable pressure. } Questions } 1. What are the advantages/disadvantages of Schottky over cold FE. } 2. I have personally dealt with conventional filament replacement (W), I } gather } that FE emitter replacement is much more involved than W filament } replacement. } What are practicalities of FE (thermal or cold) emitter replacement with } respect } to who performs the work and the cost of the emitter. } 3. Is there any reason that variable pressure will preclude the use of the } scope } for beam etching. } Thanks to all } Bruce } Bruce Cutler } Director, Microscopy & Electronic Imaging Lab } University of Kansas }
I was involved in preparation, and microscopy, of lead solders for some time at my last job, and if you wish to contact me I will be able to put you in touch with the group that I was working with.
Hello I would like to subscribe to your service, please let me know if there is any service charge. Please pass on my problem to your subscribers. I am a research lab that does a lot of hard tissue processing, I as well as our EM departments have been using SPURR as our plastic of choice. I have been recently told by my supplier (Marivac) that in six months one of the components of SPURR will be discontinued. The component is ERL (Vinylcyclohexene Dioxide). I would like to know if others are using SPURR and if so have they encountered the same problem. Please forward your information to mmendes-at-mtsinai.on.ca. Thank-you in advance for any help offered.
Greetings. I am a final student of the department of Electrical Electronics, Obafemi Awolowo University, Ile-Ife, Nigeria. I am doing a project titled "Low-Frequency Surface Microscopy". My supervisor wants me to use an electric field-probe to detect changes in the capacitance of the surface as cracks, boundaries, etc. are encountered. I would be very grateful if you can send me any information on the topic.
My e-mail address is: femi_adeluyi-at-yahoo.com and my address is: Olufemi Adeluyi Department of Electronic and Electrical Engineering Obafemi Awolowo University, Ile-Ife, Nigeria.
Dear all, We have been asked to visualize the manufacturers stamped numbers on a rifle which have been removed by grinding. So far secondary and backscatter images don't tell us much. We are told that the metal under the stamping is compressed for a significant depth. Any ideas on how to image this compressed metal. Cheers, Eric Hines Microscopy Centre CSIRO Entomology Canberra OZ
you can use the macro etching techniques and macro photography; more details in the book Vander Voort - Metallography, Principles and practise, McGraw-Hill
best regards
KJ Huebner
{hubner-at-IOd.krakow.pl} :-)
On Mon, 6 Nov 2000, Eric Hines wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear all, } We have been asked to visualize the manufacturers stamped numbers on a } rifle which have been removed by grinding. So far secondary and backscatter } images don't tell us much. } We are told that the metal under the stamping is compressed for a } significant depth. Any ideas on how to image this compressed metal. } Cheers, } Eric Hines } Microscopy Centre } CSIRO Entomology } Canberra OZ } } } }
As promised, the only reference for SEM visualization of damaged serial numbers in gun metal I was able to quickly find was "Visualization of a Restored Serial Number Using Scanning Electron Microscopy (SEM)" by Amy Mongan of Forensic Analytical Specialties. The article was published as a case report in the Journal of Forensic Sciences [J Forensic Sci 1996;41(6):1074-1076]. There are more than likely other references but I do not have them immediately available.
Good luck!
S. Frank Platek, M.S. Forensic Chemistry Center U.S. Food and Drug Administration 6751 Steger Drive Cincinnati, OH 45237-3097 (513) 679-2700 X254 (513) 679-2761 FAX fplatek-at-ora.fda.gov
Disclaimer: Opinions/recommendations stated are solely mine and do not necessarily represent those of the US Food and Drug Administration or any other Federal Agency.
There is a special etch that "highlights" the stampings, forget SEM methods. I do not recall the exact mix, but Police forensics lab know it. I believe it is a mild form of aqua reiga with one added item but forgot. I consulted to a forensics lab and did it once and we got a few digits off an engine block.
Good luck
Robert Sherman Applied Surface Technologies www.co2clean.com
} } Dear all, } We have been asked to visualize the manufacturers stamped numbers on a } rifle which have been removed by grinding. So far secondary and backscatter } images don't tell us much. } We are told that the metal under the stamping is compressed for a } significant depth. Any ideas on how to image this compressed metal. } Cheers, } Eric Hines } Microscopy Centre } CSIRO Entomology } Canberra OZ
Some forensic scientists have been successful in detecting stamped serial numbers by etching the part in a solution of 5 -10% HNO3 in water or alcohol for 10 to20 seconds. The disturbed metal will etch more darkly than the surrounding metal.
Best regards and good luck,
Sam Purdy Tecnical Center National Steel Corp. Trenton MI spurdy-at-nationalsteel.com
The immuno units available are becoming many and varied. However, the main consideration is flexibility of the unit and secondarily cost of the reagents. Dako and Cytlologix are the most reliable and flexible available for routine and special procedures. I believe both can be programmed for dual staining or to separate protocols can be used on tissue for the result. These units allow you to choose your reagents, antibodies and procedures. The Ventana is an excellent unit with one huge drawback you are forced to use their secondaries and limited on primaries to some extent. The cost is very high for the reagents to run the unit. I am a histologist who also does EM and can speak from experience of working with the units in the field as an observer and user. Pam Marcum
-----Original Message----- } From: simon watkins [mailto:swatkins+-at-pitt.edu] Sent: Friday, November 03, 2000 11:35 AM To: microscopy
Folks, heres a new thread.... We are curious about how well automated ICC/IHC machines work How easy are they to use? How flexible are the programs? How much time do they save? Can you program sophisticated multicolor protocols? (of course the inflammatory question) which is the best machine? Whats the reagent usage like? etc
Looking forward to hearing from you all Simon
--------------------------- Simon C. Watkins Ph.D. MRC Path Associate Professor, Cell Biology and Physiology Director: Center for Biologic Imaging BSTS 225 University of Pittsburgh Pittsburgh PA 15261 Tel:412-648-3051 Fax:412-648-8330 http://sbic6.sbic.pitt.edu -----------------------------------
Maria, We are not discontinuing Spurr's at Polysciences. We do have ERL and the other components available now and in the future.
Pamela A Marcum Histology/Microscopy Product Development Manager Polysciences, Inc. 400 Valley Road Warrington, PA 19876 Phone: 800-523-2575 Ext 167 Fax: 215-343-0214 E-Mail: pmarcum-at-polysciences.com {mailto:pmarcum-at-polysciences.com}
-----Original Message----- } From: Mendes, Maria [mailto:mmendes-at-mtsinai.on.ca] Sent: Sunday, November 05, 2000 8:34 AM To: Microscopy-at-sparc5.microscopy.com
Hello I would like to subscribe to your service, please let me know if there is any service charge. Please pass on my problem to your subscribers. I am a research lab that does a lot of hard tissue processing, I as well as our EM departments have been using SPURR as our plastic of choice. I have been recently told by my supplier (Marivac) that in six months one of the components of SPURR will be discontinued. The component is ERL (Vinylcyclohexene Dioxide). I would like to know if others are using SPURR and if so have they encountered the same problem. Please forward your information to mmendes-at-mtsinai.on.ca. Thank-you in advance for any help offered.
It has been many years since I worked in a Crime Lab, but the technique that we used to raise serial numbers on guns did not involve the SEM at all. First, we polished the surface where the serial number had been ground off using successively finer grades of sandpaper. When we had achieved a highly polished appearance to the surface we then brushed the area with a dilute solution of nitric acid using a cotton tipped swab. This was done very carefully, inspecting the surface after each wipe with the swab using a collimated light beam held at a shallow angle with respect to the surface. The numbers will appear due to the fact, as you mentioned, they are compressed more than the surrounding metal and therefore are etched at a different rate. When the numbers are visible, the surface is flushed with water and a non-volatile liquid like glycerine is applied to keep the surface wet until the numbers can be recorded and photographed. A hand lens or a stereomicroscope were the only microscopical tools necessary. I know that this description is sketchy, but it should give you a starting point. Also, keep in mind that if the person who was trying to remove the serial number knew his business, he (or she) may have ground deeply enough to obliterate any trace of the number and no method will restore it.
Regards, Bill Roberts
erich-at-ento.csiro.au (Eric Hines) on 11/06/2000 02:46:04 AM
To: Microscopy-at-sparc5.microscopy.com cc: (bcc: William H Roberts/US/FILM/DPT)
Dear all, We have been asked to visualize the manufacturers stamped numbers on a rifle which have been removed by grinding. So far secondary and backscatter images don't tell us much. We are told that the metal under the stamping is compressed for a significant depth. Any ideas on how to image this compressed metal. Cheers, Eric Hines Microscopy Centre CSIRO Entomology Canberra OZ
Light optical metallographic techniques, rather than electron optical, might serve you in this case.
If you polish the region - removing as little material as possible, but through 6 micron diamond anyway - and etch with 2% nital (2% nitric acid in alcohol) - you ought to reveal the underlying numbers. Other people may suggest more effective etchants for this purpose; nital is just a general purpose old standby.
Regards, Andrew T. Werner Shaped Charge Research - Metallurgy Laboratory Schlumberger Reservoir Completions Technology Center 14910 Airline Road, Rosharon, TX 77583-1590 Voice (281) 285-5272 Fax (281) 285-5273
"We shoot the hippopotamus with bullets made of platinum 'cause if we used the leaden ones his hide would surely flatten 'em" - Author Unknown
At 06:46 PM 11/06/2000 +1100, Eric Hines wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
One of my users has just asked me for references regarding the technique of acquiring electron induced color CL images by obtaining RGB channels via red-blue-green filters. One of those procedures we have simply been taking for granted, but now want to acknowledge earlier work, especially with respect to geologic materials and quartz.
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/
If you can submerge the specimen under a liquid it will greatly enhance the captive effects from 3 or so for most fluids to 70 for water due to the dielectric properties of the liquid.
I have no idea if this is possible but if it is you can get a great deal of improvement in resolution in differences in height. Cracks in particular would be highlighted.
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00 } From: "Tim Akinbo" {takinbo-at-onebox.com} } } Greetings. } I am a final student of the department of Electrical Electronics, } Obafemi Awolowo University, Ile-Ife, Nigeria. } I am doing a project titled "Low-Frequency Surface Microscopy". My } supervisor wants me to use an electric field-probe to detect changes in } the capacitance of the surface as cracks, boundaries, etc. are } encountered. I would be very grateful if you can send me any } information on the topic. } } My e-mail address is: femi_adeluyi-at-yahoo.com and my address is: } Olufemi Adeluyi } Department of Electronic and Electrical Engineering } Obafemi Awolowo University, } Ile-Ife, Nigeria. } } Thanking you in anticipation. } }
Maria Mendes wrote: ========================================== I am a research lab that does a lot of hard tissue processing, I as well as our EM departments have been using SPURR as our plastic of choice. I have been recently told by my supplier (Marivac) that in six months one of the components of SPURR will be discontinued. The component is ERL (Vinylcyclohexene Dioxide). I would like to know if others are using SPURR and if so have they encountered the same problem. Please forward your information to mmendes-at-mtsinai.on.ca. ========================================= This posting generated some worried communcations with customers asking about the continued availability of ERL 4221 (Vinylcyclohexene Dioxide).
The manufacturer reports to us no plans to discontinue ERL 4221 and there seems to be enough in the distribution pipelines to last some years into the future. ERL 4221 is currently available from SPI Supplies as I suspect it is available also from the other major suppliers of chemicals to the microscopy world.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
Depending upon the steel the receiver is made from a Fry's or Modified Fry's reagent (a cupric Chloride in HCl solution) works much better than the HNO3 but both alternated works still better. As I said in a previous post on this the NASA publication has quite a good rundown on all of the appropriate solutions. Hatcher, Jury and Weller a less detailed but acceptable section on it. The details of how to prepare the metal is as important as the solutions chosen if it is to work. Drop me a note and I'll write the procedure up if you can't find the NASA publication.
Jim Roberts
James L. Roberts Firearm and Toolmark Examiner Ventura Co. Sheriff's Lab (805) 654-2308 James.Roberts-at-mail.co.ventura.ca.us
} } } "Purdy, Sam" {SPurdy-at-nationalsteel.com} 11/06/00 05:42AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Eric:
Some forensic scientists have been successful in detecting stamped serial numbers by etching the part in a solution of 5 -10% HNO3 in water or alcohol for 10 to20 seconds. The disturbed metal will etch more darkly than the surrounding metal.
Best regards and good luck,
Sam Purdy Tecnical Center National Steel Corp. Trenton MI spurdy-at-nationalsteel.com
Experienced Confocal Microscopy/Electron Microscopy Technologist to work in Research Laboratory of Department of Pathology
The person we seek will be responsible for organization of a new facility that includes a confocal microscope and an electron microscope. The position includes overall management of the microscopy facility, user training, and user supervision. Requirements for the position include experience with light and transmission electron microscopy. This individual will oversee all aspects of specimen accession and processing, operation of the microscopes, photography, record keeping, and supervision of a technician. Knowledge of EM, biology, and pathology, as well as photographic procedures is required. The individual will work with only minimum supervision.
Position Requirements
Experience with confocal and digital imaging techniques, microinjection, visualization of living cells containing fluorescent probes, photobleaching, and fluorescence in situ hybridization.
Minimum of five years experience working in an electron microscopy laboratory with hands-on experience in tissue processing, dark room photography, and operation and maintenance of electron microscope.
Two years experience working on electron microscopy of human tissue specimens with knowledge of histology and pathology.
Excellent interpersonal and organizational skills are essential.
BachelorUs degree in science desirable.
Desirable Experience
Expertise in training in the operation of confocal microscope systems is a distinct advantage.
Familiarity with light microscopy methods, immunofluorescent staining, use of fluorescent probes for physiologic measurements and the general principles of cell biological research are critical. Significant facility with computers is desired.
Responsibilities
Serve as the technical manager of the facility and be responsible for the operation and maintenance of the confocal and EM microscope facility.
Perform routine transmission EM, including tissue processing, ultramicrotomy, and examination; do preventative maintenance on the equipment; maintain the lab, order supplies, schedule instruments, and oversee billing.
Image analysis at the light, confocal and electron microscopic levels and preparation of micrographs for publication.
Send resume to: Suzanne Mirra, M.D., Chair Department of Pathology Box 25 SUNY-Brooklyn 450 Clarkson Av. Brooklyn, NY 11203-2098
----------------- Submitted by Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
Experienced Histotechnologist to work in Neuropathology Research Laboratory of Department of Pathology
The person we seek will be responsible for maintaining a neuropathology research laboratory in the Department of Pathology at SUNY Downstate Medical Center. Knowledge of immunohistochemistry, special stains, and histopathology is required. Previous experience in handling central nervous system tissue preferred. The individual will work with only minimum supervision.
Position Requirements
Minimum of five years experience working in a laboratory with hands-on experience in tissue processing, histochemistry, immunohistochemistry, and operation of a light microscope. Experience with technical aspects of neuropathology and performing special stains including silver (Bielschowsky) and myelin stains.
Two years experience working on immunohistochemistry of CNS tissue specimens with knowledge of immunoreagents and experimental protocols.
BachelorUs degree in science desirable.
Laboratory skills including communications, ability to comply with safety and laboratory regulations, maintenance of laboratory equipment and resources, operation of computers and office equipment.
Advanced computer skills (word processing and database management) essential.
Desirable Experience
Confocal microscopy desirable.
Salary commensurate with experience
Responsibilities
Purchasing supplies and equipment, budget reports, laboratory maintenance and brain banking.
Will operate all microscopic, photographic and computer equipment, and keep accurate records of all laboratory experiments and procedures. Light and fluorescent microscopy; tissue processing for paraffin embedding, sectioning and slide stainer for immunohistochemical procedures; computer imaging (PhotoShop); general photography; and library and web searches
Familiarity with computer software for keeping records, report preparation and table/figure construction using Microsoft Office software (Word, Excel, PowerPoint) as well as Endnote and Adobe PhotoShop.
Send resume to:
Suzanne Mirra, M.D., Chair Department of Pathology Box 25 SUNY-Brooklyn 450 Clarkson Av. Brooklyn, NY 11203-2098
------------------- Submitted by Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
Experienced Electron Microscopy Technologist to work in Research Laboratory of Department of Pathology
The person we seek will be responsible for the overall operation of the EM laboratory in the Department of Pathology at SUNY Downstate Medical Center. This individual will oversee all aspects of specimen accession and processing, operation of the microscope, photography, record keeping, and supervision of a technician. Knowledge of EM, biology, and pathology, as well as photographic procedures is required. The individual will work with only minimum supervision.
Position Requirements
Minimum of five years experience working in an electron microscopy laboratory with hands-on experience in tissue processing, dark room photography, and operation and maintenance of electron microscope.
Two years experience working on electron microscopy of human tissue specimens with knowledge of histology and pathology.
BachelorUs degree in science desirable.
Laboratory management skills including effective written/verbal communication skills to interact with a diverse group, ability to comply with safety and laboratory regulations, maintenance of laboratory equipment and resources, and operation of computers and office equipment.
Desirable Experience
Previous experience in confocal microscopy highly desirable.
Previous experience in performing immunocytochemical staining and advanced computer skills usage (e.g. image analysis) is also desirable.
Salary commensurate with experience
Responsibilities
Maintain electron microscope in operating condition. Process clinical and research tissues for "thick" and "thin" sectioning. Darkroom management of photographic printing.
Send resume to: Suzanne Mirra, M.D., Chair Department of Pathology Box 25 SUNY-Brooklyn 450 Clarkson Av. Brooklyn, NY 11203-2098
----------------- Submitted by: Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
In many of the microscope rooms here at ANL. we use a product called SONEX by illbruck (http://www.illbruck.com). It is both fire rated and works reasonably well. Log into the TPM WWW site (http://tpm.amc.anl.gov) and you can see it . It is an acoustic foam panel which glues to the walls of the lab. The only problem is that being a foam product it tends to easily tear/rip if it is hit by objects.
Nestor
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
================================================================== Dr. 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 !
} We are going to install a curtain for sound absorption } in our TEM Lab. Is there any special curtain for this } purpose?
What sort of noise? For mid- to high-frequencies, a heavy fabric drape, interlined with an absorbent material, will do a pretty good job. For low frequencies, no curtain will do as you need a membrane with a seal.
On Mon, 6 Nov 2000, Nestor J. Zaluzec wrote: } In many of the microscope rooms here at ANL. we } use a product called SONEX by illbruck (http://www.illbruck.com). It is both fire } rated and works reasonably well. Log into the TPM WWW site } (http://tpm.amc.anl.gov) and you can see it . It is an acoustic foam panel which } glues to the walls of the lab. The only problem is that being a foam } product it tends to easily tear/rip if it is hit by objects.
This will do nothing to keep sound out. It will absorb high frequencies within the room.
Better sites for information and materials are Acoustic Sciences Corporation and Auralex.
Hello everyone, I was wondering if anyone had suggestions on a protocol for the fixation of either aquatic plant tissue or fish tissue to be viewed in the SEM. My interest is to study the sorption mechanisms of heavy metals (Cr, Cd and Pb) by aquatic plants and the bioaccumulation in fishes. I have no experience on SEM, so any information will be very usefull for me. Thanks, Regards!
Hi all, I would appreciate any suggestions or comments on how to suppress fluorescence of collagen in frozen sections that have been subjected to indirect immunofluorescence using Alexa conjugated antibodies. I have used normal goat and normal horse serum to block non-specific staining as well as 4 % fish gelatin but the collagen glows brightly every time. the samples were fixed in cold acetone before blocking. Incubation with sodium borohydride did not reduce fluorescence.
Thanks,
Cora Bucana Corazon D. Bucana, Ph.D. Dept. Cancer Biology UT M.D.Anderson Cancer Center 1515 Holcombe Blvd, Box 173 Houston, Texas 77030 Phone: 713-792-8106 FAX 713-792-8747
Kal could you expand on this a little? What type of membrane, arranged how?, etc. What range of frequencies can be controlled this way? I am looking for a relatively straightforward and inexpensive way of controlling particularly the low end of the spectrum of acoustic frequencies (generated by human voice, RVPs, etc.) in an FESEM lab. Chris
} For low frequencies, no curtain will do } as you need a membrane with a seal. } } Kal } }
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 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
I have found that if I keep some scraps around after the original installation of almost anything you can mix up a little glue wiht it and make a passable fix. Some times you have to do a little painting and make a creative dirty spot.
You will always be able to see it but no one else will.
If you are worried about getting the texture right use cheap wall paper paste and you can wash it out if you don't like it but it has enough water resistance to resist minor spills.
Test it where it won't show first.
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00 } } In many of the microscope rooms here at ANL. we } use a product called SONEX by illbruck (http://www.illbruck.com). It is both fire } rated and works reasonably well. Log into the TPM WWW site } (http://tpm.amc.anl.gov) and you can see it . It is an acoustic foam panel which } glues to the walls of the lab. The only problem is that being a foam } product it tends to easily tear/rip if it is hit by objects. } } Nestor } } } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Dear Listers, } } } } We are going to install a curtain for sound absorption in our TEM Lab. Is there any special curtain for this purpose? } } } } Regards, } } } } Kun Li } } } } } } Get your FREE Email at http://www.mailcityasia.com } } } ================================================================== } Dr. 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 ! } } ================================================================== } } }
We are experimenting with apoptose. Now, we want to view apoptotic cells under the SEM. Has anyone ideas what technique, solutions,....we have to use ? The sample is canine ovary.
Thx.
Bart De Pauw Ghent University Faculty of Veterinary medicine Department Morphology Salisburylaan 133 9820 Merelbeke Belgium
Description of Duties: Works with the Coordinator of Research Programs/Services in the management of the day-to-day activities of the Biological Science Imaging Resource, a campus wide imaging laboratory. This facility is responsible for assisting faculty and students in the preparation of biological images for research, publication and instruction. The Senior Biological Scientist works closely with the Coordinator to identify facility needs, establish priorities and develop policies within the laboratory as well as instruct users in the operation of the electron and light, including confocal microscopes. The incumbent must stay abreast of research standards and practices to maintain quality control and advise users on appropriate protocols. Knowledge in the use of MetaMorph, MetaFluor, Photoshop and PowerPoint along with MAC and PC platforms is a plus.
Contact Kim Riddle, 119 Bio Unit I, Department of Biological Science, Florida State University, Tallahassee, FL 32306-4370, riddle-at-bio.fsu.edu 850.644.6519 Imaging Resource web page http://bio.fsu.edu/~taylor/imaging On-line applications available at http://personnel.fsu.edu/emply/homepage.html
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Kimberly A. Riddle Florida State University tel: 850.644.6519 Biological Science Imaging Resource 119 Bio Unit I, 4370 fax: 850.644.0481 Tallahassee, FL 32306 riddle-at-bio.fsu.edu http://bio.fsu.edu/~taylor/imaging ~~~~~~~~~~~~~~~~~~~~~~~~~~~~
The Center for Materials Research and Analysis (CMRA) at the University of Nebraska at Lincoln has a matching fund in connection with a NSF award for a person to work with Prof. Y. Liu and his co-PIs on image processing of high resolution electron micrographs. The method developed will be applied to the TEM characterization of various nanostructured materials, such as TiN coatings, nanowires, super-conducting materials, recording media and permanent magnets. The initial appointment will be one year and can be renewed the second year, depending upon progress and availability of funding. Computer skills, knowledge of Fourier transform and image processing are required. Interested persons should email a letter of interest, resume and three persons of references to yliu-at-unlserve.unl.edu-- ******************************************************************* Yi Liu Department of Mechanical Eng. and CMRA 104 N Walter Scott Engineering Center University of Nebraska-Lincoln Lincoln, NE 68588-0656 Tel. (402) 472-7759 (Office) Tel. (402) 472-8762 (EM lab) Fax (402) 472-1465 Email: yliu-at-unlserve.unl.edu *******************************************************************
} could you expand on this a little? What type of membrane, arranged } how?, etc. What range of frequencies can be controlled this way? } I am looking for a relatively straightforward and inexpensive way of } controlling particularly the low end of the spectrum of acoustic } frequencies (generated by human voice, RVPs, etc.) in an FESEM } lab.
Assuming that you want to keep external noise out, you need to block the window since the glass transmits a lot. Hanging drapes, of any kind, will be most effective at higher frequencies where they can absorb energy transmitted by the glass.
If you can seal over the window with a membrane that will absorb at low frequencies (and you must seal it for it to be effective at low frequencies), take a look at SheetBlok at http://www.auralex.com/. This is effective down into the range of about 100Hz. Alternatively, you can brick up the window.
We also use Sonex foam products in most of our laboratories, but are now looking at purchasing an "enclosure" to put around a noisy water chiller. A company called Sound Seal (www.soundseal.com) makes such enclosures (at *very* reasonable prices), and has fabric curtains and panels for sound absorption purposes. Check them out...
Larry
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You might need to expand a bit on your goal. It seems that you are indicating that human speech is degrading your resolution. I find that hard to believe. Pumps, maybe. All these sources would depend on their manner of coupling to the scope. I don't think that speech would be that effectively coupled to be a problem.
Have you tried collecting some images where part of the image has the source present and part does not? I would think you could even shut down your rotary pumps for long enough to determine their influence on imaging. Take appropriate precautions.
Like the previous discussion on mu metal shielding, it seems that you might want to isolate the problem better and/or try some cheap and crude solutions before investing the bigger bucks.
FWIW, Warren Straszheim
At 07:57 AM 11/7/2000 +0000, you wrote:
} Kal } could you expand on this a little? What type of membrane, arranged } how?, etc. What range of frequencies can be controlled this way? } I am looking for a relatively straightforward and inexpensive way of } controlling particularly the low end of the spectrum of acoustic } frequencies (generated by human voice, RVPs, etc.) in an FESEM } lab. } Chris } } } For low frequencies, no curtain will do } } as you need a membrane with a seal. } } } } Kal } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } 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
I am interested in acquiring software for 3D image reconstruction and measurement (distance/volume) in 3D space. What is the current state of the market in this area? I am familiar with only one company that provides such a package (Vital Images: VoxelView), but I bet there are others. No need to evangelize - just let me know what other products I should examine. Money is not a big obstacle, I do not want to write code myself (I want a turnkey solution), and I do not require image acquisition or 2D analysis features. Thanks for the input. Robert J. Palmer Jr., Ph.D. Natl Inst Dental Craniofacial Res - Natl Insts Health Oral Infection and Immunity Branch Bldg 30, Room 308 30 Convent Drive Bethesda MD 20892 ph 301-594-0025 fax 301-402-0396
Speech can indeed be a problem with FESEMs. In the case of our instrument, the shrouding around the ion pumps behind the column was acting as a drum, transmitting speech particularly well. I removed the shrouding and the problem was eliminated. Plus now the scope looks much more interesting and impresses visitors! In newer models from the same company the shrouding is now perforated to minimize the problem.
} You might need to expand a bit on your goal. It seems that you are } indicating that human speech is degrading your resolution. I find that hard } to believe. Pumps, maybe. All these sources would depend on their manner of } coupling to the scope. I don't think that speech would be that effectively } coupled to be a problem.
Aloha, Tina
Mid-80s F, sunny with tropical breezes
**************************************************************************** * 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* ****************************************************************************
Yes I am indicating exactly that. Certain frequencies of the human voice at normal conversational amplitudes are picked up by FESEM columns and can induce saw-tooth edges on sharp-edged features (such as gold crystals on carbon) at magnifications in the x100k region. Trust me on this. It is a real problem. If one is contemplating high resolution work with an FESEM a hushed environment is essential.
Chris
} It seems that you are } indicating that human speech is degrading your resolution. I find that hard } to believe. Pumps, maybe. All these sources would depend on their manner of } coupling to the scope. I don't think that speech would be that effectively } coupled to be a problem. } } Have you tried collecting some images where part of the image has the } source present and part does not? I would think you could even shut down } your rotary pumps for long enough to determine their influence on imaging. } Take appropriate precautions. } } Like the previous discussion on mu metal shielding, it seems that you might } want to isolate the problem better and/or try some cheap and crude } solutions before investing the bigger bucks. } } FWIW, } Warren Straszheim } } At 07:57 AM 11/7/2000 +0000, you wrote: } } } Kal } } could you expand on this a little? What type of membrane, arranged } } how?, etc. What range of frequencies can be controlled this way? } } I am looking for a relatively straightforward and inexpensive way of } } controlling particularly the low end of the spectrum of acoustic } } frequencies (generated by human voice, RVPs, etc.) in an FESEM } } lab. } } Chris } } } } } For low frequencies, no curtain will do } } } as you need a membrane with a seal. } } } } } } Kal } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } } 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 } }
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 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
I have 2 Pfeiffer's no-oil vacuum pumps for 8m3/min to offer . They have only 50hours of work. The original price the last year was $6400. Any offer negotiable. Marek Malecki, M.D., Ph.D. Director and Principal Investigator
Molecular Imaging Laboratories University of California, San Diego
Warren E Straszheim: } } You might need to expand a bit on your goal. It seems that you are } indicating that human speech is degrading your resolution. I find that hard } to believe.
Dear Warren, I still remember Ken Downing demonstrating that his IVEM image would vibrate whenever one talked at the microscope--in fact, at a DOE site visit, the head of the team, a Dr. Happer, seemed singularly unimpressed until he was shown that clapping in the IVEM room would cause the image to vibrate. He was apparently quite amused, and the site visit went well after that. The IVEM was known for a time as the "Happer clapper". Yours, Bill Tivol Wadsworth Center Albany NY
I have to back Chris up on this. We have often seen voice-induced vibration in our FESEM. No talking during high-mag, high-res work.
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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Yes I am indicating exactly that. Certain frequencies of the human voice at normal conversational amplitudes are picked up by FESEM columns and can induce saw-tooth edges on sharp-edged features (such as gold crystals on carbon) at magnifications in the x100k region. Trust me on this. It is a real problem. If one is contemplating high resolution work with an FESEM a hushed environment is essential.
Chris
} It seems that you are } indicating that human speech is degrading your resolution. I find that hard } to believe. Pumps, maybe. All these sources would depend on their manner of } coupling to the scope. I don't think that speech would be that effectively
} coupled to be a problem. } } Have you tried collecting some images where part of the image has the } source present and part does not? I would think you could even shut down } your rotary pumps for long enough to determine their influence on imaging.
} Take appropriate precautions. } } Like the previous discussion on mu metal shielding, it seems that you might } want to isolate the problem better and/or try some cheap and crude } solutions before investing the bigger bucks. } } FWIW, } Warren Straszheim } } At 07:57 AM 11/7/2000 +0000, you wrote: } } } Kal } } could you expand on this a little? What type of membrane, arranged } } how?, etc. What range of frequencies can be controlled this way? } } I am looking for a relatively straightforward and inexpensive way of } } controlling particularly the low end of the spectrum of acoustic } } frequencies (generated by human voice, RVPs, etc.) in an FESEM } } lab. } } Chris } } } } } For low frequencies, no curtain will do } } } as you need a membrane with a seal. } } } } } } Kal } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } } 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 } }
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 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
} Yes I am indicating exactly that. Certain frequencies of the human } voice at normal conversational amplitudes are picked up by FESEM } columns and can induce saw-tooth edges on sharp-edged features } (such as gold crystals on carbon) at magnifications in the x100k } region. Trust me on this. It is a real problem. If one is contemplating } high resolution work with an FESEM a hushed environment is } essential.
OK. Where are these voices? In the room? If not, the issue is to keep them out. Perhaps you should contact a number of the specialist firms and consult with them via their websites. I have gotten very useful advice and information without (or prior to) purchase of sound treatment materials.
Kal
} } It seems that you are } } indicating that human speech is degrading your resolution. I find that hard } } to believe. Pumps, maybe. All these sources would depend on their manner of } } coupling to the scope. I don't think that speech would be that effectively } } coupled to be a problem. } } } } Have you tried collecting some images where part of the image has the } } source present and part does not? I would think you could even shut down } } your rotary pumps for long enough to determine their influence on imaging. } } Take appropriate precautions. } } } } Like the previous discussion on mu metal shielding, it seems that you might } } want to isolate the problem better and/or try some cheap and crude } } solutions before investing the bigger bucks. } } } } FWIW, } } Warren Straszheim } } } } At 07:57 AM 11/7/2000 +0000, you wrote: } } } } } Kal } } } could you expand on this a little? What type of membrane, arranged } } } how?, etc. What range of frequencies can be controlled this way? } } } I am looking for a relatively straightforward and inexpensive way of } } } controlling particularly the low end of the spectrum of acoustic } } } frequencies (generated by human voice, RVPs, etc.) in an FESEM } } } lab. } } } Chris } } } } } } } For low frequencies, no curtain will do } } } } as you need a membrane with a seal. } } } } } } } } Kal } } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } } } 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 } } } } } } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } 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 07710 585 401 } FAX: +44 (0) 131 653 6248 } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ }
Well, I guess I learned something today. I usually show operators that loud noises will show up as low energy noise in the EDS spectra. I had no idea that it also affected the images that much. We do not do that much work above 10 kx. I will have to bear that in mind the next time we venture into that realm and turn down the 1812 Overture in the background. I may have to quiet our pumps down too.
Warren
At 07:59 PM 11/7/2000 +0000, you wrote: } Yes I am indicating exactly that. Certain frequencies of the human } voice at normal conversational amplitudes are picked up by FESEM } columns and can induce saw-tooth edges on sharp-edged features } (such as gold crystals on carbon) at magnifications in the x100k } region. Trust me on this. It is a real problem. If one is contemplating } high resolution work with an FESEM a hushed environment is } essential. } } Chris
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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, 7 Nov 2000, Chris Jeffree wrote: } } } Yes I am indicating exactly that. Certain frequencies of the human } } voice at normal conversational amplitudes are picked up by FESEM } } columns and can induce saw-tooth edges on sharp-edged features } } (such as gold crystals on carbon) at magnifications in the x100k } } region. Trust me on this. It is a real problem. If one is contemplating } } high resolution work with an FESEM a hushed environment is } } essential. } } OK. Where are these voices? In the room? If not, the } issue is to keep them out. Perhaps you should contact a } number of the specialist firms and consult with them via } their websites. I have gotten very useful advice and } information without (or prior to) purchase of sound } treatment materials. } } Kal } (snip)
I'd like to jump into this one by adding my $0.02. I'm familiair with Ken Downing's results related to vibrations. We tried it ourselves as well and microphonics are indeed a very real problem when one does HR work. The effects are indeed very clearly visible on the TV screen at high (} = 100kx) mag. Ken (together with Bob Glaeser) and we (perhaps others have done something similar) have taken this a step further and constructed boxes made of styrofoam that fit around the cryo-holder (side entry types). The idea was to keep microphonics away from the holder including those created by us talking or on the phone or listening to a radio, but also those originating from pumps, ventilation and/or air conditioning systems.
I operate a Noran Voyager 3.3 EDS system, which is a UNIX based system and operates on a Sun Sparc Station 5 with SunOS 5.3. The problem is the local IT department is reluctant to place this on the network which is all PC based. The IT staff feels it would be more cost efficient use of their time, if I found any product which will replace the Unix box/software and utilize the present hardware (detector, amplifiers, etc.). Anyone have any experience with this?
TIA,
Dave Audette OSRAM Sylvania 71 Cherry Hill Drive Beverly, MA 01915 david.audette-at-sylvania.com
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Does anybody know of any used CCD camera and EDS in good condition that are up for sail? I would like to purchase these to use on a used JEOL 200CX TEM.
Your help would be greatly appreciated.
Regards, Thearith
_________________________
Thearith Ung, Ph.D. Quantum Dot Corporation 26136 Research Road Hayward CA 94545 Tel: +1-510-887-8775 (x4125) Fax:+1-510-783-9729 www.qdots.com
} I operate a Noran Voyager 3.3 EDS system, which is a } UNIX based system and operates on a Sun Sparc Station } 5 with SunOS 5.3. The problem is the local IT } department is reluctant to place this on the network } which is all PC based. The IT staff feels it would be } more cost efficient use of their time, if I found any } product which will replace the Unix box/software and } utilize the present hardware (detector, amplifiers, } etc.). Anyone have any experience with this?
Your IT department is over simplifying the cost. They most likey think it's only 1 or 2k to get a WinOS computer. They are very wrong.
Cost is really going to be from ~10k to ~20k to replace the Voyager depending if you go with a Noran (Quest) replacement or some other like us. You will be able to keep the detector (although maybe a preamp change) but everything else is replaced. Also you will have to learn new EDS software (your time at two to three weeks). In addition, all your old data and analysis might or might not be movable to the new EDS system (how much is your data worth).
Tell your managment that your IT department needs to to support you rather than you supporting them, after all, what is the function of the TI department. The Voyager is a plain UNIX box, if no one in the IT department understands UNIX enough to network it, they are pretty sorry. If that does not work, then tell then you will be glad to replace the UNIX with a WinOS equivalent provided they pony up the funds. Also remind managment that you will get no real work done while you learn the new software. Add the fact that previous data results might be forever lost to access. Let them figure that the IT deparment is going to cost the company 10-20k plus the cost of your time because they can't seem to do their job which is to support the computing needs of the workers. Bottom line is that it's going to cost the company 40-60k direct and indirect costs however if you can't move the data, then it could cost the company big time if they need to access it in the future. Now what is really more cost efficient?
Sorry for the rant, but it really bugs me when IT departments operate in such a priesthood fashion, we run into it all the time with our MacOS products so much that we came out with WinOS products for those that have to have WinOS because of stupid IT restrictions.
Scott ----------------------------------------------------------------------- Scott D. Davilla Phone: 919 489-1757 ext 13 (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
Huh? What PCs? If they are all DOS-based, I can identify with their concerns. If the PCs are running client TCP/IP, the addition of the Sun should be trivial.
How are LAN IPs assigned? If there is a LAN NAT server, then just assign the Sun a within-scope IP address. If it is DHCP, just do it.
I don't see what the big deal is. Except....I'll venture to say that the PC jockeys have no Unix experience. That would be a good reason to keep the Sun away from the PC LAN.
gg
At 04:36 PM 11/7/00, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} From: "Scott D. Davilla" {davilla-at-4pi.com} } } } I operate a Noran Voyager 3.3 EDS system, which is a } } UNIX based system and operates on a Sun Sparc Station } } 5 with SunOS 5.3. The problem is the local IT } } department is reluctant to place this on the network } } which is all PC based. The IT staff feels it would be } } more cost efficient use of their time, if I found any } } product which will replace the Unix box/software and } } utilize the present hardware (detector, amplifiers, } } etc.). Anyone have any experience with this? =========== Just tell them you can't get fries wiht that and do their job. And don't hold your breath. If you ask around there may be a couple working wiht Linux and they can figure it out.
Just borrow some one with unix experiance to set it up from another department for a favor to be anouced later and let the IT department spin on it. If they are windows based the box will weld it's self shut by molecular defusion before they take the time to figure out unix.
Hiring a consultant is a reasonable soluting if a college is near. Once it is set up it can be maintained from any place in the world. Unless you are adding stuff a couple of hours a month will do the job.
The best answer is some one in the shop learn to do it. It will take some time but it will pay off in he long run.
Good luck You need it. Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
} From: "William Tivol" {tivol-at-wadsworth.org} } } Warren E Straszheim: } } } } You might need to expand a bit on your goal. It seems that you are } } indicating that human speech is degrading your resolution. I find that hard } } to believe. } } Dear Warren, } I still remember Ken Downing demonstrating that his IVEM } image would vibrate whenever one talked at the microscope--in fact, } at a DOE site visit, the head of the team, a Dr. Happer, seemed } singularly unimpressed until he was shown that clapping in the } IVEM room would cause the image to vibrate. He was apparently } quite amused, and the site visit went well after that. The IVEM } was known for a time as the "Happer clapper".
Noise is a problem of inches. The best solution is a corn field 5 miles from any where in a 10 foot basment on a still night and a active noise canceler makes it better. We put our insterments in buildings with people and macinery. I was testing a simple load cell with a 12 bid A/D converter. It was no trick to measure a car drive by 30 yards away a truck 100 yards a way a fellow walking by in the hall. And a load cell with a 12 bit a/d converter is not a very sensitive insterment.
There are good solutions down to 4 or 5 Hz below that you have trouble. Floating it on an air table gets x and y but not z. floating it in liqid work until you get a resonate situation. Active cancelation works the best and costs the most. The things tried on this list sows that it is a difficult problem.
You live or die by the contact with your coustomers so you lab has to be near them. That usualy means near traffic and a lot of people. and a lot of vibration and noise. Call your favirate noise reduction vendor and say fixit I have this much money or go to you customers hat in hand and get what it takes to do it right.
Good luck Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
I had the same system when I worked with ESA. I don't remember much difficulty with SunOS 5.3. We just gave it a name & IP address on our LAN, checked a few boxes in the set-up screen & off we went. I don't think we even needed to call in the IT people.
If your IT people can set up a DOS box, they can set up SunOs 5.3. Should be a 5-10 minute job, longer if they have to RTFM.
Regards
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/
-----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] Sent: Wednesday, November 08, 2000 6:24 AM To: Dave Audette Cc: MSA listserver
Huh? What PCs? If they are all DOS-based, I can identify with their concerns. If the PCs are running client TCP/IP, the addition of the Sun should be trivial.
How are LAN IPs assigned? If there is a LAN NAT server, then just assign the Sun a within-scope IP address. If it is DHCP, just do it.
I don't see what the big deal is. Except....I'll venture to say that the PC jockeys have no Unix experience. That would be a good reason to keep the Sun away from the PC LAN.
gg
At 04:36 PM 11/7/00, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} From: "Gary Gaugler" {gary-at-gaugler.com} } } Huh? What PCs? If they are all DOS-based, I can identify } with their concerns. If the PCs are running client TCP/IP, } the addition of the Sun should be trivial. } } How are LAN IPs assigned? If there is a LAN NAT } server, then just assign the Sun a within-scope IP address. } If it is DHCP, just do it. } } I don't see what the big deal is. Except....I'll venture } to say that the PC jockeys have no Unix experience. } That would be a good reason to keep the Sun away } from the PC LAN.
No it is a good reason to keep it away from the people runing the LAN. The Lan it's self doesn't care what is hooked to it. The fool that trys to hook it up has a lot to do wiht it. Back up everything before the come and after they leave. So you can go back to a working state not connected to the lan.
I can't get Unix jocks for my own projects good luck on yours. And one is my son.
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
I would echo the sentiments you have already received. It seems like your IT folks just have no experience with UNIX. (But neither do I.) It would take them some time to learn it, even if they are willing.
It absolutely should be a trivial matter to put a Unix box on a TCP/IP network, and that is all you should need. If they are thinking that they have to run Novell or some other protocol on your Unix system, that could be a very difficult prospect. But that is not what you would want. All you need is TCP/IP and Unix is where TCP/IP got its start.
I would suggest that you get the rules for setting up an additional PC node on the network. Let them give you an IP number and name or tell you that your net runs a DHCP server to hand out IP numbers. Hey, you could even give them an old, dummy PC and say hook this up to the net and call it such and so. Then you could have someone who does know Unix and Windows come in, look over the setup, transfer the setting to the Unix box, and dispose of the dummy PC.
In other words, it really should not be that tough. I bet you have someone around that could do it. That is the way I helped another lab on campus get their PDP-based Kevex EDS system onto the net. I had already done it with ours.
Good luck.
Warren S.
At 04:36 PM 11/7/2000 -0800, you wrote: } Hello listers, } } I operate a Noran Voyager 3.3 EDS system, which is a } UNIX based system and operates on a Sun Sparc Station } 5 with SunOS 5.3. The problem is the local IT } department is reluctant to place this on the network } which is all PC based. The IT staff feels it would be } more cost efficient use of their time, if I found any } product which will replace the Unix box/software and } utilize the present hardware (detector, amplifiers, } etc.). Anyone have any experience with this? } } TIA, } } Dave Audette } OSRAM Sylvania } 71 Cherry Hill Drive } Beverly, MA 01915 } david.audette-at-sylvania.com } } } } __________________________________________________ } Do You Yahoo!? } Thousands of Stores. Millions of Products. All in one Place. } http://shopping.yahoo.com/
Our experience is exactly the same as Tim's. Ask your IT guys what it is that is peculiar about your network that precludes being able to run a unix box on it. We followed instructions provided by some user friendly unix experts and set up our Voyagers on the network ourselves. I've got the notes on how to do it and could send them to you if you want. The instructions are pretty straightforward,ie the commands to use to get to the setup screen, optional files to create and edit if you want additional networking functions, and so on.
Maybe best if you can persuade the IT guys to assign you with a permanent IP address. From there you shouldn't really need them for anything else. Any other site specific info that you need should be obtainable by looking at the network setup of any PCs (or Macs!) running nearby.
Maybe it's a 15 minute job to do once, 5 minutes if you can avoid the damn FM.
Regards, Arthur
} } I had the same system when I worked with ESA. } I don't remember much difficulty with SunOS 5.3. We just gave it a name & IP } address on our LAN, checked a few boxes in the set-up screen & off we went. } I don't think we even needed to call in the IT people. } } If your IT people can set up a DOS box, they can set up SunOs 5.3. Should be } a 5-10 minute job, longer if they have to RTFM. } } Regards } } 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/ } } -----Original Message----- } } From: Gary Gaugler [mailto:gary-at-gaugler.com] } Sent: Wednesday, November 08, 2000 6:24 AM } To: Dave Audette } Cc: MSA listserver } Subject: Re: EDS networking } } At 04:36 PM 11/7/00, 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 listers, } } } } I operate a Noran Voyager 3.3 EDS system, which is a } } UNIX based system and operates on a Sun Sparc Station } } 5 with SunOS 5.3. The problem is the local IT } } department is reluctant to place this on the network } } which is all PC based. The IT staff feels it would be } } more cost efficient use of their time, if I found any } } product which will replace the Unix box/software and } } utilize the present hardware (detector, amplifiers, } } etc.). Anyone have any experience with this? } } } } TIA, } } } } Dave Audette } } OSRAM Sylvania } } 71 Cherry Hill Drive } } Beverly, MA 01915 } } david.audette-at-sylvania.com } } } } } } } } __________________________________________________ } } Do You Yahoo!? } } Thousands of Stores. Millions of Products. All in one Place. } } http://shopping.yahoo.com/
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/
At 04:36 PM 11/7/00 -0800, Dave Audette wrote: } I operate a Noran Voyager 3.3 EDS system, which is a } UNIX based system and operates on a Sun Sparc Station } 5 with SunOS 5.3. The problem is the local IT } department is reluctant to place this on the network } which is all PC based.
If you can find anyone there adept in Unix, they would be able to install Samba, a networking package that will make your box look like every other PC in the office in terms of file sharing and printing. You can see their drives or they can see yours.
http://us1.samba.org/samba/samba.html is the start, and http://us5.samba.org/samba/ftp/Binary_Packages/solaris/Sparc/ has binaries or source code for your platform.
Without much information on the networking setup for your lab/company, it shouldn't be that difficult to add it to the existing windows network. You could do it with or without your IT department.
If you have a network line, install a switch/hub, add in your Sun computer to the network, setup its configuration, and you should be able to do simple file transfers via ftp. You could also see if Samba is available for your Sun OS, and then share files more easily.
More information about the network configuration that exists for your company would be needed to give more precise suggestions.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ 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, 7 Nov 2000, Dave Audette wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello listers, } } I operate a Noran Voyager 3.3 EDS system, which is a } UNIX based system and operates on a Sun Sparc Station } 5 with SunOS 5.3. The problem is the local IT } department is reluctant to place this on the network } which is all PC based. The IT staff feels it would be } more cost efficient use of their time, if I found any } product which will replace the Unix box/software and } utilize the present hardware (detector, amplifiers, } etc.). Anyone have any experience with this? } } TIA, } } Dave Audette } OSRAM Sylvania } 71 Cherry Hill Drive } Beverly, MA 01915 } david.audette-at-sylvania.com } } } } __________________________________________________ } Do You Yahoo!? } Thousands of Stores. Millions of Products. All in one Place. } http://shopping.yahoo.com/ } }
And Hertz. Our HVEM column is very sensitive to 20 Hz, but almost completely insensitive to frequencies a few Hz away on either side. Once you have someone sit at the scope and watch the image as a sound generator is swept through a range of frequencies, you will know what to be concerned about, and then you can get the appropriate shielding.
The best solution is a corn field 5 miles from any where in a 10 foot basment on a still night and a active noise canceler makes it better.
If you build it, will they come? ;-)
We put our insterments in buildings with people and macinery. I was testing a simple load cell with a 12 bid A/D converter. It was no trick to measure a car drive by 30 yards away a truck 100 yards a way a fellow walking by in the hall. And a load cell with a 12 bit a/d converter is not a very sensitive insterment.
There are good solutions down to 4 or 5 Hz below that you have trouble. Floating it on an air table gets x and y but not z. floating it in liqid work until you get a resonate situation. Active cancelation works the best and costs the most. The things tried on this list sows that it is a difficult problem.
There is no free lunch. Floating the instrument renders it sensitive to sound in the room, and mounting it solidly to the floor makes transmission through the ground a problem. If the soil mechanics are such that the frequencies transmitted to the instrument are not those to which it is sensitive, but the sound emitted by the pumps is at the sensitive frequencies, it can be better to bolt the instrument to the floor. As you said, an environment completely free of vibration is best, and active cancellation is good. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
I would like to say thanks to all for the information provided on preparing TEM cross sections of DLC thin films on steel. Also I would like to note that I do have access a FIB tool at Madison Area Technical College, so anyone who has any info on specialized techniques for the TEM prep of DLC thin films using the FIB would greatly be appreciated. Again, thanks to all
Daniel L. Flatoff student Madison Area Technical College
Does anyone have any experience with instituting ISO compliance of SEM's? How do you keep your SEM calibrated and how often are calibrations performed? What type of support does your mfg. offer?
Thank you to those who replied and let me know that their cameras and EDS systems are available for sale. I forgot to ask also what are some of the best vendors that sell CCD cameras and EDS systems? This information will be valuable in helping me to apply for grants to purchase these instruments.
Thank you in advance.
Regards, Thearith _________________________
Thearith Ung, Ph.D. Quantum Dot Corporation 26136 Research Road Hayward CA 94545 Tel: +1-510-887-8775 (x4125) Fax:+1-510-783-9729 www.qdots.com
At 04:36 PM 11/7/2000 -0800, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} From Dave's posting, we can't tell. Before he starts any battles with his IT department, he needs to learn (by his own research or with the help of an advocate) what the reasons are for the IT advice. Maybe he already knows this and didn't include it in his message (but I suspect not, for why would he have posted his message). Without this knowledge, though, the various responses (excepting Scott's) that have been made are not really very helpful.
I know I'm on a soapbox here, and in any forum like this, a major degree of *caveat emptor* has to apply. It is certainly true that, if the OSRAM/Sylvania network is TCP/IP, then the UNIX box is easily networked. If not, though, then Dave will have been misguided, rather than helped, by the responses he has received.
Tony.
} Hello listers, } } I operate a Noran Voyager 3.3 EDS system, which is a } UNIX based system and operates on a Sun Sparc Station } 5 with SunOS 5.3. The problem is the local IT } department is reluctant to place this on the network } which is all PC based. The IT staff feels it would be } more cost efficient use of their time, if I found any } product which will replace the Unix box/software and } utilize the present hardware (detector, amplifiers, } etc.). Anyone have any experience with this? } } TIA, } } Dave Audette } OSRAM Sylvania } 71 Cherry Hill Drive } Beverly, MA 01915 } david.audette-at-sylvania.com } } } } __________________________________________________ } Do You Yahoo!? } Thousands of Stores. Millions of Products. All in one Place. } http://shopping.yahoo.com/ }
** Anthony J. Garratt-Reed ** MIT Room # 13-1027 ** 77 Massachusetts Avenue ** Cambridge, MA 02139-4307 ** USA ** ** Phone: (+) 1-617-253-4622 ** Fax: (+) 1-617-258-6478 **
We are QS9000 certified which is the automotive equivalent to ISO. Basically, it really is up to you how often you want to do it. We have some instruments set up for twice a year and some on an annual basis. Basically, the ISO is a paper work tracking system that insures that you do what you say you do in your SOP's or in our case, SLP's (System Level Procedures). You decide what you want to do and stick to it. You write the specifications that you need to follow and document them. A suggestion, don't write them too stringent. You can use the ISO process to insure that your instrumentation is maintained properly. For example if you write a specification that your EDS detector should have a particular energy resolution for an element and if it is out, that the detector is repaired, then your company has no options, it must have it repaired or have it taken out of service. However, you should be careful in writing your procedures if you don't want it out of service. For example, you don't want the normal resolution degradation in performance with lifetime of a detector shutting you down.
We have the manufacturer's or our 3rd party's service engineers verify the calibration as part of the annual or semi-annual PM visit on our service contracts on all our scanning instruments. On our TEM, I just do the annual calibration verification myself and record it. If I do anything that requires a new calibration before the year is up, I just put the new calibration data in the records book. It doesn't hurt to do it more often. It is a real pain if you find out that it was out of calibration and you were sending data to customers. Then you have to contact them all and tell them that their results were done while it was not in calibration.
-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: Jesse Rodrigues [mailto:Jesse_Rodrigues-at-kopin.com] } Sent: Wednesday, November 08, 2000 4:02 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: ISO calibration of an 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/MicroscopyListse} rver/FAQ.html } } } -------------------------------------------------------------- } ---------. } } } } } Hi, } } Does anyone have any experience with instituting ISO } compliance of SEM's? } How do you keep your SEM calibrated and how often are calibrations } performed? What type of support does your mfg. offer? } } Any information would be helpful. } } Thank you, } } Jesse Rodrigues } Kopin Corporation } 695 Myles Standish Blvd. } Taunton, MA 02780 } } }
Dear Listers, A test I have heard of for the acoustic resonence frequency of your TEM sample holder and stage is the "whistle test". While looking at a sample with good high res. detail through the binocular eyepieces, mag. 100,000 to 200,000 X, whistle starting low and going up. You should see a frequency in there where the sample vibrates. Never move or speak while taking a high res. photo. At 01:01 PM 11/8/00 -0500, you wrote: } Dear Gordon, } } Noise is a problem of inches. } } And Hertz. Our HVEM column is very sensitive to 20 Hz, but almost } completely insensitive to frequencies a few Hz away on either side. Once you } have someone sit at the scope and watch the image as a sound generator is swept } through a range of frequencies, you will know what to be concerned about, and } then you can get the appropriate shielding. } } The best solution is a corn field 5 miles } from any where in a 10 foot basment on a still night and a active noise } canceler makes it better. } } If you build it, will they come? ;-) } } We put our insterments in buildings with people } and macinery. I was testing a simple load cell with a 12 bid A/D } converter. It was no trick to measure a car drive by 30 yards away a truck } 100 yards a way a fellow walking by in the hall. And a load cell with a 12 } bit a/d converter is not a very sensitive insterment. } } There are good solutions down to 4 or 5 Hz below that you have trouble. } Floating it on an air table gets x and y but not z. floating it in liqid } work until you get a resonate situation. Active cancelation works the best } and costs the most. The things tried on this list sows that it is a } difficult problem. } } There is no free lunch. Floating the instrument renders it sensitive to } sound in the room, and mounting it solidly to the floor makes transmission } through the ground a problem. If the soil mechanics are such that the } frequencies transmitted to the instrument are not those to which it is } sensitive, but the sound emitted by the pumps is at the sensitive frequencies, } it can be better to bolt the instrument to the floor. As you said, an } environment completely free of vibration is best, and active cancellation is } good. } Yours, } } Bill Tivol } Wadsworth Center } Albany NY } (518) 473-7399 WFT02-at-health.state.ny.us } 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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I seem you went to reflective mode of work. Sometimes it happens when charge does not flow from sample for some reasons. In this case you can see in backscattered electrons the image of the detector at polepiece (and some halves or quadrants will change contrast depending on what mode you choose), and in secondary electrons - all specimen chamber from inside. This effect can be used for example for testing of BSE signal path.
Advises: 1. Test the electrical path between the sample and microscope case (if you have absorbed current amplifier or meter the resistance may be large). 2. Check this effect on metal test sample. 2. Sputter non-metal sample by conductive film. 3. Work in low vacuum mode if you have it.
Hope it helps. Regards
Victor Sidorenko, ANTRON Co. Ltd., Moscow, Russia.
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That is a good question and I'm sure, one which will generate a high degree of variability of responses.
My (un-named) FESEM company does calibrations based on when the preventive maintenance (PM) routines are performed. These are supposed to be done at six month intervals. Actually, they get done only if I have a hard failure of the system and then when that problem is fixed, a PM is done....if there is time to do it.
In reality, I can do the key measurements myself to ensure that the SEM is in spec. Gun brightness, image symmetry, mag accuracy, etc.
I suppose that it depends on how much you want or need the maker to do the calibration versus what you can do or are allowed to do yourself.
gg
At 01:01 PM 11/8/00, you wrote:
} Hi, } } Does anyone have any experience with instituting ISO compliance of SEM's? } How do you keep your SEM calibrated and how often are calibrations } performed? What type of support does your mfg. offer? } } Any information would be helpful. } } Thank you, } } Jesse Rodrigues } Kopin Corporation } 695 Myles Standish Blvd. } Taunton, MA 02780
from: "Anthony Garratt-Reed" {tonygr-at-mit.edu} } Despite what many respondants to this message have said/implied, it is } entirely possible that Dave's IT department does know what it is talking } about. A Department/Plant network need not be implemented using TCP/IP, } and if it is all PC-based, and doesn't require routing, then NetBEUI might } be a sensible choice. IPX/SPX (Novell) networking, or even Lantastic or } another proprietary system may alternatively be in use.
Then let them build a gateway to put the Unix box on what ever they have. Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
} From: "William F. Tivol" {wft03-at-health.state.ny.us} , } Dear Bill,
} Noise is a problem of inches.
How true and the solution is often found by Edisonian iteration. } } And Hertz. Our HVEM column is very sensitive to 20 Hz, but almost } completely insensitive to frequencies a few Hz away on either side. Once you } have someone sit at the scope and watch the image as a sound generator is swept } through a range of frequencies, you will know what to be concerned about, and } then you can get the appropriate shielding. ======== Have you tried to find what is resonate at 20 Hz and do somting to break it up. That is high enough you might have a small chance of doing something about it. } } The best solution is a corn field 5 miles } from any where in a 10 foot basement on a still night and a active noise } canceler makes it better. } } If you build it, will they come? ;-) } We really do a lot of radio frequency noise testing in remote fields. Motorola has a 3 million dollar RF test chamber to test and certify some wireless devices for noise output on the receiving frequency. The company that owns the net work has a particular cornfield in Iowa they use for the same tests for considerably less money. John Deere tests the computers on their tractors for noise output and immunity by driving them out in the middle of a field using battery operated test instruments.
I built some sensors used on Ag machinery. It is the quietest environment I have ever seen The only noise was from the alternator on the diesel engine and the computers. NO 60 or 120 CPS noise at all.
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
} from: "Anthony Garratt-Reed" {tonygr-at-mit.edu} } Despite what many respondants to this message have said/implied, it is } entirely possible that Dave's IT department does know what it is talking } about. A Department/Plant network need not be implemented using TCP/IP, } and if it is all PC-based, and doesn't require routing, then NetBEUI } might be a sensible choice. IPX/SPX (Novell) networking, or even } Lantastic or } another proprietary system may alternatively be in use. } } Then let them build a gateway to put the Unix box on what ever they have. } Gordon } Gordon Couger gcouger-at-couger.com } Stillwater, OK www.couger.com/gcouger } 405 624-2855 GMT -6:00
I guess that I just don't see what the big deal is here. Get a static IP, agreed-upon host name and set the Unix box up.
If the system is on a LAN, I don't see why routing is an issue. That will/should be transparent as long as the main router is set at the default gateway. If a bridge is involved, I can see that things might be more complicated--but not insurmountable.
Unix/Linux to PCs and Macs are done all the time. No big deal. My own LAN is exactly like this. Mac G4/400, 3 P-3/800, SGI O2, SGI Indigo 2, Sun Ultra 5, P2-400 Linux. All on 10BaseT running TCP/IP. PCs use Microsoft TCP/IP client. Mac runs DAVE. SGI runs IRIX 6.5. Sun runs SunOS/Solaris.
I simply do not see the big deal here. The key is to get a static IP on the branch LAN, know the gateway IP, and provide DNS IPs. That ought to do it.
I am in need of a Monte Carlo electron scattering programme (with or without EDS) for SEM scattering simulations. Is there one for downloading for a PC, freeware or otherwise?
David Cockayne Department of Materials University of Oxford Parks Road Oxford OX1 3PH England
Union Carbide is the manufacturer of ERL 4206, which is a component of of the Spurr Kit. They have discontinued the production of ERL 4206, but they do supply ERL 4221 which does not have the low viscosity of the ERL 4206. We continue to have ERL 4206 and Spurr Kits in stock and we are confident that we will have an ideal substitute for this product very shortly.
John Arnott We continue --
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Mendes, Maria wrote:
} Hello I would like to subscribe to your service, please let me know if there } is any service charge. } Please pass on my problem to your subscribers. } I am a research lab that does a lot of hard tissue processing, I as well as } our EM departments have been using SPURR as our plastic of choice. I have } been recently told by my supplier (Marivac) that in six months one of the } components of SPURR will be discontinued. The component is ERL } (Vinylcyclohexene Dioxide). I would like to know if others are using SPURR } and if so have they encountered the same problem. Please forward your } information to mmendes-at-mtsinai.on.ca. } Thank-you in advance for any help offered. } } Maria
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I just visited with Lucille Giannuzzi at Univ. Central Florida where she taught me a few things about FIB. I did ask her about carbon. She told me that they have a water injector to help with carbon films such as resists on semiconductors. Based on my experiences with DLC films, I suspect that you will need to use that to get reactive etching going to cut through the DLC. You will probably need to cut the samples normally, but you will also will need to watch out for channeling affects. Again, I learned more about this from Lucille.
I'm not the expert here. You might consider contacting one.
-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: Daniel L Flatoff [mailto:dflatoff-at-stu.madison.tec.wi.us] } Sent: Wednesday, November 08, 2000 3:32 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: TEM sample prep of steel } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: 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 } } } -------------------------------------------------------------- } ---------. } } } I would like to say thanks to all for the information } provided on preparing TEM cross sections of DLC thin films } on steel. Also I would like to note that I do have access } a FIB tool at Madison Area Technical College, so anyone who } has any info on specialized techniques for the TEM prep of } DLC thin films using the FIB would greatly be appreciated. } Again, thanks to all } } } Daniel L. Flatoff } student } Madison Area Technical College } } } }
The other day while on a consulting job that entailed photographing a set of ceramic tiles in a range of colors, I discovered an interesting quirk of at least this Olympus C-2500L digital camera. I suspect that other consumer/prosumer cameras would suffer the same fate.
There were four tiles ranging from, say, very light purple to dark purple. We'd put them on a black velvet background. We had tremendous trouble getting a accurate color. The two light tiles would be too bright and washed-out, and the two dark tiles would be too dark and not quite right. For that matter, the black wasn't black.
I recalled an article I'd read about the custom chips in this Olympus camera, and how it was much more an active system than just a CCD pixel bucket. It tries to make a nice picture based on what it saw.
My hypothesis was to add other objects to the scene - sure enough, after adding a few other colorful objects at the margins of the frame, the appearance of the tiles changed dramatically for the better. Now they were a range. It's as if it needs a sacrificial white and black and other colors in order to produce a decent image.
Certainly this has implications for microscope use of today's digital cameras. I don't see a way to turn off this feature, at least on this camera.
Greetings, Does anyone know of software that will run a digitizing tablet for the macintosh? I am thinking of something like "sigmascan" which runs on the PC. We just need to measure lengths and angles, nothing to fancy.
Thanks for the help. Tobias Baskin -- _ ____ __ ____ Tobias I. Baskin / \ / / \ / \ \ 109 Tucker Hall / / / / \ \ \ Biological Sciences /_ / __ /__ \ \ \__ University of Missouri / / / \ \ \ Columbia, MO USA / / / \ \ \ 65211-7400 / / ___ / \ \__/ \ ____ voice: 573-882-0173 fax: 573-882-0123
Does anyone have references and recommendations(or caveats)for dehydration of tissue using tBGE? We routinely prep (testis and ovary) with PO through Epon812 and want something less toxic that still gives good ultrastructure.
==================================== Jennifer Palmer ARBL Colorado State Univ Ft. Collins, CO 80523-1683 , USA voice:(970)491-1770 fax:(970)491-3557
Why not just simply digitize the image and then use NIH Image to measure distances and angles, this will produce a nicely documented measurement as you'll have the image, and all the numerical values stored.
I do this all the time. Of course, it presumes that you have a digitized image to start with.
Nestor Your Friendly Neighborhood SysOp
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The box said "This program requires Win 95/98/NT or better..." So I bought a G3 Mac !
Does anyone know a good reference for the origin of the top-bottom effect? TIA. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
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Hi all, I am looking for a sample to calibrate magnifications of a TEM from 50,000X up. I have used catalase in the past. However the periodicity of catalase is ~8.7nm. That is too large for my present purposes as I am looking at structures in the 2-4nm range and want to reduce the margin of error in the measurements as much as possible. I would also like a sample that is easy to used by inexperienced microscopists in the hopes of encouraging more users to calibrate instrument magnification on a regular basis. It also needs to be relatively in expensive so the loss of a grid is not a catastrophy. I would like a standard that would have ~1.0nm lattice. I thought I had found the perfect sample in copper phthalocyanine which has a spacing in this range. Unfortunately the only source of a grid with this crystal has discontinued carrying it. I am back to square one. My options are to prepare my own grid but need advise on how to crystallize the copper phthalocyanine (available from Sigma in powder form) or find another crystalline lattice in this size range. I would appreciate suggestions for alternative crystals and/or methods for making the copper phthalocyanine crystals to use for this purpose. Thanks in advance. Debby
Debby Sherman Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: sherman-at-btny.purdue.edu 1057 Whistler Building West Lafayette, IN 47907-1057
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Hi Debbie,
What about crocidolite 0.9nm (020) and 0.45nm (021) spacings. It's a long time since I used it but it is still in catalogues and cheap (Agar Scientific at £7 sterling each and probably others).
Good luck, Ron
On 09 Nov 2000 16:43:30 -0500 Debby Sherman {sherman-at-btny.purdue.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 } -----------------------------------------------------------------------. } } } Hi all, } I am looking for a sample to calibrate magnifications of a TEM from 50,000X up. I have used catalase in the past. However the periodicity of catalase is ~8.7nm. That is too large for my present purposes as I am looking at structures in the 2-4nm range and want to reduce the margin of error in the measurements as much as possible. I would also like a sample that is easy to used by inexperienced microscopists in the hopes of encouraging more users to calibrate instrument magnification on a regular basis. It also needs to be relatively in expensive so the loss of a grid is not a catastrophy. } I would like a standard that would have ~1.0nm lattice. I thought I had found the perfect sample in copper phthalocyanine which has a spacing in this range. Unfortunately the only source of a grid with this crystal has discontinued carrying it. I am back to square one. My options are to prepare my own grid but need advise on how to crystallize the copper phthalocyanine (available from Sigma in powder form) or find another crystalline lattice in this size range. } I would appreciate suggestions for alternative crystals and/or methods for making the copper phthalocyanine crystals to use for this purpose. } Thanks in advance. } Debby } } Debby Sherman Phone: 765-494-6666 } Life Science Microscopy Facility FAX: 765-494-5896 } Purdue University E-mail: sherman-at-btny.purdue.edu } 1057 Whistler Building } West Lafayette, IN 47907-1057 } }
---------------------- 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 have routinely used an ethanol series straight into Epon (or its various replacement incarnations) for over 20 years now, with never any poor embedding issues. Truly dry EtOH is miscible with the epons, but requires throrough mixing and additional graded steps. For standard tissues blocks, try 3x in 100% EtOH, followed by 2:1 EtOH:epon, 1:1 EtOH:epon 1:2 EtOH:epon, and at least 2 more pure epon prior to embedding. The 3 EtOH:epon steps should be at least 45 min each (1 have used up to 1 hr for dense tissues) and the pure epon steps can be 30 to 60 min each. I can't speak to the t-BGE issue, but I developed a skin sensitivity to PO a long time ago, and worked out the EtOH method to avoid the PO (I originally used acetone for the dehydration, but had a problem maintaining dry acetone--molecular sieves work but they can introduce particulate contaminants). Hope this helps.
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc.
On Thu, 9 Nov 2000 11:33:32 -0700 (Mountain Standard Time), Jennifer Palmer 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 have references and recommendations(or caveats)for } dehydration of tissue using tBGE? We routinely prep (testis and } ovary) with PO through Epon812 and want something less toxic that } still gives good ultrastructure. } } ==================================== } Jennifer Palmer } ARBL } Colorado State Univ } Ft. Collins, CO 80523-1683 , USA } voice:(970)491-1770 } fax:(970)491-3557 } } jpalmer-at-cvmbs.colostate.edu } ==================================== } }
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I have discussed John McCaffrey's Mag-I-Cal sample at length in the past. You can contact South Bay Technology, Electron Microscopy Sciences, or SPI. I believe all of them have it described on their web sites. It will do exactly what you want it for and then some.
Try it you'll like 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: Debby Sherman [mailto:sherman-at-btny.purdue.edu] } Sent: Thursday, November 09, 2000 4:44 PM } To: message to: MSA list } Subject: TEM calibration sample } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: 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 } } } -------------------------------------------------------------- } ---------. } } } Hi all, } I am looking for a sample to calibrate magnifications of a } TEM from 50,000X up. I have used catalase in the past. } However the periodicity of catalase is ~8.7nm. That is too } large for my present purposes as I am looking at structures } in the 2-4nm range and want to reduce the margin of error in } the measurements as much as possible. I would also like a } sample that is easy to used by inexperienced microscopists in } the hopes of encouraging more users to calibrate instrument } magnification on a regular basis. It also needs to be } relatively in expensive so the loss of a grid is not a catastrophy. } I would like a standard that would have ~1.0nm lattice. } I thought I had found the perfect sample in copper } phthalocyanine which has a spacing in this range. } Unfortunately the only source of a grid with this crystal has } discontinued carrying it. I am back to square one. My } options are to prepare my own grid but need advise on how to } crystallize the copper phthalocyanine (available from Sigma } in powder form) or find another crystalline lattice in this } size range. } I would appreciate suggestions for alternative crystals } and/or methods for making the copper phthalocyanine crystals } to use for this purpose. } Thanks in advance. } Debby } } Debby Sherman Phone: 765-494-6666 } Life Science Microscopy Facility FAX: 765-494-5896 } } Purdue University E-mail: } sherman-at-btny.purdue.edu } 1057 Whistler Building } West Lafayette, IN 47907-1057 } }
We have a Physical Electronics 570 used Auger/ESCA/SIMS surface analysis instrument that we must part with. We used this primarily as a scanning Auger, but also did XPS and some SIMS. This instrument is in working order and has been under a service contract until recently. All offers or inquiries are welcome.
} ... } I discovered an interesting quirk of at least this } Olympus C-2500L digital camera. I suspect that other } consumer/prosumer cameras would suffer the same fate. } } There were four tiles ranging from, say, very light purple } to dark purple. We'd put them on a black velvet background. } We had tremendous trouble getting a accurate color. ... } } ... } } My hypothesis was to add other objects to the scene - sure } enough, after adding a few other colorful objects at the } margins of the frame, the appearance of the tiles changed } dramatically for the better. ... } } Certainly this has implications for microscope use of } today's digital cameras. I don't see a way to turn } off this feature, at least on this camera.
Could the camera be mis-judging the "white balance"?? On my camera I can disable "automatic", and force "incandescent", "fluorescent", "daylight", etc.
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 recently requested, on this site, advice on programmes for Monte Carlo simulations of electron scattering. I have been asked by Nestor to summarise them, to save others asking the same question (and clogging the site). Here is a summary of many replies I received. I have not accessed many of them, and pass them on without comment.
2 site allowing to do online simulations as well as the possibility of as well as the possibility of downloading the program for SEM (and TEM) scattering simulations. http://callisto.my.mtu.edu/soft/mc/ss_java.html http://callisto.my.mtu.edu/soft/mc/ (this address for downloading)
6 Try Cnet (www.cnet.com) and enter Monte Carlo into the "search" box. A page will come up with a list of downloadable programs divided into purchaseable programs and freeware. No idea whether any of them works as I haven't tried any. I'm just familiar with Cnet as a source of downloable software.
7 Try Electron Flight Simulator by Small World, (500)447-3340
Microscopists, A researcher here at the University of Alaska is interested in paying to have some thin sections made from her silicate glass samples. Are there any labs out there that will contract to do ion milling? She is not on the list so please reply to Jessica Faust, her E-mail address is faust-at-gi.alaska.edu Thanks once again. Kim DeRuyter
University of Pittsburgh Department of Materials Science and Engineering
Postdoctoral Research Position in Transmission Electron Microscopy
An additional vacancy for 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 Technology. 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 transmission 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 from December 2000. 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 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
The Microcharacterization laboratory in the Department of Materials Science and Engineering of the University of Pittsburgh has an opening for a full-time technician in a multiple user facility. The facility's main instrumentation consists of two Xpert XRD, an XL30FEGSEM with EDS, BSE and EBSD, and two JEOL TEM's. The successful applicant will have a minimum of a Bachelor¹s degree and 3 years experience working with scanning electron microscopes and X-ray diffraction instruments. Responsibilities will include daily operation and maintenance of the SEM and XRD instrumentation including the training of users (including students), specimen preparation and documentation of experimental data. The successful applicant must demonstrate the necessary organizational, management and communication skills to efficiently operate in an academic multi-user environment. Applicants should submit a cover letter describing their experience with different instrumentation, a resume and contact information for three references. Salary will be commensurate with the candidate¹s experience.
Resumes should be sent to; Professor G. H. Meier, Interim Chair Department of Materials Science and Engineering 848 Benedum Hall University of Pittsburgh Pittsburgh, PA 15261
The University of Pittsburgh is an equal opportunity and affirmative action employer.
At 07:24 AM 11/10/00 -0800, michael shaffer wrote: } Could the camera be mis-judging the "white balance"?? } On my camera I can disable "automatic", and force } "incandescent", "fluorescent", "daylight", etc.
No, I'd already preset and locked the color temperature against a white card in the same environment. These other color adjustments were beyond simple white balance problems.
I know that this has been posted before but I lost the link. Does anyone have a recommendation for WEB scheduling software. Thanks. ______________________________ Roberto Garcia NCSU/Analytical Instrumentation Facility Campus Box 7531 Room 318 EGRC 1010 Main Campus Dr. Raleigh NC 27695-7531 P: (919) 515-8628 F: (919) 515-6965 rgarcia-at-unity.ncsu.edu http://spm.aif.ncsu.edu/aif http://spm.aif.ncsu.edu/asm
This might be of use to those of you wanting to convert your older stereo and compound microscopes to digital imaging capability. We recently explored the possibilities of mounting a Nikon CoolPix 990 (CCD=3.34 million pixels) to our 14 year old Zeiss stereoscope and Standard microscopes. Neither has a "C" mount, which is what the Nikon adapter fits onto. There were several very expensive (} $1,500) possible solutions, but the most satisfying was one proposed by Spectra Tech (716-265-4320; e-mail Michael Specht {mspecht-at-frontiernet.net} ). For about $250, they were able to provide an adapter which screws directly into the camera and fits either in the ocular tube or phototube of the microscopes. Adapters can be purchased for different tube diameters. Contact them for further information if interested.
I have no financial interest.
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
Question: I am working on a final year project titled "low frequency surface microscopy". My project supervisor wants me to use a probe that consists of a concentric cylinder with a pin in between. This is to concentrate the field on the surface to be scanned. With the occurence of cracks, boundaries, etc. on the surface, the electric field and hence the capacitance is expected to change. The reflected voltage in the bridge circuit implementation gives a reflection of the surface characteristic.
My questions go thus (sorry for the preambles): 1. How do I design a probe that can help me to achieve this( what materials should be used, what is the expected capacitance value, what are the relevant equations) 2. At what frequencies can this function properly 3. Can you let me into the theory of this method and other similar methods? 4. Can you refer me to some links and/or send some reference materials to me?
At the moment, i am doing a physics assignment for my HSC assessment on the following topic:
"Compare the resolving powers of light and electrom microscopes and assess the impact of their development".
I you could help by sending some information, I would be very grateful. Thank you.
Sujitha
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Doug, does this adapter have a relay lens? What zoom to you use and is there any vignetting of the field?
I'm buying adapters for a 990 right now.
Regards, Glen
On Fri, 10 Nov 2000, Douglas Keene 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 might be of use to those of you wanting to convert } your older stereo and compound microscopes to digital } imaging capability. We recently explored the possibilities } of mounting a Nikon CoolPix 990 (CCD=3.34 million pixels) to } our 14 year old Zeiss stereoscope and Standard microscopes. } Neither has a "C" mount, which is what the Nikon adapter } fits onto. There were several very expensive (} $1,500) } possible solutions, but the most satisfying was one } proposed by Spectra Tech (716-265-4320; e-mail Michael } Specht {mspecht-at-frontiernet.net} ). For about $250, they } were able to provide an adapter which screws directly into } the camera and fits either in the ocular tube or phototube } of the microscopes. Adapters can be purchased for } different tube diameters. Contact them for further } information if interested. } } I have no financial interest. } } 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 have some epon-araldite blocks to section with tiny wallaby embryos embedded in. Unfortunately the blocks are very soft and the one micron sections are sticky and very hard to pick up does anyone have a solution please as I am getting very frustated Thanks in anticipation Joan Clark
Depending on the 'scope, it may have an intermediate lens/optics unit. I am using the CP 990 on Axioskop and Olympus and the common adapter unit has a lens.
gg
At 05:57 PM 11/12/00, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Can anyone advise what voltage and polarity should be applied to the BNC plug on the short co-axial cable labelled "OM1" which issues from the flange of the JEOL OM on an 840?
It's something to do with an electron deflector or collector.
tia
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
I have read here that you can put the blocks back in the oven at a higher temperature. Why not try this with an unwanted portion of a block?
Dave
On Sun, 12 Nov 2000 23:45:57 -0600 Joan Clark {j.clark-at-zoology.unimelb.edu.au} 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 some epon-araldite blocks to section with tiny wallaby embryos } embedded in. Unfortunately the blocks are very soft and the one micron } sections are sticky and very hard to pick up does anyone have a solution } please as I am getting very frustated } Thanks in anticipation } Joan Clark } } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
Try placing the blocks back in the oven at anything up to 100C for several hours, or even overnight. Remember that the blocks harden after removal from the oven for up to two hours; not just the couple of minutes it takes to physically cool. 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 ABN: 99 724 136 560 www.proscitech.com
On Monday, November 13, 2000 3:46 PM, Joan Clark [SMTP:j.clark-at-zoology.unimelb.edu.au] 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 some epon-araldite blocks to section with tiny wallaby embryos } embedded in. Unfortunately the blocks are very soft and the one micron } sections are sticky and very hard to pick up does anyone have a solution } please as I am getting very frustated } Thanks in anticipation } Joan Clark } }
} This might be of use to those of you wanting to convert } your older stereo and compound microscopes to digital } imaging capability. We recently explored the possibilities } of mounting a Nikon CoolPix 990 (CCD=3.34 million pixels) to } our 14 year old Zeiss stereoscope and Standard microscopes. } Neither has a "C" mount, which is what the Nikon adapter } fits onto. There were several very expensive (} $1,500) } possible solutions, but the most satisfying was one } proposed by Spectra Tech ... For about $250, they } were able to provide an adapter which screws directly into } the camera and fits either in the ocular tube or phototube } of the microscopes. Adapters can be purchased for } different tube diameters. ...
When you say your were "satisfied", what features/characteristics would you care to mention? Did the amount of field-of-view vary? How much of the field were you able to capture? (If satisfaction essentially boils down to price/quality, a similarly priced and quality eyepiece adapter is also available from Optem International {www.optemintl.com} ... however, I am testing their C-mount adapter).
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've been looking at some of these sites lately for web-based calendars. I don't necessarily recommend any of them. You'll see that each has shortcomings.
1. http://www.webevent.com (Can be run on all sorts of operating systems. Commercial software. Lower prices for non-profit institutions.)
2. http://srv.emunit.unsw.edu.au (Excellent, although it is probably only for Windows servers. Some accounting capabilities. Not commercial software. About $500 US.)
3. http://www.webprog.com/appoint/freetest.html (About $60 US. Requires a Windows server. Basic.)
4. http://cmmserv.mrl.uiuc.edu/calendar (UserNumber = 2399, password = DEMO, instrument = Zeiss DSM-960) (Windows server required.) } -----------------------------------------------------------------------. } } I know that this has been posted before but I lost the link. Does anyone } have a recommendation for WEB scheduling software. Thanks. } ______________________________ } Roberto Garcia } NCSU/Analytical Instrumentation Facility } Campus Box 7531 Room 318 EGRC } 1010 Main Campus Dr. } Raleigh NC 27695-7531 } P: (919) 515-8628 } F: (919) 515-6965 } rgarcia-at-unity.ncsu.edu } http://spm.aif.ncsu.edu/aif } http://spm.aif.ncsu.edu/asm
---------------------------------------------------------------------- Russell E. Cook, Ph.D. Electron Microscopy Center 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
We have been asked to embed some human skin biopsies for electron microscopy. We used our routine protocol for processing kidney biopsies which includes glutaraldehyde fixation, dehydration through ethanol and acetone and embeddment in PolyBed 812. Our protocol resulted in good embeddment all the layers of the skin except for the stratum corneum where there were numberous holes in the PolyBed. Is it possible to get good embeddment of this keratinized zone?
Thanks for any suggestion,
John
John M. Basgen Department of Pediatrics University of Minnesota Mayo Mail Code 491 420 Delaware Street SE Minneapolis, MN 55455 USA Phone: 612-625-7979 FAX: 612-626-2791 E-mail: basgen-at-tc.umn.edu
Does anyone have experience with the 990 under low light conditions? We normally shoot brightfield on our inverted metallograph, but there is the occasional need for darkfield and DIC.
Thanks in advance, Jim Hyres
-----Original Message----- } From: michael shaffer [mailto:epmalab-at-darkwing.uoregon.edu] Sent: Monday, November 13, 2000 10:40 AM To: DRK-at-shcc.org; Microscopy-at-sparc5.microscopy.com
Doug writes ...
} This might be of use to those of you wanting to convert } your older stereo and compound microscopes to digital } imaging capability. We recently explored the possibilities } of mounting a Nikon CoolPix 990 (CCD=3.34 million pixels) to } our 14 year old Zeiss stereoscope and Standard microscopes. } Neither has a "C" mount, which is what the Nikon adapter } fits onto. There were several very expensive (} $1,500) } possible solutions, but the most satisfying was one } proposed by Spectra Tech ... For about $250, they } were able to provide an adapter which screws directly into } the camera and fits either in the ocular tube or phototube } of the microscopes. Adapters can be purchased for } different tube diameters. ...
When you say your were "satisfied", what features/characteristics would you care to mention? Did the amount of field-of-view vary? How much of the field were you able to capture? (If satisfaction essentially boils down to price/quality, a similarly priced and quality eyepiece adapter is also available from Optem International {www.optemintl.com} ... however, I am testing their C-mount adapter).
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 am looking for manuals for a Kevex 770 XRF. In particular we would be interested in copies of the circuit diagrams and documentation for the XRF unit and the Detector/Pulse Processor/ADC interface. The Kevex DELTA computer and software is not really of interest, since we would like to change it to a PC platform.
Also, any suggestions of other listservers, etc. to look would be appreciated.
Thanks in Advance, Jim Mabon
__________________________________________ James C. Mabon, Ph.D. Research Electron Microscopist Materials Research Laboratory University of Illinois at Urbana-Champaign 104 S. Goodwin Avenue Urbana, IL 61801 http://ntweb.mrl.uiuc.edu/cmm __________________________________________
OK, I'll bite. What the heck is color scanning EM, as referenced in the title of the following article?
TIA
Bob
Histochemistry of food tissue by colour scanning electron microscopy Mitsuhiko Yamada, Masako Nishimura, Takeo Suzuki, Shigeru Kawamata, Eisaku Oho, and Toshiaki Kimura, pp. 503-507. Journal of Electron Microscopy vol 49(3)
Robert R. Wise, Ph.D. Associate Professor of Plant Physiology Department of Biology and Microbiology University of Wisconsin Oshkosh Oshkosh, WI 54901 tele: (920) 424-3404 fax: (920) 424-1101 wise-at-uwosh.edu http://www.uwosh.edu/departments/biology/wise/wise.html
I am interested in finding an IGP for a Philips (T)EM410 that can be rebuilt / refurbished. Anybody got one lying around?
TIA Ron Veil
Veil Electron Instrument Lab Customer Services Tel: (209) 521-3332 FAX: (209) 521-3033 e-mail: veilcs-at-earthlink.net ________________________________________________________________ YOU'RE PAYING TOO MUCH FOR THE INTERNET! Juno now offers FREE Internet Access! Try it today - there's no risk! For your FREE software, visit: http://dl.www.juno.com/get/tagj.
I am interested in removing the cytoplasm from plant cells for viewing cell walls by SEM. the cells are epidermal cells of bean cotyledons, and I was wanting a simple and rapid method to get rid of the cytoplasm which doesn't harm the cell wall.
Also, I was wondering if anyone knew of an SEM image analysis program that can count things, do nearest-neighbour analysis, density measurements and so on. The things I want to measure are quite variable in shape and size, so it might be a little difficult.
Hi Mark, We use a commercially available washing powder called Ariel (Procter and Gamble) to remove the cytoplasm. Use a 5% (w/v) aqueous solution - it contains a Bacillus subtilis derived protease. References: Honegger, R. 1985. Scanning electron microscopy of the fungus-plant interface: a simple preparative technique. Trans Br. Mycol. Soc. 84: 530-533.
Mims et al. 1989. Ultrastructure of the haustorium of the peanut late leaf spot fungus Cerosporidium personatum. Can. J. Bot. 67: 1198-1202.
It worked quite well for us...good luck, Beth
} Hi everyone, } } I am interested in removing the cytoplasm from plant cells for viewing cell } walls by SEM. the cells are epidermal cells of bean cotyledons, and I was } wanting a simple and rapid method to get rid of the cytoplasm which doesn't } harm the cell wall.
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
hello, for life cell microscopy of fp-tagged cells we consider to buy a TILL PHOTONICS imaging system in combination with a Olympus microscope. We would appreciate any comments on this system or possible alternatives. Beside single channel recording we are also interested in multiple color imaging including FRET. We would like to hear comments on general system performance, usability, flexibility, support, and problems with the system.
} Does anyone have experience with the 990 under low light } conditions? We normally shoot brightfield on our } inverted metallograph, but there is the } occasional need for darkfield and DIC. } ...
With ISO senstivity selectable (100,200,400), and manual maximum aperture and a manual selectable shutter up to 8sec (including bulb), should allow anything you can "see" thru the eyepieces. But I do suspect an increase in noise as you approach the limits, and you might want a trinoc head which can throw 100% of the light at the camera.
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/
Electron Microscopy Facility at Fox Chase Cancer Center is seeking a motivated individual for a Technician/Research Assistant position.
Required qualifications: B.S. or M.S. in biology, 2+ years of experience in biological electron microscopy.
Responsibilities include sample preparation and electron microscopy (TEM/SEM), dark room work, computer image processing, report preparation. The great variability of work due to collaborations with large number of laboratories provides exceptional opportunity for professional growth.
We offer a competitive salary commensurate with experience, an excellent benefit package (health/dental insurance, pension plan, paid vacation) and a very friendly working environment.
For confidential consideration, please send a CV including a statement of experience to:
Dr. Michael Jarnik Fox Chase Cancer Center EM Facility 7701 Burholme Avenue Philadelphia, PA 19111 e-mail: m_jarnik-at-fccc.edu
We are looking for a replacement belt for the control unit of an LKB Ultrotome III 8800. Any ideas what-so-ever? It's only 25+ yrs old! Thanks in advance.
John Hardy City of Hope Medical Center Duarte, CA (626) 301-8265
If you have a mechanical/instrument shop at your institution, ask the gurus there for belt material. All you need to be careful of is matching the diameter of the new belt with the old one. I've done this in the past and it works really well. If there is no shop, go to a trusted hardware/parts store with the old belt and ask for a similar replacement.
Giant rubber bands do not work......(but it was worth a try!)
Tamara Howard CSHL
On Tue, 14 Nov 2000, John Hardy wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } We are looking for a replacement belt for the control unit of an LKB } Ultrotome III 8800. Any ideas what-so-ever? It's only 25+ yrs old! Thanks } in advance. } } John Hardy } City of Hope Medical Center } Duarte, CA } (626) 301-8265 } } jhardy-at-coh.org } } } }
We have a ten year old Panametrics Hyscan scanning acoustic microscope that we must part with. This includes an old computer, controller, scan drive, and tank. Does anyone have an interest in the system or part of it?
Salve, sono Emma Bonino, e le propongo un "sondaggio" e 7 iniziative online. Grazie e buona... azione!
Quali alleanze per le elezioni politiche ? "SONDAGGIO": http://www.radicali.it/action/sondaggio/
Azioni online:
ABORTO E CONTRACCEZIONE Per introdurre in Italia pillola abortiva RU486 http://www.radicali.it/action/ru486/
FREE SOFTWARE Petizione europea contro la brevettabilita' del software http://www.radicali.it/action/freesoft/
LEGALIZZAZIONE DROGHE Sostegno a Pannella e ai radicali sotto processo per disobbedienza civile http://www.radicali.it/action/legalize/
FIRMA DIGITALE E VOTO ELETTRONICO Richiesta al Governo per la legalita' delle elezioni politiche http://www.radicali.it/action/digital/
CONTRO LA TELEVISIONE DI STATO Per abolire la concessione unica radiotelevisiva http://www.radicali.it/action/rai/
CONTRO IL SINDACATO DI STATO Per l'abolizione del sistema di rinnovo automatico dell'iscrizione sindacale http://www.radicali.it/action/sindacato/
LIBERALIZZAZIONE TELECOMUNICAZIONI Per l'abolizione del canone Telecom http://www.radicali.it/action/telecom/
Su questi ed altri temi si confronteranno tra pochi giorni anche le liste delle elezioni radicali online. Per partecipare e votare, clicca qui : http://www.radicali.it/action/register
*** PS Questa email e stata inviata nel rispetto della legge sulla privacy; se desidera maggiori informazioni o se intende cancellarsi, puo cliccare qui: http://www.radicali.it/action/privacy/
Salve, sono Emma Bonino, e le propongo un "sondaggio" e 7 iniziative online. Grazie e buona... azione!
Quali alleanze per le elezioni politiche ? "SONDAGGIO": http://www.radicali.it/action/sondaggio/
Azioni online:
ABORTO E CONTRACCEZIONE Per introdurre in Italia pillola abortiva RU486 http://www.radicali.it/action/ru486/
FREE SOFTWARE Petizione europea contro la brevettabilita' del software http://www.radicali.it/action/freesoft/
LEGALIZZAZIONE DROGHE Sostegno a Pannella e ai radicali sotto processo per disobbedienza civile http://www.radicali.it/action/legalize/
FIRMA DIGITALE E VOTO ELETTRONICO Richiesta al Governo per la legalita' delle elezioni politiche http://www.radicali.it/action/digital/
CONTRO LA TELEVISIONE DI STATO Per abolire la concessione unica radiotelevisiva http://www.radicali.it/action/rai/
CONTRO IL SINDACATO DI STATO Per l'abolizione del sistema di rinnovo automatico dell'iscrizione sindacale http://www.radicali.it/action/sindacato/
LIBERALIZZAZIONE TELECOMUNICAZIONI Per l'abolizione del canone Telecom http://www.radicali.it/action/telecom/
Su questi ed altri temi si confronteranno tra pochi giorni anche le liste delle elezioni radicali online. Per partecipare e votare, clicca qui : http://www.radicali.it/action/register
*** PS Questa email e stata inviata nel rispetto della legge sulla privacy; se desidera maggiori informazioni o se intende cancellarsi, puo cliccare qui: http://www.radicali.it/action/privacy/
We still have a Kevex 7000 system connected to a LSI11/23 computer. Now, we have problem with booting the computer when the analyser is connected to the PDP-bus. We think, but we don't know, if we ahve a problem with the Kevex Q23 board,which interfaces the analyser to the bus. However, do anybody have other suggestions? Or even better, is there a Q23 board out there?
Ypurs Sincererly Gunnar Kopstad
Vennlig Hilsen dr.ing Gunnar Kopstad overingeniør Avd f Patologi, Rit
Salve, sono Emma Bonino, e le propongo un "sondaggio" e 7 iniziative online. Grazie e buona... azione!
Quali alleanze per le elezioni politiche ? "SONDAGGIO": http://www.radicali.it/action/sondaggio/
Azioni online:
ABORTO E CONTRACCEZIONE Per introdurre in Italia pillola abortiva RU486 http://www.radicali.it/action/ru486/
FREE SOFTWARE Petizione europea contro la brevettabilita' del software http://www.radicali.it/action/freesoft/
LEGALIZZAZIONE DROGHE Sostegno a Pannella e ai radicali sotto processo per disobbedienza civile http://www.radicali.it/action/legalize/
FIRMA DIGITALE E VOTO ELETTRONICO Richiesta al Governo per la legalita' delle elezioni politiche http://www.radicali.it/action/digital/
CONTRO LA TELEVISIONE DI STATO Per abolire la concessione unica radiotelevisiva http://www.radicali.it/action/rai/
CONTRO IL SINDACATO DI STATO Per l'abolizione del sistema di rinnovo automatico dell'iscrizione sindacale http://www.radicali.it/action/sindacato/
LIBERALIZZAZIONE TELECOMUNICAZIONI Per l'abolizione del canone Telecom http://www.radicali.it/action/telecom/
Su questi ed altri temi si confronteranno tra pochi giorni anche le liste delle elezioni radicali online. Per partecipare e votare, clicca qui : http://www.radicali.it/action/register
*** PS Questa email e stata inviata nel rispetto della legge sulla privacy; se desidera maggiori informazioni o se intende cancellarsi, puo cliccare qui: http://www.radicali.it/action/privacy/
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Carol, Thanks for the post. Since I want the grid for calibration, not resolution, I needed something with a lattice which was visible at magnifications from about 50,000X up. I may have found a couple of possiblilities. Esbestos fiber grids with 1.3nm periodicity are available commercially. Also Ted Pella has arranged to obtain Cu-phthalocyanine grids (which had been in their catalog but were discontinued) with a 1.0nm periodicity. I think that either or both of these should do the trick. Debby
Microscopical Optical Consulting (MOC) Inc. Helmut Patzig P.O Box 586 Valley Cottage, NY 10989 (914) 268-6450 MOCLeica-at-aol.com
They had picked up the Leica service and were able to help with my old LKB system.
Good Luck!
George R. Munzing Jr Strategic Technologies Group Engelhard Corporation 25 Middlesex/Essex Tpk. Iselin, NJ 08830 (732) 205-7030
"John Hardy" {jhardy-at-coh.org} on 11/14/2000 02:26:18 PM
To: "Microscopy" {Microscopy-at-sparc5.microscopy.com} cc:
We are looking for a replacement belt for the control unit of an LKB Ultrotome III 8800. Any ideas what-so-ever? It's only 25+ yrs old! Thanks in advance.
John Hardy City of Hope Medical Center Duarte, CA (626) 301-8265
MAS SPECIAL TOPICS WORKSHOP Dale Newbury and Ryna Marinenko
The NIST Microanalysis Research Group and the Microbeam Analysis Society will co-sponsor an MAS Special Topics Workshop, "Understanding the Accuracy Barrier in Quantitative Electron Probe Microanalysis and the Role of Standards," that will be held at NIST in Gaithersburg, Maryland October 15-18, 2001. This event is part of the 2001 celebration of the NIST Centennial. The limited attendance workshop will deal with the experimental and theoretical factors that currently limit the accuracy of quantitative results to approximately 2% relative. The topics will include sample and instrument parameters, counting statistics, correction procedures, individual matrix correction parameters, etc. In addition, one or more sessions will be specifically devoted to discussions on microanalysis standards. Invited speakers will be expected to provide a short (6 pages, maximum) manuscript of their paper at the time of the meeting. These manuscripts will be subsequently published in a monograph. The intent of the meeting and monograph is to provide detailed information on accuracy limits and on acceptable standards procedures in quantitative microprobe analysis that is presently not readily available in the literature.
Below is a proposed outline of presentations. Please feel free to suggest possible speakers (don't be afraid to call your own number!) and additional topics. We don't expect to be able to address all significant topics, but rather hope to initiate an ongoing dialogue that will be addressed by periodic special workshops on this topic.
We are now preparing a list of invited speakers, and we are actively soliciting volunteers who would like to be considered to present a talk, especially on microanalysis standards. Not all types or groups of microanalysis standards have been considered in this list below. We would like to cover as much of the periodic table as possible.
In addition, if you are simply interested in attending the meeting, please send your name, address, telephone number, email, and FAX to receive future information.
Factors that play a significant role in the 2% relative accuracy barrier … Experimental factors … Instrument stability: how good is it really? … Electron beam performance … WDS peak reproducibility … EDS performance … The role of the specimen … Preparation issues … Stability (beam damage) … Charging … Spectral processing … Peak deconvolution in WDS … Extracting intensities from EDS spectra (peak overlaps) … Quantitative matrix corrections … How well do matrix correction schemes work on data outside their native databases? … What is missing? Where is research needed? Who will do it?
Standards in Quantitative EPMA … Procedures for Preparation of Bulk Standards specimens for Quantitative EPMA … Sources, Characterization, and Validation of Standard Materials … Commercial, private, institutional sources … Composition, Microheterogeneity, Stability, etc. … Traceability, certification, intercomparisons, round robins … Industry/Technology Specific Standards … Mineralogy and Geology - sources of standard specimens and preparation procedures … Pure Element and Oxide Standards … Useful Stoichiometric Compounds - crystalline, amorphous, sintered … Sulphides, selenides, and tellurides … Rare Earth Elements … Halides … Alkali Earth Elements … Low-atomic number elements, C, N, B, Be … Glass standards
My last contact information for LKB belts and service is:
Norm Woodside for belts at about $50 apiece. Fax: 410-744-8522.
Pat Capogrosi for service. Telephone: 410-744-1580.
This information is over a year old, but worth a try. Also, the drive belts can be almost duplicated by belts that cost about $8-10 each, if you have a good machine or instrument repair shop with comprehensive belt catalogs. Here are some specs you can try:
Belt thickness: 0.5mm. Outer circumference: 18cm. Width: 5mm. Number of teeth: 39. Size of teeth: 1mm by 5mm. Height of teeth: 2mm. (Thanks to Preston Stogsdill for this information.)
I've never been able to find one that EXACTLY fits, but I've gotten close enough that the microtomes work fine, if you don't mind a little "bump" as you turn the knob to raise and lower the specimen arm. The problem is finding the exact number of teeth. The bump doesn't translate into choppy cuts, but you might feel it in the knob itself.
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/
Here is a protocol I developed over a few years of trying to examine stratum corneum (it was based of course on published protocols and suggestions from this listserve -- nothing novel). The only big differences between this and Michael Delannoy's reply are the extended dehydration and infiltration times, along with the use of ruthenium tetroxide. I found the latter to be a must if you are interested in preserving the intercellular lipids of the stratum corneum. Also I found the use of a dermatome led to increased splitting off of the outermost layers of corneocytes, so simply excising small pieces of skin was better. Hope this helps,
Karen Zaruba 3M Company, St. Paul, MN kszaruba1-at-mmm.com
Preparation of Skin for TEM examination of Stratum Corneum
At necropsy, excise the skin with a minimum of stretching or force. Pin out skin in containers of half-strength Karnovsky?s fixative, which is 2% glutaraldehyde + 2% paraformaldehyde in cacodylate Buffer (0.15M sodium cacodylate, pH 7.4, with 0.1 M sucrose). After about 1 hr. the outer tissue should be trimmed away. The central areas should be divided into final embedding sizes (1 x 1 x 3 mm) by cutting over a piece of dental wax using a Personna razor blade. Continue fixation for a total of 2-12 hr., then wash in buffer 3 x 5 min. Post-fix in 1% osmium tetroxide in buffer for 30 min. Wash in buffer without sucrose 2 x 5 min., then briefly in distilled water. Post-fix in 0.2% ruthenium tetroxide (with or w/o 0.25% potassium ferricyanide) in 0.1 M cacodylate buffer without sucrose for time indicated. Rinse in buffer or distilled water, 3 x 5 min. (Use buffer if being held overnight at this stage.) Note: RuO4 reacts violently with filter paper and alcohols. Also reacts with benzene rings and organics. Dehydrate in ethanol series, 25-35 min. each step from 30% through 95%, then 3 x 20 min. in 100%. Incubate in tert-Butyl Glycidyl Ether, 2 x 30 min. Infiltrate in a mixture of 2 parts t-BGE to 1 part Spurr?s resin for 1-2 hrs. Then follow with 1 part t-BGE to 2 parts Spurr?s resin overnight. Continue to infiltrate in Spurr?s resin alone, 3 changes over 6-8 hrs. Embed in fresh resin in flat molds, orienting to allow cross-sectioning of the epidermis. Polymerize at approximately 58 C for 1-3 days. To visualize lipid lamellae, do not post-stain. For best contrast of desmosomes and cellular detail, post-stain with standard Tannin/Uranyl Acetate stain followed by Reynold?s Lead Citrate.
----------------------------------------------------------------------- John Basgen wrote:
To those doing EM on skin biopsies:
We have been asked to embed some human skin biopsies for electron microscopy. We used our routine protocol for processing kidney biopsies which includes glutaraldehyde fixation, dehydration through ethanol and acetone and embeddment in PolyBed 812. Our protocol resulted in good embeddment all the layers of the skin except for the stratum corneum where there were numberous holes in the PolyBed. Is it possible to get good embeddment of this keratinized zone?
Thanks for any suggestion,
John
John M. Basgen Department of Pediatrics University of Minnesota Mayo Mail Code 491 420 Delaware Street SE Minneapolis, MN 55455 USA Phone: 612-625-7979 FAX: 612-626-2791 E-mail: basgen-at-tc.umn.edu
As part of a hazard review on our new ESEM, many questions were raised regarding the observation of biohazards. We won't be looking at actual human pathogens (although maybe others do?? CDC/NIH??). However, just looking at unfixed human skin puts us under the Class 2 category. Also we'll be observing non-pathogenic (opportunisitic) bacteria. Here are the questions:
Are there conditions inside the ESEM that would favor the creation of an aerosol from the sample? Assuming any of the sample becomes detached/airborn inside the chamber, would pumping down to higher vacuum prior to opening the door remove them (seems logical that it would)? How do you dispose of pump oil containing microbes/biohazards? Would anything capable of infecting humans even survive in pump oil? (Remember we're talking about viruses as well as bacteria here). Finally, would the chamber walls, etc. need to be decontaminated periodically?
If anyone has gone through this sort of safety review and generated a protocol, we'd love to benefit from your hard work!! Otherwise we'd appreciate any suggestions/opinions.
Thanks as always, Karen Zaruba 3M Company, St. Paul, MN kszaruba1-at-mmm.com
Hi- We are analyzing experimental charges of magnetite + ilmenite in a glass of rhyolitic composition. The problem we have run into is that the ilmenite grains are very small and thin, typicaly {10um in length, such that nearly all ilmenite analyses overlap the glass. My question is: Does anyone have any experience/know-how/ideas that would help in calculating the composition of the ilmenite from an analysis that is from a mixture of ilmenite + glass.
Analyses are with a JEOL 733 microprobe at 15kV and 5nA. We have not yet tried a lower kV that would potentially reduce the beam penitration depth. Our method of recalculation thus far has been to adjust the final analysis for density differences between glass and ilmenite following the method of Warren (LPSC ?1997). The fraction of glass is then subtracted from the analysis based on the amount of Sio2 (all SiO2 in the glass, no SiO2 in the ilmenite). We then adjust the result by noting that the ZAF correction applied to the original analysis is a function of the amount of glass overlap and extrapolate this relationship for each element in ilmenite back to a ZAF at 0% Sio2.
TEM EDS is a last resort as the sample would have to be mined out of a polished microprobe mount.
************************************************ ### Free the Psoas ### **************
Dr. Scott Kuehner kuehner-at-u.washington.edu Dept. of Geological Sciences ph.206-543-8393 Box 351310 Fax 206-616-6873 The University of Washington Seattle, Washington 98195-1310 ************************************************
On Wed, 15 Nov 2000 kszaruba1-at-mmm.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 fellow microscopists, } } As part of a hazard review on our new ESEM, many questions were raised regarding } the observation of biohazards. We won't be looking at actual human pathogens } (although maybe others do?? CDC/NIH??). However, just looking at unfixed human } skin puts us under the Class 2 category. Also we'll be observing non-pathogenic } (opportunisitic) bacteria. Here are the questions: } } Are there conditions inside the ESEM that would favor the creation of an } aerosol from the sample? } Assuming any of the sample becomes detached/airborn inside the chamber, would } pumping down to higher vacuum prior to opening the door remove them (seems } logical that it would)? } How do you dispose of pump oil containing microbes/biohazards? Would } anything capable of infecting humans even survive in pump oil? (Remember } we're talking about viruses as well as bacteria here). } Finally, would the chamber walls, etc. need to be decontaminated } periodically? } } If anyone has gone through this sort of safety review and generated a protocol, } we'd love to benefit from your hard work!! Otherwise we'd appreciate any } suggestions/opinions. } } Thanks as always, } Karen Zaruba } 3M Company, St. Paul, MN } kszaruba1-at-mmm.com } } } } }
We are in the market for buying a automated tissue processor for electron microscopy and a paper printer for the dark room. I will appreciate any recommendations for certain makes and models.
Does anyone know if there is a protein stain you can apply in vapour form in order not to change the structure of very delicat samples when using liquids (as for e.g. starch and fat)?
We've had an ESEM for about 7 years now, but we've never looked at anything remotely human in origin in it. However, I think I have a pretty good idea of what conditions are really like in the chamber of an ESEM, so I'll contribute my (fairly uneducated) guesses/impressions. Here goes:
} Are there conditions inside the ESEM that would favor the creation of an aerosol from the sample?
Don't think so - unless your sample was easily evaporated...
Assuming any of the sample becomes detached/airborn inside the chamber, would pumping down to higher vacuum prior to opening the door remove them (seems logical that it would)?
You're right....that does seem logical...
} How do you dispose of pump oil containing microbes/biohazards? Would } anything capable of infecting humans even survive in pump oil? (Remember } we're talking about viruses as well as bacteria here).
Being at an oceanographic institute, there are these really big blue vats out on the dock, put there to receive waste oil from our ships. When I change the oil in our rotary pumps, that's where it goes. I kind of doubt that any human pathogens could live in pump oil (but that's a real guess....)
} Finally, would the chamber walls, etc. need to be decontaminated } periodically? } Probably not, but if it was deemed necessary, I suppose a rinse with some kind of antiseptic wouldn't be a bad idea - as long as you kept the fluids away from the stage electrics :-)
Good luck with the new ESEM - they're pretty nice, well-built instruments.
Frank Thomas MicroAnalysis Facility Geological Survey of Canada (Atlantic) Bedford Institute of Oceanography Dartmouth, Nova Scotia Canada
For safety reasons we fix human cell cultures and catheters before examination in our ESEM. Unfixed human and animal cells suffer beam damage in tungsten filament ESEMs so we do not feel we are missing out. It would be nice to look at the catheter biofilms unfixed but we plumped for a safety first approach.
Dave
On Wed, 15 Nov 2000 13:26:22 -0600 "kszaruba1-at-mmm.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 fellow microscopists, } } As part of a hazard review on our new ESEM, many questions were raised regarding } the observation of biohazards. We won't be looking at actual human pathogens } (although maybe others do?? CDC/NIH??). However, just looking at unfixed human } skin puts us under the Class 2 category. Also we'll be observing non-pathogenic } (opportunisitic) bacteria. Here are the questions: } } Are there conditions inside the ESEM that would favor the creation of an } aerosol from the sample? } Assuming any of the sample becomes detached/airborn inside the chamber, would } pumping down to higher vacuum prior to opening the door remove them (seems } logical that it would)? } How do you dispose of pump oil containing microbes/biohazards? Would } anything capable of infecting humans even survive in pump oil? (Remember } we're talking about viruses as well as bacteria here). } Finally, would the chamber walls, etc. need to be decontaminated } periodically? } } If anyone has gone through this sort of safety review and generated a protocol, } we'd love to benefit from your hard work!! Otherwise we'd appreciate any } suggestions/opinions. } } Thanks as always, } Karen Zaruba } 3M Company, St. Paul, MN } kszaruba1-at-mmm.com } } } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
Microscopy folks, Anyone out there have any info on the Tracor-Northern 6100 diode array detector as used in the TN spectrophotometers? Feel free to point me to another list that is more appropriate for this question if you know of one. Thanks, Greg M. -- Gregory Mulhollan, Ph.D. Extreme Devices Inc. 101 West 6th Street Suite 200 Austin, TX 78701 (512)479-7740 x2231 voice (512)479-7750 fax mulhollan-at-extremedevices.com
} We are analyzing experimental charges of magnetite + } ilmenite in a glass of rhyolitic composition. } The problem we have run into is that the } ilmenite grains are very small ... My question is: } Does anyone have any experience/know-how/ideas } that would help in calculating the composition of the } ilmenite from an analysis that is from a mixture of } ilmenite + glass. } } ...
This will indeed be a problem ... not only do x-rays generated outside the ilmente reach the detectors, but they also travel through different compositions on their way to the spectrometers which are located differently relative to the ilmenites orientation in the glass. One spectro may measure Fe largely absorbed by the ilmenite, and another may measure Ti largely absorbed by glass. You would have to model each based on simplistic assumptions which may be unjustified.
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/
There is a position for an EM technician at the Electron Microscopy Core lab of the Biotechnology Program at the University of Florida in Gainesville. The laboratory is mostly a fee-for-service lab serving the needs of biological, biomedical and agricultural scientists at the university. Applicants must be skilled in the preparation of biological samples for both scanning and transmission electron microscopy. Experience with both PC and Mac as well as website management is desirable. Experience with digital imaging and fluorescence microscopy also a plus. This is a full time, permanent position with standard benefits of State of Florida employees.
For further details, reply to this message or contact Greg Erdos at the number below.
Gregory W. Erdos, Ph.D. Assistant Director, Biotechnology Program PO Box 110580 University of Florida Gainesville, FL 32611
} Does anyone know if there is a protein stain you can apply in vapour form in } order not to change the structure of very delicat samples when using liquids } (as for e.g. starch and fat)?
} Thanks for your help. Gudrun
Dear Gudrun, I don't know what you intend to do, LM, EM, or other, but I once used a stain to detect peptides on chromatography paper (it was a long time ago). 1) Place the protein specimen in an enclosed box in a well-ventillated hood. 2) Mix equimolar solutions of HCl and KMnO4 (in the hood, of course), place the beaker containing the mix in the box alongside the specimen, and seal the box. The Cl-atoms generated will react with the peptide bonds to produce chloropeptides. 3) Add iodide (I added KI solution, but HI or ICl would probably work.). Instead of the last step, you could add anything which reacts with the chloropeptides and gives a signal which can be detected by whatever type of microscopy you want to use. This stain is extremely sensitive, because it can label each peptide bond, so a 100-residue protein can have 100 labels attached. Good luck; please let me know if this technique works for you. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
I don't have any specific experience with the analytes and the matrix that you reference, but have you tried using backscattered electron imaging? There's a difference of a little over six in the average atomic number of ilemite and magnetite with an even greater difference between the ilemite and the volcanic glass matrix, so atomic number contrast should be no problem. Most EDX systems offer routines for measuring phase area based on atomic number contrast which is fairly easily translated into volume concentration. You should have no problem with overlap using backscattered electrons as opposed to x-rays. In addition, with the help of an image analysis package, you can get size, shape, orientation and other information at the same time. Just a thought.
Bill
"S. Kuehner" {kuehner-at-u.washington.edu} on 11/15/2000 03:13:46 PM
To: "kszaruba1-at-mmm.com"-at-sparc5.microscopy.com cc: Microscopy-at-sparc5.microscopy.com (bcc: William H Roberts/US/FILM/DPT)
Hi- We are analyzing experimental charges of magnetite + ilmenite in a glass of rhyolitic composition. The problem we have run into is that the ilmenite grains are very small and thin, typicaly {10um in length, such that nearly all ilmenite analyses overlap the glass. My question is: Does anyone have any experience/know-how/ideas that would help in calculating the composition of the ilmenite from an analysis that is from a mixture of ilmenite + glass. Analyses are with a JEOL 733 microprobe at 15kV and 5nA. We have not yet tried a lower kV that would potentially reduce the beam penitration depth. Our method of recalculation thus far has been to adjust the final analysis for density differences between glass and ilmenite following the method of Warren (LPSC ?1997). The fraction of glass is then subtracted from the analysis based on the amount of Sio2 (all SiO2 in the glass, no SiO2 in the ilmenite). We then adjust the result by noting that the ZAF correction applied to the original analysis is a function of the amount of glass overlap and extrapolate this relationship for each element in ilmenite back to a ZAF at 0% Sio2. TEM EDS is a last resort as the sample would have to be mined out of a polished microprobe mount. ************************************************ ### Free the Psoas ### ************** Dr. Scott Kuehner kuehner-at-u.washington.edu Dept. of Geological Sciences ph.206-543-8393 Box 351310 Fax 206-616-6873 The University of Washington Seattle, Washington 98195-1310 ************************************************
This is slightly off the EM mark, but still in the ballpark for this List...
We recently received a donated xray diffractometer for which we need service and installation assistance. Does anyone know of a local rep or contact info??
It is a Philips APD 3720 Automated Power Diffraction System with a Philips XRG 3100 X-Ray Generator, vintage ca. mid-80's.
Thanks, all.
Ann Hein Lehman
---------------- Ann Hein Lehman EM Facility Manager Trinity College Hartford, CT 06106 v. 860-297-4289 f. 860-297-2538 e. ann.lehman-at-trincoll.edu w. http://www.trincoll.edu/~alehman
I realize that you are pretty far away, but we offer an electron microscopy course here at USU, each spring semester, which includes a large segment on SEM.
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: O. O. Ilori [mailto:sojilori-at-oauife.edu.ng] Sent: Thursday, November 16, 2000 7:47 AM To: microscopy-at-sparc5.microscopy.com
Hello Guys, I am looking for a place,an institute or something where I can be trained to operate an SEM. Any suggestions? Thanks.
Mr. O. O. ILORI DEPARTMENT OF ELECTRONIC/ELECTRICAL ENGINEERING OBAFEMI AWOLOWO UNIVERSITY, ILE-IFE, OSUN STATE NIGERIA.
I don't have any specific experience with the analytes and the matrix that you reference, but have you tried using backscattered electron imaging? There's a difference of a little over eleven in the average atomic number of ilmenite and magnetite with an even greater difference between the ilmenite and the volcanic glass matrix, so atomic number contrast should be no problem. Most EDX systems offer routines for measuring phase area based on atomic number contrast which is fairly easily translated into volume concentration. You should have no problem with overlap using backscattered electrons as opposed to x-rays. In addition, with the help of an image analysis package, you can get size, shape, orientation and other information at the same time. Just a thought.
Bill
"S. Kuehner" {kuehner-at-u.washington.edu} on 11/15/2000 03:13:46 PM
To: "kszaruba1-at-mmm.com"-at-sparc5.microscopy.com cc: Microscopy-at-sparc5.microscopy.com (bcc: William H Roberts/US/FILM/DPT)
Hi- We are analyzing experimental charges of magnetite + ilmenite in a glass of rhyolitic composition. The problem we have run into is that the ilmenite grains are very small and thin, typicaly {10um in length, such that nearly all ilmenite analyses overlap the glass. My question is: Does anyone have any experience/know-how/ideas that would help in calculating the composition of the ilmenite from an analysis that is from a mixture of ilmenite + glass. Analyses are with a JEOL 733 microprobe at 15kV and 5nA. We have not yet tried a lower kV that would potentially reduce the beam penitration depth. Our method of recalculation thus far has been to adjust the final analysis for density differences between glass and ilmenite following the method of Warren (LPSC ?1997). The fraction of glass is then subtracted from the analysis based on the amount of Sio2 (all SiO2 in the glass, no SiO2 in the ilmenite). We then adjust the result by noting that the ZAF correction applied to the original analysis is a function of the amount of glass overlap and extrapolate this relationship for each element in ilmenite back to a ZAF at 0% Sio2. TEM EDS is a last resort as the sample would have to be mined out of a polished microprobe mount. ************************************************ ### Free the Psoas ### ************** Dr. Scott Kuehner kuehner-at-u.washington.edu Dept. of Geological Sciences ph.206-543-8393 Box 351310 Fax 206-616-6873 The University of Washington Seattle, Washington 98195-1310 ************************************************
Does anyone know of a way to stain for collagen in Epon embedded cardiac muscle for viewing by light microscopy?
Thanks for any suggestions.
Dotty
Dotty Sorenson Microscopy and Image Analysis Laboratory Department of Cell and Developmental Biology University of Michigan Medical School Ann Arbor, Michigan dsoren-at-umich.edu (734)763-1170
Of course immunolabeling might work. I've had success with collagen types III and IV in Epon and Spurr's resin, definitely not processed with immuno in mind.
On the other hand if you just want something simple to help highlight the collagen a little, I've seen Methyl Green staining collagen more than other components in skin cross sections.
Good luck, Karen
--------------------------------------- Hello all,
Does anyone know of a way to stain for collagen in Epon embedded cardiac muscle for viewing by light microscopy?
Thanks for any suggestions.
Dotty
Dotty Sorenson Microscopy and Image Analysis Laboratory Department of Cell and Developmental Biology University of Michigan Medical School Ann Arbor, Michigan dsoren-at-umich.edu (734)763-1170
Come join us at the joint meetings of Florida Microscopy Society, FL AVS, and Surface Science 2001
March 12-16, 2001
University of Central Florida, Orlando, FL
THERE IS NO REGISTRATION FEE TO ATTEND THE MEETING (but please register at http://natasha.eng.usf.edu/gilbert/avs/anouncement/registrationform.html so we get an accurate head count for meals)
- An FIB afternoon session followed by:
- An early evenning session devoted to an FIB workshop and users group meeting (open to all users of FIBs and all FIB manufacturers)
- There will be 2 FIB-related short courses one on theory (see www.vacuum.org for fees and information) FIB will be included in a comprehensive TEM specimen preparation course March 14-16, 2001 (contact lag-at-mail.ucf.edu)
To submit an abstract for the meeting or for more information follow the FL local affiliates link to FL from: www.microscopy.org
or contact Lucille Giannuzzi, lag-at-mail.ucf.edu
******************************************************************* 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 ********************************************************************
Does anyone have an experience that they would like to share involving the SVMicro digital camera by Sound Vision? I'm hoping to learn anything at all, from delivery, support, service, image quality, product reliability, etc. Thanks in advance.
We have one. It is about two years old. It can provide good images, but it is tough to work with and you have to work too hard to get a good image. The TWAIN software that I have is tough to set up the contrast, brightness, and exposure. The person to whom I gave this camera didn't like it, gave up and went back to film. I then traded him my Pixera camera for the SVMicro and he was much happier. I don't know if the software on the SVMicro has improved or not.
We don't do really critical work with either microscope that these are put on and we usually don't do color.
I would trade the SVMicro for another El Cheapo Pixera for the ease of use.
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: William H Roberts [mailto:William.H.Roberts-at-usa.dupont.com] Sent: Thursday, November 16, 2000 4:38 PM To: Microscopy-at-sparc5.microscopy.com Subject: SVMicro Digital Microscope Camera
--------------------------------------------------------------- --------- The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserv er/FAQ.html
Does anyone have an experience that they would like to share involving the SVMicro digital camera by Sound Vision? I'm hoping to learn anything at all, from delivery, support, service, image quality, product reliability, etc. Thanks in advance.
We have a SVMicro attached to an old Zeiss Ultraphot. We do not do anything critical with it; just BF/DF and DIC at medium mags on materials samples. I would say it works quite well. We use the Photoshop interface for the Mac; the interface is quite easy to work with. One caveat, the sensor is quite large (~25mm) so you need good transfer optics to get a good image over the whole sensor. At the time (~2yrs bp), the performance to price ratio was very high. The whole package was around $1100. I do not know how it compares now. Just my $0.02.
Ray D. Twesten, PhD. Center for Microanalysis of Materials Seitz Materials Research Laboratory 104 S. Goodwin Ave., Urbana, IL 61801 +1 217 244-6177 (Fax -2278)
-----Original Message----- } From: Walck, Scott D. [mailto:walck-at-ppg.com] Sent: Thursday, November 16, 2000 5:07 PM To: 'William H Roberts' Cc: 'Microscopy'
We have one. It is about two years old. It can provide good images, but it is tough to work with and you have to work too hard to get a good image. The TWAIN software that I have is tough to set up the contrast, brightness, and exposure. The person to whom I gave this camera didn't like it, gave up and went back to film. I then traded him my Pixera camera for the SVMicro and he was much happier. I don't know if the software on the SVMicro has improved or not.
We don't do really critical work with either microscope that these are put on and we usually don't do color.
I would trade the SVMicro for another El Cheapo Pixera for the ease of use.
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: William H Roberts [mailto:William.H.Roberts-at-usa.dupont.com] Sent: Thursday, November 16, 2000 4:38 PM To: Microscopy-at-sparc5.microscopy.com Subject: SVMicro Digital Microscope Camera
--------------------------------------------------------------- --------- The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserv er/FAQ.html
Does anyone have an experience that they would like to share involving the SVMicro digital camera by Sound Vision? I'm hoping to learn anything at all, from delivery, support, service, image quality, product reliability, etc. Thanks in advance.
Hi, We are looking for opinions or reviews on the the Casio QV 3000 digital camera, with a 340 MB microdrive and macro focusing down to 6 cm. any help would be greatly appreciated. thanks, Beth
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
I would appreciate comments and suggestions on automatic sample processors for EM. Vendors welcome. Marek Malecki, M.D., Ph.D. Director and Principal Investigator
Molecular Imaging Laboratories University of California, San Diego
Is there any other stain reagents, other than UA and LC, that are used in TEM? If so, do they have comparable results, are they ETOH or water based, what are there disposal precautions, and what are their staining protocols.
Any information will be appreciated.
Thanks in advance.
Donald G. Awbrey, HT(ASCP) QIHC Electron Microscopy / Image Analysis DonaldAwbrey-at-TexasHealth.org donaldawbrey-at-hotmail.com (817)-878-5647
I am in the process of publishing a brochure that advertises the AVS short courses and symposium in Florida. Since this is joint meeting would it be possible to rent/exchange a mailing list from the Society for this purpose. The meeting is March 2001.
Della
_____________________________
Della Miller AVS West 1265 El Camino Real, Ste. 109 Santa Clara CA 95050
I have walked into a laboratory that has had an Edwards E306A Vacuum Coater purchased in 1991. It has been used very rarely since, and by no one presently working in the laboratory. The instruction manuals help to pump down the system and get it ready for the application, but I have yet to get the carbon source deposited on the sample we are trying to coat for SEM analysis (I can get the carbon rods glowing, but nothing else happens).
I am wondering if anyone out there has this system and knows how to operate it with the carbon evaporation source. Any help to what I could be doing wrong, or not doing at all would be appreciated. Also perhaps if I could call someone who has this system and they could walk me though the steps would be great.
- Jonathan
(I have contacted the manufacturer and they have not made this coater in years so know little about it as well)
and do an ALL groups, all history search for "casio 3000." I did this and found 800 hits. Most are as expected in the rec.photo.digital usenet forum. I did not immediately see any threads regarding its use on a microscope. So, this may or may not be of value to you.
gary g.
At 07:09 AM 11/17/00, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
At 10:27 AM -0600 11/17/00, Awbrey, Donald wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Yes there are other TEM stains, although UrAc and Lead Citrate are by far the most commonly used. You can also use Bismuth in lieu of lead. Kits are commercially available and come with instructions. Its a similar protocol to that used for Pb, but with concetration and time differences. For a comprehensive review, you can refer to Vol. 5 Part I: Staining Methods for Sectioned Material. by PR Lewis and DP Knight in the "Practical Methods on Electron Microscopy series edited by Audrey Glauert. This covers general as well as cytochemical methods.
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 came across a used Blazers Electon Beam Evaroration unit with a 552 gun BAF 300 Freeze Etch unit BAE 300 Coating unit 300, Crystal thin film monitor QSG 202, dual rotary vane vacuum pumps and what appears to be all documentation. .
The unit was removed from a working lab but it was not decommissioned by experienced professionals. It did appear to be carefully taken down. I think it was taken for a decommissioned lab because many their notes on procedures are included
It is for sale by a friend of mine who I won't plug on the list. But it looks like it might be a good deal for some one that need a set up like this and he is not very proud of it. I don't normally do this but I hate to see something that appears to be this good go for scrap. Even though I would like to have the vacuum chamber.
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
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Good Evening, all:
We have an old LKB Ultramicrotome III (series 8800) that has developed a problem in the past few weeks. Apparently the cantilever arm is not going down far enough on the cutting stroke to activate the kickback on the knife holder, resulting in wet blocks nearly every stroke. Needless to say, this is a huge pain in the butt. We're kind of fond of the old machine, so we're interested in fixing it. My question is: does anyone know if there is an adjustment to change at what point the relay for the knife holder kicks in? Or, is there some way to get the arm to travel farther down? Thanks much!
-Chris
(Sorry....I don't have a fancy title yet...I'm still just a student!)
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Hi folks, Anyone out there have any info on the Tracor-Northern 6100 diode array detector as used in the TN spectrophotometers? Feel free to point me to another list that is more appropriate for this question if you know of one. Thanks, Greg M. -- Gregory Mulhollan, Ph.D. Extreme Devices Inc. 101 West 6th Street Suite 200 Austin, TX 78701 (512)479-7740 x2231 voice (512)479-7750 fax mulhollan-at-extremedevices.com
For microscopists in the state of Virginia. Please contact Lou Solebello at microls1297-at-mindspring.com if you are interested in getting involved, or would attend regular meetings held in Richmond. I would like to start up a Virginia Microscopical Society if there is enough interest to do so. Professionals, hobbyists, teachers all welcome. Let me know if you are interested, and what types of activities and presentations you would like to see. Lou Solebello
Dear listers, I've inherited an old (1970s?) Leitz Orthoplan microscope in great need of a cleaning and possibly some new objectives. Can anyone give advice on who can help me rehabilitate it? I'm in the Atlanta-Birmingham-Montgomery (Alabama) area. Thanks Missy Josephson
Eleanor M. Josephson, D.V.M., Ph.D. Assistant Professor Department of Anatomy, Physiology and Pharmacology College of Veterinary Medicine 111 Greene Hall Auburn University, AL 36849-5518 Ph. (334) 844-5423 FAX (334) 844-4542 josepem-at-vetmed.auburn.edu
} We have an old LKB Ultramicrotome III (series 8800) that has } developed a problem in the past few weeks. } My question is: does anyone know if there is an adjustment to change } at what point the relay for the knife holder kicks in?
Yes, there is and I have the relevant LKB service note somewhere. I'll be happy to fax it to you if you email me a fax number that will reach you.
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 **
Dear Sir/Madam , JEM3010 was manufactured in 1999.it has got 1.500.000x mag and digital camera system JSM5600 has got 300.000xmag and EDS system. we have two kind of chose for prices. If we study together (I mean we preapare a paper together), we reseach samples without any price. On the other hand we get approximately 30$ per each sample. But if you have many samples we can make a discount. Thanks for your interests.. I am waiting for your emails
********************************** ********************************** ** 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 ** ********************************** **********************************
Hi Beth, I'm using the Casio QV 3000 privately and I'm very satisfied with it. High resolution, light sensitive CCD, easy to use functions and easily understandable menus. Nearly all my images are correctly exposed, and the macro setting works great, with really high resolution. It uses standard batteries, or as I do rechargeables, that can be bought everywhere. The camera body is slightly larger than most of the competitors, but this makes it so much easier to hold with a steady grip. All this together with the possibility of storing 244 images at high resolution with the 340 mB drive makes it one of the best choices. The Photoloader software package is easy to work with and makes the handling of your images in the PC fast and well arranged. I'm using a Lexar card reader, and image transfer to my PC is really fast, although USB usage is even faster. The only disadvantage I´ve found is that the software does not save images in TIFF- format, only JPEG. It's a good alternative to my Hasselblad and a lot easier to carry around outdoors. Now, you did not state what you need it for, ordinary photography is as you can see OK, but I think it would be hard to get adaptors to use it, for example, as a microscope camera .
Yours sincerely
Per Hörstedt Department of Medical Biosciences Pathology Unit for Electron Microscopy University of Umeå S-90187 Umeå Sweden
We have two Edwards 306 coaters in our lab. As with all carbon rod based evaporators you will need to experiment with the spring pressure holding the rods together. Too much pressure and the rods will glow and not arc properly (most likely the situation you described) or, too little pressure and the arc will cease prematurely. On the 306 coaters the spring pressure is adjusted by lengthening or shortening the carbon rods - first loosen the clamping collars and slide the rods through.
Hope this is some help.
David Vowles Electron Microscope 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
On Fri, 17 Nov 2000, Dunlap, Jonathan C. wrote:
} I have walked into a laboratory that has had an Edwards E306A Vacuum Coater } purchased in 1991. It has been used very rarely since, and by no one } presently working in the laboratory. The instruction manuals help to pump } down the system and get it ready for the application, but I have yet to get } the carbon source deposited on the sample we are trying to coat for SEM } analysis (I can get the carbon rods glowing, but nothing else happens).
there are two possibilities that spring to mind: 1. you've probably checked this but if the cutting range is set too high the arm might not fall far enough to engage the retract mechanism. 2. something in the retract circuit may have blown. It happened once on an LKBI and the engineer found a blown capacitor. I assume that there must be a fairly big current activating the electromagnet.
Have you watched the cutting cycle through the binoculars? Does it appear to try to move at the bottom of the cutting stroke or make a clicking noise? How far below the mid knife position does the specimen arm travel (it should be several millimetres)?
Good luck
Malcolm Haswell e.m. unit University of Sunderland UK
christopher 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 Evening, all: } } We have an old LKB Ultramicrotome III (series 8800) that has developed a } problem in the past few weeks. Apparently the cantilever arm is not going } down far enough on the cutting stroke to activate the kickback on the knife } holder, resulting in wet blocks nearly every stroke. Needless to say, this } is a huge pain in the butt. We're kind of fond of the old machine, so we're } interested in fixing it. } My question is: does anyone know if there is an adjustment to change at } what point the relay for the knife holder kicks in? Or, is there some way } to get the arm to travel farther down? } Thanks much! } } -Chris } } (Sorry....I don't have a fancy title yet...I'm still just a student!)
We used to use 1%KMNO4 in phosphate buffer, pH 6.5 instead of UA. We stained for 5min. It is supposed to be good for membranes. We flushed it down the sink.
Dave
On Fri, 17 Nov 2000 10:27:29 -0600 "Awbrey, Donald" {DonaldAwbrey-at-texashealth.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 TEM netters, } } Is there any other stain reagents, other than UA and LC, that are } used in TEM? If so, do they have comparable results, are they } ETOH or water based, what are there disposal precautions, and what } are their staining protocols. } } Any information will be appreciated. } } Thanks in advance. } } } } Donald G. Awbrey, HT(ASCP) QIHC } Electron Microscopy / Image Analysis } DonaldAwbrey-at-TexasHealth.org } donaldawbrey-at-hotmail.com } (817)-878-5647 } } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
I have responded to these queries before, so here goes again. The unit I have used for years is the Lynx Tissue Processor (currently available from EM Sciences). The original unit I purchased (from the manufacturer) was marvelous. A second unit (purchased when Leica was marketing it) took over 5 years to get it into reliable working condition. My guess is that EMS will provide a reliable product. The only other unit I know of is sold by RMC (recently cycled through Ventana and now Boekeler -- Tucson, AZ) that is an updated version of the old LKB design. I know of very happy users for this product, but I honestly do not know the current status under the new owners. I have attempted to get a demo of the RMC device, but RMC never followed through, and the last incarnations have occurred so quickly that I still can't provide a personal comparison.
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc. On Fri, 17 Nov 2000 00:19:10 -0800, Marek Malecki 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 appreciate comments and suggestions on automatic sample processors } for EM. Vendors welcome. } Marek Malecki, M.D., Ph.D. } Director and Principal Investigator } } Molecular Imaging Laboratories } University of California, San Diego } } address: 1500 Suite Bonner Hall, 9500 Gilman Drive, La Jolla, CA92093-0368 } phone - office: 8588223373 } phone - cell: 8583443347 } fax: 8588223715 } e.mail: mmm-at-ucsd.edu } pager: 8586161420 } e.mail: mmm-at-ucsd.edu } e.pager: 1620024619-at-alphapage.airtouch.com } www site: http://mil.ucsd.edu/ } ftp site: mil1.ucsd.edu } } }
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In addition to the Glauert series, there are the Hayat books (one specifically on stains, and volume one of the EM series -- both may be out of print now, but they are all available through interlibrary loan) and any of the basic biological TEM books also should cover (at least mentioning) stains other than UrAc and Pb Citrate. Bismuth is the only one I have used, and it is really a bear to work with. Potassium permanganate has also been used as have lanthanum, phosphotungstic acid and ruthenium. The Hayat book is "Stains and Cytochemical Methods", Plenum Press, 1993. The other one is "Principles and Techniques of Electron Microscopy; Biological Appications, Volume I". Wiley has a huge volume on EM methods (it is a loose-leaf beast that comes with updates--or it used to) that includes all of these methods as well.
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc. On Fri, 17 Nov 2000 10:27:29 -0600, Awbrey, Donald 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 TEM netters, } } Is there any other stain reagents, other than UA and LC, that are } used in TEM? If so, do they have comparable results, are they } ETOH or water based, what are there disposal precautions, and what } are their staining protocols. } } Any information will be appreciated. } } Thanks in advance. } } } } Donald G. Awbrey, HT(ASCP) QIHC } Electron Microscopy / Image Analysis } DonaldAwbrey-at-TexasHealth.org } donaldawbrey-at-hotmail.com } (817)-878-5647 } } }
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Jonathan: Don't feel all alone. I purchased a 306A about 16 years ago when Edwards was manufacturing and marketing the units. Even then, they didn't seem to know all that much about the units. I had problems with the main valve, and got less than no help. I don't think I EVER got a decent coat of carbon out of the unit. I did manage to melt down a set of carbon rod holders and burned out a transformer, but it never did provide a decent carbon coat. It did provide a good metal (read, gold, for SEM) coat, and I had purchased it because of access to electrodes and the great rotary table provided with it. However, I eventually went back to the old reliable Denton for everything but the gold coating, and then managed to get a sputter coater, so the 306A was left standing, abandoned. Justice at last.
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc. On Fri, 17 Nov 2000 14:24:30 -0500, Dunlap, Jonathan C. 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 have walked into a laboratory that has had an Edwards E306A Vacuum Coater } purchased in 1991. It has been used very rarely since, and by no one } presently working in the laboratory. The instruction manuals help to pump } down the system and get it ready for the application, but I have yet to get } the carbon source deposited on the sample we are trying to coat for SEM } analysis (I can get the carbon rods glowing, but nothing else happens). } } I am wondering if anyone out there has this system and knows how to operate } it with the carbon evaporation source. Any help to what I could be doing } wrong, or not doing at all would be appreciated. Also perhaps if I could } call someone who has this system and they could walk me though the steps } would be great. } } - Jonathan } } (I have contacted the manufacturer and they have not made this coater in } years so know little about it as well) }
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We are looking for antibodies to mitochondrial proteins , for a immunocytochemical study on the effect of parasites in the development of youngs birds . Does anyone or company have mitochondrial antibodies ?
Beth...I have owned the Casio QV-3000 since April 2000. I think it a totally awesome for Macro work. A picture is worth a 1000 words...please go to my online photo album and look for yourself. Some of the pictures have a reference scale in them.
http://communities.msn.com/CactusCritters
Best,
Al Coritz Sales & Service Manager RMC-Boeckeler Instruments 4650 S. Butterfield Dr. Tucson, AZ 85714 Voice: 520-745-0001 Cell: 520-465-3598 Fax: 520-745-0004 Email:Al-at-Boeckeler.com Website:RMCProducts.com
-----Original Message----- } From: Beth Richardson [mailto:beth-at-dogwood.botany.uga.edu] Sent: Friday, November 17, 2000 8:10 AM To: microscopy-at-sparc5.microscopy.com
Hi, We are looking for opinions or reviews on the the Casio QV 3000 digital camera, with a 340 MB microdrive and macro focusing down to 6 cm. any help would be greatly appreciated. thanks, Beth
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
I've used one to mitochondrial hsp70 from Affinity BioReagents that works well. Their number is: (800) 278-2424. The catalogue number for the antibody is MA3-028. In our hands it works in human and mouse cells....never tried it on birds or inverts.
The other option would be to use one of the MitoTracker dyes from Molecular probes.
No financial interest in either company, unfortunately!
Tamara Howard CSHL
On Mon, 20 Nov 2000, Gilles Grondin 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 antibodies to mitochondrial proteins , for a } immunocytochemical study on the effect of parasites in the development of } youngs birds . Does anyone or company have mitochondrial antibodies ? } } Thank you very much . } } Gilles Grondin } } ggrond01-at-courrier.usherb.ca } } }
This is for the person asking about alternative post stains. Instead of using lead after uranyl acetate I have been very happy with bismuth (I have been looking at various biological samples embedded in Spurrs).
STOCK SOLUTION: In 10ml of DH2O add 0.2g of NaOh, 400mg of sodium tartrate, and 200mg of bismuth subnitrate. Shake until dissolved and store up to 2 weeks in the refrigerator.
WORKING SOLUTION: add 20 micro liters of stock to 1ml of DH2O and float the grids on 50 micro liter drops of this for 5 minutes and rinse with H2O.
Good Luck, Tim
Timothy G. Schneider Director of Electron Microscopy Department of Pathology Room 229 Jefferson Hall Thomas Jefferson University 1020 Locust St. Philadelphia Pa. 19107 215-503-4798 work 610-613-8170 cellular timothy.schneider-at-mail.tju.edu
Years ago I worked in a lab that had a small forced (and filtered) film dryer (with heater). It was just large enough to hold two racks of TEM negatives. I cannot find one in any of the catalogues. I can only find film dryers for 35 mm roll film. Could anyone please recommend a source of a similar dryer? (Vendors, please feel free to respond directly to me).
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
I am trying to find out information on a good imaging system(photographic only) that could be attached to my SEM, which is an older ABT-55. I do not have the funds to buy a newer microscope, so I would appreciate any input you all have. The cost of Polaroid film is extremely expensive, so I am trying to find a system that I could send digital images through the e-mail and/or print them instead of using this costly film. Thank you in advance.
Cathy Kelloes Microscopy Technician bp Fabrics & Fibers Unit 260 The Bluffs Austell, GA. 30168 (770) 941-1711, Ext. 3255
My friend's son is writing a report on oxytocin and called me to ask if I had or could find any images of it. Its the typical "call X, s/he is a scientist" type referal. Does anyone out there have any images, or does anyone know of a good web site I can send him to? I assume that polarization optics images of the crystalized molecule would be pretty, but anything would be more than he currently has.
Please respond to me off-list, and I will forward the info to him.
TIA, 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
The 34th Annual NESM (New England Society for Microscopy) Symposium will be held at the Newton Marriott, Newton, Massachusetts on Friday, December 1st from Noon to 9:30 pm.
The meeting will consist of 2 sessions. SESSION I (1-2:30 pm) will focus on Raman and FT-IR Spectroscopy. Speakers will include Barbara Foster-"Bridging the Microscopy-Spectroscopy Chasm", John Reffner-"Resolving Molecular Chemistry with FT-IR Microspectroscopy" and Patrick Treado-"Chemical Imaging: A Powerful Tool for Molecular Microscopy".
A Poster Session/Contest will then run concurrent with the coffee break (2:30- 3:15 pm) (details below).
SESSION II (3:15-4:45 pm) will focus on Genechip Technology. Speakers for that session will include: Eiman H. Al-Mutari-"The Human Genome Study: cDNA Micro- arrays and Signal Amplification", Joseph Paulauskis-"Microarray Technologies: Promise and Problems", and Steven Lott-"Microarray Technology: Changing the Face of Functional Genomics".
The annual business meeting/election of new officers will convene at 5:00 pm, followed by a Cocktail Hour (6-7 pm) and dinner (7-8 pm). The after dinner speaker will be Paul Hlava, MAS Tour Speaker, from Sandia National Labs whose talk will be: "Causes of Color in Minerals and Gemstones".
The DEADLINE for advance registration (and dinner reservations) is Friday, November 24th. Payment must accompany advance registration form if you wish to have dinner. The choices for dinner are: herbed crusted chicken, swordfish, and vegetarian (please choose one). Please contact Mary McCann, Treasurer, at (617) 484-7865 or by email: mccanns-at-tiac.net for further information. Send registration forms WITH payment to NESM, c/o Mary McCann, 161 Claflin Street, Belmont, MA 02478. (Registration will be held at the meeting from 12Noon-1pm, but will NOT include dinner).
Poster abstracts should be sent to: Christopher Santeufemio, Biological Director at: 57 Bancroft Street, Pepperell, MA 01463 by November 24th. Any questions, contact Chris at (978) 433-8031 or by email: csanteufemio-at-epion.com. Prizes will be awarded for Best Poster, Second and Third Place.
We hope to see you there at this most interesting meeting!
Peggy Sherwood Corresponding Secretary, NESM
-- Peggy Sherwood Lab Associate, Photopathology Wellman Laboratories of Photomedicine (W224) Massachusetts General Hospital 50 Blossom Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) sherwood-at-helix.mgh.harvard.edu
I've been asked to do some TEM on hookworms. I've never even seen a hookworm before, let alone do EM on one. I've heard that they're tough customers when it comes to fixation.
Does anybody out there have a protocol that is fairly simple but results in good utlrastructural preservation?
There is a company called California Stainless that made film dryers that some of the EM supply houses sold. I bought one from them a number of years ago. Sorry that I don't have the contact info, but I'm sure that this is just what you want.
-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: Donald Lovett [mailto:lovett-at-tcnj.edu] Sent: Monday, November 20, 2000 9:51 AM To: Microscopy.lst Subject: Film dryers
--------------------------------------------------------------- --------- The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserv er/FAQ.html
Years ago I worked in a lab that had a small forced (and filtered) film dryer (with heater). It was just large enough to hold two racks of TEM negatives. I cannot find one in any of the catalogues. I can only find film dryers for 35 mm roll film. Could anyone please recommend a source of a similar dryer? (Vendors, please feel free to respond directly to me).
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
On Mon, 20 Nov 2000 09:33:34 -0500 Timothy Schneider {Timothy.Schneider-at-Mail.TJU.EDU} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } This is for the person asking about alternative post stains. Instead of } using lead after uranyl acetate I have been very happy with bismuth (I have } been looking at various biological samples embedded in Spurrs). } } STOCK SOLUTION: In 10ml of DH2O add 0.2g of NaOh, 400mg of sodium tartrate, } and 200mg of bismuth subnitrate. Shake until dissolved and store up to 2 } weeks in the refrigerator. } } WORKING SOLUTION: add 20 micro liters of stock to 1ml of DH2O and float the } grids on 50 micro liter drops of this for 5 minutes and rinse with H2O. } } Good Luck, Tim } } Timothy G. Schneider } Director of Electron Microscopy } Department of Pathology } Room 229 Jefferson Hall } Thomas Jefferson University } 1020 Locust St. } Philadelphia Pa. 19107 } 215-503-4798 work } 610-613-8170 cellular } timothy.schneider-at-mail.tju.edu } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
See if an active or passive digital control & capture unit would work on your SEM. Soft Imaging makes ADDA which does active and passive very nicely. There are others which do active and some which only do passive. I prefer active. But if your system can't be adapted to active, perhaps it will support passive. If not, maybe active.
Several of the suppliers are on this list. Perhaps they will contact you. I use ADDA and like it very much. It is actually more than just SEM image capture. I use it to capture chamber TV camera video and SEM TV scan images as well as up to 4096x4096 pixel active scan SEM image files (TIFF).
gary
At 07:28 AM 11/20/00, you wrote:
} Hi, } } I am trying to find out information on a good imaging system(photographic } only) that could be attached to my SEM, which is an older ABT-55. I do not } have the funds to buy a newer microscope, so I would appreciate any input } you all have. The cost of Polaroid film is extremely expensive, so I am } trying to find a system that I could send digital images through the e-mail } and/or print them instead of using this costly film. Thank you in advance. } } Cathy Kelloes } Microscopy Technician } bp Fabrics & Fibers Unit } 260 The Bluffs } Austell, GA. 30168 } (770) 941-1711, Ext. 3255
The RMC "EMP 5160" EM tissue processor is available through Boeckeler Instruments as is the entire RMC Instrument catalog. For more information visit our website: www.rmcproducts.com
Al Coritz Sales & Service Manager RMC-Boeckeler Instruments 4650 S. Butterfield Dr. Tucson, AZ 85714 Voice: 520-745-0001 Cell: 520-465-3598 Fax: 520-745-0004 Email:Al-at-Boeckeler.com Website:RMCProducts.com
-----Original Message----- } From: Roger Moretz [mailto:rcmoretz-at-excite.com] Sent: Monday, November 20, 2000 6:00 AM To: Marek Malecki; Microscopy-at-sparc5.microscopy.com
Mareck:
I have responded to these queries before, so here goes again. The unit I have used for years is the Lynx Tissue Processor (currently available from EM Sciences). The original unit I purchased (from the manufacturer) was marvelous. A second unit (purchased when Leica was marketing it) took over 5 years to get it into reliable working condition. My guess is that EMS will provide a reliable product. The only other unit I know of is sold by RMC (recently cycled through Ventana and now Boekeler -- Tucson, AZ) that is an updated version of the old LKB design. I know of very happy users for this product, but I honestly do not know the current status under the new owners. I have attempted to get a demo of the RMC device, but RMC never followed through, and the last incarnations have occurred so quickly that I still can't provide a personal comparison.
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc. On Fri, 17 Nov 2000 00:19:10 -0800, Marek Malecki 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 appreciate comments and suggestions on automatic sample processors } for EM. Vendors welcome. } Marek Malecki, M.D., Ph.D. } Director and Principal Investigator } } Molecular Imaging Laboratories } University of California, San Diego } } address: 1500 Suite Bonner Hall, 9500 Gilman Drive, La Jolla, CA92093-0368 } phone - office: 8588223373 } phone - cell: 8583443347 } fax: 8588223715 } e.mail: mmm-at-ucsd.edu } pager: 8586161420 } e.mail: mmm-at-ucsd.edu } e.pager: 1620024619-at-alphapage.airtouch.com } www site: http://mil.ucsd.edu/ } ftp site: mil1.ucsd.edu } } }
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Thanks again for all of the responses. They were very informative.
We primarily perform TEM on renal biopsies particularly the Bowman's capsule and glomerulus. The main ultra- structure we concentrate on is the basement membranes of the glomerulus. Less frequently we perform TEM on tumor tissues. This is done in a clinical setting.
Any other information out there in the "Netter Land" will be appreciated.
Thanks Again,
Donald G. Awbrey, HT(ASCP) QIHC Electron Microscopy / Image Analysis DonaldAwbrey-at-TexasHealth.org donaldawbrey-at-hotmail.com (817)-878-5647
At 16:37 16/11/00 -0500, you wrote: } Dear Listers, } Does anyone have an experience that they would like to share involving the } SVMicro digital camera by Sound Vision? I'm hoping to learn anything at } all, from delivery, support, service, image quality, product reliability, } etc. Thanks in advance. } Bill Roberts
We have an SVMicro attached to a Nikon Labophot-Pol petrographic microscope, with a Nikon 1x relay lens in the adaptor. We also use it with a Zeiss Jenaval. Alternatively you can just screw a lens on it and point it at something (with a tripod, preferably). The camera runs off a SCSI card in a PC.
Our experience is that the Twain interface software is a little tricky to use, and it's difficult for new users to get good images until you have done some tweaking. You usually need to turn the illumination down a bit, or use a neutral density filter. The colour balance needs to be corrected -- I use a neutral density filter as the specimen to set the neutral gray balance -- but a pale blue filter may also work. The images tend to be very contrasty unless you set the gamma level (the middle triangle on the intensity histogram) a long way to the left. Adjusting the levels tends to be very touchy, as compared to say Photoshop. However once you get the exposure time right you can get very nice RGB images, at a true pixel resolution of about 1000x800.
Thor Bostrom Analytical EM Facility Faculty of Science Queensland University of Technology (QUT) PO Box 2434, Brisbane, QLD 4001 AUSTRALIA
I may have a copy of a paper somewhere which advocated hot fixatives for nematodes because t worked better than room temp/cold fixatives. Purely on penetration? Getting through the cuticle? Come back if you're still desperate - I think it is in my cellar somewhere at home!
A colleague here is trying to determine the presence and size of microcrystallites in a glass ceramic. It is an alkali-lime borosilicate glass, which appears to be amorphous by XRD. However it has a translucent appearance which suggests that it may contain microcrystallites or some other defects, though these are not evident by optical microscopy.
The suggestions so far are: (a) light etching of a polished surface, followed by examination in a FESEM; or (b) preparation of an ion beam thinned sample for TEM.
We would welcome any other suggestions. For example, would EBSD work? And if we tried (a), is there a suitable etchant (dilute HF?) for this material?
With thanks,
Thor Bostrom Analytical EM Facility Faculty of Science Queensland University of Technology (QUT) PO Box 2434, Brisbane, QLD 4001 AUSTRALIA Email: t.bostrom-at-qut.edu.au
=-=-=-=-=-=-=-=-=-=-=-=-=-= Dr Thor Bostrom Ext. 2351 Analytical EM Facility, Faculty of Science, QUT t.bostrom-at-qut.edu.au =-=-=-=-=-=-=-=-=-=-=-=-=-=
Does anyone have suggestions how to add a label/element to Lowicryl HM20. We would like to differentiate/locate the Lowicryl in some porous structures using EDX.
thanks,
Johan Hazekamp
************************************************** Johan Hazekamp Central Analytical Science Unilever Research Vlaardingen P.O. Box 114, 3130 AC Vlaardingen The Netherlands Tel.: (31) 10-4605530 Fax: (31) 10 4605671 Email: Johan.Hazekamp-at-unilever.com
TEM examination is best and would allow identification of the crystallites. Ion milling silicates is potentially a problem. I'd suggest choosing a method that doesn't involve much, if any, ion milling, such as cleaving, crushing, or tripod polishing.
Ron Anderson, IBM, Hopewell Jct., New York, USA. anderron-at-us.ibm.com IBM Analytical Services; http://www.chips.ibm.com/services/asg
Thor Bostrom {t.bostrom-at-qut.edu.au} on 11/21/2000 12:42:07 AM
To: microscopy-at-sparc5.microscopy.com cc:
A colleague here is trying to determine the presence and size of microcrystallites in a glass ceramic. It is an alkali-lime borosilicate glass, which appears to be amorphous by XRD. However it has a translucent appearance which suggests that it may contain microcrystallites or some other defects, though these are not evident by optical microscopy.
The suggestions so far are: (a) light etching of a polished surface, followed by examination in a FESEM; or (b) preparation of an ion beam thinned sample for TEM.
We would welcome any other suggestions. For example, would EBSD work? And if we tried (a), is there a suitable etchant (dilute HF?) for this material?
With thanks,
Thor Bostrom Analytical EM Facility Faculty of Science Queensland University of Technology (QUT) PO Box 2434, Brisbane, QLD 4001 AUSTRALIA Email: t.bostrom-at-qut.edu.au
=-=-=-=-=-=-=-=-=-=-=-=-=-= Dr Thor Bostrom Ext. 2351 Analytical EM Facility, Faculty of Science, QUT t.bostrom-at-qut.edu.au =-=-=-=-=-=-=-=-=-=-=-=-=-=
In the early 1980s, we followed someone's suggestion to dissolve Iodoform in epoxy resin to raise its average atomic number to provide contrast with coal particles. It also provided elemental contrast.
It worked with epoxy; it setup just fine. But I am not sure how it would work Lowicryl. It does list a number of precautions listed on the MSDS.
At 11:24 AM 11/21/2000 +0100, you wrote:
} Dear All, } } Does anyone have suggestions how to add a label/element to Lowicryl HM20. } We would like to differentiate/locate the Lowicryl in some porous structures } using EDX. } } thanks, } } Johan Hazekamp } } ************************************************** } Johan Hazekamp } Central Analytical Science } Unilever Research Vlaardingen } P.O. Box 114, 3130 AC Vlaardingen } The Netherlands } Tel.: (31) 10-4605530 } Fax: (31) 10 4605671 } Email: Johan.Hazekamp-at-unilever.com }
Johan Polysciences http://www.polysciences.com/ list barium methacrylate, lead methacrylate and lead acrylate, all of which can be used to impart x-ray / electron opacity to acrylic polymers. I don't know how you would formulate Lowicryl to take advantage of this, but it looks like a possible starting point. I would be very interested to know of specific formulations for using these compounds in EM grade embedding, because I have a related application.
Best wishes Chris
Date sent: Tue, 21 Nov 2000 11:24:32 +0100 } From: "Johan Hazekamp" {Johan.Hazekamp-at-unilever.com}
Microcrystallites can be identified using back scattered electron image on a polished surface of the sample. If the microcrystallites are big enough, e.g., a few microns or larger, its chemical composition can be determined on a polished surface using electron microprobe. In some cases, the shapes and chemical composition of the phase will be enough to determine whether the phase is crystalline.
At 04:42 PM 11/21/00 +1100, Thor Bostrom wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
To everyone who sent me information on SEM image systems, I want to tell you how much I appreciate it. I have received so many good suggestions and references that I now will be able to make a more knowledgeable decision. Once again, thank you so much.
Cathy Kelloes Microscopy Technician bp Fabrics & Fibers Unit 260 The Bluffs Austell, GA. 30168 (770) 941-1711, Ext. 3255
I was wondering if anyone can help me find a tiltable and rotatable specimen cartridge for a Zeiss EM 109 TEM. Vendors quite welcome!
Thanks very much for your help
Kindest Regards
Stephen Wood Meridian Scientific Services Inc. Ottawa Canada Tel: 613-836-6749 Fax: 613-836-5880 e-Fax:413-460-3007 e-mail: stephenwood-at-meridiansci.com web site: http://meridiansci.com/
Any one have a reference on the effect of pH on the osmication reaction? My attempts at a literature search on this topic have not been much success. Happy Thanksgiving.
-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
Dear Cathy, You may find that Quartz PCI, sold by Hitachi Instruments, is most suitable for passively capturing images from an older SEM. You get an image that is very close to the Poaroid image, stored on the computer in your choice of 18 different formats. Visit them at: www.quartzimaging.com. At 09:28 AM 11/20/00 -0600, you wrote: } } Hi, } } I am trying to find out information on a good imaging system(photographic } only) that could be attached to my SEM, which is an older ABT-55. I do not } have the funds to buy a newer microscope, so I would appreciate any input } you all have. The cost of Polaroid film is extremely expensive, so I am } trying to find a system that I could send digital images through the e-mail } and/or print them instead of using this costly film. Thank you in advance. } } Cathy Kelloes } Microscopy Technician } bp Fabrics & Fibers Unit } 260 The Bluffs } Austell, GA. 30168 } (770) 941-1711, Ext. 3255
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 saw your messages in the microscopy list and was so happy to "hear" from you. I don't know if you'll remember me, I used to be a student at the Hort Department with Dr. Hazel Wetzstein and took both EM courses at the EM center in Athens.
I'm back in Brazil since 1994 and I've been back to Athens a few times after that. I visited most of the people I've known and missed seeing you there. I knew you're in Atlanta from Dr. Farmer.
How are things going with you? Do you still have horses? I see you are still working with EM, which is good!
Well, whenever you have a chance, let me know how things are going. I'm going to the US in December January and will spend some time in Athens and also will visit some friends in Atlanta. Maybe I'll have a chance to meet you again.
Take care, regards,
Adriana
Adriana P. M. Rodriguez Laboratório de Biotecnologia Vegetal CENA, Universidade de São Paulo adriana-at-cena.usp.br
Being new to this system, I don't know how this is going to work. However, I purchased an ISI-SS40 a few year ago, but I did't get any manuals with it. It appears to work ok, but there are a lot of questions. If anynoe
Knows where I can get manuals for this particular instrument I would greatly appreciate hearing from you. You may e-mail me at oconnell-at-ltu.edu or call 734-668-3309and I will get back with you.
I don't see how examination in a FESEM will be helpful. I doubt that EBSD will work because of the volume required to produce a pattern. I think that TEM is the only way to get the information. ..Russ Cook } ----------------------------------------------------------------------- } } A colleague here is trying to determine the presence and size of } microcrystallites in a glass ceramic. It is an alkali-lime borosilicate } glass, which appears to be amorphous by XRD. However it has a translucent } appearance which suggests that it may contain microcrystallites or some } other defects, though these are not evident by optical microscopy. } } The suggestions so far are: (a) light etching of a polished surface, } followed by examination in a FESEM; or (b) preparation of an ion beam } thinned sample for TEM. } } We would welcome any other suggestions. For example, would EBSD work? And } if we tried (a), is there a suitable etchant (dilute HF?) for this material? } } } With thanks, } } Thor Bostrom } Analytical EM Facility } Faculty of Science } Queensland University of Technology (QUT) } PO Box 2434, Brisbane, QLD 4001 } AUSTRALIA } Email: t.bostrom-at-qut.edu.au } } =-=-=-=-=-=-=-=-=-=-=-=-=-= } Dr Thor Bostrom } Ext. 2351 } Analytical EM Facility, } Faculty of Science, QUT } t.bostrom-at-qut.edu.au } -----------------------------------------------------------------------
---------------------------------------------------------------------- Russell E. Cook, Ph.D. Electron Microscopy Center 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
We need another specimen holder for a Philips /FEI CM 120. Gatan made a perfect holder with ±60 tilt & 360° of rotation. I am not looking for a cryoholder. Hope someone has one of these Gatan gems on their shelf that they would like to sell to us. thanks...Tom Reese, NIH
************************************************ Final Call for Papers ************************************************
This international conference will focus on the latest developments in the study of the structural and electrical properties of semiconductors by the application of transmission and scanning electron microscopy, scanning probe microscopy and X-ray techniques.
The state-of-the-art in all important subject areas will be addressed, including the characterisation of bulk and thin film as-grown materials, the study of lattice defect and impurity behaviour and the investigation of advanced semiconductor processing procedures.
Special conference sessions will concentrate on recent developments in high-resolution imaging and analytical electron microscopy, advances in SEM and SPM applications, the characteristics of epitaxial layers (including III-V nitrides), quantum wells, wires and dots, the effects of device processing treatments (including, especially, advanced silicon technology) and metal-semiconductor contacts and silicides. Prominent invited speakers will introduce each topic area.
The Proceedings of the conference will be published and the final call for papers has now been issued. Abstracts (deadline 1 DECEMBER 2000) should be submitted by E-mail to: jenny-at-rms.org.uk
Full conference information (with the invited speaker listing, etc) can be found at the conference Web site http://www.rms.org.uk/currentevents2.htm#MSMXII
The conference Chairmen are Prof Tony Cullis (a.g.cullis-at-sheffield.ac.uk) and Dr John Hutchison (john.hutchison-at-materials.ox.ac.uk) who may be contacted with any general enquiries.
************************************************ Final Call for Papers ************************************************
This international conference will focus on the latest developments in the study of the structural and electrical properties of semiconductors by the application of transmission and scanning electron microscopy, scanning probe microscopy and X-ray techniques.
The state-of-the-art in all important subject areas will be addressed, including the characterisation of bulk and thin film as-grown materials, the study of lattice defect and impurity behaviour and the investigation of advanced semiconductor processing procedures.
Special conference sessions will concentrate on recent developments in high-resolution imaging and analytical electron microscopy, advances in SEM and SPM applications, the characteristics of epitaxial layers (including III-V nitrides), quantum wells, wires and dots, the effects of device processing treatments (including, especially, advanced silicon technology) and metal-semiconductor contacts and silicides. Prominent invited speakers will introduce each topic area.
The Proceedings of the conference will be published and the final call for papers has now been issued. Abstracts (deadline 1 DECEMBER 2000) should be submitted by E-mail to: jenny-at-rms.org.uk
Full conference information (with the invited speaker listing, etc) can be found at the conference Web site http://www.rms.org.uk/currentevents2.htm#MSMXII
The conference Chairmen are Prof Tony Cullis (a.g.cullis-at-sheffield.ac.uk) and Dr John Hutchison (john.hutchison-at-materials.ox.ac.uk) who may be contacted with any general enquiries.
Hi Y'all: We're looking to buy or trade for an annealing holder for a 100CX/200CX/1200 EX TEM's (fits all of these and probably others). We hope someone has one that they haven't used in years and would like to get some money out of it Please let us know. Regards, Mike Coviello Lab Manager UT Arlington 817 272-5496
Does anyone have information about a supplier of amber colored carboy containers. We need these carboys to be about 1-2 gallon in size. They need to made of a plastic polymer that is resistant to hazardous chemicals. They are to be used to store stock film processing fluids. Any info would be greatly appreciated.
Thanks in advance.
Donald G. Awbrey, HT(ASCP) QIHC Electron Microscopy / Image Analysis DonaldAwbrey-at-TexasHealth.org donaldawbrey-at-hotmail.com (817)-878-5647
Ahoy fellow enthusiasts! Has anyone out there used a comparison microscope before? I got a bridge unit and dropped it on top of 2 AO 10 scopes, there seems to me a knob on the front that does nothing. I get one scope in once side of the field and the image from the other scope on the other side.....
I suspect the knob makes them overlap (like we have seen on the police shows but it does not seem to move the image. when they compare bullets...)
The original volume of Hayat's Principles & Techniques of Electron Microscopy (vol. 1), now long out of print, discusses this in chapter 1. He references a paper by Trump and Ericsson (Lab Invest 14:1245, 1965) as a primary reference. In fact, if you were to review Ben Trump's literature, you would find a wealth of information on the role of pH, osmolarity, fixative concentrations and types, etc. Most of this has been summarized by Hayat in various volumes and in the Glauert series. A quick on-line search at PubMed will give you enough references to bury you! However, using interlibrary loan you should be able to at least get the Glauert series, altho' I think that precious few libraries (including most EM labs) have the Hayat books. Although I can't lay my hands on a lot of books right now, I would also have thought that any basic book on TEM would cover your question, so if you have access to any of the recent books, check out the chapter on fixatives and fixation.
Roger Moretz, Ph.D. Dept. of Toxicology Boehringer Ingelheim Pharmaceuticals
On Tue, 21 Nov 2000 11:59:23 -0600, Tom Phillips 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 } -----------------------------------------------------------------------. } } } Any one have a reference on the effect of pH on the osmication } reaction? My attempts at a literature search on this topic have not } been much success. Happy Thanksgiving. } } -- } 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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Hi everybody, hope someone can help me: I need to stick each other two pieces of coated metallic samples to make a XTEM observation; then I need to stick them (already together bonded!) on the lapping tool in order to thin them. I have tried to use superglue (cyanoacrylic glue) for the first operation and thermoplastic glue for the second. Unfortunately I haven't found nothing to dissolve the thermoplastic but the superglue.
Does anyone can suggest me two different glue that can be suitable and can be dissolved selectively?
Thank you in advance, Edoardo.
Dr. Eng. Edoardo Bemporad, Ph. D. Assistant Professor of Materials Science University of Rome "Roma Tre" (Italy) Dipartimento di Ingegneria Meccanica e Industriale (Department of Mechanical and Industrial Engineering) Via Vasca Navale 79 - 00146 Rome, Italy Tel: +39 06 5517.3293 Fax: +39 06 5517.3256 LIME Lab Tel: +39 06 5517.3200 LIME Web Site: http//materials.dimi.uniroma3.it/lime E-Mail:bemporad-at-uniroma3.it
I do XTEM of metal films, and have found Gatan's G-1 epoxy to be effective in creating the "sandwich". I use crystal bond low-melting wax to bond the sandwich to the polishing stub for thinning. The former is resistant to most solvents; the latter dissolves readily in acetone.
I also hear that M-bond 610 adhesive is good for gluing the halves of the sandwich together, though I have not tried this brand.
The material you should use to stick the pieces together is Epotek 353ND which is produced by Epoxy Technology. You can find information on the product on their website at www.epotek.com/optical.html. Their distributor in Italy is:
Anna Giallo Kontek Comatel S.P.A. Via Rio Vallone #5 20050 Mezzago. Milan Italy
The acetone soluble wax that you can use it our QuickStick 135. This comes in a package with 20 small sticks and is ideal for this application. I hope this helps!
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 "Bemporad, Edoardo" } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi everybody, hope someone can help me: I need to stick each other two pieces of coated metallic samples to make a XTEM observation; then I need to stick them (already together bonded!) on the lapping tool in order to thin them. I have tried to use superglue (cyanoacrylic glue) for the first operation and thermoplastic glue for the second. Unfortunately I haven't found nothing to dissolve the thermoplastic but the superglue.
Does anyone can suggest me two different glue that can be suitable and can be dissolved selectively?
Thank you in advance, Edoardo.
Dr. Eng. Edoardo Bemporad, Ph. D. Assistant Professor of Materials Science University of Rome "Roma Tre" (Italy) Dipartimento di Ingegneria Meccanica e Industriale (Department of Mechanical and Industrial Engineering) Via Vasca Navale 79 - 00146 Rome, Italy Tel: +39 06 5517.3293 Fax: +39 06 5517.3256 LIME Lab Tel: +39 06 5517.3200 LIME Web Site: http//materials.dimi.uniroma3.it/lime E-Mail:bemporad-at-uniroma3.it
Dear Thor, EBSP might work, if the micro-crystallites are larger than the e-beam size. The specimen preparation is the same as for rocks for EBSP: polish down to 0.01 micron or Syton (colloidal silica) level. Best to ask someone who has done it. Try the web sites of HKL (www.channel.dk) or TSL. At 04:42 PM 11/21/00 +1100, you wrote: } } A colleague here is trying to determine the presence and size of } microcrystallites in a glass ceramic. It is an alkali-lime borosilicate } glass, which appears to be amorphous by XRD. However it has a translucent } appearance which suggests that it may contain microcrystallites or some } other defects, though these are not evident by optical microscopy. } } The suggestions so far are: (a) light etching of a polished surface, } followed by examination in a FESEM; or (b) preparation of an ion beam } thinned sample for TEM. } } We would welcome any other suggestions. For example, would EBSD work? And } if we tried (a), is there a suitable etchant (dilute HF?) for this material? } } } With thanks, } } Thor Bostrom
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 all, I am looking at real-time X-ray radiography systems and scanning acoustic microscopes for evaluation of solder bonds in optoelectronic packages. If anyone knows of suppliers of such systems - or even better has one they like (or don't), I would really like to hear from you. This includes manufacturers too - I'm not sure I have a comprehensive list yet.
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It depends on who's bridge you have. The two most common are the Leitz and the AO design. An AO type would say AO, Reichart or Leica on the manufactures logo depending on how new or old it is. Cambidge decided to use the Leica name on both once they owned both designs so the Leitz could also say Leica. There are other designs but these two make up probably more than 90% of all comparison bridges. I haven't worked with any of the others in over 20 years and then only a bit.
The knob in the center of the front makes it sound more likely to be an AO type. If it is, that knob should move the center prism set so that with it turned fully clock-wise you would see all of the left scope and turned fully counter-clock-wise you would see all of the right scope. With the knob centered you would see half of each field. In some other position you would see a smaller part of one field and a larger part of the other. If it is an AO style bridge it sounds to me as if the rack and gear might be damaged so the prism is not moving. The thin dividing line you see between the two fields should move as you turn the knob to give you a smaller percentage of one field and a larger percentage of the other. This lets us compare the pattern of stria over a length of the toolmark, a bullet comparison is a type of toolmark comparison. Side by side comparison is much more useful in Firearm and toolmark comparison than overlapping fields are.
If it is a Leitz design some do not let you move the dividing line off center but have other knobs that overlap the two fields or let you view just one. These bench top Leitz bridges don't have a knob in the center of the front so this probably isn't what you have.
In modern bridges the bridges that attach to separate scopes are designed for hair and fiber comparisons and the like. Bullet ("Universal Forensic") scopes have the objectives attached into the bridges in scopes that have been built since well before I entered the field over a 1/4 century ago. They are designed for much lower power and much longer working distances. The bridges for hair and fiber comparison scopes (yes they can be used for all manor of comparison but in forensic science labs that is their primary job and the way they are usually refereed to) are quite similar to bullet scope bridge designs but are designed to be attached to separate scopes that are basically conventional compound microscopes with sub-stage lighting.
If you tell me more about the bridge you are trying to use I may be able to tell you more that can help you set it up and see if it is working properly. AO then Reichart and now Leica have always said that users should not open up bridges to try repairs, as prism alignment is critical and easily disturbed. Rebuilds are available basically only at the factory, no one else has the alignment equipment to do it right. When other places have told me they could rebuild one I found that their version of a rebuild amounted to a real good cleaning job not prism replacement or realignment. If you do have an AO type and the knob is not moving the rack the center prism is on you may need such a rebuild. I did have a knob set screw come loose on an old one quite a number of years ago. I got away with going inside and tightening it up but I would not recommend it to anyone else as these are expensive pieces of equipment and rebuilds are several thousand dollars (around 12K to 15K last time I checked several years ago).
Jim
James L. Roberts Firearm and Toolmark Examiner Ventura Co. Sheriff's Lab (805) 654-2308 James.Roberts-at-mail.co.ventura.ca.us
} } } {"COURYHOUSE-at-aol.com"-at-sparc5.microscopy.com} 11/21/00 10:43PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Ahoy fellow enthusiasts! Has anyone out there used a comparison microscope before? I got a bridge unit and dropped it on top of 2 AO 10 scopes, there seems to me a knob on the front that does nothing. I get one scope in once side of the field and the image from the other scope on the other side.....
I suspect the knob makes them overlap (like we have seen on the police shows but it does not seem to move the image. when they compare bullets...)
Well I guess I should read the subject line as well as the body of the note and I would have seen that it was an AO bridge. Sorry for the info on the Leitz design. As indicated in my earlier message the knob should move the prism set so that you see part of one field and part of the other and what percentage of each is determined by the position of the knob. In the AO they do not "overlap."
If the knob is not moving the prism set you may have a loose set screw or you may need a bridge rebuild. Again I would not recommend going into the bridge yourself as alignment of the left, right and center prism is critical. Jim
James L. Roberts Firearm and Toolmark Examiner Ventura Co. Sheriff's Lab (805) 654-2308 James.Roberts-at-mail.co.ventura.ca.us
} } } {"COURYHOUSE-at-aol.com"-at-sparc5.microscopy.com} 11/21/00 10:43PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Ahoy fellow enthusiasts! Has anyone out there used a comparison microscope before? I got a bridge unit and dropped it on top of 2 AO 10 scopes, there seems to me a knob on the front that does nothing. I get one scope in once side of the field and the image from the other scope on the other side.....
I suspect the knob makes them overlap (like we have seen on the police shows but it does not seem to move the image. when they compare bullets...)
I have been waiting for a response from your technical support concerning mapping . Can you have them call me.
Thank you Larry Howard
} From: "Mary Mager" {mager-at-interchange.ubc.ca} To: "Kelloes, Cathy L" {KELLOECL-at-bp.com} Cc: {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, November 21, 2000 1:07 PM
NIST used to offer a SEM calibration standard SRM-2069. I don't see this any longer. If one needs to calibrate or qualify a SEM's resolution at 250A or better, what is an appropriate method?
I've used Au on C "standards" but these are more qualitative than quantitative. The question is how to make a quantitative measurement of SEM resolution?
Earlier this month I requested information to help network an EDS system, which is a UNIX based system and operates on a Sun Sparc Station 5 with SunOS 5.3. I really appreciate all the info people have sent me and I wanted to let you know that the system has been placed on the network (TCPIP). The local IT people suggested I use an outside expert, so we did arrange for an ex-Sun employee to come in and do the job. It did take, as one suggested, 15 minutes to place the Sun on the network, but it did take another hour to (re)write some missing files (not his exact words), which allows someone on the pc network (me) to be able to ftp or take files from the Sun and bring to the pc environment. It was worth the work as we don't have to scan in the data for our reports etc. So the process does work and is pretty simple, but later I may explore some of the Windows programs suggested to facilitate these file transfers in a drag and drop style.
Thanks again,
Dave Audette david.audette-at-sylvania.com
__________________________________________________ Do You Yahoo!? Yahoo! Shopping - Thousands of Stores. Millions of Products. http://shopping.yahoo.com/
suggests to me that she is with the University of British Columbia, Canada and not part of Quartz/Hitachi. Are you having trouble with her or Quartz/Hitachi?
gary g.
At 10:58 AM 11/22/00, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Gary I agree with you that an objective test is hard to come by and that an industry-standard approach is lacking. I make the following observations, for what they're worth:
There is no straightforward way of making a specimen that tests the resolution of all SEMs, under all operating conditions and for all specimen types. Gold on carbon is the best choice if that is what you look at for a living. If you look at fine carbon structures on carbon it's a poor test of practical resolution, although it may indicate the general health of the instrument.
Makers differ in their approaches to resolution testing. Most employ gold on carbon, but sometimes gold on magnetic tape or dendritic crystalline gold or silver is preferred. Some measure the smallest particles visible, others measure the smallest gaps between larger particles. One practical difficulty in the former approach is that the particles need to be most abundant in a size range close to the dimensions you want to test for. SEM resolutions span at least an order of magnitude from {0.5nm for in-lens fegsem to } 5nm for tungsten sem, making production of a single universal specimen difficult. At the limits of FEGSEM resolution, ~1nm, suitably small gold particles on carbon or mag tape can be as rare as hen's teeth. Another issue is that at the limit of an instrument's resolution the image of a small particle has a bell-shaped gaussian distribution of signal, fading imperceptibly into the background. The problem here is to determine where the edges of the feature are located. Pixel size is relevant here. If the feature is only two or three pixels wide, typical of a x100k image at 1kx1k pixels, then pixel size has a dominant effect on the measured size increments - ten pixels wide or more begins to yield an approximation to continuous data, so the image size must be large or the magnification high. The standard test of resolution in microanalysis, where peaks are again approximately normal curves, is to measure peak width at half height, and one can approximate this approach in measuring a gold particle by expanding the image contrast until local background is 0 and max signal is 255, setting the threshold to 127 and measuring mean feret diameter of the feature. I think you'll find that if you apply this type of criterion you will get a more conservative estimate of resolution than is claimed by the manufacturer.
A variant of gold on carbon which we have tried is to deposit 1 nanometre gold colloid onto formvar/carbon, with 10nm gold to provide something to focus on. This is a specimen that can be characterised by TEM before examination in the SEM, so you can determine what the actual particle size distribution is (difficult to do that with gold on mag tape). FEGSEMs can "see" the 1nm gold particles because there is strong signal against low background, but as in light microscopy, where 25nm colloidal gold is readily seen under reflected illumination, may image them to a larger diameter. This larger diameter, measured using peak width at half height, thus constitutes an estimate of instrument resolving power. Another approach in principle (harder in practice) is to create perfectly square, perfectly nanometre-sharp square edges in a perfectly smooth surface of the element of your choice - silicon e.g., or a layer of another element deposited thereon - and to measure the peak width at half height of the edge transition.
Finally, theoretically it is possiible objectively to determine the limiting dimensions recorded in any image by looking at its fourier power spectrum. Biomolecular structure people regularly report such data for TEM images, so presumably the technology is availalble. Any observations on where, how, and the limitations of applying it to sem resolution measurment would be gratefully received.
Best wishes Chris
} NIST used to offer a SEM calibration standard SRM-2069. } I don't see this any longer. If one needs to calibrate or } qualify a SEM's resolution at 250A or better, what is } an appropriate method? } } I've used Au on C "standards" but these are more } qualitative than quantitative. The question is how to } make a quantitative measurement of SEM resolution? } } Vendor responses are welcome. } } Thanks, } gary g. } }
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 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
Gary I agree with you that an objective test is hard to come by and that an industry-standard approach is lacking. I make the following observations, for what they're worth:
There is no straightforward way of making a specimen that tests the resolution of all SEMs, under all operating conditions and for all specimen types. Gold on carbon is the best choice if that is what you look at for a living. If you look at fine carbon structures on carbon it's a poor test of practical resolution, although it may indicate the general health of the instrument.
Makers differ in their approaches to resolution testing. Most employ gold on carbon, but sometimes gold on magnetic tape or dendritic crystalline gold or silver is preferred. Some measure the smallest particles visible, others measure the smallest gaps between larger particles. One practical difficulty in the former approach is that the particles need to be most abundant in a size range close to the dimensions you want to test for. SEM resolutions span at least an order of magnitude from {0.5nm for in-lens fegsem to } 5nm for tungsten sem, making production of a single universal specimen difficult. At the limits of FEGSEM resolution, ~1nm, suitably small gold particles on carbon or mag tape can be as rare as hen's teeth. Another issue is that at the limit of an instrument's resolution the image of a small particle has a bell-shaped gaussian distribution of signal, fading imperceptibly into the background. The problem here is to determine where the edges of the feature are located. Pixel size is relevant here. If the feature is only two or three pixels wide, typical of a x100k image at 1kx1k pixels, then pixel size has a dominant effect on the measured size increments - ten pixels wide or more begins to yield an approximation to continuous data, so the image size must be large or the magnification high. The standard test of resolution in microanalysis, where peaks are again approximately normal curves, is to measure peak width at half height, and one can approximate this approach in measuring a gold particle by expanding the image contrast until local background is 0 and max signal is 255, setting the threshold to 127 and measuring mean feret diameter of the feature. I think you'll find that if you apply this type of criterion you will get a more conservative estimate of resolution than is claimed by the manufacturer.
A variant of gold on carbon which we have tried is to deposit 1 nanometre gold colloid onto formvar/carbon, with 10nm gold to provide something to focus on. This is a specimen that can be characterised by TEM before examination in the SEM, so you can determine what the actual particle size distribution is (difficult to do that with gold on mag tape). FEGSEMs can "see" the 1nm gold particles because there is strong signal against low background, but as in light microscopy, where 25nm colloidal gold is readily seen under reflected illumination, may image them to a larger diameter. This larger diameter, measured using peak width at half height, thus constitutes an estimate of instrument resolving power. Another approach in principle (harder in practice) is to create perfectly square, perfectly nanometre-sharp square edges in a perfectly smooth surface of the element of your choice - silicon e.g., or a layer of another element deposited thereon - and to measure the peak width at half height of the edge transition.
Finally, theoretically it is possiible objectively to determine the limiting dimensions recorded in any image by looking at its fourier power spectrum. Biomolecular structure people regularly report such data for TEM images, so presumably the technology is availalble. Any observations on where, how, and the limitations of applying it to sem resolution measurment would be gratefully received.
Best wishes Chris
} NIST used to offer a SEM calibration standard SRM-2069. } I don't see this any longer. If one needs to calibrate or } qualify a SEM's resolution at 250A or better, what is } an appropriate method? } } I've used Au on C "standards" but these are more } qualitative than quantitative. The question is how to } make a quantitative measurement of SEM resolution? } } Vendor responses are welcome. } } Thanks, } gary g. } }
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 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
The knob on the front will not affect the image, just move across the field to expose a portion or all of either the right or left image. This is used in searching the surface areas of bullets or tool marks in forensic comparisons. You're doing it right so far. Have fun.
Phil Aviles Forensic Microanalyst
} -----Original Message----- } From: "COURYHOUSE-at-aol.com"-at-sparc5.microscopy.com } [SMTP:"COURYHOUSE-at-aol.com"-at-sparc5.microscopy.com] } Sent: Tuesday, November 21, 2000 11:43 PM } To: MICROSCOPY-at-sparc5.microscopy.com } Subject: Has anyone out there used an AO comparison microscope } before? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Ahoy fellow enthusiasts! } Has anyone out there used a comparison microscope before? } I got a bridge unit and dropped it on top of 2 AO 10 scopes, there seems } to } me a knob on the front that does nothing. I get one scope in once side of } the field and the image from the other scope on the other side..... } } I suspect the knob makes them overlap (like we have seen on the police } shows but it does not seem to move the image. } when they compare bullets...) } } any input? } } thanks Ed Sharpe archivist for SMECC } } }
Thank you for the clarification, but im still confused, your business card states that you are the EDX Product Development person. then would you be able to answer my questions? Why is it I cant get an answer about imaging/elemental mapping. ( See attached email)
Larry Howard
In a message dated 11/22/00 11:04:36 PM Eastern Standard Time, mmager-at-qrtz.com writes:
{ { Dear Larry,
I'm sorry if our technical support team did not respond to your request for information on our mapping option. The passive, full position-tagged mapping option is just now finished and the team is working very hard to get the information out to all the customers that requested it. I will be happy to phone you and answer any questions that you may have, if you will send me your phone number by return e-mail.
In answer to the subject of your e-mail, I work as a microscopist for the University of British Columbia and conduct any correspondance with the MSA Listserver in that capacity. I am also an EDX technical consultant for the X-ray Division of Quartz Imaging Corp. I have little to do with the Quartz PCI product except as a user and friend of the owner of Quartz.At 01:58 PM 11/22/00 -0500, you wrote: } Mary, } } I have been waiting for a response from your technical support concerning } mapping . Can you have them call me. } } Thank you } Larry Howard
Best regards, Mary
Mary Mager EDX Product Specialist Quartz Imaging Corp. 810 - 1112 West Pender Street Vancouver, B.C. Canada V6E 3X5 tel: 604-488-3911 fax: 604-488-3922 e-mail: mmager-at-qrtz.com --------------------
Dear Larry,
I'm sorry if our technical support team did not respond to your request for information on our mapping option. The passive, full position-tagged mapping option is just now finished and the team is working very hard to get the information out to all the customers that requested it. I will be happy to phone you and answer any questions that you may have, if youwill send me your phone number by return e-mail.
In answer to the subject of your e-mail, I work as a microscopist for the University of British Columbia and conduct any correspondance with theMSA Listserver in that capacity. I am also an EDX technical consultant for the X-ray Division of Quartz Imaging Corp. I have little to do with the Quartz PCI product except as a userand friend of the owner of Quartz.At 01:58 PM 11/22/00 -0500, youwrote: } Mary, } } I have been waiting for a response from your technical supportconcerning } mapping . Can you have them call me. } } Thank you } Larry Howard
Best regards, Mary
Mary Mager EDX Product Specialist Quartz Imaging Corp. 810 - 1112 West Pender Street Vancouver, B.C. Canada V6E 3X5 tel: 604-488-3911 fax: 604-488-3922 e-mail: mmager-at-qrtz.com
----------------------- Headers -------------------------------- Return-Path: {mmager-at-qrtz.com} Received: from rly-yd04.mx.aol.com (rly-yd04.mail.aol.com [172.18.150.4]) by air-yd02.mail.aol.com (v77.14) with ESMTP; Wed, 22 Nov 2000 23:04:36 -0500 Received: from www.quartzimaging.com (www.quartzimaging.com [205.206.193.13]) by rly-yd04.mx.aol.com (v76_r1.19) with ESMTP; Wed, 22 Nov 2000 23:04:13 1900 Received: from CR1001314-A (unverified [24.113.41.19]) by www.quartzimaging.com (EMWAC SMTPRS 0.83) with SMTP id {B0000039292-at-www.quartzimaging.com} ; Wed, 22 Nov 2000 20:12:15 -0800 Message-Id: {4.1.20001122195359.00a11a30-at-www.quartzimaging.com} X-Sender: mmager-at-www.quartzimaging.com X-Mailer: QUALCOMM Windows Eudora Pro Version 4.1 Date: Wed, 22 Nov 2000 20:03:28 -0800 To: Lhconsulting-at-aol.com From: Mary Mager {mmager-at-qrtz.com} Subject: Re: Does Mary Mager work for Quartz PCI or University of British Columbia In-Reply-To: {18.5496f0e.274d715b-at-aol.com} Mime-Version: 1.0 Content-Type: multipart/alternative; boundary="=====================_1931615516==_.ALT" X-Antirelay: Good relay from local net4 24.113.41.19/32
the way people with TEMs measure the resolution is pretty straight forward: They take an image and do a Fourier transform (either through software FFT or with an optical diffractometer). The FFT then shows essentially the frequencies that are on the image. For a TEM you can also calculate the Contrast Transfer Function (CTF), which essentially gives you the same information. By comparing the two, you can thus find out, what the resolution of the microscope is. Since the CTF is a fairly complex function, there are different "resolutions": Point resolution (corresponds to the frequency where the CTF becomes zero the first time), or the "lattice resolution" (where the intensity of the oscillations goes below a certain threshold).
For an SEM, I assume there is only a gradually declining FFT (Intensity vs. spatial frequency). Unless someone "defines" where the cut-off is (for example where the intensity has gone down to 20%), I don't think you can really determine the resolution with certainty. The sample also needs to be defined (for example a sharp metal edge, as any charging may affect the resolution. Also the material itself may have to be defined, as edge effects may influence the measurement. Finally, the sample must show structure below the resolution limit of the microscope, otherwise you measure the "resolution" of your sample. In TEMs that is fairly simple: use an amorphous material. For SEM samples I don't know what could be used. Something with atomic structure on the surface? A surface reconstructed semiconductor?
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: Thursday, November 23, 2000 2:24 AM To: Gary Gaugler Cc: microscopy-at-sparc5.microscopy.com
Gary I agree with you that an objective test is hard to come by and that an industry-standard approach is lacking. I make the following observations, for what they're worth:
There is no straightforward way of making a specimen that tests the resolution of all SEMs, under all operating conditions and for all specimen types. Gold on carbon is the best choice if that is what you look at for a living. If you look at fine carbon structures on carbon it's a poor test of practical resolution, although it may indicate the general health of the instrument.
Makers differ in their approaches to resolution testing. Most employ gold on carbon, but sometimes gold on magnetic tape or dendritic crystalline gold or silver is preferred. Some measure the smallest particles visible, others measure the smallest gaps between larger particles. One practical difficulty in the former approach is that the particles need to be most abundant in a size range close to the dimensions you want to test for. SEM resolutions span at least an order of magnitude from {0.5nm for in-lens fegsem to } 5nm for tungsten sem, making production of a single universal specimen difficult. At the limits of FEGSEM resolution, ~1nm, suitably small gold particles on carbon or mag tape can be as rare as hen's teeth. Another issue is that at the limit of an instrument's resolution the image of a small particle has a bell-shaped gaussian distribution of signal, fading imperceptibly into the background. The problem here is to determine where the edges of the feature are located. Pixel size is relevant here. If the feature is only two or three pixels wide, typical of a x100k image at 1kx1k pixels, then pixel size has a dominant effect on the measured size increments - ten pixels wide or more begins to yield an approximation to continuous data, so the image size must be large or the magnification high. The standard test of resolution in microanalysis, where peaks are again approximately normal curves, is to measure peak width at half height, and one can approximate this approach in measuring a gold particle by expanding the image contrast until local background is 0 and max signal is 255, setting the threshold to 127 and measuring mean feret diameter of the feature. I think you'll find that if you apply this type of criterion you will get a more conservative estimate of resolution than is claimed by the manufacturer.
A variant of gold on carbon which we have tried is to deposit 1 nanometre gold colloid onto formvar/carbon, with 10nm gold to provide something to focus on. This is a specimen that can be characterised by TEM before examination in the SEM, so you can determine what the actual particle size distribution is (difficult to do that with gold on mag tape). FEGSEMs can "see" the 1nm gold particles because there is strong signal against low background, but as in light microscopy, where 25nm colloidal gold is readily seen under reflected illumination, may image them to a larger diameter. This larger diameter, measured using peak width at half height, thus constitutes an estimate of instrument resolving power. Another approach in principle (harder in practice) is to create perfectly square, perfectly nanometre-sharp square edges in a perfectly smooth surface of the element of your choice - silicon e.g., or a layer of another element deposited thereon - and to measure the peak width at half height of the edge transition.
Finally, theoretically it is possiible objectively to determine the limiting dimensions recorded in any image by looking at its fourier power spectrum. Biomolecular structure people regularly report such data for TEM images, so presumably the technology is availalble. Any observations on where, how, and the limitations of applying it to sem resolution measurment would be gratefully received.
Best wishes Chris
} NIST used to offer a SEM calibration standard SRM-2069. } I don't see this any longer. If one needs to calibrate or } qualify a SEM's resolution at 250A or better, what is } an appropriate method? } } I've used Au on C "standards" but these are more } qualitative than quantitative. The question is how to } make a quantitative measurement of SEM resolution? } } Vendor responses are welcome. } } Thanks, } gary g. } }
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 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
I would like to ask you how it is possible to identify the inflammatory cells in paraffin embedded human liver specimens by means of immunohistochemistry technique. Is there a better antibody?
I don't see why this is being conducted on the list.
Can't you guys sort it out in private?
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
Thanks to James and all others that replied and helped shed some light on this matter
What it appears we have is bridge for an AO hair and fiber comparison scope. It seems to be fixed in the split mode, which is fine, as it will show a half for each scope. We have 2 model 10 AO scopes in the Museum's parts stash and I put enough stuff together to make 2 with the exception that I am missing the parts for both of the mechanical stages... Any one have some AO carcasses out there we can raid for parts?
thanks Ed Sharpe archivist for SMECC
{ { Subj: Re: Has anyone out there used an AO comparison microscope before? Date: 11/22/00 1:33:22 PM US Mountain Standard Time From: James.Roberts-at-mail.co.ventura.ca.us (James Roberts) To: COURYHOUSE-at-sparc5.microscopy.com, 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 -----------------------------------------------------------------------.
It depends on who's bridge you have. The two most common are the Leitz and the AO design. An AO type would say AO, Reichart or Leica on the manufactures logo depending on how new or old it is. Cambidge decided to use the Leica name on both once they owned both designs so the Leitz could also say Leica. There are other designs but these two make up probably more than 90% of all comparison bridges. I haven't worked with any of the others in over 20 years and then only a bit.
The knob in the center of the front makes it sound more likely to be an AO type. If it is, that knob should move the center prism set so that with it turned fully clock-wise you would see all of the left scope and turned fully counter-clock-wise you would see all of the right scope. With the knob centered you would see half of each field. In some other position you would see a smaller part of one field and a larger part of the other. If it is an AO style bridge it sounds to me as if the rack and gear might be damaged so the prism is not moving. The thin dividing line you see between the two fields should move as you turn the knob to give you a smaller percentage of one field and a larger percentage of the other. This lets us compare the pattern of stria over a length of the toolmark, a bullet comparison is a type of toolmark comparison. Side by side comparison is much more useful in Firearm and toolmark comparison than overlapping fields are.
If it is a Leitz design some do not let you move the dividing line off center but have other knobs that overlap the two fields or let you view just one. These bench top Leitz bridges don't have a knob in the center of the front so this probably isn't what you have.
In modern bridges the bridges that attach to separate scopes are designed for hair and fiber comparisons and the like. Bullet ("Universal Forensic") scopes have the objectives attached into the bridges in scopes that have been built since well before I entered the field over a 1/4 century ago. They are designed for much lower power and much longer working distances. The bridges for hair and fiber comparison scopes (yes they can be used for all manor of comparison but in forensic science labs that is their primary job and the way they are usually refereed to) are quite similar to bullet scope bridge designs but are designed to be attached to separate scopes that are basically conventional compound microscopes with sub-stage lighting.
If you tell me more about the bridge you are trying to use I may be able to tell you more that can help you set it up and see if it is working properly. AO then Reichart and now Leica have always said that users should not open up bridges to try repairs, as prism alignment is critical and easily disturbed. Rebuilds are available basically only at the factory, no one else has the alignment equipment to do it right. When other places have told me they could rebuild one I found that their version of a rebuild amounted to a real good cleaning job not prism replacement or realignment. If you do have an AO type and the knob is not moving the rack the center prism is on you may need such a rebuild. I did have a knob set screw come loose on an old one quite a number of years ago. I got away with going inside and tightening it up but I would not recommend it to anyone else as these are expensive pieces of equipment and rebuilds are several thousand dollars (around 12K to 15K la! st time I checked several years ago).
Jim
James L. Roberts Firearm and Toolmark Examiner Ventura Co. Sheriff's Lab (805) 654-2308 James.Roberts-at-mail.co.ventura.ca.us
} } } {"COURYHOUSE-at-aol.com"-at-sparc5.microscopy.com} 11/21/00 10:43PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html -----------------------------------------------------------------------.
Ahoy fellow enthusiasts! Has anyone out there used a comparison microscope before? I got a bridge unit and dropped it on top of 2 AO 10 scopes, there seems to me a knob on the front that does nothing. I get one scope in once side of the field and the image from the other scope on the other side.....
I suspect the knob makes them overlap (like we have seen on the police shows but it does not seem to move the image. when they compare bullets...)
Looking for some information on references (websites,books,journals, etc.) and/or SOPs/Procedures for the analysis of explosives using SEM/EDS. Thanks.
Steve Wintonick _____________________________________________________________________________________ Get more from the Web. FREE MSN Explorer download : http://explorer.msn.com
Scope Fullerenes, nanotubes and related nanomaterials are receiving great interest due to the new mechanical, physical and chemical properties and potential applications in various areas. A workshop is designed to bring together research scientists and engineers working in various disciplines in the broad area of nanotube and Fullerene -related materials and to provide an opportunity to exchange new ideas and results. Postgraduate students are particularly encouraged to present their thesis work.
Program The program consists of invited lectures from both Australian and international leading scientists as well as oral and poster contributions.
It will be divided into the following sessions (but not limited to):
Thermodynamics and Modeling Synthesis and Processing Characterization Properties and Applications
Partial list of invited speakers
Professor David Tomanek, Michigan State University, USA
Professor H. M. Cheng Institute of Metal Research, Chinese Academy of Science, China Title: Carbon nanotube synthesis and hydrogen storage
Professor M. Wilson, University of Technology, Sydney Title: Changing the diameter of carbon nanotubes
Professor G. Wallace, University of Wollongong Title: Carbon Nanotube/Conducting polymer Composites
Dr. L.M. Dai, Molecular Science, CSIRO Title:Controlled growth and surface modification of carbon nanotubes
Venue The workshop will be held in the seminar room of the link building (58) of
the Research School of Physical Science and Engineering, Mills Road, The Australian National University, Canberra, Australia
Abstract submission Please send your abstracts and registration form by email attachment to awnf2001-at-anu.edu.au Acceptance and presentation type (invited, oral or poster) will be informed by e-mail. Example of abstract format and registration forms can be found at: http://www.rsphysse.anu.edu.au/nanotube/awnf2001/
Correspondence Dr Ying Chen Department of Engineering, FEIT &Department of Electronic Materials Engineering Research School of Physical Science and Engineering Australian National University Canberra, ACT 0200 Australia E-mail: ying.chen-at-anu.edu.au, or awnf2001-at-anu.edu.au Telephone: 61 02 6249 0380, ( 6125 0380 after 31-12-2000) Fax: 61 02 6279 8338 ( 6125 6279 after 31-12-2000)
PhD scholarships on nanotube research are available at ANU and information
can be found at group webpage: http://www.rsphysse.anu.edu.au/nanotube/
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
Yes it is very possible to identify such cells. Do you want to identify all cells that might be involved in the inflammatory/immune response or subsets of these. In the first case you could use a broad spectrum antibody to the 'leukocyte common antigen/s' CD45 family or if you want to identify the subcategories eg macrophages, neutrophils, T/B lymphocytes etc then there is also a wide range available.
If you want further details of supply companies mail back to me and I can send you some web addresses - I have no connection to any companies - I am just a user.
Have you talked to your local hospital pathology department?
At 20:32 2000-11-23 +0100, Grizzi Fabio ICH wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Med vänliga hälsningar/With best regards
Gareth
e-mail Gareth.Morgan-at-impi.ki.se
http://www.ki.se/biomedlab/index_se.html
Tel +46 8 728 3734 Fax +46 8 728 3688
"Words are the seeds of misunderstanding - use them carefully."
"Give us the wisdom to know and not to feel that not knowing is less than wisdom itself."
Gareth Morgan MPhil MSc FIBMS, Institutionen för Mikrobiologi, Patologi och Immunologi(IMPI), H5, Karolinska Institutet, Avdelningar för biomedicinsk laboratorievetenskap och biomedicinska ämnen, Box 127 73, S 112 96, Stockholm Sverige
Mike What software do you recommend for this? Does AnalySIS have this capability? Chris
} From: Mike Bode {mb-at-Soft-Imaging.com} To: "'Microscopy-at-MSA.Microscopy.Com'" {Microscopy-at-sparc5.microscopy.com}
Dear All,
TNT (Trends in Nanotechnology) Weekly, a free, weekly e-mail round up of happenings in the world of nanotechnology will be launched on 5th December 2000. More information and a sample issue can be found at www.cientifica.com
If anyone on the list would like to contribute, or comment on the newsletter, or indeed help us to enhance is usefulness to the microscopy community, please contact me directly.
Regards
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/
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A general message to answer a number questions I have been receiving.
The eActive Mail dated Nov. 16th is SPAM/JUNK mail. Neither MSA nor the Microscopy Listserver has sold your address to anyone. I keep hold all addresses private and do not disclose them to anyone.
The mail you all received was just a junk mailing, which occassionally gets through the filters.
I have added eActive.Com to the SPAM filter so they should not be bothering us again.
Cheers... Nestor Your Friendly Neighborhood SysOp.
Hi Jonathan, I sent a preliminary message to your private e-mail address but got an 'undeliverable' note. Given a working address I can send you a copy of my own instruction sheet, photos of set up, etc. used weekly for 20 yrs with total success. Please contact me direct on: chris.smith-at-bbsrc.ac.uk EM Unit, IACR-Rothamsted, Harpenden, UK
Dear Sir/Madam , JEM3010 was manufactured in 1999.it has got 1.500.000x mag and digital camera system JSM5600 has got 300.000xmag and EDS system. we have one kind of chose for prices. If we study together (I mean we preapare a paper together), we reseach samples without any price. Thanks for your interests.. I am waiting for your emails
********************************** ********************************** ** 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 hate to seem unkind, but isn't once enough for this?
cheers
rtch
} Date: Mon, 27 Nov 2000 19:43:34 +0200 (EET) } From: Erdem Yasar {yasar-at-turkuaz.kku.edu.tr} } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: ABOUT JSM5600 SEM AND JEM3010 TEM
} -------------------------------------------------------------------- } ---- The Microscopy ListServer -- Sponsor: The Microscopy Society 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/Madam , } JEM3010 was manufactured in 1999.it has got 1.500.000x mag and } digital } camera system JSM5600 has got 300.000xmag and EDS system. } we have one kind of chose for prices. If we study together (I mean } we preapare a paper together), we reseach samples without any price. } Thanks for your interests.. } I am waiting for your emails } } ********************************** } ********************************** } ** 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 ** } ********************************** } ********************************** } } Home Page: http://turkuaz.kku.edu.tr/~yasar } } } } } 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, We have a Balzer's 301 Freeze fracture etch unit that worked for some time with no problems. However, one day the unit was overheating so I changed the oil in the rotary pump. It worked fine until recently. I found that one fuse had blown in the Pumping unit control TCS 100. Changing the fuse did not help, and I noticed the transformer in the pumping unit was discolored.
I was wondering if anyone has a spare transformer or TCS 100 Pumping unit control they are not using? If not, can anyone suggest why the transformer chose to die on us at this moment? Previously we kept the unit pumping constantly. Due to a vacuum leak, I shut it off until it was to be used for coating. Unfortunately, now it won't start up. 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
Can anyone answer this question . Remember Ask-A-Microscopist is a forum that is used by NON-MICROSCOPISTS looking for help so keep try to your technical details at the right level.
Email: jekstrom-at-mediaone.net Name: Jim Ekstrom School: Phillips Exeter State: NH
Question: I picked up an objective lens that is made in the US. It has Apergon (in italics) with 200X below it. The lens which is roughly the size of a normal 10X objective had a different thread than a normal objective. Can you tell me anything about it?
} we have one kind of chose for prices. If we study together (I mean we } preapare a paper together), we reseach samples without any price.
If you study alone - how much for a paper? (Joking ... sorry couldn't stop myself)
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 ---------------------------------------------------------------------
Claire Blue is best To a rough approximation the resolving power is limited to half the wavelength of the light used. Thus blue -at-480nm = 240 nm, green -at- 540nm = 270 nm Chris
Date sent: Tue, 28 Nov 2000 10:27:20 +0000 } From: "C.L.Healey" {MBA00CLH-at-sheffield.ac.uk} Organization: The University of Sheffield To: microscopy-at-sparc5.microscopy.com
If its a good microscope, then under oil immersion the blue would give better resolution because blue light has a shorter wavelengths than green, but . . . Some people may find blue harder on the eye and less subjectively, it depends on the colours of the specimen too. If highest resolution is the object then the specimen should be very thin, say 2 um or less and at that point contrast becomes an issue and its useful to know that the filter will render the complimentary colours within the specimen more darkly. So you could have fun and draw an Oswald colour circle (rainbow colours arranged in a circle with complimentary colours opposite) and figure out which filter would render which colours in the specimen darker or lighter. Thus specimen contrast could be enhanced and green will be more useful to optimize contrast with more specimens. Such fancy colour considerations apply to viewing and B+W photography, whereas in colour photography most people would prefer realistic colours and that, with a daylight type film requires the right blue filter and the transformer set to the bulb's voltage. There is also an argument to be made that all of that is too complicated and that a digital camera is more fun. 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 ABN: 99 724 136 560 www.proscitech.com
On Tuesday, November 28, 2000 8:27 PM, C.L.Healey [SMTP:MBA00CLH-at-sheffield.ac.uk] wrote: } } Hi, } } I just wondered whether it would be at all possible to ask you a } question, } } "Is the resolution of an optical microscope better with green or blue } light?". } } Thanks for your help, } } Claire } mba00clh-at-sheffield.ac.uk
Press Release for Fall 2000 Microscopy Society of America Undergraduate Research Scholarship Program
With this year's call for applications the MSA Undergraduate Research Scholarship Program begins its 13th year providing funding for undergraduate research. To date over 65 projects covering a wide range of topics in the physical and biological sciences have received support through this program. Over the years nearly all the scholarship recipients have maintained a strong interest in imaging sciences and have gone on to graduate school, professional school, teaching, or industry positions.
The program, which is funded by MSA, is able to support approximately 30% to 40% of applicants. The maximal award from MSA is currently $3000 and helps to provide student stipends, supply costs, and limited travel expenses associated with the research. Additional support in the form of instrument use time, equipment purchases, etc. is generally provided by the studentís supervisor and/or through the sponsoring institution. Abstracts reporting the research results, are prepared by scholarship awardees, and published in "Microscopy and Microanalysis".
The program actively seeks external sources of matching funds in order to maintain the favorable levels of support both in terms of the number of projects supported and the level of support for each. We are extremely grateful for the matching support provided by MSA sustaining members and individuals. Their support has enabled the program to increase both the number of awards and the maximum amount of each award to $3000.
The MSA Undergraduate Research Scholarship Program is currently soliciting applications from students interested in conducting a research project which involves the use of any microscopy technique. Students should be sponsored by a member of MSA. The maximal award is $3000. **The application deadline is Dec 31, 2000.** Applications can be obtained from the MSA Business Office, businessoffice-at-msa.microscopy.com or call toll free at (800) 538-3672. If you have any questions or require additional information regarding the program please contact either:
Dr. Ralph Albrecht, University of Wisconsin 1656 Linden Drive, Madison, WI 53706 (608) 263-3952 or 263-4162; (608) 262-7420 FAX; albrecht-at-ahabs.wisc.edu
Dr. Richard Ornberg, Monsanto Company Analytical Sciences Center, 800 N. Lindbergh Blvd., St. Louis, MO 63167 (314) 694-1184; (314) 694-6727 FAX; rlornb-at-ccmail.monsanto.com
A Job position for a Materials Scientists with a background in electron beam microcharacterization has opened today and has been posted on the US DoE BES web site:
An appointee will be required to provide verification of U. S. citizenship and/or employment eligibility under the Immigration Reform and Control Act of 1986 (Public Law 99-603). If selected, a male applicant born after December 31, 1959, must confirm his selective service registration status
Applications will be accepted through January 26, 2001.
Cheers... Nestor Your Friendly Neighborhood SysOp
P.S. Contact DoE at BES-at-science.doe.gov for additional information
I am looking for an adapter to mount an Olympus D340 digital camera to a standard stereo microscope eyepiece. Does anyone have a possible solution for my problem? Thanks. Barry Kromer kromerb-at-cellutissue.com
We have a JSM-845 SEM with a fairly old water recirculator. Because of the age, the water recirculator does not work properly and needs to be replaced. Could anybody help me locate a used chiller for the SEM? The chiller itself has to be air cooled. Thanks very much.
Chao-Ying Ni Rodel Inc. Materials Development Center 451 Bellevue Road Newark, DE 19713
Yes, you are right. The resolution of a microscope is proportional to the wave length of the light you use for illumination. The "green" or "blue" light has shorter wave length compared with other lights/colors, hence has higher resolving capability, e.g., point-to-point resolution. Also, "white light" is composed of all of "the lights" in the visible range of the light spectrum, and it may cause so-called chromatic aberration for microscopes.
-cy Rodel
-----Original Message----- } From: C.L.Healey [mailto:MBA00CLH-at-sheffield.ac.uk] Sent: Tuesday, November 28, 2000 5:27 AM To: microscopy-at-sparc5.microscopy.com
Hi,
I just wondered whether it would be at all possible to ask you a question,
"Is the resolution of an optical microscope better with green or blue light?".
1) Detector sensitivity. As a rule, the human eye is more sensitive to green than blue. Other detectors (cameras, film, etc) will have their own wavelength dependence.
2) Sample character. A living cell is more likely to be hurt/stimulated by blue than green light.
Just a few more tangential remarks on this thread.
I just wondered whether it would be at all possible to ask you a question,
"Is the resolution of an optical microscope better with green or blue light?".
Thanks for your help, Dear Claire, In theory, the best attainable resolution with blue light would be slightly better than that with green light; however, in practise, the lenses for a given microscope might be designed to have lower spherical aberration for green light (if, for example, the scope were designed to use all wavelengths of visible light, green would be a better compromise than blue), and the properties of the specimen would probably play a bigger role in attainable resolution than the wavelength of light used to observe it, for example, a specimen damaged by the shorter wavelength light would be better observed using the longer wavelength. Also, the medium used to observe or record the image might be capable of better resolution with green light. Now that I've probably told you more than you wanted to know, I can say that it is always possible to ask a question, as long as you are willing to put up with the answer. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
In response to a question, I would like to add this bit to the MSA undergraduate scholarship announcement:
This is not limited to students in the USA or North America. Any undergraduate student anywhere is eligible to apply for the award, provided the person supervising their research is a member of MSA.
Phil -- }}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{ Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
Hallo Folks, We're fiddling around with some rather Fe-rich amphiboles here. Analysis by WD on our trusty JEOL 733. Totals are coming out a bit high for an amphibole, and we've started discussing the problem a bit. One of us has had a bell rung with regards to ZAF correction on Fe-rich silicates. Anyone have any more ideas on this, and how we might do a bit of believable scientific fudging? 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: m.roberts-at-ru.ac.za SOUTH AFRICA
We in the micropaleontology business (or formerly in it) have for years been staining Foraminiferal and Ostracode shells with green food colouring to enhance their appearance in binocular microscopes. It seems to bring out the contrast a bit, and the dye tends to settle in, and accentuate, morphological features which aid in identification. Green seems to work a bit better than blue, but as Tobias points out, that may have more to do with the sensitivity of the human eye. A pretty cheap little trick, too - you wouldn't believe how many foram shells you can dye with one little bottle.....
Frank Thomas Geological Survey of Canada (Atlantic) Bedford Institute of Oceanography Dartmouth, Nova Scotia Canada
It should just dissolve in water or buffer if you soak the slides for a while. Then recoat with more dilute MeCell if you want.
On Tue, 28 Nov 2000, ken blight wrote:
} Date: Tue, 28 Nov 2000 09:18:45 +0000 } From: ken blight {blight-at-icrf.icnet.uk} } To: Microscopy-at-sparc5.microscopy.com } Subject: MethylCellulose } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 method for removing hardened methycellulose from } cryocections when the film is too thick? } } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
We have a Reichert-Jung Cryocut 1800 cryostat. The motor which moves the specimen in and out is stuck in the STOP, fully advanced position and will not retract. I know there is a manual way to get the specimen holder back to its Home position, but I have forgotten, and it is not in my manual. Can anyone help me with this?
Mahalo!
Dr. David T. Webb University of Hawaii at Manoa Dept. of Botany dave-at-hawaii.edu
You may email him directly, or I will forward any replies.
High 70s F, windy, intermittant clouds. Beautiful November light.
**************************************************************************** * 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* ****************************************************************************
To all, I just found out that Kodak has discontinued the manufacture of its transparent sleeves, CAT 150 3812. Does anyone know of a suitable replacement for this product and where it can be purchased, preferably in either Canada or the N. Eastern U.S.?
Thanks for your help. Paul
Paul J. Gerroir Microscopy Materials Characterization Xerox Research Centre of Canada 2660 Speakman Drive Mississauga, Ontario L5K 2L1
Blue, but to such a small extent that it really can't make much difference. The shorter the wavelength, the higher the resolution. However, for some black and white films used for photomicrography, such as Technical Pan, a green filter gives better results. Hope this helps.
Lesley Weston.
On Tue, 28 Nov 2000, C.L.Healey 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 just wondered whether it would be at all possible to ask you a } question, } } "Is the resolution of an optical microscope better with green or blue } light?". } } Thanks for your help, } } Claire } mba00clh-at-sheffield.ac.uk } }
Hello Jonathan, Tried to send message but got undeliverable message. If you want to contact me direct I could give you some tips if that is any use. I have used one very regularly for carbon coating for some years, with a good degree of success. cheers. Richard Black
To: ListServer Re: Please post the message below. Thank you.
Would like to purchase (now unobtainable) Leitz Wetzlar long working distance objective originally manufactured for a Universal Stage. 1or 2 off. Manufacturer Part number : 559123 Barrell details: 160/- L25/0.22 (P) (UT40/0.34).
Any information on possible source would be welcome.
-- Dr Andrew P. Jephcoat +44-(0)1865-272067 Department of Earth Sciences Parks Road +44-(0)1865-272000 (Dept.) Oxford OX1 3PR +44-(0)1865-272072 (FAX) UK andrew-at- earth.ox.ac.uk http://www.earth.ox.ac.uk/Research/interior.htm
Chuck Berger wrote: } Take into account that the eye is most sensitive in the green } } region & what color is your objective corrected for?
Yes, but sensitivity is a detector property, not a property of the optics.
Correct me if I am wrong, but the corrections made to an apochromat are primarily to bring different wavelengths to the same plane of focus, not primarily to optimise for resolution at a particular wavelength
Lesley Weston wrote: } Blue, but to such a small extent that it really can't make much } difference. The shorter the wavelength, the higher the } resolution. However, for some black and white films used for } photomicrography, such as Technical Pan, a green filter gives } better results. Hope this helps.
Again this is a detector-sensitivity issue which does not alter the fundamental fact that diffraction-limited resolving power of light optics is higher in blue light than in green.
One of the reasons why users of Panchromatic films prefer to work with a green filter is that it provides more natural-looking contrast and grey-scale values, for the very reason that Chuck Berger is referring to - that the eye has better brightness and contrast sensitivity in green. Technical Pan has red sensitivity extended into the near infrared, and contributions to the image from these wavelengths may certainly degrade resolution of the recorded image a) because they may not focus at the same plane as green light, and b) because these longer wavelengths cannot form diffraction limited image details as small as the shorter blue and green wavelengths. The relationship between wavelength and resolution becomes important here - because the near infra red (~900-1000nm) has twice the wavelength of blue/blue-green (450- 500nm) the diffraction limited resolution is poorer by a factor of two.
Incidentally, such considerations are also important when using monochrome CCD cameras. Many of these (such as Pulnix TM6, for example) have spectral sensitivity extended into the infrared (considerably further than Technical Pan). IR forms a large fraction of the illumination intensity of a light microscope unless an IR filter is fitted. The IR component degrades both resolution and contrast, and the problem is in this case not adequately resolved by using a green filter alone, since most transmit IR. Use of a heat-absorbing glass filter, or even a petri dish with a layer of dilute copper sulphate solution, will improve performance in such systems quite dramatically.
(PS this will not help improve the performance of your colour ccd camera, so don't ask :-)
Best wishes Chris ========================================== Dr. Chris Jeffree University of Edinburgh BIOSEM - Biological Sciences Electron Microscope Facility Institute of Cell and Molecular Biology Daniel Rutherford Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JH, Scotland, UK Tel. #44 (0) 131 650 5345 FAX. #44 (0) 131 650 6563 Mobile 07710 585 401 email c.jeffree-at-ed.ac.uk =========================================
Research Assistant Samuel Roberts Noble Foundation A research assistant position is available to join the microscopy support group. The research assistant will be assigned to work on various projects using microscopy techniques. This will involve immunocytochemistry (light and electron), in situ hybridization and confocal microscopy. A B.S. degree in Biology and previous experience in light and electron microscopy of plant material is required. Organizational skills and the ability to work collaboratively with various research groups are essential. Salary is commensurate with experience ($28,000 to $39,200.00). Application and job description obtainable from the Noble Foundation website, www.noble.org {../index.html} . To apply, send a letter outlining research interests and experience, CV and names of 3 references to:
Jane Nance, Human Resources Department RE: Position #PltBio30700-EB147 Samuel Roberts Noble Foundation PO Box 2180 Ardmore, OK 73402, USA.
You're not wrong, but there is a subtle point here: the optics are designed around green light, typically 550nm, and then corrected for blue and red (if apochromatic). This means the the optics work optimally in green light, and therefore would resolve green light best. Theoretically, shorter, blue light has higher resolution, but the theory isn't taken advantage of unless the optics are designed for blue light. Note that the filter in phase contrast microscopes is typically green, because the phase annulus in the objectives is designed for 550nm, because the (uncorrected) optics are designed for 550nm. +/- of course.
Phil
} Chuck Berger wrote: } } Take into account that the eye is most sensitive in the green } } } region & what color is your objective corrected for? } } Yes, but sensitivity is a detector property, not a property of the } optics. } } Correct me if I am wrong, but the corrections made to an } apochromat are primarily to bring different wavelengths to the same } plane of focus, not primarily to optimise for resolution at a particular } wavelength -- }}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{ Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
Robert No need to apologize! On the contrary, many thanks for your most informative comments. You clearly know rather more about it than I do! Best wishes Chris
Date sent: Wed, 29 Nov 2000 09:25:16 -0500 (EST) } From: Robert Wieland {wieland-at-ME.UDel.Edu} To: c.jeffree-at-ed.ac.uk
Paul,
I don't know the size or cost of those Kodak sleeves, but I have used Print File archival sleeves from Get Smart Products in Manhasset, NY. According to my 1999 catalog:
Phone: (800) 827-0673 Fax: (516) 365-0673
Hope this helps! Matt
Matthew J. Lynn, Ph.D. Center for Advanced Microscopy University of Miami (305)284-4736 mlynn-at-miami.edu
On Tuesday, November 28, 2000 3:51 PM, Gerroir, Paul J [SMTP:Paul.Gerroir-at-crt.xerox.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 } -----------------------------------------------------------------------. } } } November 28, 2000 } } To all, } I just found out that Kodak has discontinued the manufacture of its } transparent sleeves, CAT 150 3812. Does anyone know of a suitable } replacement for this product and where it can be purchased, preferably in } either Canada or the N. Eastern U.S.? } } Thanks for your help. } Paul } } Paul J. Gerroir } Microscopy } Materials Characterization } Xerox Research Centre of Canada } 2660 Speakman Drive } Mississauga, Ontario L5K 2L1 } } Phone: (905) 823-7091, ext. 216 } FAX: (905) 822-7022 } email: paul.gerroir-at-crt.xerox.com
Do you have any advice regarding environmental controls for a small room to be used for ultrathin sectioning?
I have been asked for recommendations regarding things like air filters, temperature control, and humidity.
It will be a small room, maybe 6' x 9', in a modern university science building. The researcher is going to put a new ultramicrotome in there and will want to do serial sectioning on conventional resin blocks. Cryo may come later, so if there is anything special needed for that, now is the time to add it to the list.
I have always been OK with standard sort of rooms, regular building air filters etc, if there is anything special I have overlooked, please let me know so I can pass it along.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
I put in a call to the photographic equipment supplier in the Chicago area through whom I've ordered the sleeves in the past. He contacted Kodak, and was told that Kodak is no longer carrying this item (apparently it's not actually manufactured by Kodak?), but that the sleeves are available from Tiffen, a maker of filters and other photographic equipment. I was told that the next time I place an order, I just need to let the purchasing agent know that the supplier is Tiffen, rather than Kodak. I haven't yet been able to get contact information for Tiffen (they have a website, but I'm blocked from accessing it at work), or to verify this by placing an order, but I thought I'd pass along what I've learned so far. If I find that this is a dead end, I'll let you know.
Elaine Schumacher UOP Des Plaines, IL 847-391-3403 efschuma-at-uop.com
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I put in a call to the photographic equipment supplier in the Chicago area through whom I've ordered the sleeves in the past. He contacted Kodak, and was told that Kodak is no longer carrying this item (apparently it's not actually manufactured by Kodak?), but that the sleeves are available from Tiffen, a maker of filters and other photographic equipment. I was told that the next time I place an order, I just need to let the purchasing agent know that the supplier is Tiffen, rather than Kodak. I haven't yet been able to get contact information for Tiffen (they have a website, but I'm blocked from accessing it at work), or to verify this by placing an order, but I thought I'd pass along what I've learned so far. If I find that this is a dead end, I'll let you know.
Elaine Schumacher UOP Des Plaines, IL 847-391-3403 efschuma-at-uop.com
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I put in a call to the photographic equipment supplier in the Chicago area through whom I've ordered the sleeves in the past. He contacted Kodak, and was told that Kodak is no longer carrying this item (apparently it's not actually manufactured by Kodak?), but that the sleeves are available from Tiffen, a maker of filters and other photographic equipment. I was told that the next time I place an order, I just need to let the purchasing agent know that the supplier is Tiffen, rather than Kodak. I haven't yet been able to get contact information for Tiffen (they have a website, but I'm blocked from accessing it at work), or to verify this by placing an order, but I thought I'd pass along what I've learned so far. If I find that this is a dead end, I'll let you know.
Elaine Schumacher UOP Des Plaines, IL 847-391-3403 efschuma-at-uop.com
} Hi everyone: } I would like the society's views on the Spot RT-SLider digital camera. How } long does it take to capture an image (fluorescence) and any hardware / } software bugs experienced by users? } } S. Srinidhi, } Materials Scientist } gJohn F. Welch Technology Center } _______________________________________ } Inorganic Materials Laboratory } GEITC, Upper ground floor. Unit IV, } Innovator building, International Tech Park, } Whitefield Road, Bangalore - 560066. } Phone: 8410702 / 03 / 08 / 09 / 841 1580 x: 1176 } Fax:8410704. } e-mail: srinidhi.sampath-at-geind.ge.com } }
************************************************ Plan Now to Participate FOCUS ON MICROSCOPY 2001 ************************************************
Focus on Microscopy 2001 (FOM 20001) is a world leading international conference on advanced microscopy. It is the joint meeting of the 13th International Conference on Confocal Microscopy and the 13th International Conference on 3D Image Processing in Microscopy.
FOM 2001 will be held on April 1-4, 2001 at the Academic Medical Centre (AMC) of the University of Amsterdam, Amsterdam, The Netherlands.
For further information: http://www.focusonmicroscopy.org/ ----------------------------------------------------------
Important Dates: ---------------- Camera-ready Abstract Due Date: February 1, 2001 Early Registration Due Date: February 15, 2001 Hotel Reservation Due Date: February 15, 2001
Location: --------- The Academic Medical Centre (AMC) of the University of Amsterdam is situated only 15 minutes from the Amsterdam city centre where you will find all sorts of restaurants, museums, concert halls historical buildings and nice pubs. The organisation provides you with a public transport card, so you can easily find your way from your hotel, through the city centre, to the conference.
Hotel Accommodation: -------------------- Hotels can be reserved through the RAI Hotel Service. See hotelform on the website (or ask at the Conference Office). The Conference Office does not make reservations. You can also make you own (cheaper) reservation through on-line hotel reservation sites such as http://www.medialink.nl/amsterdam .
Conference Topics: ------------------ Theory, Instrumentation, 4D-Imaging of living cells, Multi-photon excitation, Recovery after photobleaching (FRAP, FLIP), Fluorescence energy transfer (FRET), Non-linear optics (CARS, SHG, THG), Time-resolved imaging (FLIM), Green fluorescent protein (GFP), 4D- and 3D-Image processing, reconstruction and analysis, Virtual Reality (VR), X-Ray microscopy, Near field microscopy, Industrial applications in lithography and data storage, and many other techniques and applications.
This year, there will be special attention for the issue: "Microscopy of living cells and tissue"
International Organizing Commitee: ---------------------------------- G.J. Brakenhoff, University of Amsterdam, The Netherlands; A.C. Boccara, ESPCI, France; P.C. Cheng, SUNY at Buffalo, USA; C. Cogswell, University of Sydney, Australia; J. Dobrucki, Jagiellonian University, Poland; R. Van Driel, University of Amsterdam, The Netherlands; M. Gu, Swinburne University of Technology, Australia; K.S. Ha, Korea Basic Science Institute, Korea; S.W. Hell, MPI for Biophysical Chemistry, Germany; B. Hermann, University of Texas HSC at San Antonio, USA; F.J. Kao, National Sun Yat-Sen University, Taiwan; S. Kawata, Osaka University, Japan; A. Kriette, University of Giessen, Germany; E.M.M. Manders, University of Amsterdam, The Netherlands; M. Müller, University of Amsterdam, The Netherlands; C. Saloma, University of Philippines, Philippines; C.J.R. Sheppard, University of Sydney, Australia; E.H.K. Stelzer, EMBL, Germany; T. Wilson, University of Oxford, UK.
if cryo will be used later, it might be worth considering whether there will be enough ventilation to prevent a build up of nitrogen gas and subsequent oxygen depletion. If nothing else is done then an oxygen depletion monitor should be top of the list before liquid nitrogen is used or stored in such a small room.
The sums for calculating oxygen loss when liquid nitrogen is evaporated are fairly straight forward and I would always assume the worst scenario.
In the last two years there have been two fatal incidents, that I know of, which have involved oxygen depletion so even if the use of liquid nitrogen is in the future and a remote possibility it's worth considering now.
On a more trivial note it might be worth specifying floor and benches which don't crack with constant liquid nitrogen spillages.
I hope this helps.
Malcolm Haswell e.m. unit School of Sciences University of Sunderland UK
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: } } Do you have any advice regarding environmental controls for a small room to } be used for ultrathin sectioning? } } I have been asked for recommendations regarding things like air filters, } temperature control, and humidity. } } It will be a small room, maybe 6' x 9', in a modern university science } building. The researcher is going to put a new ultramicrotome in there and } will want to do serial sectioning on conventional resin blocks. Cryo may } come later, so if there is anything special needed for that, now is the } time to add it to the list. } } I have always been OK with standard sort of rooms, regular building air } filters etc, if there is anything special I have overlooked, please let me } know so I can pass it along. } } Thanks } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu
Jon: As you have said, the standard conditions and filters are really all that are necessary. The only thing I have really had a problem with are air currents. I generally ended up setting up a cublicle type area just barely large enough for the microtome and me, with a curtain (the "plastic strip" type used in cold rooms works)to minimize air currents. One other thing that needs to be monitored/controlled is humidity (more than temperature, even). Too dry or too humid, and forget sectioning! Of course, I was doing massive serial sectioning, so controlling the environment became an obsession. Hope this helps.
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals
On Wed, 29 Nov 2000 11:57:22 -0800, 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: } } Do you have any advice regarding environmental controls for a small room to } be used for ultrathin sectioning? } } I have been asked for recommendations regarding things like air filters, } temperature control, and humidity. } } It will be a small room, maybe 6' x 9', in a modern university science } building. The researcher is going to put a new ultramicrotome in there and } will want to do serial sectioning on conventional resin blocks. Cryo may } come later, so if there is anything special needed for that, now is the } time to add it to the list. } } I have always been OK with standard sort of rooms, regular building air } filters etc, if there is anything special I have overlooked, please let me } know so I can pass it along. } } Thanks } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } } }
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Jonathan, Assuming the building is completed, the simplist modification to a "typical" room is to install a drip ventilation ceiling rather than one with a normal type of air vent. The drip ceilings are made up of ceiling tiles that have many small slits throughout which allow air to filter down into the room. This type of ventilation virtually eliminates drafts which, aside from vibration, can be a major problem in microtomy. If you are lucky enough to be designing a room in a building under construction, the ideal situation for microtome and microscope rooms is a "floating " foundation. This is when the room floor (obviously in the lowest level) is on concrete supports that are isolated from the main building foundation. That way vibrations from the building as a whole are not transfered directly to the floor of the room in question. Going along with this is the use of flexible morter where internal walls join outside walls.
Debby
Debby Sherman Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu 1057 Whistler Building West Lafayette, IN 47907-1057 --------------------------------------
Greetings:
Do you have any advice regarding environmental controls for a small room to be used for ultrathin sectioning?
I have been asked for recommendations regarding things like air filters, temperature control, and humidity.
It will be a small room, maybe 6' x 9', in a modern university science building. The researcher is going to put a new ultramicrotome in there and will want to do serial sectioning on conventional resin blocks. Cryo may come later, so if there is anything special needed for that, now is the time to add it to the list.
I have always been OK with standard sort of rooms, regular building air filters etc, if there is anything special I have overlooked, please let me know so I can pass it along.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
I arrived in post to find that a brand new Ultracut S had been installed in a converted cleaners cupboard measuring (4' x 10') located several metres from the building's main plant room. Despite standing on an anti-vibration table the resonance through the walls from the plant room mean that I have to time my sectioning work so that it doesn't coincide with the machinery next door. In addition, there was a cool air ventilation shaft directly above the Ultracut that made the water in the knife boat resemble something out of 'A Perfect Storm'!!!. ( I subsequently had that blocked off). One other problem that you may encounter in such a small room is the build up of static electricity due to the dry air. I find that it helps the fill the sink with water when cutting. I also suggest that you consider very carefully if it is wise to work with cryo in such a small area as, I believe that this could be a major hazard.
These are my personal experiences and I do not speak on behalf of my organisation, but I hope that they are of some use to you in this matter.
First of all, I would like to thank people who responded to my previous post regarding problems on cultured hippocampal EM. Special thanks to JoAnn Buchanan, Dorothy Sorenson and Michael Plociniak and others who provided valuable suggestion that lead to the fantastic results.
Here are the major modification on my previous protocol:
1. Make sure the culture is healthy. 2. Shorten fixation and dehydration time. 3. Prolong the uranyl acetate staining to 30-40 min. 4. Section at 50-60 nm thickness (setting). 5. Observe cells at 60 kV rather than a higher accelerating voltage.
Hope this helps with people who have the similar problems as I had before.
Xinran
******************************************** Xinran Liu, M.D., Ph.D. Center for Basic Neuroscience The University of Texas Southwestern Medical Center at Dallas 6000 Harry Hines Blvd., NA4.214A Dallas, TX 75390-9111 Phone: (214) 648-1830 Fax: (214) 648-1801 E-mail: xinran.liu-at-utsouthwestern.edu
My name is Giselle. I am a graduate student working on my thesis. My thesis topic is "Methods of preparation of petrographic thin sections for Electron Microscopy". I have a collection of thin sections (chert) and I am preparing the sections for analysis by a TEM 1200. The thin sections are initially prepared by methods Ultramicrotomy and Ion Milling. TEM parameters used will be electron diffraction and elemental composition to obtain identification of the mineral and microstructures. I need more information (references) on recent literature (1998-2000) on my subject or similiar work done using these methods for analyis by TEM.
I need your help!!!
Thank you for your time.
_______________________________________________________ Tired of slow Internet? Get -at-Home Broadband Internet http://www.home.com/xinbox/signup.html
Could anyone provide me with a phone number, e-mail or fax number for the company "RAITH USA" It is a Germany Company and I thought there was a distrbutor in the US
Thanks
D. Sicard --
LADD RESEARCH 131 Dorset Lane Williston, VT 05495 USA
web site http://www.laddresearch.com
TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail sales-at-laddresearch.com
My boss wants to sell one of our projection microscope systems to someone she knows. I have been given the job of suggesting a price and would appreciate suggestions from anyone on the listserver. To prevent clutter on the listserver, please respond to my e-mail address: Louise_Harner-at-albint.com
The projection microscope system has Bausch & Lomb optics. It is basically an inverted monocular microscope and projects an image onto the tabletop - magnification of 500X at the table. The top of the lamp house is about 42 inches above the table. The light goes through a condenser, then through the slide, into a 20X objective, and out the monocular eyepiece. The normal focusing and x-y stage controls are supplemented by hand control extensions (x, y, and z) and by foot control extensions (x and y). There is a catalog number on the lamp housing: B&L cat. # 31-32-61, but I'm not sure if that is for the lamp only or the whole system. The system was used primarily to measure fiber diameters.
Thanks in advance for your help!
Louise Harner Albany International Research Co. Mansfield, MA
I work for a public school system in Houston, TX. We acquired a Philips TEM as a donation. It is now installed. We are not set up to prepare specimens and won't be (if ever) at least until next year. } } Am in serach of prepared grids you may not need anymore for our high } school students to look at. Descriptions of grid would be beneficial, } i.e. what specimen is and what might be occurring. } } Seeking specifically: } plant tissue (leaves, stems, & roots) } animal cells } prokaryotic cells } samples from various kingdoms } dividing cells } human tissue } } Please advise if you can help! THANKS! } } Denise Martin } Cy-Fair ISD - Science Resource Center } 11206 Telge Rd. } Cypress, TX 77429 } 281-897-4695 }
In my experience, in most climatic zones a room for ultramicrotomy should be air-conditioned. This frequently creates drafts and these must be eliminated. The frequently used louvered airduct ceiling outlets are unsuitable and if they are blocked this would unbalances a complex AC systems. The cheapest, highly effective way to make such an outlet draft-free is to mount a 600mm sheet metal inverted "Chinese hat" below the outlet so that a 150mm gap is maintained. This hat will break the draft and direct the airflow along the ceiling and then down the walls. Effectively the outlet area is greatly increased and the air speed slowed and re-directed. 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 ABN: 99 724 136 560 www.proscitech.com
On Thursday, November 30, 2000 10:42 PM, Roger Moretz [SMTP:rcmoretz-at-excite. 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 } -----------------------------------------------------------------------. } } } Jon: } As you have said, the standard conditions and filters are really all that } are necessary. The only thing I have really had a problem with are air } currents. I generally ended up setting up a cublicle type area just barely } large enough for the microtome and me, with a curtain (the "plastic strip" } type used in cold rooms works)to minimize air currents. One other thing } that needs to be monitored/controlled is humidity (more than temperature, } even). Too dry or too humid, and forget sectioning! Of course, I was doing } massive serial sectioning, so controlling the environment became an } obsession. Hope this helps. } } Roger Moretz, Ph.D. } Dept of Toxicology } Boehringer Ingelheim Pharmaceuticals } } On Wed, 29 Nov 2000 11:57:22 -0800, 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: } } } } Do you have any advice regarding environmental controls for a small room } to } } be used for ultrathin sectioning? } } } } I have been asked for recommendations regarding things like air filters, } } temperature control, and humidity. } } } } It will be a small room, maybe 6' x 9', in a modern university science } } building. The researcher is going to put a new ultramicrotome in there } and } } will want to do serial sectioning on conventional resin blocks. Cryo may } } come later, so if there is anything special needed for that, now is the } } time to add it to the list. } } } } I have always been OK with standard sort of rooms, regular building air } } filters etc, if there is anything special I have overlooked, please let } me } } know so I can pass it along. } } } } Thanks } } } } Jonathan Krupp } } Microscopy & Imaging Lab } } University of California } } Santa Cruz, CA 95064 } } (831) 459-2477 } } jmkrupp-at-cats.ucsc.edu } } } } } } } } } } } } _______________________________________________________ } Tired of slow Internet? Get -at-Home Broadband Internet } http://www.home.com/xinbox/signup.html }
Here are two adresses of Raith GmbH, in the USA and in Germany : Raith INC. 6 Beech Road Islip NY 11751-4907 Phone (516)224-1764 Fax (516)224-2620 E-mail 73164.1330-at-compuserve.com
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We have tested these two FE-SEM and we want to have other opinions about the differences between these two apparatus. Did you see a big difference in resolution at low energy (1-2 keV). What about the practical use of the "In Lens" detector of the S4700 ? Does it really bring more information than the others ? I think there are some 6700F in Far East (and in the States ?), but no one in Europe. We are interested in all you can say about these two SEMs.
Sincerely Yours
J. Faerber IPCMS-GSI 23, rue de Loess 67037 Strasbourg CEDEX FRANCE
Jacques We have also evaluated these two microscopes, and I would be interested to compare notes with you, but not online. Chris
} } Dear collègues } } We have tested these two FE-SEM and we want to have other opinions about } the differences between these two apparatus. Did you see a big difference } in resolution at low energy (1-2 keV). What about the practical use of the } "In Lens" detector of the S4700 ? Does it really bring more information } than the others ? I think there are some 6700F in Far East (and in the } States ?), but no one in Europe. We are interested in all you can say } about these two SEMs. } } Sincerely Yours } } J. Faerber } IPCMS-GSI } 23, rue de Loess } 67037 Strasbourg CEDEX } FRANCE } } Tel 00 33(0)3 88 10 71 01 } Fax 00 33(0)0 88 10 72 48 } E-mail Jacques.Faerber-at-ipcms.u-strasbg.fr } }
========================================== Dr. Chris Jeffree University of Edinburgh BIOSEM - Biological Sciences Electron Microscope Facility Institute of Cell and Molecular Biology Daniel Rutherford Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JH, Scotland, UK Tel. #44 (0) 131 650 5345 FAX. #44 (0) 131 650 6563 Mobile 07710 585 401 email c.jeffree-at-ed.ac.uk =========================================
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Thanks to everyone who responded to my cry for help.
To summarize my findings and your suggestions/comments; Kodak has discontinued the sale of transparent sleeves, CAT 150 3812, however, that item with the same catalogue number is now being sold by Tiffen (1-800-368-6257). Of course these sleeves may also be available from other outlets, eg. National Graphic Supply (1-800-223-7130). A number of microscopists are using and have suggested I try the polyview sheets which hold 6 TEM negatives and fit nicely into a 3-ring binder. These sheets, cat. #1310/3406 are sold by Negafile (1-888-881-6435) but again may also be available from other outlets, eg. Ted Pella (916-243-2200), in Canada (1-800-243-7765).
Regards, Paul
Paul J. Gerroir Microscopy Materials Characterization Xerox Research Centre of Canada 2660 Speakman Drive Mississauga, Ontario L5K 2L1
Senior technician/manager for Electron Microscopy Laboratory Managerial position for state-of-the-art electron microscopy facility within the Center for Biologic Imaging, University of Pittsburgh Medical School. Duties include overseeing and conducting the processing and preparation of samples for Transmission and Scanning Electron Microscopy and specialized techniques including cryosectioning and immunoelectron microscopy. Incumbent will help with training and assisting students and faculty on equipment, oversee general lab procedures including budget, supply, purchase orders, photography and quality control. Prior experience in electron microscopy essential. Bachelor’s or Master’s degree or equivalent desired. The successful applicant will be responsible for directing the 3 other staff in the EM component of the center The Center is a nationally recognized imaging resource using and developing a complete array of imaging technologies using both light and electron optics. More details about the center can be found at our web site (http://sbic6.sbic.pitt.edu):. This position is within the EM component of the CEnter. We have a full array of prep equipment from freeze fracture to ultracryomicrotomes. Our microscope base includes two transmission and two SEM scopes including a new field emission gun SEM. If you would like further information please contact Dr. Donna Beer Stolz or myself. Please send applications electronically to dstolz+-at-pitt.edu
--------------------------- Simon C. Watkins Ph.D. MRC Path Associate Professor, Cell Biology and Physiology Director: Center for Biologic Imaging BSTS 225 University of Pittsburgh Pittsburgh PA 15261 Tel:412-648-3051 Fax:412-648-8330
Does anyone know what happened to MSA Certification? I stopped at their booth in August at MSA 2000 and was assured that things getting "back on track" but so far I've heard nothing from them.
J. Faerber wrote: ================================== To D. Sicard
Here are two adresses of Raith GmbH, in the USA and in Germany : Raith INC. 6 Beech Road Islip NY 11751-4907 Phone (516)224-1764 { { {NOT CORRECT Phone (631) 224-1764 { { {CORRECT Fax (516)224-2620 { { {NOT CORRECT FAX (631)224-2620 { { { {CORRECT E-mail 73164.1330-at-compuserve.com ====================================== There has been an area code change and the old numbers absolutely won't work !
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
Does anyone already have a prepared procedure according to paragraphs 2.2 and 2.3 of MIL-STD-883E, Method 2018.3 they would share with me? What I am wondering is whether this procedure is detailed (this knob, that button, etc.) or more general in nature?
Based on the responses I did not receive from my earlier post about quantitative measurement of SEM resolution, it looks like paragraph 2 cannot be met. It states a resolution of 250A or less. Semantically, I'm sure it meant 250A or a lesser number. A resolution of 250A or less would tend to mean a poorer resolution than 250A and hence a larger number. Probably picky on my part.
Would appreciate any inputs regarding this MIL-STD.
I've recently started lowicryl (K4M) processing of mouse tissue. I am having good success with skeletal muscle, heart, kidney and lung but stomach, spleen, pancreas and adrenal had large holes and cut poorly. In addition, a yeast pellet that was resuspended in 2% agarose prior to processing was not well infiltrated.
Processing procedure: 3% paraformaldehyde/0.5% glut. fixation for 60 minutes with rotation (RT) Wash 3X for tem minutes each with PBS buffer Dehydrate 30% ETOH for 30 min. at 0C 50% ETOH for 60 min. at -20C 80% ETOH for 60 min. at -35C 100% ETOH (freshly opened bottle) for 60 min. at -35C 100% ETOH for 60 min. at -35C ETOH/K4M 1:1 for 60 min. at -35C ETOH/K4M 1:2 for 60 min. at -35C K4M for 60 min. at -35CC K4M overnight at -35C K4M 6-8 hrs at -35C
Embed with fresh resin Specimens are constantly rotated in freezer. All alcohols and resins are prechilled prior to use. Resin is mixeed by gently bubbling Argon gas to dissolve initiator. Polymerize at -35C overnight using UV light Raise temp to 0C and polymerize additional two days with UV Polymerize at room temp. for two days using UV
Any thoughts on what I'm doing wrong or things that I can change would be greatly appreciated.
Tom Januszewski Senior Electron Microscopist UT Southwestern Medical Center at Dallas Dallas, TX 75390-9039 214-648-7291 FAX: 214-648-6408 Email: tom.januszewski-at-utsouthwestern.edu
I have an old business card (how old I don't know) for Raith. The contact name is George Lanzarotta at (516) 293-0870. The address is (was?) 70C Carolyn Boulevard, Farmingdale, NY 11735.
Good luck, Paul
Paul J. Gerroir Microscopy Materials Characterization Xerox Research Centre of Canada 2660 Speakman Drive Mississauga, Ontario L5K 2L1
-----Original Message----- } From: Ladd Research [mailto:sales-at-laddresearch.com] Sent: Thursday, November 30, 2000 4:26 PM To: microscopy-at-sparc5.microscopy.com
Could anyone provide me with a phone number, e-mail or fax number for the company "RAITH USA" It is a Germany Company and I thought there was a distrbutor in the US
Thanks
D. Sicard --
LADD RESEARCH 131 Dorset Lane Williston, VT 05495 USA
web site http://www.laddresearch.com
TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail sales-at-laddresearch.com
Just got a new vacuum evaporator after many years of using our Balzers 400 for the same purpose. Course now we don't have e guns and must use a little tungsten basket and a bit of Pt wire. I did this many years ago but now I am having a problem I don't recall encountering back then. The tungsten basket breaks just as the wire melts. I suspect the energy on phase transition is cooling the tungsten and causing it to break but I could be way off there. Anyone else see this problem and know the solution?
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
Giselle You probably should check out the Journal of Fuel or "Fuel". "Carbon" occasionally has articles linking the mineral content of the feedstock with the resultant char. The Northern Carbon Research Laboratories, U. of Newcastle-upon-tyne used to publish regularly in this field. Penn. State also has a very active Fuel Science Dept. Good Luck.
J. Roy Nelson, Ph.D. Material Testing Laboratory mtl-at-njcc.com
giselle melville wrote:
} } Good afternoon } } My name is Giselle. I am a graduate student working on my thesis. My } thesis topic is "Methods of preparation of petrographic thin sections for } Electron Microscopy". I have a collection of thin sections (chert) and I am } preparing the sections for analysis by a TEM 1200. The thin sections are } initially prepared by methods Ultramicrotomy and Ion Milling. TEM } parameters used will be electron diffraction and elemental composition to } obtain identification of the mineral and microstructures. I need more } information (references) on recent literature (1998-2000) on my subject or } similiar work done using these methods for analyis by TEM. } } I need your help!!! } } Thank you for your time.
I am helping a friend find a scope. I came across a Zetopan that may fit his budget. I use a Nachet DIC for viewing living organisms and find it very satisfactory. Could anyone help him with the versatility of a DIC Zetopan. I know my Nachett works as a bright field, DIC and sort of polarizing scope lacking a rotating stage and 1/4 and 1/2 wave plates. It is the scope I use all the time. Unless I am doing darkfield.
Having never been around a Zetopan I can't really answer his questions.
Thanks Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger
} } I my quest to find a better 'scope I have come across (with the help } } of one of the list members) a Reichert Zetopan DIC scope that *might* } } be available at in my price range. Previously I was just looking for } } a nice phase contrast job. I have worked with phase before but not } } DIC and would like to solicit other opinions as to whether this is } } the right scope for me. This is going to pretty much blow my budget } } so I want to be sure. } } } } I will be using this primarily for observing living organisms } } (protozoa, algae, etc). I have been looking at a lot of photos of } } protozoa taken using Nomarski DIC. While the photos are striking, I } } keep asking myself "but what does it really look like". The problem } } is the false 3D effect. I'm never sure what the part of the shape is } } real and what is generated. Doesn't this ever bother you? } } } } I have been told that it is possible to adjust out the 3-D effect but } } I'm not sure what kind of image you are left with. I would assume } } something in-between bright field and DIC. Anyone have any experience } } with this? } } } } I think if the Zetopan is unable to easily switch between a DIC image } } and a bright (or dark) field image then I may pass on it. I need the } } more traditional illumination as well to see what the real shape is. } } It really makes the Zetopan too special purpose for someone like } } myself. I think I would probably be happier with a less } } sophisticated scope that can do phase, bright, and dark field (and } } maybe oblique lighting as well). } } } } I would appreciate any thoughts you all may have. } } } } Bill } } -- } } Bill Tschumy } } Otherwise -- Austin, TX } } bill-at-otherwise.com
Rick A. Harris wrote: ================================================================ Just got a new vacuum evaporator after many years of using our Balzers 400 for the same purpose. Course now we don't have e guns and must use a little tungsten basket and a bit of Pt wire. I did this many years ago but now I am having a problem I don't recall encountering back then. The tungsten basket breaks just as the wire melts. I suspect the energy on phase transition is cooling the tungsten and causing it to break but I could be way off there. Anyone else see this problem and know the solution? ================================================================ There really is no perfect "solution". The precious group metals tend to alloy quite readily with tungsten, Au and Pd very quickly, and I believe Pt is quite similar in behavior.
You can reduce the magnitude of the "problem" by making sure that when you install the tungsten basket, the wire itself is under zero stress. This might mean a bit more time adjusting the holders and posts than you might be doing now. Also, ball up the Pt wire and press into the very bottom of the basket, this will enable you to get the most evaporated in the shortest possible time period, before the wire basket breaks. And you want to use more of a "flash" evaporation than a long slow continuous kind of evaporation.
Another approach, but one we would not recommend, is to use a thicker wire basket but many EM samples could be heat damaged from the additional radiant heat. 20 mil wire is our recommended basket wire diameter.
Hope this information will be helpful.
Disclaimer: SPI Supplies manufactures tungsten wire baskets for vacuum evaporation.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
Does anyone out there in microscopy land know of a computer database program ( preferably PC) that has the scientist in mind and not the business man. I have Excel, Access, and Project 2000 but they are orientated to calculate dollars and cent not milliliters and millimeters if you know what I mean.
Ron Austin Dept of Pathology LSU Medical Center Shreveport, LA rla-at- mindspring.com 318-675-4557
yes I am using databases from dbase II on cp/m systems
ricardo
----- Original Message ----- } From: "Vr. Richard Bejsak-Collorado-Mansfeld" {ricardo-at-ans.com.au} To: "Ronald Austin" {rla-at-mindspring.com} Sent: Sunday, December 03, 2000 11:16 PM
Greetings, Nomarski optics are widely considered appropriate for viewing living cells. While it is true that the "3-D" look might or might not be present in the cell (more about that later), one must remember that in phase contrast there is the famous "halo", representing an out of focus reverse-contrast image that certainly is not present in the cell.
Nomarski optics provide contrast by means of a gradient in optical path (= sample thickness times refractive index). That is anywhere in the sample where the local gradient in optical path is different than the background (which typically would have a zero gradient) the intenstity of the image is different than the background. When the gradient is positive the intensity is brighter than background, when the gradient is negative then the intensity is darker than background. The most obvious gradients of optical path for a living cell suspended in an aqueous medium is the change in thickness that happens at the cell's edges: at one edge the cell is getting thicker (= postive gradient, hence bright) and at the opposite edge the cell gets thinner (= negative gradient, hence dark). Consquently, the topographic appearance does relate to the sample in a real, predictable way.
Nomarski optics can be harder to adjust properly than phase contrast. In phase, you have the phase ring in the condenser and this has to be centered with the phase ring in the objective. That's all you have to do. For Nomarski, you have to have first crossed polarizers, you have to know where they are and be able to rotate at least one of them. THen, the system also has two Wallason prisms, one in the condenser and one above the objective. The Wallaston prism has an optical axis and this axis of each Wallaston must be parallal and at 45 degrees to the analyzer/polarizer alignment. Different implementations of Nomarski differ vastly in the accessibility of the elements and the ease of their aligment. In the good designs, it is no trouble at all. But of course, one should know where each one is, what it does and how it should be aligned.
Changing between nomarki and brightfied is no trouble, you just remove the analyzer or polarizer from the optical path.
One important fact about Nomarski arises from the optical axis of the Wallastons. This defines the direction used to assess the gradient in optical path. The optics cannot detect gradients that run perpendicular to the Wallaston's optical axis. This means that features revealed in the image depend on the orientation of the cell. A rotatable stage is therefore very desirable. What is more, depending on your purpose, and the types of cells your are looking at, the directional properties of Nomarski will be more or less of a problem.
I have no experience with a Zetopan. If at all possible have demonstration before you buy. Finally, in many scopes you can exchange conderser elements so that you can do either phase or Nomarski. Thus, you could expand your repertoire later, as needs arise. I would be useful to find out if the Zetopan lets you do that.
Hope this helps, Tobias Baskin
} } I am helping a friend find a scope. I came across a Zetopan that may fit } his budget. I use a Nachet DIC for viewing living organisms and find it } very satisfactory. Could anyone help him with the versatility of a DIC } Zetopan. I know my Nachett works as a bright field, DIC and sort of } polarizing scope lacking a rotating stage and 1/4 and 1/2 wave plates. It } is the scope I use all the time. Unless I am doing darkfield. } } Having never been around a Zetopan I can't really answer his questions. } } Thanks } Gordon } Gordon Couger gcouger-at-couger.com } Stillwater, OK www.couger.com/gcouger } } } } } I my quest to find a better 'scope I have come across (with the help } } } of one of the list members) a Reichert Zetopan DIC scope that *might* } } } be available at in my price range. Previously I was just looking for } } } a nice phase contrast job. I have worked with phase before but not } } } DIC and would like to solicit other opinions as to whether this is } } } the right scope for me. This is going to pretty much blow my budget } } } so I want to be sure. } } } } } } I will be using this primarily for observing living organisms } } } (protozoa, algae, etc). I have been looking at a lot of photos of } } } protozoa taken using Nomarski DIC. While the photos are striking, I } } } keep asking myself "but what does it really look like". The problem } } } is the false 3D effect. I'm never sure what the part of the shape is } } } real and what is generated. Doesn't this ever bother you? } } } } } } I have been told that it is possible to adjust out the 3-D effect but } } } I'm not sure what kind of image you are left with. I would assume } } } something in-between bright field and DIC. Anyone have any experience } } } with this? } } } } } } I think if the Zetopan is unable to easily switch between a DIC image } } } and a bright (or dark) field image then I may pass on it. I need the } } } more traditional illumination as well to see what the real shape is. } } } It really makes the Zetopan too special purpose for someone like } } } myself. I think I would probably be happier with a less } } } sophisticated scope that can do phase, bright, and dark field (and } } } maybe oblique lighting as well). } } } } } } I would appreciate any thoughts you all may have. } } } } } } Bill } } } -- } } } Bill Tschumy } } } Otherwise -- Austin, TX } } } bill-at-otherwise.com
I'm pleased to announce that a revised, up-to-date version of "Children's Microscopy: a Bibliography" will be mailed as a supplement to the December issue of Microscopy Today; watch for it. It has brief reviews and ordering information on books, videos, CD-ROMs and websites. If you can't use it yourself, PLEASE give it to a teacher, or a parent, or a Scout leader, or a librarian - anyone who can use it to introduce children to the microworld.
If you aren't a Microscopy Today subscriber {microtoday-at-mindspring.com} you can read the same text soon on Project MICRO's website (URL below). But it's a tightly packed 16 pages, so it will be easier to browse the printed version.
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
Does anyone out there have suggestions on histochemical stains (or other techniques) to reduce the autofluorescence of lignin? I'm using DAPI staining on wood (xylem) sections, which have highly lignified cell walls, and I would like to reduce background fluorescence. Thanks.
Rachel
****************************************** Rachel Spicer Biological Laboratories 3119 Organismic and Evolutionary Biology Harvard University 16 Divinity Avenue Cambridge, MA 02138
It is relatively easy to program in Microsoft's Visual Basic 6 using databases. It is one of that program's strengths. You can tailor any properties that you want. It is easy to set it up to be compatible with Microsoft's Access, but you can have it set up for any ODBC database. You can actually set up a working database program by just using the Form Wizard that comes with VB6.
-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: Ronald Austin [mailto:rla-at-mindspring.com] Sent: Saturday, December 02, 2000 10:28 PM To: Microscopy Society of America Subject: a data base program
--------------------------------------------------------------- --------- The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserv er/FAQ.html
Does anyone out there in microscopy land know of a computer database program ( preferably PC) that has the scientist in mind and not the business man. I have Excel, Access, and Project 2000 but they are orientated to calculate dollars and cent not milliliters and millimeters if you know what I mean.
Ron Austin Dept of Pathology LSU Medical Center Shreveport, LA rla-at- mindspring.com 318-675-4557
I've started wondering about what actually the point resolution of a TEM in phase-contrast imaging is.
} From Raleigh's criterion I would conclude that for a typical 120 kV TEM with a numerical aperture of about 20 mrad distances larger than about 2 Angstrom or 'density waves' with a wavelength longer than 2 Angstrom simply cannot be resolved.
In Scherzer defocus the Phase Contrast Transfer Function (PCTF) has its first zero at about the same resolution but there seems to be information at higher frequencies (even if it is inverted in contrast).
Which is the true resolution limit?
Yours sincerely,
Philip
-- Philip Koeck Karolinska Institutet Dept. of Bioscience Novum S-14157 Huddinge Sweden Tel.: +46-8-608 91 86 Fax.: +46-8-608 92 90 Email: Philip.Koeck-at-csb.ki.se http://www.csb.ki.se/em
Trying to cut serial sections of Spurr for 3D reconstruction. Anyone know of any tricks to make the sections stay together in long ribbons? Ellen Morgan
In the field of high-resolution transmission electron microscopy, the "resolution" is defined as the first zero in the phase contrast transfer function (PCTF) at Scherzer (or optimum) defocus as you point out. This definition is chosen because any higher-frequency information is phase inverted (as you also point out). However, it results in defining the resolution in terms of the spatial frequency at which transfer goes to zero, rather than in terms of the highest transferred spatial frequency.
I have suggested a definition in which the resolution is defined at the 70% pass limit within the Scherzer passband, giving d = 0.67 Cs^1/4 lambda^3/4 at an "extended" optimum defocus of sqrt(1.5Cs.lambda) instead of Scherzer's d = 0.707 Cs^1/4 lambda^3/4 at an optimum defocus of sqrt(Cs.lambda). Current practice seems to use the extended defocus with the crossover (zero-transfer) frequency to get d = 0.64 Cs^1/4 lambda^3/4 at the "extended" optimum defocus.
The information beyond the Scherzer cross-over (zero-transfer) frequency can be utilized to provide structural information. Because of the misphasings, this information is difficult to interpret directly. Originally, the way to get at this higher-frequency (smaller-spacing) information was to compare focal series of experimental images with images simulated from possible models. In this way, we "saw" the small tunnels in Nb12O29 (with a 2.5Angstrom spacing) with a JEOL 100B (Scherzer resolution of 3.5A) in a focal series obtained by Sumio Iijima (see "Resolution-limiting effects in electron microscope images", G.R. Anstis and M.A. O'Keefe, In 34th Ann. Proc. EMSA, Miami Beach (1976) 480-481). I used HRTEM image simulation programs such as my SHRLI (simulated high-resolution lattice image) code, or my later interactive TEMPaS (TEM processing and simulation) code (ported to the Mac as MacTempas) with great success, but only when the model was known or limited to several possible ones (for example "Inversion domains in GaN grown on sapphire", L.T. Romano, J.E. Northrup and M.A. O'Keefe, Appl. Phys. Lett. 69 (1996) 2394-2396). Currently, post-Scherzer information can be accessed by image reconstruction algorithms from focal series, or tilt series, or holography. I use the Philips/Brite-Euram software for focal-series reconstruction by Coene and Thust in my one-Angstrom microscope (OAM) project to extend the resolution of a modified Philips CM300FEG/UT from its native resolution of 1.7Angstrom to a super-resolution of 0.89Angstrom that can image the "dumbbells" in [110] diamond (see “The NCEM One-Ångstrom Microscope project reaches 0.89Å resolution”, M. A. O'Keefe in 58th Ann. Proc. MSA, Philadelphia, Pennsylvania (2000) 1192-1193).
For the equations that belong with a discussion of resolution, see "Resolution in high-resolution electron microscopy", M.A. O'Keefe, Ultramicroscopy 47 (1992) 282-297. For a (simple) explanation of the use of focal-series to obtain super-resolution see "Push TEM limits with super-resolution", Michael A. O'Keefe (1999), R&D Magazine October 1999, p79 or {http://www.rdmag.com/archives/basics/10microscopy.htm} . For details of the Philips/Brite-Euram software for focal-series reconstruction by Coene and Thust, see W.M.J. Coene, A. Thust, M. Op de Beeck and D. Van Dyck, Ultramicroscopy 64 (1996) 109-135 and A. Thust, W.M.J. Coene, M. Op de Beeck and D. Van Dyck, Ultramicroscopy 64 (1996) 211-230. In the above, I have dealt only with linear terms in the transfer of information from the specimen (more properly, the exit-surface wave from the specimen) into the HREM image. Non-linear terms can give rise to even-higher spatial frequencies in the image. However, the question of whether these can be termed "resolution" is doubtful (see "Resolution-damping functions in non-linear images", M.A. O'Keefe, in 37th Ann. Proc. EMSA, San Antonio, Texas (1979) 556-557).
Mike O'Keefe
Philip Koeck wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hallo everybody, } } I've started wondering about what actually the point resolution of } a TEM in phase-contrast imaging is. } } } From Raleigh's criterion I would conclude that for a typical 120 kV } TEM with a numerical aperture of about 20 mrad distances larger than } about 2 Angstrom or 'density waves' with a wavelength longer than } 2 Angstrom simply cannot be resolved. } } In Scherzer defocus the Phase Contrast Transfer Function (PCTF) has its } first zero at about the same resolution but there seems to be } information } at higher frequencies (even if it is inverted in contrast). } } Which is the true resolution limit? } } Yours sincerely, } } Philip } } -- } Philip Koeck } Karolinska Institutet } Dept. of Bioscience } Novum } S-14157 Huddinge } Sweden } Tel.: +46-8-608 91 86 } Fax.: +46-8-608 92 90 } Email: Philip.Koeck-at-csb.ki.se } http://www.csb.ki.se/em
-------------------------------------------------------- Your mail has been rejected for the following reason(s): -------------------------------------------------------- OK, Nestor -- this time it's in plain text. -Mike
At 11:55 AM -0500 12/4/00, Ann Dvorak wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
************************ Hi Ellen, I vaguely remember reading something about coating the sides of the block that will become the leading and trailing faces with something slightly tacky (like the adhesive dissolved off of 3M Magic Scotch tape). Once that has dried, the edges of the sections are supposed to stick together. I've never actually tried it, and I wonder how you do manage to separate the sections where you need to.
Does anyone out there remember this? Has anyone tried it?
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
The Microscopy Society of America is seeking an Archivist to manage the historical documents of our Society. This position is appointed by MSA Council for a period of three years.
We are looking for someone who is experienced in the storage and cataloging of (primarily) paper documents. Although this is a voluntary position (non-paying), the Society would provide funding for expendibles and day-to-day operations.
If interested (or if you know of a qualified person), please contact me at:
-- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ##############################################################
A predecessor in our laboratory came back from the MSA conference a few years back with a sample from a vendor of something called Tacki Wax, which was supposed to serve this purpose. I think the idea was that the sections would be gently held together, but not so strongly that you couldn't work them apart as needed while collecting them. I've never tried it myself, as I don't need to do serial sectioning, but having a product name to go by might help your search.
Elaine Schumacher UOP 25 East Algonquin Rd. Des Plaines, IL 60017-5017 phone: 847-391-3403 fax: 847-391-3719 efschuma-at-uop.com
I am pleased to announce that the Live-cell Course continues to grow and prosper. Over 130 students from 23 countries have taken the course over the last 5 years.
Now I am organizing the next 11-day summer course on "3D Microscopy of Living Cells" for June of 2001. As in the past it will be at the University of British Columbia, in Vancouver, BC. The Fifth Workshop on 3D Image Processing will be held June 30 - July 2.
Please check out http://www.cs.ubc.ca/spider/ladic/course/bulletin.html if you would like to have a feeling for last years's course.
The information for 2001 is below.
Thanks for your help,
Jim Pawley
Sixth Annual INTERNATIONAL 11-Day Short Course on
3D Microscopy of Living Cells June 18 - 28, 2001
Fifth, Post-course Workshop on
3D Image Processing, June 30 - July 2, 2001
Organized by Prof. James Pawley, (University of Wisconsin-Madison) (SEE 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, 2001 Deposit due April 15, 2001 Registration 5:00 - 7:00 PM Sunday, June 17, 2001 First Lecture 7:30 PM Sunday, June 17, 2001 Live-cell Course ends, noon Thursday, June 28, 2001 3D Image Processing Course, June 30, - July 2, 2001
APPLICATIONS DUE BY MARCH 1, 2001
APPLICATIONS Applicants must complete a questionnaire to assess knowledge level, field of interest and proposed personal, live-cell, project. Enrollment will be limited to about 24 participants (exact number depends on number of 3D Systems available). Selection will be made on the basis of background and perceived need. Those without previous LM experience will be provided with access to basic texts to read before the course begins. Application forms requesting information on field of interest and level of experience may be down-loaded from the WWW site at http://www.cs.ubc.ca/spider/ladic/course/reg1.htm, or obtained from:
Dr. Elaine Humphrey, Biosciences EM Facility Biosciences Building Univ. of British Columbia 6270 University Blvd. Vancouver, BC, V6T-1Z4
Additional information is available from: http://www.cs.ubc.ca/spider/ladic/course/bulletin.html
We expect to have at least 11, 3D microscope workstations for student use and there will be an international faculty of 17.
Application deadlines:
Application forms must be received for screening by March 1, 2000. Successful applicants will be notified by April 1, and a deposit of 50% must be received by April 15, 2000. In general, refunds of the deposit will not be possible. The remainder is due before Registration.
3D Course tuition (includes lunches and snacks): $2150 (US) Workshop Tuition (includes lunches and snacks): $850 (US)
Room / board about $40/day (US)
I hope that this includes all of the information that you need, but if not, please get back to me. -- **************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 223, Zoology Research Building, FAX 608-265-5315 1117 Johnson Ave., Madison, WI, 53706 JBPAWLEY-at-FACSTAFF.WISC.EDU "A scientist is not one who can answer questions but one who can question answers." Theodore Schick Jr., Skeptical Enquirer, 21-2:39
Thanks for offering to put my 3D rendering question on the discussion group. My lab currently acquires and processes images using IPlab and a SPOT RT camera. We have MicroTome by VayTek for deconvolution, but this isn't much good without 3D rendering. Our primary application will be measuring the volume of nuclei and cells which have been fluorescently stained. Which 3D programs for a PC might work well for this?
Hello all, Any one with on advice on who in the Baltimore area can do chemlumanesence (spelling?) on live cells, realtime and time series? What equipment do we need, which company or labs are doing this.
I'll take a stab at this. Please correct me if I am wrong:
The point resolution refers to the resolution limit of a TEM for (as the name implies) two point like objects. A pointlike object is mathematically a delta-function and has a Fourier spectrum that is constant for all frequencies. So the question becomes: how many of the Fourier components can be transmitted without changing them, which in turn determines the minimum size of the object. The answer is then basically given by the first zero of the CTF. Until this point, the spatial frequencies are transmitted with the same phase and almost the same amplitude. For higher frequencies, the amplitude of the CTF oscillates, so that no reliable transmission can be achieved. The setting, where the first zero is furthest out in Fourier space is the Scherzer defocus. Thus the point resolution is determined by the first zero of the CTF at Scherzer defocus.
Line resolution refers to a periodic structure with very few Fourier components. It is therefore not necessary to transmit the entire Fourier spectrum, but only one specific component. Thus by changing the CTF through changing the focus, one can find a setting, where that specific frequency is transmitted and shows up in the image. The question therefore becomes: How well can I detect the structure. This is then in principle determined by the amplitude of the CTF. So the line resolution is then determined by the CTF envelope going below a certain threshold (I believe it is 20%, but I am not sure). That threshold is of course more or less arbitrary, but should give a reasonable estimate of the line resolution and allow comparison of the microscope resolutions.
Disclaimer: Please use this with caution. I have not been involved with EM theory for a few years. For a good account of high-resolution TEM, check out the book(s) by John Spence.
Michael Bode
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: Philip Koeck [mailto:philip.koeck-at-csb.ki.se] Sent: Monday, December 04, 2000 8:45 AM To: 3dem list; MSA mailing list
Hallo everybody,
I've started wondering about what actually the point resolution of a TEM in phase-contrast imaging is.
} From Raleigh's criterion I would conclude that for a typical 120 kV TEM with a numerical aperture of about 20 mrad distances larger than about 2 Angstrom or 'density waves' with a wavelength longer than 2 Angstrom simply cannot be resolved.
In Scherzer defocus the Phase Contrast Transfer Function (PCTF) has its first zero at about the same resolution but there seems to be information at higher frequencies (even if it is inverted in contrast).
Which is the true resolution limit?
Yours sincerely,
Philip
-- Philip Koeck Karolinska Institutet Dept. of Bioscience Novum S-14157 Huddinge Sweden Tel.: +46-8-608 91 86 Fax.: +46-8-608 92 90 Email: Philip.Koeck-at-csb.ki.se http://www.csb.ki.se/em
For Light microscopy, we used a mixture of Weldwood contact cement from the local hardware store and toluene mixed 50/50. Place it on the leading and trailing edges of the block using a pin. We used a tool to pick up the sections I don't remember its name. It looked like a small metal trough.
For EM I used to make the block face area as small as possible focusing on a specific area of interest. Then I would cut the the leading and trailing edges parallel and then put the block on the ultra microtome and polish the edges with a glass knife set on zero advance. If the leading and trailing edges are smooth the sections stick together when coming off the block. I would make groups of ten sections. pick up with a tungsten wire loop (premade loops work better from EM venders of your choice) and place on a formvar coated slot grid.
Good Luck
Jon
{color} {param} 0100,0100,0100 {/param} On 4 Dec 00, at 15:23, Leona Cohen-Gould wrote:
} } Trying to cut serial sections of Spurr for 3D reconstruction. Anyone
} } know of any tricks to make the sections stay together in long
} } ribbons? Ellen Morgan
}
} ************************
} Hi Ellen,
} I vaguely remember reading something about coating the sides of the
} block that will become the leading and trailing faces with something
} slightly tacky (like the adhesive dissolved off of 3M Magic Scotch
} tape). Once that has dried, the edges of the sections are supposed to
} stick together. I've never actually tried it, and I wonder how you do
} manage to separate the sections where you need to.
}
} Does anyone out there remember this? Has anyone tried it?
}
} 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
}
}
{nofill} 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
I would like to know whether there are any commercial labs that can provide up to 3000 um x 3000 um contact profilometry image maps such as the Veeco Dektak Profiler or the KLA-Tencor Profiler can provide.
-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
Go to Rivlin and Raymond, 1987, Journal of Neuroscience Methods, 20:23-33, also see Raymond and Rivlin, 1987, Developmental Biology, 122:120-138. There you will find reference to the technique originally published by W. Fahrenbach, 1984, J. Electron. Microsc. Tech., 1:387-398. In the Rivlin paper you will find that a solution of Weldwood cement (rubber cement) was applied to the top and bottom edges of the trapezoid block face. This glue can be use undiluted or diluted in the appropriate solvent to obtain the desired workable concentration.
Linda Barthel, M.S. Research Associate II University of Michigan Department of Cell and Developmental Biology 4607 Medical Science Building II Ann Arbor, MI 48109-0616
There may be a position opening up in the Dallas/Ft. Worth Area for an experienced microscopist with polymers-TEM experience. If you might be interested please send me a resume. Jeff Day/wa5ekh-at-juno.com ________________________________________________________________ GET INTERNET ACCESS FROM JUNO! Juno offers FREE or PREMIUM Internet access for less! Join Juno today! For your FREE software, visit: http://dl.www.juno.com/get/tagj.
Does anyone have suggestions as to how to polish silicon wafers in cross section. I have a wafer (~0.5mm thick ) which has been randomly fractured with about a micron of metal on the surface and want to determine the metal thickness. We are trying various methods - first choice is by RBS but presently limited in max. energy so can not penetrate layer.
In my first couple of attempts at polishing (embedded wafer in thermoset) the silicon breaks quite easily producing a very rough surface. I assume I am being too aggressive and am trying slower more gentle procedure. Does anyone have a procedure for polishing cross sections of such brittle material?
The following comments are for the person who is trying to cut serial sections with Spurrs. I routinely cut serial sections with Spurrs without resorting to any kind of glue. Probably the most important thing is how the block is faced. I cut the trapezoid with a really long (.6 to .9 mm base) and the top of the trapezoid is almost the same length. What's really critical here is that those two cuts are parallel. The two sides of the trapezoid are very short (.1mm). I cut the sections with a Diatome knife and on a Reichert Ultracut E with a cutting speed of 0.8mm per second and a thickness setting of 80nm. I am doing this in a small room with a blasting air vent in the ceiling. I have cut up a cardboard box and stuck it into the ceiling tiles in such a way that the air is diverted away from the microtome. I can pick up 15 to 25 sections in a row on a 0.4 X 2mm copper slotted grid coated with just formvar or formvar/carbon. I have never been any good at making the support films my self, so I buy the slotted grids precoated from Ted Pella (catalog #s 01706, 01806, or 01816). Before picking up the ribbon, I dip a stick into chloroform and wave that over the sections, and they expand (or relax, depending on your point of view). I have a bunch of self locking tweezers and after picking up the sections, I leave the grid with the sections on it locked in the tweezers until they dry (if you put a wet slotted grid down on filter paper it might break the film). Good luck, Tim
Timothy G. Schneider Director of Electron Microscopy Department of Pathology Room 229 Jefferson Hall Thomas Jefferson University 1020 Locust St. Philadelphia Pa. 19107 215-503-4798 work 610-613-8170 cellular timothy.schneider-at-mail.tju.edu
} Trying to cut serial sections of Spurr for 3D reconstruction. Anyone know } of any tricks to make the sections stay together in long ribbons? Ellen } Morgan
Ellen -
I assume that you're talking about EM. not LM. Make sure that the top and bottom of your block face are REALLY smooth and clean - and as small as is consistent with the reconstruction. Viewed thru the microtome binocular, they should look glassy, not like sandpaper. Change your trimming tool (glass knife? Weck razor blade?) often.
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
Has anyone used a HP Laserjet IIIp with an EDAX PV9900, or know what drivers are needed, or where we can acquire them?
We are trying to use the IIIp because it has serial and parallel connections needed by the EDAX and by the Philips XL40 SEM with which we use it and we cant afford to buy a new one.
Reply in confidence to chris.smith-at-bbsrc.ac.uk if you don't want to clog up the list.
Many thanks, Chris, Plant Path., IACR-Rothamsted, UK.
Ed, I represent Allied High Tech Products. We have detailed procedures for preparing semiconductor device cross-sections. We also offer all the necessary products to accomplish it. I would be happy to speak to you and provide you with detailed procedures on how to prepare these samples. Since it can be quite detailed it would be best to do this offline. Please provide me with your phone number and I will call you.
Sincerely,
Ed
************************************************* 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: Ed Kurz [mailto:ekurz-at-mail.ims.uconn.edu] Sent: Tuesday, December 05, 2000 8:32 AM To: MICROSCOPY BB
Does anyone have suggestions as to how to polish silicon wafers in cross section. I have a wafer (~0.5mm thick ) which has been randomly fractured with about a micron of metal on the surface and want to determine the metal thickness. We are trying various methods - first choice is by RBS but presently limited in max. energy so can not penetrate layer.
In my first couple of attempts at polishing (embedded wafer in thermoset) the silicon breaks quite easily producing a very rough surface. I assume I am being too aggressive and am trying slower more gentle procedure. Does anyone have a procedure for polishing cross sections of such brittle material?
We have a two year old PGT Prism EDS detector/preamp/dewar assembly which we are thinking of selling. But I don't have much of an idea of the value of such a device.
Any thoughts?
Anyone interested?
Richard Shalvoy Arch Chemicals Cheshire, CT 203-271-4394
This is not a job application. Would like to seek your advice on how to analyse a polymer samples using TEM. We have tried on Poly-carbonate injection molded lense with hard coating. The purpose is to see the interface bonding of the hard coat on the lense. Unfortunately, when we microtome the samples, the sample curve up and we cannot manage to see anything. Would appreciate if you can share your experience on the sample preparation with us.
Thanks in advance of your help.
Regards, YH Koh (MQE Engineer) Motorola CGISS Penang, Bayan Lepas FIZ, Phase 3, 11900 Penang, Malaysia. Tel : 60-4-8504089 Fax: 60-4-6124903
-----Original Message----- } From: charles j day [mailto:wa5ekh-at-juno.com] Sent: Sunday, December 03, 2000 12:50 PM To: Microscopy-at-sparc5.microscopy.com
There may be a position opening up in the Dallas/Ft. Worth Area for an experienced microscopist with polymers-TEM experience. If you might be interested please send me a resume. Jeff Day/wa5ekh-at-juno.com ________________________________________________________________ GET INTERNET ACCESS FROM JUNO! Juno offers FREE or PREMIUM Internet access for less! Join Juno today! For your FREE software, visit: http://dl.www.juno.com/get/tagj.
"james patrick birrell" {jbirrell-at-students.uiuc.edu} wrote in message news: {Pine.GSO.4.10.10012041635310.22505-100000-at-ux12.cso.uiuc.edu} ... } I was wondering if anybody would know where I could obtain software to } facilitate the acquisition/analysis of images to be used to perform } wavefront reconstruction using images from a TEM. Specifically, I'm } looking for any software compatable with a JEOL 4000ex microscope with a } Gatan CCD camera. } Thanks, } James Birrell
Ammonium molybdate (AM) is suitable in the cases when uranyl acetate (UA) is bad (mostly if pH is a critical factor, UA pH = 5-5.5). In most cases AM gives less contrast than UA. AM gives you "pure" negative staining because do not chemically interfere with most biological samples. UA from another hand sometimes gives you mixed contrast, positive (DNA/RNA)&negative(proteins). For instance, ribosomes and many viruses shown mixed staining with UA. It's not bad because it may even enhance some structural details, but you have to remember about that. My own experience indicated that in most cases UA or other uranium salts (U-formiate or oxalate) after conditions adjusting (concentration, time, temperature, freshness, support film) has shown perfect results. If you have a problem with stain spreading, you may try different support films (carbon, formvar, parlodion). Very good results you may obtain using "double carbon" technique - this may solv even very worse cases. I also has have a good results with poly-lysine treatment of the grids prior sample adsorption. Personally, I don't like the glow-discharge. But it works in some way.
Sergey.
At 11:52 AM 12/5/00 -0800, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
EDAX has the HP laser printer driver for old DEC based systems. Call EDAX, or www.edax.com
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax
This message is made of 100% recycled electrons. -----Original Message----- } From: chris smith (IACR-RES) {chris.smith-at-bbsrc.ac.uk} To: 'Microscopy-at-MSA.Microscopy.Com' {Microscopy-at-sparc5.microscopy.com}
I found your post interesting. How do you make your edges parallel? Do you use a razor blade?
Dave
On Tue, 5 Dec 2000 09:47:29 -0500 Timothy Schneider {Timothy.Schneider-at-Mail.TJU.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 } -----------------------------------------------------------------------. } } } The following comments are for the person who is trying to cut serial } sections with Spurrs. I routinely cut serial sections with Spurrs without } resorting to any kind of glue. Probably the most important thing is how the } block is faced. I cut the trapezoid with a really long (.6 to .9 mm base) } and the top of the trapezoid is almost the same length. What's really } critical here is that those two cuts are parallel. } The two sides of the trapezoid are very short (.1mm). I cut the sections } with a Diatome knife and on a Reichert Ultracut E with a cutting speed of } 0.8mm per second and a thickness setting of 80nm. I am doing this in a } small room with a blasting air vent in the ceiling. I have cut up a } cardboard box and stuck it into the ceiling tiles in such a way that the air } is diverted away from the microtome. I can pick up 15 to 25 sections in a } row on a 0.4 X 2mm copper slotted grid coated with just formvar or } formvar/carbon. I have never been any good at making the support films my } self, so I buy the slotted grids precoated from Ted Pella (catalog #s 01706, } 01806, or 01816). Before picking up the ribbon, I dip a stick into } chloroform and wave that over the sections, and they expand (or relax, } depending on your point of view). I have a bunch of self locking tweezers } and after picking up the sections, I leave the grid with the sections on it } locked in the tweezers until they dry (if you put a wet slotted grid down on } filter paper it might break the film). Good luck, Tim } } Timothy G. Schneider } Director of Electron Microscopy } Department of Pathology } Room 229 Jefferson Hall } Thomas Jefferson University } 1020 Locust St. } Philadelphia Pa. 19107 } 215-503-4798 work } 610-613-8170 cellular } timothy.schneider-at-mail.tju.edu } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
{ { Would like to seek your advice on how to analyse a polymer sample using TEM. We have tried on Poly-carbonate injection molded lens with hard coating. The purpose is to see the interface bonding of the hard coat on the lens. Unfortunately, when we microtome the samples, the sample curves up and we cannot manage to see anything. Would appreciate if you can share your experience on the sample preparation with us. } }
We have looked at surface layers in polymer samples by cutting open a surface, then etching and looking, NOT at a cut section, but at the cross-sectional view of the remaining surface, either by SEM or by replication/TEM. Usually we etch our specimens, so before cutting open, a protective layer is applied over the outer surface to prevent this being etched downwards. Mainly we have looked at polyethylene or polypropylene specimens, using our permanganic etching technique, and different reagents would be needed for polycarbonate. However, it might be possible to replicate the cut surface directly and get useful information.
+------------------------------------------------------------------------+ | 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 | +------------------------------------------------------------------------+
The question was: how do I make the top and bottom edges parallel? I do it by eye. I use Gem single edge industrial blades from Ted Pella Cat# 121-1 (I'm sure these are available from many sources). One important point is that I spray them with 70% ETOH before using and wipe them off with a Kimwipe EXL. A word about the way I described the trapezoid face: it doesn't look like a trapezoid. What it looks like is a very thin shiny line (heck, I'm talking about putting 15 to 25 of these guys on a 2mm slot, you do the math). More good luck, Tim
Timothy G. Schneider Director of Electron Microscopy Department of Pathology Room 229 Jefferson Hall Thomas Jefferson University 1020 Locust St. Philadelphia Pa. 19107 215-503-4798 work 610-613-8170 cellular timothy.schneider-at-mail.tju.edu
Thanks to all who responded to my question regarding conductive adhesives for salmon skin. I have ordered many of the products that were suggested and am looking forward to success (finally) :)
Thanks again and have a great holiday season!
Carla Aiwohi Western Fisheries Research Center Seattle, WA
We routinely use HEPES for our fixations with paraformaldehyde, glutaraldehyde and osmium. We see little or no difference from classical cacodylate buffers. I chose HEPES a long time ago and in retrospect, PIPES might have been a slightly better choice since HEPES has a pKa of 7.5 and PIPES has a pKa of 6.76. Since one generally uses aldehyde buffers at pH 7.0 to 7.4, either would seem appropriate but since I read that during fixation, H+ ions are generated and the solution tends to acidify, then having a buffer on the high side of the pKa would be better since you would have greater buffering power. Despite this "improved" logic, I have remained loyal to HEPES and never had a problem. Good luck.
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-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
Would anyone happen to have the actual recipe for Gurr's buffer? I think it is some phosphate formulation - we've been hunting the recipe and have found some refs, but will have to interlibrary the journals/books and we thought we'd give the 'net a try, first.
I know we can buy the pre-made buffer, but we need such a tiny amount that we'd rather make it from scratch (plus we want to know what it is!).
I would be a bit concerned about your use of chloroform to flatten sections, especially in a small microtome room. We have used heat pens for the last decade or so, because apparently the chloroform we use these days is a lot more toxic than it used to be (as well as formaldehyde, glutaraldehyde, carbon tetrachloride etc).
Malcolm Haswell e.m. unit School of Sciences University of Sunderland UK
Timothy Schneider 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 following comments are for the person who is trying to cut serial } sections with Spurrs. I routinely cut serial sections with Spurrs without } resorting to any kind of glue. Probably the most important thing is how the } block is faced. I cut the trapezoid with a really long (.6 to .9 mm base) } and the top of the trapezoid is almost the same length. What's really } critical here is that those two cuts are parallel. } The two sides of the trapezoid are very short (.1mm). I cut the sections } with a Diatome knife and on a Reichert Ultracut E with a cutting speed of } 0.8mm per second and a thickness setting of 80nm. I am doing this in a } small room with a blasting air vent in the ceiling. I have cut up a } cardboard box and stuck it into the ceiling tiles in such a way that the air } is diverted away from the microtome. I can pick up 15 to 25 sections in a } row on a 0.4 X 2mm copper slotted grid coated with just formvar or } formvar/carbon. I have never been any good at making the support films my } self, so I buy the slotted grids precoated from Ted Pella (catalog #s 01706, } 01806, or 01816). Before picking up the ribbon, I dip a stick into } chloroform and wave that over the sections, and they expand (or relax, } depending on your point of view). I have a bunch of self locking tweezers } and after picking up the sections, I leave the grid with the sections on it } locked in the tweezers until they dry (if you put a wet slotted grid down on } filter paper it might break the film). Good luck, Tim } } Timothy G. Schneider } Director of Electron Microscopy } Department of Pathology } Room 229 Jefferson Hall } Thomas Jefferson University } 1020 Locust St. } Philadelphia Pa. 19107 } 215-503-4798 work } 610-613-8170 cellular } timothy.schneider-at-mail.tju.edu
LM - Need help on staining semithin sections embedded in LR-white
I am doing anatomical studies on the seed coat and currently use toluidine blue with sodium hypochlorite prestaining and iodine/potassium iodite poststaining, described by Gutmann, M. : Improved staining procedures for photografic documentation of phenolic deposits in semithin sections of plant tissue. Journal of Microscopy 179:277-281 (1995) Can anyone suggest a better stain, specially one that is suitable for staining lignin ?
1. Apply a thin layer of M-bond on the surfaces of two pieces of your wafer and put these two surfaces together. 2. Use a clip to press these pieces of wafers as close as possible (not to break them). 3. Leave the sample to try (24 hours in air? or 1 hour at 120 C degree on a hot plate).
This is a good way to protect the thin "film" (your metal features) I think. The smaller the gap between these two pieces, the better. In this way you can get two cross section surfaces. Even after step 3, you may apply another thin layer of M-bond on the outside surfaces of the sample, so that you may get 4 surfaces for examination.
4 Embed the sample in a suitable epoxy as you did before, and grinding and polishing the cross sections carefully from coarse to fine.
When you observe the cross sections, it is easy to see what is the M-bond and what is the other thing you want to see.
Just a suggestion.
Good luck!
Zhenquan Liu
I think you may try the following way:
1. Apply a thin layer of M-bond on the surfaces of two pieces of your wafer and put these two surfaces together. 2. Use a clip to press these pieces of wafers as close as possible (not to break them). 3. Leave the sample to try (24 hours in air? or 1 hour at 120 C degree on a hot plate).
This is a good way to protect the thin "film" (your metal features) I think. The smaller the gap between these two pieces, the better. In this way you can get two cross section surfaces. Even after step 3, you may apply another thin layer of M-bond on the outside surfaces of the sample, so that you may get 4 surfaces for examination.
4 Embed the sample in a suitable epoxy as you did befoe, and grinding and polishing the cross sections carefully from coarse to fine.
When you observe the cross sections, it is easy to see what is the M-bond and what is the other thing you want to see.
Just a suggestion.
Good luck!
Zhenquan Liu
=================
Does anyone have suggestions as to how to polish silicon wafers in cross section. I have a wafer (~0.5mm thick ) which has been randomly fractured with about a micron of metal on the surface and want to determine the metal thickness. We are trying various methods - first choice is by RBS but presently limited in max. energy so can not penetrate layer.
In my first couple of attempts at polishing (embedded wafer in thermoset) the silicon breaks quite easily producing a very rough surface. I assume I am being too aggressive and am trying slower more gentle procedure. Does anyone have a procedure for polishing cross sections of such brittle material?
Thanks, Ed
------------------------------------ Zhenquan Liu (Dr.) Arizona State University Room 146A, CSSS Tempe, AZ 85287 Tel (480) 965 4544 (o) (480) 775 7428 (h) Fax (480) 965 9004 (o) Email zhenquan.liu-at-asu.edu ------------------------------------
I have used PIPES as a substitute for cacodylate in most routine procedures simply because it has none of the precipitation problems of phosphate buffers. I understand that there may be more extraction with cacodylate in TEM so I would have thought that it may be a better buffer for SEM (less chance of shrinkage?). I would assume that HEPES was similar but have no experience with it.
These are my own feelings and I have seen no scientific evidence to support this - has anyone else?
Malcolm Haswell e.m. unit School of Sciences University of Sunderland UK
Bart De Pauw wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello listers, } } We use cacodylate buffer for our SEM samples. Is HEPES or PIPES buffer } an alternative for the toxic cacodylate ? Our samples are mainly animal } tissue. } } Bart De Pauw } Ghent University } Faculty of Veterinary Medicine } Department Morphology } Belgium
} To: Timothy.Schneider-at-Mail.TJU.EDU } From: Sergey Ryazantsev {sryazant-at-ucla.edu} } Subject: Re: MORE SERIAL SECTIONS } } If you are not limited by particular area in your sample, you may do the } trick, which warranty the parallelness of the edges. This trick did show } to me Prof. Borovyagim many years ago. The trick is: you do not form a } trapezoid, instead you make two cuts by razor blade, which owerlapping, so } you will have "/\"-shape without flat top. The top should be formed by } two overlapped cuts. There is no flat space on the top, which we usually } called "trapezoid". This two cuts will form a "line" which will be } paralle to the knife edge later. You have to form side fases of the } piramid as well. Finall, you will have something looks pretty like } triangle prizm. Not perfect prizm, of coarse. When you start cutting, } you have to carefully cut avay the top of the "prizm" to form } trapezoid. The final shape of the trapezoid will depends from orientation } of your cuts, but owerlapped cuts will form perfectly parallel sides. } } Another choice I friequently use: to use glass knife and trim on the } goniometer-head when you may tilt and rotate the sample. Make one face, } rotate 180 deg and make second face. } } At 09:24 AM 12/6/00 -0500, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
I am currently in the process of setting up a histology/microbiology/EM facility. Does anyone have comments or suggestions on a good rotary tissue processor for light microscopy? My main concerns are cost, service availability, ease in operation(changing of fluids, etc.) and reliability. I may process 10 - 60 specimens a month, more as the lab is established. I appreciate any info.
I'm currently in the process of setting up a Histology/Microbiology/EM Lab. Having been away from histology for almost 30 years (been doing EM), I've gotten away from instrumentation associated with histology. Does anyone have a good recommendation for a rotary tissue processor? I've got info on Shandon and Tissue-Tek. Are there others out there? Thanks for any info.
} Date: Wed, 06 Dec 2000 15:16:31 -0800 } To: Timothy.Schneider-at-Mail.TJU.EDU } From: Sergey Ryazantsev {sryazant-at-ucla.edu} } Subject: Re: MORE SERIAL SECTIONS } } If you are not limited by particular area in your sample, you may do the } trick, which warranty the parallelness of the edges. This trick did show } to me Prof. Borovyagin many years ago. The trick is: you do not form a } trapezoid, instead you make two cuts by razor blade, which overlapping, so } you will have "/\"-shape without flat top. The top should be formed by } two overlapped cuts without "free" space between. There is no flat space } on the top, which we usually called "trapezoid". This two cuts will form } a "line" which will be parallel to the knife edge later. You have to } form side faces of the pyramid as well. Finally, you will have something } looks pretty like triangle prism. Not perfect prism, of coarse. When you } start cutting, you have to carefully cut away the top of the "prism" to } form trapezoid. The final shape of the trapezoid will depends from } orientation of your cuts, but overlapped cuts will form perfectly parallel } sides. This technique works if sample will permit. } } Another choice I frequently use: to use glass knife and trim on the } goniometer-head when you may tilt and rotate the sample. Make one face, } rotate 180 deg and make second face. If I have enough time, I prefer this } way, almost gives me the perfect pyramid. } } Sergey } } At 09:24 AM 12/6/00 -0500, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
I'm 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 usually glue (with superglue or thin epoxy) 2-4 wafers together, metal coated surfaces facing each other. Then embed them or mount in microwise, cut with low speed diamond saw and polish, preferably with diamond. If you do not need nice pictures, just measurements, do not use fine polish, you will more or less ruin edges with it.
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: Ed Kurz [mailto:ekurz-at-mail.ims.uconn.edu] } Sent: Tuesday, December 05, 2000 8:32 AM } To: MICROSCOPY BB } Subject: Wafer Cross sections } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } 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 suggestions as to how to polish silicon } wafers in cross } section. I have a wafer (~0.5mm thick ) which has been } randomly fractured } with about a micron of metal on the surface and want to } determine the metal } thickness. We are trying various methods - first choice is by RBS but } presently limited in max. energy so can not penetrate layer. } } In my first couple of attempts at polishing (embedded wafer } in thermoset) } the silicon breaks quite easily producing a very rough } surface. I assume I } am being too aggressive and am trying slower more gentle } procedure. Does } anyone have a procedure for polishing cross sections of such } brittle material? } } Thanks, } Ed }
} Would anyone happen to have the actual recipe for Gurr's buffer? I think } it is some phosphate formulation ... } I know we can buy the pre-made buffer, but we need such a tiny amount that } we'd rather make it from scratch (plus we want to know what it is!).
Quite right too! (For making and wanting to know) I haven't heard of it so can't tell you the answer, BUT:
Does it matter what the exact formulation is? It is unlikely that either of the late Gurr Bros (Edward, and George T.) invented an original buffer. They were vendors of stains in Britain, about whom some older HistoNetters may well have entertaining anecdotes.
If a buffer does its job of stabilizing the pH at the correct value, and doesn't contain anything that would react with the other ingredients of the mixture, it shouldn't matter what you use. Phosphate buffers make a precipitate with many metal salts, including those of Ca, Co, Cu, Fe, Mg, and others that have insoluble phosphates. All the techniques books published since about 1955 have sets of tables for making buffers to cover a wide range of pH.
John A. Kiernan, Department of Anatomy & Cell Biology, The University of Western Ontario, LONDON, Canada N6A 5C1
Ted Pella Poly-L-Lysin 0.1% w/v aqueous solution, Cat # 18026 Float your grid with film on small drop of the solution for 1 min and than remove excess of the liquid by filter paper and put on the drop of water for 5-10 sec, blot it and immediately move it on the drop with your sample. Temperature - as necessary for your sample. It also helps to adsorb tuff samples. Surprisingly, it seems to me that it does not affect background.
Concerning "double-carbon" technique. It is more tricky. The idea is to sandwich your sample between two carbon films just before you finish the staining. I prefer the following technique. The approx. 1 ml plastic cap (have no idea what is that, you may use any suitable container) 0.7 cm diameter I filled with 0.5-1% aqueous UA in the way to make flat meniscus, so it will reflect the light. Than I will cut 3x3 mm piece of mica with carbon film and float carbon on the UA solution. Reflecting light will makes it visible under some particular angle (you have to practice a little bit). So, you have the plastic cap with UA and floated piece of carbon film on it and you can see carbon easily. Than you will adsorb sample on the regular EM grid with carbon. Than you have to move your grid into the cap with stain (avoid contact to the floated carbon, soak grid into the cap away from carbon). Looking through the carbon film, still floated on the UA, move the grid just under the film. So, you will see the grid through floated film. Than using one smooth movement you have to move grid vertically up picking up the carbon film. So, you will put grid just under the film (deep under the surface) and than move grid vertically up. The film will holds over the grid with drop of the stain solution. The last most important step - to remove excesses of staining solution: fix grid securely in the tweezer by rubber O-ring (or use "reverse-action" tweezer), small triangle shape piece of filter paper (I am using Whatmann 1M paper) insert between tweezer's "legs" (sorry, guys, I have no idea how it is called) as closer to the grid as possible. Filter paper chip should contact with excess of liquid holds by capillary force between the tweezer's "legs". Keep tweezer with grid and filter paper until grid will dry completely (at least 5 min). Enjoy the perfect spreading of the stain! There is a number of disadvantages in this method: -"double-carbon" will decrease the resolution; -sandwich will slightly flat the particles, expect to see 10-15% bigger size because of that.
Advantages: -sandwich will protect your sample from radiation damage; -such samples usually are more stable from all points of view; -because of thick carbon, you may use 400 mesh naked EM grids without "holey" support film (I still prefer to use the grids with my home-made "holey" film).
PTA vs AM. In my point of view, PTA does not have any serious advantages over the AM. Personally, I prefer AM. In some particular cases PTA may work better, I guess. If there is no chance to use UA, I would try U-formiate first and oxalate - next. U-formiate (UF) has pH around 6 or even higher (I don't remember the exact number). It gives you a very nice fine staining (I love it better than UA). The disadvantage of this stain - it is not stable. You have to prepare fresh solution every time. It is light-sensitive also (general room illumination is OK for 20-30 min, no direct light, please!). You could store it in the dark in the ice for couple of hours. U-oxalate (UO) is less stable than UF, you have to use it immediately. The beauty of UO - you may use it at pH 7. People's unsuccess with UF or UO mostly related to the storage problem, I believe. Most people don't understand how unstable those substances are. By the way, UA is light-sensitive also.
My staining conditions for all stains are: +4oC (if sample permits), adsorption time - 1-2 min; staining 0.5-1% aqueous solution - 1-2 min. I find, that it is better to adjust sample concentration rather than to play with adsorption time. Extended staining time will not dramatically improve the staining but may harm the sample's integrity (it depends, for IgG staining I am using 4 min 1% UA). For UA staining I also prefer to use my favorite "EM" buffer based on ammonium acetate with additives if necessary (50-100 mM ammonium acetate, pH 7.8 in simplest case). It is highly compatible with UA and gives good stain's distribution.
I am sorry if my description is not clear. Please, feel free to call me if any question. It's much simpler to do than read.
God luck Sergey
At 11:08 AM 12/6/00 -0500, you wrote: } Sergey, } Would you mind giving me the details of your treatment of grids with } poly-lysine and the double-carbon technique that you refer to in your } E-mail. I usually go to PTA if PH is a problem. How do you compare PTA } with AM? I am always looking for new methods to deal with some of the } negative stain. problems. } Thanks very much, } Debby } } Debby Sherman Phone: 765-494-6666 } Life Science Microscopy Facility FAX: 765-494-5896 } Purdue University E-mail: } dsherman-at-purdue.edu } 1057 Whistler Building } West Lafayette, IN 47907-1057 } } } On Tuesday, December 5, 2000 10:43 PM, Sergey Ryazantsev } {sryazant-at-ucla.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 } } -----------------------------------------------------------------------. } } } } } } Ammonium molybdate (AM) is suitable in the cases when uranyl acetate (UA) } } is bad (mostly if pH is a critical factor, UA pH = 5-5.5). In most cases AM } } gives less contrast than UA. AM gives you "pure" negative staining because } } do not chemically interfere with most biological samples. UA from another } } hand sometimes gives you mixed contrast, positive } } (DNA/RNA)&negative(proteins). For instance, ribosomes and many viruses } } shown mixed staining with UA. It's not bad because it may even enhance } } some structural details, but you have to remember about that. My own } } experience indicated that in most cases UA or other uranium salts } } (U-formiate or oxalate) after conditions adjusting (concentration, time, } } temperature, freshness, support film) has shown perfect results. If you } } have a problem with stain spreading, you may try different support films } } (carbon, formvar, parlodion). Very good results you may obtain using } } "double carbon" technique - this may solv even very worse cases. I also } } has have a good results with poly-lysine treatment of the grids prior } } sample adsorption. Personally, I don't like the glow-discharge. But it } } works in some way. } } } } Sergey. } } } } At 11:52 AM 12/5/00 -0800, you wrote: } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } Dear all , } } } Does any body use Ammonium Molybdate for Negative staining and also by } } } this chemical visualize structure better then Uranyl acetate. } } } } _____________________________________ } } } } 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 } } } } } } } }
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
We use the Shandon Citadel 1000. It's a good processor, but there's one disadvantage : there's evaporation of your fluids. There's no fume safeguard on the processor, so you have to put it in a fume hood.
Phillip Rutledge schreef:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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
I did not publish this technique yet. So, you may refer to our ListServer. If you need more details - call me.
Sergey.
At 07:52 AM 12/7/00 +0000, you wrote: } Brilliant, Sergey. Have you published these methods anywhere? If } so, I would be interested to have the references. } Best wishes } Chris } } Date sent: Wed, 06 Dec 2000 22:18:27 -0800 } To: Debby Sherman {dsherman-at-purdue.edu} } From: Sergey Ryazantsev {sryazant-at-ucla.edu} } Subject: Re: Negative stain } Copies 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 } } -----------------------------------------------------------------------. } } } } } } Hello Debby } } } } Ted Pella Poly-L-Lysin 0.1% w/v aqueous solution, Cat # 18026 } } Float your grid with film on small drop of the solution for 1 min and than } } remove excess of the liquid by filter paper and put on the drop of water } } for 5-10 sec, blot it and immediately move it on the drop with your } sample. } } Temperature - as necessary for your sample. It also helps to adsorb tuff } } samples. Surprisingly, it seems to me that it does not affect background. } } } } Concerning "double-carbon" technique. It is more tricky. The idea is to } } sandwich your sample between two carbon films just before you finish the } } staining. I prefer the following technique. The approx. 1 ml plastic cap } } (have no idea what is that, you may use any suitable container) 0.7 cm } } diameter I filled with 0.5-1% aqueous UA in the way to make flat meniscus, } } so it will reflect the light. Than I will cut 3x3 mm piece of mica with } } carbon film and float carbon on the UA solution. Reflecting light will } } makes it visible under some particular angle (you have to practice a } little } } bit). So, you have the plastic cap with UA and floated piece of carbon } film } } on it and you can see carbon easily. Than you will adsorb sample on the } } regular EM grid with carbon. Than you have to move your grid into the cap } } with stain (avoid contact to the floated carbon, soak grid into the cap } } away from carbon). Looking through the carbon film, still floated on the } } UA, move the grid just under the film. So, you will see the grid through } } floated film. Than using one smooth movement you have to move grid } } vertically up picking up the carbon film. So, you will put grid just } under } } the film (deep under the surface) and than move grid vertically up. The } } film will holds over the grid with drop of the stain solution. The last } } most important step - to remove excesses of staining solution: fix grid } } securely in the tweezer by rubber O-ring (or use "reverse-action" } tweezer), } } small triangle shape piece of filter paper (I am using Whatmann 1M paper) } } insert between tweezer's "legs" (sorry, guys, I have no idea how it is } } called) as closer to the grid as possible. Filter paper chip should } } contact with excess of liquid holds by capillary force between the } } tweezer's "legs". Keep tweezer with grid and filter paper until grid will } } dry completely (at least 5 min). Enjoy the perfect spreading of the } } stain! There is a number of disadvantages in this method: } } -"double-carbon" will decrease the resolution; } } -sandwich will slightly flat the particles, expect to see 10-15% bigger } } size because of that. } } } } Advantages: } } -sandwich will protect your sample from radiation damage; } } -such samples usually are more stable from all points of view; } } -because of thick carbon, you may use 400 mesh naked EM grids without } } "holey" support film (I still prefer to use the grids with my home-made } } "holey" film). } } } } PTA vs AM. In my point of view, PTA does not have any serious advantages } } over the AM. Personally, I prefer AM. In some particular cases PTA may } } work better, I guess. If there is no chance to use UA, I would try } } U-formiate first and oxalate - next. U-formiate (UF) has pH around 6 or } } even higher (I don't remember the exact number). It gives you a very nice } } fine staining (I love it better than UA). The disadvantage of this } stain - } } it is not stable. You have to prepare fresh solution every time. It is } } light-sensitive also (general room illumination is OK for 20-30 min, no } } direct light, please!). You could store it in the dark in the ice for } } couple of hours. U-oxalate (UO) is less stable than UF, you have to } use it } } immediately. The beauty of UO - you may use it at pH 7. People's } } unsuccess with UF or UO mostly related to the storage problem, I } } believe. Most people don't understand how unstable those substances } } are. By the way, UA is light-sensitive also. } } } } My staining conditions for all stains are: +4oC (if sample permits), } } adsorption time - 1-2 min; staining 0.5-1% aqueous solution - 1-2 min. I } } find, that it is better to adjust sample concentration rather than to play } } with adsorption time. Extended staining time will not dramatically improve } } the staining but may harm the sample's integrity (it depends, for IgG } } staining I am using 4 min 1% UA). For UA staining I also prefer to use my } } favorite "EM" buffer based on ammonium acetate with additives if necessary } } (50-100 mM ammonium acetate, pH 7.8 in simplest case). It is highly } } compatible with UA and gives good stain's distribution. } } } } I am sorry if my description is not clear. Please, feel free to call } me if } } any question. It's much simpler to do than read. } } } } God luck } } Sergey } } } } } } } } At 11:08 AM 12/6/00 -0500, you wrote: } } } Sergey, } } } Would you mind giving me the details of your treatment of grids with } } } poly-lysine and the double-carbon technique that you refer to in your } } } E-mail. I usually go to PTA if PH is a problem. How do you compare PTA } } } with AM? I am always looking for new methods to deal with some of the } } } negative stain. problems. } } } Thanks very much, } } } Debby } } } } } } Debby Sherman Phone: 765-494-6666 } } } Life Science Microscopy Facility FAX: 765-494-5896 } } } Purdue University E-mail: } } } dsherman-at-purdue.edu } } } 1057 Whistler Building } } } West Lafayette, IN 47907-1057 } } } } } } } } } On Tuesday, December 5, 2000 10:43 PM, Sergey Ryazantsev } } } {sryazant-at-ucla.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 } } } } -----------------------------------------------------------------------. } } } } } } } } } } } } Ammonium molybdate (AM) is suitable in the cases when uranyl acetate } (UA) } } } } is bad (mostly if pH is a critical factor, UA pH = 5-5.5). In most } cases AM } } } } gives less contrast than UA. AM gives you "pure" negative staining } because } } } } do not chemically interfere with most biological samples. UA from } another } } } } hand sometimes gives you mixed contrast, positive } } } } (DNA/RNA)&negative(proteins). For instance, ribosomes and many viruses } } } } shown mixed staining with UA. It's not bad because it may even enhance } } } } some structural details, but you have to remember about that. My own } } } } experience indicated that in most cases UA or other uranium salts } } } } (U-formiate or oxalate) after conditions adjusting (concentration, } time, } } } } temperature, freshness, support film) has shown perfect results. If you } } } } have a problem with stain spreading, you may try different support films } } } } (carbon, formvar, parlodion). Very good results you may obtain using } } } } "double carbon" technique - this may solv even very worse cases. I also } } } } has have a good results with poly-lysine treatment of the grids prior } } } } sample adsorption. Personally, I don't like the glow-discharge. But it } } } } works in some way. } } } } } } } } Sergey. } } } } } } } } At 11:52 AM 12/5/00 -0800, you wrote: } } } } } -------------------------------------------------------------------- } ---- } } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } } } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } -------------------------------------------------------------------- } ---. } } } } } } } } } } } } } } } Dear all , } } } } } Does any body use Ammonium Molybdate for Negative staining and also by } } } } } this chemical visualize structure better then Uranyl acetate. } } } } } } } } _____________________________________ } } } } } } } } 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 } } } } } } } } } } } } } } } } } } } } _____________________________________ } } } } 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 } } } } } } } } } } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } 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 07710 585 401 } FAX: +44 (0) 131 653 6248 } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
EMBO Practical Course on Electron Microscopy, Immunocytochemistry and Stereology for Cell Biology at EMBL, Heidelberg, Germany, Aug 30 - Sep 8, 2001
This is a course about the production of thin sections of biological material and their use in studying ultrastructure in the context of molecular cell biology. The course will introduce the techniques of cryosectioning, rapid freezing methods as an alternative to chemical fixation, freeze substitution and resin embedding. It will instruct participants in the sectioning of suitably prepared material and will then concentrate on the use of these sections for localizing specific molecules within cells. Ample time will be given to hands on practical work as well as formal and informal discussion of available techniques for the localization of subcellular molecules.
Further info: http://www.EMBL-Heidelberg.DE/courses/ElectronMicroscopy01/
**************************** Sylke Helbing Course and Conference Office European Molecular Biology Laboratory Meyerhofstr. 1 D-69117 Heidelberg
A general question whether anyone knows where I can obtain information on the metallisation or backside coating (through evaporation) of silicon wafers after backgrinding ? My limited previous metals coating experience has given me the impression that true metal bonding is the result of the formation of a intermetallic/segregated region - of both metallic constituents cooling to form what is basically a weld zone with no discernible porosity interface . A mechanical bond on the other hand would simply be the 'knitting' of rough surfaces . My interest includes the goal of successful metallisation and how it is assessed as normal mechanical preparation of a cross section can tear and smear the face causing difficulty in porosity / interface evaluation . Is FIB used or are there non destructive methods for example ?
Thanks
Martyn Harris ESM LTD Cardiff Rd, Newport , Gwent South Wales , UK . Email : harrism-at-esm-semi.co.uk
I don't think the chloroform has gotten more toxic (unless I forgot about that fact, having used that technique for more years than I can remember....). Hopefully, a lot of us have gotten a tiny bit smarter about exposure to potentially harmful chemicals. Actually, over the years I have developed sensitivity to a number of chemicals in the EM Lab, and still tend to be a little careless about exposure to vapors - especially formalin, glut, osmium, xylene, acetone, EtOH. But I, too, have used the heat pens for well over 20 years (ever since the first cautery pens became available thru one of the EM suppliers), and urge individuals who do a lot of sectioning to switch to the heat pens.
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals
On Wed, 06 Dec 2000 17:20:07 +0000, Malcolm Haswell 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 } -----------------------------------------------------------------------. } } } Timothy } } I would be a bit concerned about your use of chloroform to flatten sections, } especially in a small microtome room. We have used heat pens for the last decade } or so, because apparently the chloroform we use these days is a lot more toxic } than it used to be (as well as formaldehyde, glutaraldehyde, carbon } tetrachloride etc). } } Malcolm Haswell } e.m. unit } School of Sciences } University of Sunderland } UK } } Timothy Schneider 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 following comments are for the person who is trying to cut serial } } sections with Spurrs. I routinely cut serial sections with Spurrs without } } resorting to any kind of glue. Probably the most important thing is how the } } block is faced. I cut the trapezoid with a really long (.6 to .9 mm base) } } and the top of the trapezoid is almost the same length. What's really } } critical here is that those two cuts are parallel. } } The two sides of the trapezoid are very short (.1mm). I cut the sections } } with a Diatome knife and on a Reichert Ultracut E with a cutting speed of } } 0.8mm per second and a thickness setting of 80nm. I am doing this in a } } small room with a blasting air vent in the ceiling. I have cut up a } } cardboard box and stuck it into the ceiling tiles in such a way that the air } } is diverted away from the microtome. I can pick up 15 to 25 sections in a } } row on a 0.4 X 2mm copper slotted grid coated with just formvar or } } formvar/carbon. I have never been any good at making the support films my } } self, so I buy the slotted grids precoated from Ted Pella (catalog #s 01706, } } 01806, or 01816). Before picking up the ribbon, I dip a stick into } } chloroform and wave that over the sections, and they expand (or relax, } } depending on your point of view). I have a bunch of self locking tweezers } } and after picking up the sections, I leave the grid with the sections on it } } locked in the tweezers until they dry (if you put a wet slotted grid down on } } filter paper it might break the film). Good luck, Tim } } } } Timothy G. Schneider } } Director of Electron Microscopy } } Department of Pathology } } Room 229 Jefferson Hall } } Thomas Jefferson University } } 1020 Locust St. } } Philadelphia Pa. 19107 } } 215-503-4798 work } } 610-613-8170 cellular } } timothy.schneider-at-mail.tju.edu } }
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I have an old American Optical Spencer microscope (rescued from a trash bin). It is a binoc, three objective turret (with 4x, 10x, 20x) and built-in light source scope. I have loaned the scope to a young man who is being home schooled, and he has become really interested in microscopy, and aquatic biology. He is at the point that he needs at least a 40x objective, and all I have are the 3 that came with the scope. The only numbers I can find on the scope are 1063-1AA and 1235, and neither is specifically designated as the model or serial numbers. Can anyone direct me to a source for objectives for this type of microscope? These old scopes used a standard thread and tube length, so maybe there are non-manufacturer objectives that I can put on the scope. I haven't been able to look thru an Edmund Scientific catalog (there's one around here somewhere), but I would hope that either a collector or someone at Leica could provide some additional info.
Thanks in advance.
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals
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In principle, wave front reconstruction can be done by:
1) Off-axis holography: This methods need a special biprism for overlapping of two beams (one passed the specimen, the other one passed a hole). For the wavefront construction are two commercial programs available - one from Gatan and the other one from our company. The group around Prof. Hannes Lichte in Dresden is very active in this field - you should contact him, if you want to know details.
2) Focal variation: This method reconstructs the wave front from a focal series with very small defocus steps. For the reconstruction is at least one commercial program available - you should contact Andreas Thust in Juelich, Germany, if you want to know more about it. Also the group from Prof. Dirk van Dyck in Antwerp is very active in this field. The automatic acquisition of focal series is integrated in our programs and we are supporting also the JEOL-4000 remote control, but we are supporting only our cameras, no Gatan cameras. Sorry.
I apologize if I might have forgotten some references - I remember also a poster from Michael O'Keefe in Philadelphia from this year, which was related to this matter.
Regards, Ingo
++++++++++++++++++++++++++++++++++++++++++++++++ Dr. Ingo Daberkow Tietz Video and Image Processing Systems GmbH D-82131 Gauting, Germany Tel: +49-89-8506567 FAX: +49-89-8509488 Internet: www.tvips.com Email: ingo.daberkow-at-tvips.com
-------- Original Message --------
forwarded from the imaging news group
"james patrick birrell" {jbirrell-at-students.uiuc.edu} wrote in message news: {Pine.GSO.4.10.10012041635310.22505-100000-at-ux12.cso.uiuc.edu} ... } I was wondering if anybody would know where I could obtain software to } facilitate the acquisition/analysis of images to be used to perform } wavefront reconstruction using images from a TEM. Specifically, I'm } looking for any software compatable with a JEOL 4000ex microscope with a } Gatan CCD camera. } Thanks, } James Birrell
I have always understood that we use cacodylate buffer because it penetrates tissue faster and further than other buffers, taking the fixative with it. Plus, of course, the advantages that it does not form a precipitate with osmium, and that if you need to store tissues after fixing, then it discourages the growth of microorganisms. It is toxic, but surely not as much so as glutaraldehyde or OsO4, which we can't really avoid in EM. So, I would suggest we all go on using it, and take all the proper precautions for this and the other toxic agents that we need.
Lesley Weston.
On Wed, 6 Dec 2000, Malcolm Haswell 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 } -----------------------------------------------------------------------. } } } Bart } } I have used PIPES as a substitute for cacodylate in most routine procedures } simply because it has none of the precipitation problems of phosphate } buffers. I understand that there may be more extraction with cacodylate in } TEM so I would have thought that it may be a better buffer for SEM (less } chance of shrinkage?). I would assume that HEPES was similar but have no } experience with it. } } These are my own feelings and I have seen no scientific evidence to support } this - has anyone else? } } } Malcolm Haswell } e.m. unit } School of Sciences } University of Sunderland } UK } } Bart De Pauw wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } Hello listers, } } } } We use cacodylate buffer for our SEM samples. Is HEPES or PIPES buffer } } an alternative for the toxic cacodylate ? Our samples are mainly animal } } tissue. } } } } Bart De Pauw } } Ghent University } } Faculty of Veterinary Medicine } } Department Morphology } } Belgium } } }
I recently inherited a 160L liquid nitrogen dewar that I use to supply LN2 to the scopes in our facility. Both the previous director and I feel that we do not get our monies worth, so to speak, from this dewar. I had our local supplier look at it and they say it is fine. Once it gets to 1/2 tank,the remaining LN vaporizes and bleeds off within a day. The last time I had it filled, I calculated that I used about 50L of the 160L before it went dry. Once it got down to half, I dumped the remaining LN into a 34L dewar (and it is stll there). Does everyone have pretty much the same experience with these 160L dewars?
Thanks for your help, and Merry Christmas to all microscopists (who are the salt of the earth!)
Sincerely,
Todd
Dr. Todd A. Kostman Assistant Professor of Biology and Microbiology Director, Electron Microscopy Facility University of Wisconsin Oshkosh 800 Algoma Boulevard Oshkosh, WI 54901-8640 ph: (920) 424-3069 fax: (920) 424-1101 kostman-at-uwosh.edu
Yes, I have also found 160L dewars NOT to be cost effective with only a small user base. I switched to a 35L dewar many years ago. Even with the more frequent deliveries I'm saving money over the 160L.
************************************************ ### Free the Psoas ### **************
Dr. Scott Kuehner kuehner-at-u.washington.edu Dept. of Geological Sciences ph.206-543-8393 Box 351310 Fax 206-616-6873 The University of Washington Seattle, Washington 98195-1310 ************************************************
On Thu, 7 Dec 2000, Todd Kostman 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 EMers, } } I recently inherited a 160L liquid nitrogen dewar that I use to supply LN2 } to the scopes in our facility. Both the previous director and I feel that } we do not get our monies worth, so to speak, from this dewar. I had our } local supplier look at it and they say it is fine. Once it gets to 1/2 } tank,the remaining LN vaporizes and bleeds off within a day. The last time } I had it filled, I calculated that I used about 50L of the 160L before it } went dry. Once it got down to half, I dumped the remaining LN into a 34L } dewar (and it is stll there). Does everyone have pretty much the same } experience with these 160L dewars? } } Thanks for your help, and Merry Christmas to all microscopists (who are the } salt of the earth!) } } } Sincerely, } } } Todd } } Dr. Todd A. Kostman } Assistant Professor of Biology and Microbiology } Director, Electron Microscopy Facility } University of Wisconsin Oshkosh } 800 Algoma Boulevard } Oshkosh, WI 54901-8640 } ph: (920) 424-3069 } fax: (920) 424-1101 } kostman-at-uwosh.edu } } } }
We have had a similar experience here, going to a 160L dewar after having to make a trip across campus with a 30L dewar everytime we needed LN2. The huge discount on the 160L container means we're paying about the same per liter actually used as we were before, plus we no longer have the aggravation of having to go and get it.
One discovery we made, however, is that by transferring LN2 from the 160L dewar to smaller, non-pressurized ones we are able to keep the nitrogen much, much longer. We have 10L, 50L and 30L containers in our lab, so now when a new 160L dewar is delivered, we fill up the smaller ones immediately, then use the 160L until it's gone. The smaller dewars will keep LN2 literally for weeks with minimal losses. Initially we were averaging 60 usable liters per 160 delivered, but now we are probably up way past 100 (I haven't actually figured it out.).
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: Todd Kostman [mailto:kostman-at-vaxa.cis.uwosh.edu] Sent: Thursday, December 07, 2000 12:29 PM To: Microscopy Listserver
Dear EMers,
I recently inherited a 160L liquid nitrogen dewar that I use to supply LN2 to the scopes in our facility. Both the previous director and I feel that we do not get our monies worth, so to speak, from this dewar. I had our local supplier look at it and they say it is fine. Once it gets to 1/2 tank,the remaining LN vaporizes and bleeds off within a day. The last time I had it filled, I calculated that I used about 50L of the 160L before it went dry. Once it got down to half, I dumped the remaining LN into a 34L dewar (and it is stll there). Does everyone have pretty much the same experience with these 160L dewars?
Thanks for your help, and Merry Christmas to all microscopists (who are the salt of the earth!)
Sincerely,
Todd
Dr. Todd A. Kostman Assistant Professor of Biology and Microbiology Director, Electron Microscopy Facility University of Wisconsin Oshkosh 800 Algoma Boulevard Oshkosh, WI 54901-8640 ph: (920) 424-3069 fax: (920) 424-1101 kostman-at-uwosh.edu
I don't agree. I seriously doubt that the diffusion rate of cacodylate and the "Good buffers" (i.e., the buffers like PIPES, HEPES, MES, that were designed by Dr. Good) were ever compared since they are from different eras. I am also not sure I understand how the diffusion rate of a buffer component "takes the fixative with it" at the molecular level. Arsenic buffers do inhibit bacterial growth but I have had bacteria grow in cacodylate buffers. Furthermore, if you have aldehydes or osmium in your buffer, you have all the anti-bacterial agent you need. Cacodylate buffers can give off toxic gases when acidified but more importantly, you are adding needlessly to the toxic waste stream. Aldehydes will eventually react or breakdown into non-toxic substances but the arsenic will remain a problem forever. Cacodylate was originally selected based on its pKa and non-reactivity with fixatives. Phosphate caused problems with calcium precipitation and Tris buffers had amino groups that react with the fixatives. The choices for buffers were very limited in the old days. Dr. Good made a significant contribution in his description of this family of buffers. Anatomists tend to be the most old fashioned scientists in the world.
} } } I have always understood that we use cacodylate buffer because it } penetrates tissue faster and further than other buffers, taking the } fixative with it. Plus, of course, the advantages that it does not form a } precipitate with osmium, and that if you need to store tissues after } fixing, then it discourages the growth of microorganisms. It is toxic, but } surely not as much so as glutaraldehyde or OsO4, which we can't really } avoid in EM. So, I would suggest we all go on using it, and take all the } proper precautions for this and the other toxic agents that we need. } } Lesley Weston. } } } } On Wed, 6 Dec 2000, Malcolm Haswell 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 } } -----------------------------------------------------------------------. } } } } } } Bart } } } } I have used PIPES as a substitute for cacodylate in most routine procedures } } simply because it has none of the precipitation problems of phosphate } } buffers. I understand that there may be more extraction with cacodylate in } } TEM so I would have thought that it may be a better buffer for SEM (less } } chance of shrinkage?). I would assume that HEPES was similar but have no } } experience with it. } } } } These are my own feelings and I have seen no scientific evidence to support } } this - has anyone else? } } } } } } Malcolm Haswell } } e.m. unit } } School of Sciences } } University of Sunderland } } UK } } } } Bart De Pauw wrote: } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } Hello listers, } } } } } } We use cacodylate buffer for our SEM samples. Is HEPES or PIPES buffer } } } an alternative for the toxic cacodylate ? Our samples are mainly animal } } } tissue. } } } } } } Bart De Pauw } } } Ghent University } } } Faculty of Veterinary Medicine } } } Department Morphology } } } Belgium } } } } } }
-- 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 Friends, Once a gain I need that voodoo that you do so well.
A colleague is asking how to embed, freeze and frozen-section (cross section) a rat sciatic perpheral nerve. Several attempts have failed to give adequate structural detail. Any tips and tricks are welcome.
We got similar results when we monitored nitrogen use 10 years ago, and again when we retested earlier this year. We tried two different 160 liter tanks, one of which was brand new, and never got more than about 80 liters out. I eventually gave up and went to four 50 liter tanks, which we receive at atmospheric pressure and tap with a withdrawal device that self pressurizes to about 9 psi. The device has a rubber (now teflon for better thermal properties) transfer hose, so there is little loss due to warming the hose. We usually get 40-45 liters out of these.
The losses, as I understand them, are from
1. passive losses from the tank itself (static evaporation rate) which are higher for the 160 liter tanks 2. losses due to cooling of the transfer tube (lower in the 50 liter tank with no phase separator) 3. depressurization loss, which is higher in the 160 liter tank pressurized to 20 psi than for the 50 L tank at 9 psi.
Obviously if you need high pressure or use high volumes, the 160 liter tank is better. Also, with the 50 liter tanks you have to plan ahead because it takes some time to pressurize the tanks, but otherwise the 50 liter option works for us.
Marie
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Dr. Marie E. Cantino Dept. of Physiology and Neurobiology, U-2131 University of Connecticut Storrs, CT 06269-2131 Phone: 860-486-3588 Fax: 860-486-6369
we seem to be the exception. I expect to recover about 100L from a 160L dewar over a two to three week period. I'm sure we once got a near record 110L out but occasionally fall a little below the 100. We still therefore find the 160L storage dewar more convenient because the alternative would require more space, higher delivery cost and the need for multiple trolleys for several 25 litre dewars. We can normally get nitrogen almost to the bottom of the 160L dewar, providing that we don't leave less than about 10L in it (I'm guessing). The only down-side is that it is a pressure vessel and requires regular maintenance and testing to meet UK regulations, but this means that the spring loaded pressure relief valve is regularly checked (as well as the burst disk valve of course). It does however mean that if our 160L dewar ever fails its tests that we will probably just buy more 25L dewars instead of replacing it.
Good luck
Malcolm Haswell e.m. unit School of Sciences University of Sunderland UK
Todd Kostman 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 EMers, } } I recently inherited a 160L liquid nitrogen dewar that I use to supply LN2 } to the scopes in our facility. Both the previous director and I feel that } we do not get our monies worth, so to speak, from this dewar. I had our } local supplier look at it and they say it is fine. Once it gets to 1/2 } tank,the remaining LN vaporizes and bleeds off within a day. The last time } I had it filled, I calculated that I used about 50L of the 160L before it } went dry. Once it got down to half, I dumped the remaining LN into a 34L } dewar (and it is stll there). Does everyone have pretty much the same } experience with these 160L dewars? } } Thanks for your help, and Merry Christmas to all microscopists (who are the } salt of the earth!) } } Sincerely, } } Todd } } Dr. Todd A. Kostman } Assistant Professor of Biology and Microbiology } Director, Electron Microscopy Facility } University of Wisconsin Oshkosh } 800 Algoma Boulevard } Oshkosh, WI 54901-8640 } ph: (920) 424-3069 } fax: (920) 424-1101 } kostman-at-uwosh.edu
Many thanks to all who replied to this query. I should have thought of the EDAX site, senile dementia is really kicking in now. Just as well I'm being thrown on the scrap heap at Christmas.
I do not know if I can help, but I have a an excellent 50x oil AO objective with a correction collar used on an AO MicroStar series compound microscope. This is the scope with a grey finish. The numerical aperture of this objective is 0.80 and with the correction collar exceeds the light gathering capacity and resolution of any dry 40x or 63x dry objective.
Do you have any items you can perhaps trade, i.e., Zeiss objective lenses?
Let me know what you think.
Ken ------------- Ken Tiekotter, Adjunct Professor The University of Portland Department of Biology 5000 N. Willamette Blvd. Portland, OR 97203
Contact tel. number: 503-413-5391
On Thu, 7 Dec 2000, Roger Moretz 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: } } I have an old American Optical Spencer microscope (rescued from a trash } bin). It is a binoc, three objective turret (with 4x, 10x, 20x) and } built-in light source scope. I have loaned the scope to a young man who is } being home schooled, and he has become really interested in microscopy, and } aquatic biology. He is at the point that he needs at least a 40x objective, } and all I have are the 3 that came with the scope. The only numbers I can } find on the scope are 1063-1AA and 1235, and neither is specifically } designated as the model or serial numbers. Can anyone direct me to a source } for objectives for this type of microscope? These old scopes used a } standard thread and tube length, so maybe there are non-manufacturer } objectives that I can put on the scope. I haven't been able to look thru an } Edmund Scientific catalog (there's one around here somewhere), but I would } hope that either a collector or someone at Leica could provide some } additional info. } } Thanks in advance. } } Roger Moretz, Ph.D. } Dept of Toxicology } Boehringer Ingelheim Pharmaceuticals } } } } } } _______________________________________________________ } Tired of slow Internet? Get -at-Home Broadband Internet } http://www.home.com/xinbox/signup.html } } }
I receive my liquid nitrogen in 160 L tanks from our supplier and they generally last from 24 to 30 days with me filling up the EDX dewar twice a week (about 3-4 liters). I have had some tanks that only lasted two weeks and after I told our supplier about that they said the leak valve was stuck open and they replaced or adjusted the leak valve as needed. You should get your local supplier to inspect your tank and maybe replace or adjust the leak valve. I would not adjust it myself unless you really know what you are doing since an explosion is possible without proper pressure release. Terry Ellis Hallmark Cards Inc. e-mail: tellis2-at-hallmark.com
Imagine all day sectioning, it isn't hard to do. Putting sections on bare 200 mesh grids and heat fixing under a light to boot.
Imagine all the sections coming off while you do the stain. Oh oh You have to figure out what's wrong, while racking your brain. If you all can help, you'll help me from going insane.
Any suggestions will be greatly appreciated. Thanks is advance!
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Dept. of Pathology, Ross Hall rm 505 Electron Microscope Lab 2300 Eye St. Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
We have a post-doc that is interested in looking at tight junctions in cultured endothelial cells. He has found a vague reference for using silver stain to visualize endothelial cell junctions. Does anyone have any experience with this technique? Can you provide some technical details or point us to a reference? Thank you!!
ctfExplorer has been updated. New features/improvements: 1. Now it is possible to export graphs to text (tab-delimited). 2. List-tree with the microscopes is wider now. 3. Tab stops in dialogs arranged in a logical sequence. 4. Now it calculates the Lichte-defocus. (Useful for holography). 5. Additional slider-control is added: allows to change "magnification", i.e. convergence. Just to show people why it is not always a good idea to work at 1,000,000x and why some people do high resolution work at as low as 50,000x magnification.
Here is the link: http://clik.to/ctfexplorer (always use this link. it will direct you to the right location).
Enjoy,
Max Sidorov --- (until December 31 2000) TEM Applications Specialist FEI/Philips Electron Optics Eindhoven, The Netherlands e-mail: maxsidorov-at-bigfoot.com (mailto:maxsidorov-at-bigfoot.com)
----------Additional Info---------- Dear All: I wrote a piece of software which I believe would be of interest to the microscopy community. It's a highly interactive program which calculates/displays the contrast transfer function of TEMs. I know that there are similar programs floating around but ctfExplorer does not only 1d but also 2d calculations/display with 2-fold and 3-fold astigmatism imposed. There are other unique features to it. All parameters (defocus, voltage, Cs, etc) can be changed interactively.
DISCLAIMER: I do work for FEI/Philips Electron Optics. However, this software is not an official product of FEI and FEI is not responsible for its distribution/support. This software is freeware.
To the best of my knowledge, the software does not have any nasty bugs. It is tested under Windows 95/98 and Windows NT4.
Please give it a try. Please direct your suggestions and comments to maxsidorov-at-bigfoot.com I hope you'll find the software useful.
Here is the link: http://clik.to/ctfexplorer
Enjoy,
Max Sidorov --- TEM Applications Specialist FEI/Philips Electron Optics Eindhoven, The Netherlands e-mail: maxsidorov-at-bigfoot.com
Todd- I have not calculated how much LN2 I am able to deliver from my 160 L dewar, but it must be about 100 L since it will last me a month or more. Perhaps my dewar is of a better grade as it is also certified for liquid He use. I suspect that much of my LN2 loss is in cooling down the 10 L dewar I use to transport it to the EDX dewar. Are you using a high pressure, or low pressure dewar (mine has ~24psi max)? The high pressure dewars I have had have an internal coil for warming up some liquid to maintain the head pressure. If your dewar has this, there should be valves for closing off this pressure generation coil which will extend the storage time. By the way, what does your fill gauge read when the dewar is empty? These gauges are frequently very inaccurate. The fact that you could fit the remainder in a 34 L dewar when your gauge said 1/2 full, indicates to me that your gauge probably isn't accurate. I hope that something in there helps... Matt
Todd Kostman {kostman-at-vaxa.cis.uwosh.edu} on 12/07/2000 01:29:29 PM
To: Microscopy Listserver {Microscopy-at-sparc5.microscopy.com} cc:
Dear EMers,
I recently inherited a 160L liquid nitrogen dewar that I use to supply LN2 to the scopes in our facility. Both the previous director and I feel that we do not get our monies worth, so to speak, from this dewar. I had our local supplier look at it and they say it is fine. Once it gets to 1/2 tank,the remaining LN vaporizes and bleeds off within a day. The last time I had it filled, I calculated that I used about 50L of the 160L before it went dry. Once it got down to half, I dumped the remaining LN into a 34L dewar (and it is stll there). Does everyone have pretty much the same experience with these 160L dewars?
Thanks for your help, and Merry Christmas to all microscopists (who are the salt of the earth!)
Sincerely,
Todd
Dr. Todd A. Kostman Assistant Professor of Biology and Microbiology Director, Electron Microscopy Facility University of Wisconsin Oshkosh 800 Algoma Boulevard Oshkosh, WI 54901-8640 ph: (920) 424-3069 fax: (920) 424-1101 kostman-at-uwosh.edu
Paula, I loved your poem! I too , had the same problem with sections falling off grids while staining, especially if using the thin bar grids. There's just not enough surface area for the sections to stick to. I solved the problem by putting the grids in the 60 degree oven overnight before staining. I haven't lost any sections since, and I cut both very large and small sections. The time in the oven doesn't seem to adversely affect the tissue.
Good luck, Mary Gail Engle Electron Microscopy & Imaging Facility University of Kentucky
Thanks to all who have already answered my plea. I think I need to make a few things clear. Me, in my effort to be witty forgot a few major details.
1. I'm cutting Spurr's embedded samples.
2. I'm cleaning the samples in a dilute acetic acid followed by an acetone chaser.
3. I've been heat fixing the grids/sections under a light for about 10 minutes.
4. I've been doing this for that past 10 months and things worked great until 3 weeks ago. I'm new to this area (Washington, DC) having been in California doing EM for years. Could this be a climate/weather related problem?
5. I will use formvar if I have to. Has anyone just dipped grids in formvar, instead of casting a film, to make the grids a little sticky? I inherited a bunch of filthy formvar with water & junk in it and it takes about a month to get supplies.
6. I haven't changed my supplier of grids or Spurr's components. So I think that things should be OK on the quality control end.
A lot of the suggestions I received are very good, but I have already tried most of them. Any bizarre rituals that I must do? I WILL NOT swing a dead cat over my head, but I might try other things. EM is basically voodoo, after all.
Thanks again,
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Dept. of Pathology, Ross Hall rm 505 Electron Microscope Lab 2300 Eye St. Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
{html} {head} {meta Name='keywords' Content='commtouch, pronto, mail, free email, free, branded, web based, free web based email, communications, internet, software, advertising banners, e-mail, free software'} {/head} {body } {div align='left'} {font } {blockquote} {blockquote} {TT} Hello, {BR} {BR} Please add me to the microscopy discussion group. {BR} {BR} Thanks, {BR} Darren {BR} {BR} {/TT} {br} {br} {font} {p align=left} {br} Get your Free E-mail at http://thunderdome.zzn.com {br} ____________________________________________________________ {br} Get your own FREE Web and POP E-mail Service in 14 languages at http://www.zzn.com. {br} {/blockquote} {/blockquote} {/div} {/font} {/body} {/html}
Hi Paula, It is horrible to see naked grids come off the stain. We've probably all been there. Here are my auggestions:
1. Make sure that your grids are clean to start. You can dip them in acetone or ethanol (absolute) and then water and dry thoroughly.
2. Make sure that the sections have fully dried onto the grids. If you're not rushed, just let them dry overnight. If you are rushed (probably the usual situation), you can lay the grids out on a clean glass slide that is resting on a warm (NOT HOT) hot plate (around 35-40deg.C) for a few minutes.
3. And as wierd as this seems....make sure that the water you are using to make up your stains and to wash the grids is really clean (deionized or double distilled or microfiltered). I had a friend to used to come over to my building to fill a small carboy with water because my dept. had better water treatment than hers. She swore that her sections fell off when she used their water.
4. Maybe the simplest life saver....buy a "Coat-Quick G" grid coating pen. It deposits a small amount of the "secret formula" adhesive onto the grid bars, without blocking the spaces or causing problems with dirt/stain deposition. I frequently cut enormous sections (by EM standards) and this has been a blessing. I know that Electron Microscopy Sciences sells them. Other companies probably do too. Look in you favorite vendor's catalogue.
(Disclaimer: I have no financial interests in EMS)
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
Responding to the message of {sa30a2a7.040-at-gwise.gwumc.edu} from "Paula Sicurello" {patpxs-at-gwumc.edu} :
Paula,
Imagine........ a magic solution........
GRID CLEANING SOLUTION
Clean your uncoated grids each time you set up to do sectioning, because newly cleaned copper grids will oxidize within a day and sections will no longer stick to them very well during staining and rinsing.
*ADD 40 ml CONCENTRATED HCL TO 400 ml DISTILLED WATER
*ADD 40 ml ACETONE TO THE ABOVE
*DILUTE TO 500 ml FINAL VOLUME
*SONICATE GRIDS IN THIS SOLUTION IN A 50 ml BEAKER FOR 1-2 MINUTES
*DISCARD USED SOLUTON DOWN DRAIN AND RINSE/SONICATE GRIDS FOR 30 SECONDS TWICE WITH 100% ACETONE
*DUMP GRIDS ONTO CLEAN FILTER PAPER TO DRY
* ITS TRADITIONAL TO COLLECT SECTIONS ON THE DULL SIDE OF THE GRIDS.
It works well for us here, and is used routinely every day. Good luck,
Gib
} } Happy holidays oh great wise ones of microscopy! } } (Apologies to John Lennon) } } } Imagine all day sectioning, } it isn't hard to do. } Putting sections on bare 200 mesh grids } and heat fixing under a light to boot. } } Imagine all the sections coming off } while you do the stain. } Oh oh } You have to figure out what's wrong, } while racking your brain. } If you all can help, } you'll help me from going insane. } } } Any suggestions will be greatly appreciated. Thanks is advance! } } Paula :-) } } Paula Sicurello } George Washington Univ. Medical Center } Dept. of Pathology, Ross Hall rm 505 } Electron Microscope Lab } 2300 Eye St. } Washington, DC 20037 } 202-994-2930 phone } 202-994-2518 fax
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'm looking for methods of preparing compacted potassium chloride fertilizer for the SEM. We believe that grain size is indicative of compaction quality in our process, however we're not sure of the best way to prepare the samples. The compactor flake is approximately 2"x1"x0.25". Any advice would be much appreciated.
I have a freind that need then main prism slide containing the Wollaston prism for Reichert Zetopan DIC scope. This is the slide going in the upper body of the scope. He also needs the polorizor that fits over the light soruce but that is of much less importance beause it can be easily made. While the prism is constructed of unobtainium.
He is willing to pay a reasonable price for the part.
If any of you have a spare or left over you could make a very dissapointed man a lot happier.
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
I have noticed that in some cases the kind of grid you use is quite important. I had similar problems with Gilder grids when using Spurr embedded plant tissue sections. Later I switched to Veco grids, the problem went away.
Good luck,
Soumitra
************************************************************* Soumitra Ghoshroy Ph.D. Electron Microscopy Lab and Fluorescence Imaging Facility Department of Biology Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3600 Fax: 505-646-5665 e-mail:sghoshro-at-nmsu.edu http://confocal.nmsu.edu/eml
Sandy Hancock Try the following reference; Zand, Underwood, Nunnari, Majno and Joris, Endothelium and 'silver lines'. An electron microscopic study. 1982 Virchows Archiv of Path Anat. 395: 133-144 Good luck John
Thanks to everyone for the response. It will take me a little while to fully sort through everything, but I appreciate the help.
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals
On Fri, 8 Dec 2000 02:31:47 -0800 (PST), Ken Tiekotter wrote:
} Dear Roger, } } I do not know if I can help, but I have a an excellent 50x oil AO } objective with a correction collar used on an AO MicroStar series compound } microscope. This is the scope with a grey finish. The numerical aperture } of this objective is 0.80 and with the correction collar exceeds the light } gathering capacity and resolution of any dry 40x or 63x dry objective. } } Do you have any items you can perhaps trade, i.e., Zeiss objective lenses? } } Let me know what you think. } } Ken } ------------- } Ken Tiekotter, Adjunct Professor } The University of Portland } Department of Biology } 5000 N. Willamette Blvd. } Portland, OR 97203 } } Contact tel. number: 503-413-5391 } } On Thu, 7 Dec 2000, Roger Moretz 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: } } } } I have an old American Optical Spencer microscope (rescued from a trash } } bin). It is a binoc, three objective turret (with 4x, 10x, 20x) and } } built-in light source scope. I have loaned the scope to a young man who is } } being home schooled, and he has become really interested in microscopy, and } } aquatic biology. He is at the point that he needs at least a 40x objective, } } and all I have are the 3 that came with the scope. The only numbers I can } } find on the scope are 1063-1AA and 1235, and neither is specifically } } designated as the model or serial numbers. Can anyone direct me to a source } } for objectives for this type of microscope? These old scopes used a } } standard thread and tube length, so maybe there are non-manufacturer } } objectives that I can put on the scope. I haven't been able to look thru an } } Edmund Scientific catalog (there's one around here somewhere), but I would } } hope that either a collector or someone at Leica could provide some } } additional info. } } } } Thanks in advance. } } } } Roger Moretz, Ph.D. } } Dept of Toxicology } } Boehringer Ingelheim Pharmaceuticals } } } } } } } } } } } } _______________________________________________________ } } Tired of slow Internet? Get -at-Home Broadband Internet } } http://www.home.com/xinbox/signup.html } } } } } } }
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Paula Try this cleaning procedure for your copper grids: 1) Place your grids in a clean beaker, 100ml size will do. 2) Cover the gird with Glacial Acetic Acid, approximately 20cc (the idea here is to get the grim and grease off the grids not to eat the metal away) 3) Sonicate the grids in this acid for about 5 to 10 minutes. 4) Rinse in 100% acetone until the smell of the acetic acid is gone. 5) Do a final rinse in 100% ETOH (denatured is ok) and pipette the ETOH off and invert the beaker into a glass petri dish that has a clean whatman filter paper in it and place in your 60 degree over for an hour or more. The grids will fall on to the filter paper. Cover the bottom of the glass with a top. The grids will keep without father cleaning until they are all used up. I use a glass petri dish because there is less static charge with glass.
Ron Austin Dept. of Pathology LSU Medical Center Shreveport, LA rla-at-mindspring.com 318-675-4557
I'd just try a simple first shot approach of putting some of the particles on a sticky tab, and coating it with about 60A of Au/Pd. Then, take a look at it under the SEM. Based on this imaging, other approaches may or may not be necessary.
It may or may not make a difference relative to the type of SEM that you are using (FESEM vs. thermionic). I would think that these are rather large specimens. Either system should do a nice job.
gary g.
At 01:26 PM 12/8/00, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} From time to time, someone opens a grid storage box in our lab, and because of the static that has built up on the plastic, the grids fly out and are randomized. We have a Zerostat gun next to the TEM, but our laboratory/office space is fragmented worse than a SWAP file, and one can't simply reach for the ion gun every time one opens a grid box. Does anyone (a) have any simple solutions to this problem (b) know of a supplier of grid boxes made of a different plastic that doesn't charge up so horribly?
In the longer term, would there be scope for making grid boxes out of a plastic loaded with a filling to make it conductive enough for charges to slowly leak away, so avoiding the problem altogether?
And on a similar theme, does anyone know of storage boxes for SEM stubs into which the stubs can fit easily, i.e. not having to be rammed in?
+------------------------------------------------------------------------+ | 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 | +------------------------------------------------------------------------+
Antistatic sprays are available from electronic components suppliers such as Farnell and RS Components. These are mainly intended to de-static the plastic covers of ammeters, etc. to stop them influencing the readings.
You could also try coating the interior surfaces of your box with a thin layer of carbon or gold next time you have the coater running.
Chris
Date sent: Sat, 9 Dec 2000 14:59:05 +0000 (GMT Standard Time) } From: "Robert H. Olley" {r.h.olley-at-reading.ac.uk} To: Microscopy Newsgroup {Microscopy-at-sparc5.microscopy.com} Copies to: # {r.h.olley-at-reading.ac.uk}
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Robert Olley wrote: ================================================================ } From time to time, someone opens a grid storage box in our lab, and because of the static that has built up on the plastic, the grids fly out and are randomized. We have a Zerostat gun next to the TEM, but our laboratory/office space is fragmented worse than a SWAP file, and one can't simply reach for the ion gun every time one opens a grid box. Does anyone (a) have any simple solutions to this problem (b) know of a supplier of grid boxes made of a different plastic that doesn't charge up so horribly?
In the longer term, would there be scope for making grid boxes out of a plastic loaded with a filling to make it conductive enough for charges to slowly leak away, so avoiding the problem altogether?
And on a similar theme, does anyone know of storage boxes for SEM stubs into which the stubs can fit easily, i.e. not having to be rammed in? ================================================================= There are a number of grid boxes being molded today, some of which are being molded with plastics that have had added antistatic additives. The SPI Slide-A-Grid™ storage boxes (see website below) are molded with such antistatic additives. This same box is offered (but under other names) by some of the other leading firms offering these kinds of products to EM users . However, under certain conditions, apparently humidity related, even these boxes reportedly can develop a charge.
For SEM storage boxes, there are two approaches to the "base plate" that sits in the bottom of each box, the first being a "hard" plastic such as polypropylene and the other being with a "soft" plastic, specifically an elastomeric plastic. SPI Supplies has offered storage boxes with the soft plastic for some number of years, the advantage being that the mounts do fit in easily and do not have to be "rammed" in. I do not know of anyone else who is molding mounting plates out of the soft "elastomeric plastic" (but of course I could be wrong about that).
Note: One of the biggest sources of misfit between the storage boxes and the 3/8" (9.5 mm) round mounts is that boxes molded in the USA tend to be made for 3/8" diameter mounts and boxes molded elsewhere seem to be made for 10 mm diameter mounts. The problem arises when one tries to put a 10 mm round mount into a box designed for the 3/8"round mounts. If the "ramming" problem you described could be this kind of a problem, a mixing up of boxes/mounts might be the reason for the described difficulty. The flip- side of this problem is when 3/8" mounts are put into boxes molded in Europe to take 10 mm round mounts, and the result is, the mounts fall out when the box is tipped.
Disclaimer: SPI Supplies offers boxes for the storage of both TEM grids and SEM mounts, the details of which can be found on our website below.
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
TECHNICAL OFFICER/SENIOR TECHNICAL OFFICER/CHIEF TECHNICAL OFFICER ELECTRON MICROSCOPE UNIT
If you have experience in electron microscopy and the use of computers, then we invite you to apply for this post in a fast-moving and ever-changing environment geared to servicing research needs.
The Electron Microscope Unit (EMU) primarily serves researchers in the faculties of Science, Engineering and the Built Environment and Health Sciences at UCT. The EMU aims to be the foremost provider of such services in the Western Cape. Currently, the Unit has two scanning and two transmission electron microscopes as well as a number of light microscopes and will take delivery of a cryo-TEM early next year.
Duties of the appointee will include instrument operation, sample preparation, user training and laboratory management.
The level of appointment will depend on the qualifications and the experience of the successful candidate. Therefore, the remuneration package, including benefits, is negotiable between R75 000 - R156 100 a year.
Please send a letter of application plus your CV (including the names, postal/email addresses, telephone/fax numbers of 2 referees) to Associate Professor Trevor Sewell, Director: Electron Microscope Unit, UCT, Rondebosch, 7701 by 8 January 2001; tel: (021) 650-2817; fax: 689-1528; email: sewell-at-uctvms.uct.ac.za
During my laboratory days I never worked in a room were static was that bad - and that included Canadian winters. Carpet and poor humidification can make a static problem worse. I never liked the boxes with a sliding lid for another reason: If they dropped when open from a modest height several grids may escape and that is really the same problem. Probably all of the EM suppliers carry grids boxes that hold 24 grids and have a rotating lid with two apertures. Only one grid at a time is can to escape. Presto: No more feral grids.
I'm sure that "smarter" stub boxes could be designed, but would people buy them at 2 or 3 times the price? We (and others) have a single SEM mount Storage/mailer and this is much better for holding pin type mounts. Wouldn't is be lovely if all EM manufacturers used the same (I vote for pin type) mounts. 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 ABN: 99 724 136 560 www.proscitech.com
On Sunday, December 10, 2000 12:59 AM, Robert H. Olley [SMTP:r.h.olley-at-reading.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 } -----------------------------------------------------------------------. } } } } } From time to time, someone opens a grid storage box in our lab, and } because of the static that has built up on the plastic, the grids fly out } and are randomized. We have a Zerostat gun next to the TEM, but our } laboratory/office space is fragmented worse than a SWAP file, and one } can't simply reach for the ion gun every time one opens a grid box. Does } anyone (a) have any simple solutions to this problem (b) know of a } supplier of grid boxes made of a different plastic that doesn't charge up } so horribly? } } In the longer term, would there be scope for making grid boxes out of a } plastic loaded with a filling to make it conductive enough for charges to } slowly leak away, so avoiding the problem altogether? } } And on a similar theme, does anyone know of storage boxes for SEM stubs } into which the stubs can fit easily, i.e. not having to be rammed in? } } +------------------------------------------------------------------------+ } | 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 behalf of the Microscopy Society of America and the Microbeam Analysis Society, we invite you to attend Microscopy and Microanalysis 2001, August 5-9, in Long Beach, California. The program chair, Bob Price, and Co-Chairs Edgar Voelkl (MSA) and Inga Holl Musselman (MAS), have assembled another excellent technical program that continues the tradition of exciting and current presentations at the meetings. An outstanding list of invited speakers highlights the symposia in a number of key areas. Special symposia this year again cover a wide range, including "Microscopy and Microanalysis in the Real World," a special biological symposium honoring Dr. Inou*, and "Atom Location by Channeling Enhancement of X-Ray and EELS Signals" among others, plus a wide variety of applications in biology and materials research.
Both invited and contributed abstracts are critical to the success of these symposia, so we urge you to submit your work. Remember that the abstract deadline is February 15th, 2001.
A special Pre-meeting Congress, "Imaging Life: From Cells to Whole Animals," as well as several workshops and short courses on important, basic techniques will precede the meeting. Special symposia, tutorials and presentations sponsored by the Technologists' Forum, the MSA Education Committee, and various commercial exhibitors will be held during the course of the meeting. There are also Presidential Happenings, ceremonies for Award Winners, and the world's largest display of microscopes and related equipment.
Our Local Arrangements Committee, headed by Robert Koch, has arranged an excellent venue at the Long Beach Convention Center for both the scientific sessions and the exhibits. They have also arranged a memorable program of social events including a Sunday night reception on the Queen Mary, the annual golf tournament, a harbor cruise, and a large number of options for enjoying the city and the surrounding area (don't forget the theme parks!). With its rich history, wealth of educational institutions and active industry, Long Beach provides a wonderful context for this meeting, and we urge you to come to learn, teach, share and above all to enjoy. Whatever your connection to microscopy, we look forward to welcoming you to Long Beach in August at Microscopy and Microanalysis 2001.
Ron Anderson, President Microscopy Society of America
Richard Linton, President Microbeam Analysis Society
--------------------------
The program is as usual extensive and detailed information can be found on the WWW page.
I have fought static problems in wide variety of problems and three things help. Grounding with metal, raising the humidity to 50% and spraying the area or items with antistatic products designed for laundry. Downy was the one I used. Temporary relief can be had by spraying the area with water from Windex like spray bottle. This only last for a few minutes to an hour at most.
I am sure fabric antistatic spray would not be acceptable in many places in a lab. But floors and tables might benifit from them.
Spraying carpets with antistatic spray once a month work for us in 15 to 20% humidity.
Your mileage my vary. Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
} From: "Jim at ProSciTech" {jim-at-proscitech.com} } During my laboratory days I never worked in a room were static was that bad - } and that included Canadian winters. Carpet and poor humidification can make a } static problem worse. } I never liked the boxes with a sliding lid for another reason: If they dropped } when open from a modest height several grids may escape and that is really the } same problem. } Probably all of the EM suppliers carry grids boxes that hold 24 grids and have } a rotating lid with two apertures. Only one grid at a time is can to escape. } Presto: No more feral grids. } } I'm sure that "smarter" stub boxes could be designed, but would people buy } them } at 2 or 3 times the price? } We (and others) have a single SEM mount Storage/mailer and this is much better } for holding pin type mounts. } Wouldn't is be lovely if all EM manufacturers used the same (I vote for pin } type) mounts. } Cheers } } } } } From time to time, someone opens a grid storage box in our lab, and } } because of the static that has built up on the plastic, the grids fly out } } and are randomized. We have a Zerostat gun next to the TEM, but our } } laboratory/office space is fragmented worse than a SWAP file, and one } } can't simply reach for the ion gun every time one opens a grid box. Does } } anyone (a) have any simple solutions to this problem (b) know of a } } supplier of grid boxes made of a different plastic that doesn't charge up } } so horribly? } } } } In the longer term, would there be scope for making grid boxes out of a } } plastic loaded with a filling to make it conductive enough for charges to } } slowly leak away, so avoiding the problem altogether? } } } } And on a similar theme, does anyone know of storage boxes for SEM stubs } } into which the stubs can fit easily, i.e. not having to be rammed in? } } } }
Kevex Quantum EDS system with Sesame drive is available for no charge for pick-up in Easton, PA. The detector is NOT included. System has Syquest drives and monitor is in need of repair.
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I am doing some bacterial counting work. My basic method involves taking some bacteria in a measured quantity of suspension and running it thru an anodisc (aluminum oxide) filter having .2 micron pores.
My problem is this: when mounting the filter on a slide, I find that the disc is almost dry before I can mount it. I have tried adding water and or oil below and above the filter. Water sometimes works but it is a very tricky matter because the water may redistribute the bacteria when applied. This affects my sampling since the bacteria may no longer be uniform.
The same problem occurs with oil. Furthermore, the oil, if applied to a filter with some water remaining, gives a blurry image, perhaps due to the oil-water mix.
I am also concerned with air bubbles. Is there a method by which one can prevent air bubbles under, over and in the pores of the filter?
I would *STRONGLY* recommend against using any anti-static spray on grid boxes themselves unless you are certain of the components of the spray. I remember one batch of grid boxes we had to throw away because the formulation of the plastic mix during manufactur was evidently wrong, and we got horrendous contamination of samples which had been stored in the boxes. We don't even touch the tweezers we are going to use for our samples with our bare hands! I would never spray some unknown material intended for some other, much less critical (in terms of cleanliness) application, anywhere near anything related to the samples themselves.
Tony G-R
} } Antistatic sprays are available from electronic components suppliers } such as Farnell and RS Components. These are mainly intended to } de-static the plastic covers of ammeters, etc. to stop them } influencing the readings. } } You could also try coating the interior surfaces of your box with a } thin layer of carbon or gold next time you have the coater running. } } Chris } }
} } } } } From time to time, someone opens a grid storage box in our lab, and } } because of the static that has built up on the plastic, the grids fly out } } and are randomized. We have a Zerostat gun next to the TEM, but our } } laboratory/office space is fragmented worse than a SWAP file, and one } } can't simply reach for the ion gun every time one opens a grid box. Does } } anyone (a) have any simple solutions to this problem (b) know of a } } supplier of grid boxes made of a different plastic that doesn't charge up } } so horribly? } } } } In the longer term, would there be scope for making grid boxes out of a } } plastic loaded with a filling to make it conductive enough for charges to } } slowly leak away, so avoiding the problem altogether? } } } } And on a similar theme, does anyone know of storage boxes for SEM stubs } } into which the stubs can fit easily, i.e. not having to be rammed in? } }
Apparently, Bravenet (who provides a quick link url for me) is down for updates (at least that's what they say). The quick link (http://clik.to/ctfexplorer) should start working again shortly. In a meantime you can use a direct link: http://www.ctfexplorer.f2s.com
Hope this helps, Max S
------------------- Dear All:
ctfExplorer has been updated. New features/improvements: 1. Now it is possible to export graphs to text (tab-delimited). 2. List-tree with the microscopes is wider now. 3. Tab stops in dialogs arranged in a logical sequence. 4. Now it calculates the Lichte-defocus. (Useful for holography). 5. Additional slider-control is added: allows to change "magnification", i.e. convergence. Just to show people why it is not always a good idea to work at 1,000,000x and why some people do high resolution work at as low as 50,000x magnification.
Here is the link: http://clik.to/ctfexplorer (always use this link. it will direct you to the right location).
Enjoy,
Max Sidorov --- (until December 31 2000) TEM Applications Specialist FEI/Philips Electron Optics Eindhoven, The Netherlands e-mail: maxsidorov-at-bigfoot.com (mailto:maxsidorov-at-bigfoot.com)
----------Additional Info---------- Dear All: I wrote a piece of software which I believe would be of interest to the microscopy community. It's a highly interactive program which calculates/displays the contrast transfer function of TEMs. I know that there are similar programs floating around but ctfExplorer does not only 1d but also 2d calculations/display with 2-fold and 3-fold astigmatism imposed. There are other unique features to it. All parameters (defocus, voltage, Cs, etc) can be changed interactively.
DISCLAIMER: I do work for FEI/Philips Electron Optics. However, this software is not an official product of FEI and FEI is not responsible for its distribution/support. This software is freeware.
To the best of my knowledge, the software does not have any nasty bugs. It is tested under Windows 95/98 and Windows NT4.
Please give it a try. Please direct your suggestions and comments to maxsidorov-at-bigfoot.com I hope you'll find the software useful.
Here is the link: http://clik.to/ctfexplorer
Enjoy,
Max Sidorov --- TEM Applications Specialist FEI/Philips Electron Optics Eindhoven, The Netherlands e-mail: maxsidorov-at-bigfoot.com
Microscopy Society of America Undergraduate Research Scholarship Program
With this year's call for applications the MSA Undergraduate Research Scholarship Program begins its 13th year providing funding for undergraduate research. To date over 65 projects covering a wide range of topics in the physical and biological sciences have received support through this program. Over the years nearly all the scholarship recipients have maintained a strong interest in imaging sciences and have gone on to graduate school, professional school, teaching, or industry positions.
The program, which is funded by MSA, is able to support approximately 30% to 40% of applicants. The maximal award from MSA is currently $3000 and helps to provide student stipends, supply costs, and limited travel expenses associated with the research. Additional support in the form of instrument use time, equipment purchases, etc. is generally provided by the studentís supervisor and/or through the sponsoring institution. Abstracts reporting the research results, are prepared by scholarship awardees, and published in "Microscopy and Microanalysis".
The program actively seeks external sources of matching funds in order to maintain the favorable levels of support both in terms of the number of projects supported and the level of support for each. We are extremely grateful for the matching support provided by MSA sustaining members and individuals. Their support has enabled the program to increase both the number of awards and the maximum amount of each award to $3000.
The MSA Undergraduate Research Scholarship Program is currently soliciting applications from students interested in conducting a research project which involves the use of any microscopy technique. Students must be sponsored by a member of MSA. The maximal award is $3000.
This is not limited to students in the USA or North America. Any undergraduate student anywhere is eligible to apply for the award, provided the person supervising their research is a member of MSA.
**The application deadline is Dec 31, 2000.**
Applications can be obtained from the MSA Business Office, businessoffice-at-msa.microscopy.com or call toll free at (800) 538-3672.
If you have any questions or require additional information regarding the program please contact either:
Dr. Ralph Albrecht, University of Wisconsin 1656 Linden Drive, Madison, WI 53706 (608) 263-3952 or 263-4162; (608) 262-7420 FAX; albrecht-at-ahabs.wisc.edu
Dr. Richard Ornberg, Monsanto Company Analytical Sciences Center, 800 N. Lindbergh Blvd., St. Louis, MO 63167 (314) 694-1184; (314) 694-6727 FAX; rlornb-at-ccmail.monsanto.com
FOR SALE: 1988 JEOL 100CXII with (+/-) 45degree tilt stage, dual sample holder, and Gatan camera system: a rugged, reliable transmission electron microscope in perfect working order, with water chiller: $30,000. Buyer is responsible for take-down, shipping, and set-up costs. A Kevex Delta I Class EDS analysis unit is also included, but currently not installed.
Feel free to come by for a test drive! 317 Egan Research Center Northeastern University Boston, MA 02115 For directions and to set up an appointment, call (617) 373-5909.
If interested in purchasing, please contact: Professor Terry K. Baker (617) 373-2123
RJ Lee Group's Bay Area Facility, a materials characterization and consulting laboratory, is seeking a motivated individual to actively manage projects related to the evaluation of materials and processes used in the electronics industry. Job applicants should have an MS or equivalent in physics, materials science or related discipline, at least two years of relevant work experience, and be amenable to working in a hands-on and fast-paced team environment. A working knowledge of the application and theory associated with sample preparation methods and corresponding analytical techniques such as optical microscopy, scanning electron microscopy (SEM), transmission electron microscopy (TEM), Auger spectroscopy and other surface analysis methods is critical. Strong oral and written communication skills are a must. Job duties would involve, but not be limited to: materials consulting, developing / evaluating analytical approaches to constructively investigate problems, and presenting laboratory results to a variety of technical as well as non-technical personnel. Salary is commensurate with experience. Interested candidates should forward qualifications along with salary history to: RJ Lee Group, Inc., Attn. Human Resources, 350 Hochberg Road, Monroeville, PA 15146.
RJ Lee Group is an Equal Opportunity Employer
Drew R. Van Orden, PE RJ Lee Group, Inc. 350 Hochberg Road Monroeville, PA 15146 (724) 387-1869 drew-at-rjlg.com
I am new to this group, so if this is question has been covered, please excuse. I usually do TEM on tissue from biopsies, etc., and now someone wants EM on cells grown in culture. I have done this for intact monolayers (very tedious), but these would be on trypsined-off and pelleted cells. I need to know if there is an easy way to handle them as an entire pellet, so I don't have to cfg. after each fixation/dehydration step, and so they will be compacted for sectioning. Thanks in advance for any advice. Marilyn Howton, Pathology Dept. West Virginia University
I would like to apologise for my humble trivial request to all highly qualified expert like you. I am running website www.coleoptera.org about beetles and I just open new section FAQ with info about collecting, preparation, techniques etc.
I just like to add section about mounting small insects into permanent slide. Is there anything so good as Canadian balsam?
Next question will be how to best prepare insects for SEM..
Any help deeply appreciated.
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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Below is the result of your feedback form. It was submitted by (alineves-at-unisys.com.br) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, December 11, 2000 at 14:50:32 ---------------------------------------------------------------------------
Email: alineves-at-unisys.com.br Name: Aline Neves School: Federal University of Rio de Janeiro State: Rio de Janeiro
Question: How do I measure the retardation of a specimen in polarized light microscopy. I have the Berek conpensator to achive a quantitative measure, but exactly how do I do it? Where can I get more information about that?
Hi I insect morphology and use Nikon SMZ-2T binocular stereomicroscope to study the various insect body parts. I use micrometer inside the ocular lens to take measurements. I do not know how to calibrate these measurements. It will be nice if if someone can refer literature or material on internet for calibration and mathematical calculations.If my question is incomplete please let meknow so that i can provide more nformation. sandeep gupta
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I am working with human cell lines in suspension and process the cells in a pellet.
After the usual problems and much help from this list I worked out a procedure which works for me.
It is critical that the pellet is small - in my experience not more than 250 000 cells / pellet. This means you will not or hardly see the pelleted cells until they have been in OsO4. All the dehydration steps and infiltration steps are performed on a rotary shaker.
Here it is:
I transfer the cell suspension into conical BEEM capsules and spin them in home made adapters at 900 x g. (It is essential to use a swing bucket rotor, otherwise your cells smear along the sides of the container and float off during the dehydration steps.)
After carefully removing the supernatant with fine tipped mini pastetts I overlay the pellet with fixative, remove it again with pastetts and so on. As soon as the OSO4 stains the pellet one can see it and that makes the handling easier. I use glass pipetts with OsO4 and propylenoxide and the resin- propylen mixtures.
I usually infiltrate twice with epon-propylenoxide (1+2 and 1+1 respectively) for 45 minutes, then twice with pure resin. The tips of glass Pasteur pipetts have to be cut to widen the opening otherwise it is very difficult to remove the very viscous resin.
For sectioning I carefully flatten the very fine tip of the block with a razor blade. The cells are evenly distributed, there is no need to cut away surplus plastic. Just shape the block face to a trapezoid and it can go straight into the ultramicrotome.
Good luck - if you require more details feel free to ask.
Regards Claudia
Dr. C. Hayward-Costa School of Life Sciences Kingston University Penrhyn Road, Kingston upon Thames Surrey KT1 2EE, UK 44(0)208 547 2000 x 2240 Email: c.hayward-at-kingston.ac.uk
Dear Marylyn Howton - one approach to fixing cells that have been stripped off is to spin them into a pellet(500rpm) and then take up the pellet in 2% agar - you can then take the pellet through deydration and embedding and all the cells remain in a pellet. You can fix the cells in the pellet but that is rather time consuming and it may be best to do the fixation steps first - spinning down after each and then make into a pellet. hope this helps Carole Elleman
your freind could try to contact Mr. Rasche in Germany: email: mikrovid-at-gmx.de homepage: www.mikrovid.com
best wishes, Joachim
Dr. Joachim Prutsch Product Manager EM Specimen Preparation
Leica Microsystems Hernalser Hauptstr. 219 email: Joachim.Prutsch-at-leica-microsystems.com A-1170 Vienna Tel. +43 1 48899 - 235 Austria Fax +43 1 48899 - 350 ---------------------- Weitergeleitet von Joachim Prutsch/AUVIE/LMSCentral/Leica am 12.12.2000 14:11 ---------------------------
"Gordon Couger" {gcouger-at-couger.com} am 09.12.2000 00:12:26
An: microscopy-at-sparc5.microscopy.com
Kopie: (Blindkopie: Joachim Prutsch/AUVIE/LMSCentral/Leica)
Thema: Wanted Wollaston prisms for Reichert Zetopan DIC scope
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I have a freind that need then main prism slide containing the Wollaston prism for Reichert Zetopan DIC scope. This is the slide going in the upper body of the scope. He also needs the polorizor that fits over the light soruce but that is of much less importance beause it can be easily made. While the prism is constructed of unobtainium.
He is willing to pay a reasonable price for the part.
If any of you have a spare or left over you could make a very dissapointed man a lot happier.
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
Since you work in a hospital setting, get some unneeded serum from your clinical lab, pellet the sample cells, add 50-100 ul of serum to pellet, mix well pellet cells again and then overlay with 3% glut. Next day, remove plug of cells/serum, cut into em size tissue pieces and process as you normally would.
Best of Luck,
Ed
Edward P. Calomeni
Medical College of Ohio Department of Pathology 3000 Arlington Ave. Toledo, OH 43614
} } } "Marilyn Howton" {mhowton-at-hsc.wvu.edu} 12/11/00 03:38PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am new to this group, so if this is question has been covered, please excuse. I usually do TEM on tissue from biopsies, etc., and now someone wants EM on cells grown in culture. I have done this for intact monolayers (very tedious), but these would be on trypsined-off and pelleted cells. I need to know if there is an easy way to handle them as an entire pellet, so I don't have to cfg. after each fixation/dehydration step, and so they will be compacted for sectioning. Thanks in advance for any advice. Marilyn Howton, Pathology Dept. West Virginia University
Dear Nafisa Ghori: Ammonium Molybdate is especially useful to contrast membranes as in vesicles or liposomes. It also can be titrated to neutral pH whereas UA cannot.
Don Gantz Boston University School of Medicine Biophysics Dept.
Colleagues, a pathologist asked to use our Transmission Electron Microscope(TEM) and when I was going to help him load his copper grids, he said he would do it as it was a fecal preparation for viral studies. I found out that material on formar coated copper grids was not fixed. After he viewed (no photos taken) I pulled the copper grid holder and all items touched and ultrasonicated with absolute ethanol and then acetone and then placed items without touching in a 70oC oven for an hour. I feel that is enough, but would like to know if anyone else has been in this predicament. Thanks, Teresa
I am looking for a computer programm that simulates the contrast induced
by defects in crystals. The defects I am interested in are twins and steps in the twin planes. Which programms are available in which you can simulate many different types of defects and that allow to change the parameters easily?
Marilyn, I concentrate the cells by a gentle spin, and embed them in a drop of 2% noble agar. After hardening and trimming the blocks to a size of you usual biopsy you can process them without loosing any. A}
Alice Dohnalkova S&E Assoc., Dep. of Environmental Microbiology Battelle, Pacific Northwest National Laboratory MS P7-50 Richland, WA 99352 tel. (509) 372-0692 fax (509) 376-1321
-----Original Message----- } From: Marilyn Howton [SMTP:mhowton-at-hsc.wvu.edu] Sent: Monday, December 11, 2000 12:39 PM To: Microscopy-at-sparc5.microscopy.com
I am new to this group, so if this is question has been covered, please excuse. I usually do TEM on tissue from biopsies, etc., and now someone wants EM on cells grown in culture. I have done this for intact monolayers (very tedious), but these would be on trypsined-off and pelleted cells. I need to know if there is an easy way to handle them as an entire pellet, so I don't have to cfg. after each fixation/dehydration step, and so they will be compacted for sectioning. Thanks in advance for any advice. Marilyn Howton, Pathology Dept. West Virginia University
"Re: EM on cell cultures" (Dec 12, 8:42am) References: {3A35E4DF.17130.27D219-at-localhost} X-Mailer: Z-Mail (4.0.1 13Jan97) To: Claudia Hayward-Costa {LS_S562-at-crystal.kingston.ac.uk}
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I wonder if you have heard of the Hohenberg technique described some 6 years ago in J.Microscopy; this technique makes life so much easier for people working with single cells in suspension, whether they are prokaryotes or eukaryotes.
H.Hohenberg could nicely demonstrate that the embedding and sectioning is very much facilitated and structures are much better preserved when: a) cells are taken up in cellulose capillaries, 200 micron in diameter; b) cells are high-pressure frozen (as the very first step for cryo-immobilization) c) cells are freeze-substituted but not dehydrated at RT. This technique has been used in the meanwhile by many other people with success, ... but still is not known to all working with single cells in suspensions.
Hohenberg et al (1994) J. Microscopy 175, 34-43
have fun! Reinhard Rachel
Dr. Reinhard Rachel Universitaet Regensburg Lehrstuhl fuer Mikrobiologie (Prof. Dr. K.O. Stetter) D - 93040 Regensburg Tel.: +49-941-943-4534 Fax.: +49-941-943-1824 e-mail: Reinhard.Rachel-at-biologie.uni-r.de
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Dear Marylyn Howton - one approach to fixing cells that have been stripped off is to spin them into a pellet(500rpm) and then take up the pellet in 2% agar - you can then take the pellet through deydration and embedding and all the cells remain in a pellet. You can fix the cells in the pellet but that is rather time consuming and it may be best to do the fixation steps first - spinning down after each and then make into a pellet. hope this helps Carole Elleman
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Morning Marilyn,
Since you work in a hospital setting, get some unneeded serum from your clinical lab, pellet the sample cells, add 50-100 ul of serum to pellet, mix well pellet cells again and then overlay with 3% glut. Next day, remove plug of cells/serum, cut into em size tissue pieces and process as you normally would.
Best of Luck,
Ed
Edward P. Calomeni
Medical College of Ohio Department of Pathology 3000 Arlington Ave. Toledo, OH 43614
} } } "Marilyn Howton" {mhowton-at-hsc.wvu.edu} 12/11/00 03:38PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am new to this group, so if this is question has been covered, please excuse. I usually do TEM on tissue from biopsies, etc., and now someone wants EM on cells grown in culture. I have done this for intact monolayers (very tedious), but these would be on trypsined-off and pelleted cells. I need to know if there is an easy way to handle them as an entire pellet, so I don't have to cfg. after each fixation/dehydration step, and so they will be compacted for sectioning. Thanks in advance for any advice. Marilyn Howton, Pathology Dept. West Virginia University
You can use a tabletop airfuge with a very small rotor. Each tube holds about 0.2 ml liquid. You can spin your sample for about an hour at a very high speed and it will give you a very tight pellet. I used to spin the sample in fixative and once you are done spinning, you can process the pellet and do osmication, dehydration, infiltration and embedding.
Good luck,
Soumitra
************************************************************* Soumitra Ghoshroy Ph.D. Electron Microscopy Lab and Fluorescence Imaging Facility Department of Biology Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3600 Fax: 505-646-5665 e-mail:sghoshro-at-nmsu.edu http://confocal.nmsu.edu/eml
I have been working with human red blood cells grown in cell culture that are infected with Plasmodium falciparum for the past 9 years. The way I handle them is to use normal fixatives (glut, OsO4, uranyl acetate) while they are in suspensions and spin them down in-between steps. Following fixation I spin them down in warm (45 degree C) 3% agarose (Sigma A5030 25gm, type 9 ultra low gelling temperature). When the agarose cools to room temperature the pellet becomes like Jell-O in the tip of the 15ml falcon tube. Then I simply cut off the tip and transfer the "glued together pellet" to a glass vial and continue with dehydration and infiltration steps as if I was handling a small hunk of tissue. Good luck, Tim
Timothy G. Schneider Director of Electron Microscopy Department of Pathology Room 229 Jefferson Hall Thomas Jefferson University 1020 Locust St. Philadelphia Pa. 19107 215-503-4798 work 610-613-8170 cellular timothy.schneider-at-mail.tju.edu
Another person to try would be Dave Paschke, The Dawson Co., (617)484-7900. He had some parts for our Reichert MeF2 when I spoke to him a year ago...you never know.
Diane Ciaburri General Dynamics Pittsfield, MA 01235 (413)494-2847
"joachim.prutsch-at-leica-microsystems.com" 12/12/00 08:17 AM
To: Microscopy-at-sparc5.microscopy.com cc: Subject: RE: Wanted Wollaston prisms for Reichert Zetopan DIC scope
Dear Gordon,
your freind could try to contact Mr. Rasche in Germany: email: mikrovid-at-gmx.de homepage: www.mikrovid.com
best wishes, Joachim
Dr. Joachim Prutsch Product Manager EM Specimen Preparation
Leica Microsystems Hernalser Hauptstr. 219 email: Joachim.Prutsch-at-leica-microsystems.com A-1170 Vienna Tel. +43 1 48899 - 235 Austria Fax +43 1 48899 - 350 ---------------------- Weitergeleitet von Joachim Prutsch/AUVIE/LMSCentral/Leica am 12.12.2000 14:11 ---------------------------
"Gordon Couger" {gcouger-at-couger.com} am 09.12.2000 00:12:26
An: microscopy-at-sparc5.microscopy.com
Kopie: (Blindkopie: Joachim Prutsch/AUVIE/LMSCentral/Leica)
Thema: Wanted Wollaston prisms for Reichert Zetopan DIC scope
I have a freind that need then main prism slide containing the Wollaston prism for Reichert Zetopan DIC scope. This is the slide going in the upper body of the scope. He also needs the polorizor that fits over the light soruce but that is of much less importance beause it can be easily made. While the prism is constructed of unobtainium.
He is willing to pay a reasonable price for the part.
If any of you have a spare or left over you could make a very dissapointed man a lot happier.
Gordon Gordon Couger gcouger-at-couger.com Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
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Reply to: RE: EM on cell cultures Dear Marilyn,
A good way to prepare cell monolayers for EM by embedding them as a pellet is as follows:
Fix the cells on the substrate by adding double strength fixative to the culture medium. This will contain serum which is good for the next steps. Leave the fixative on the cells for about 2-5 min and then carefully scrape off the monolayer using a small scraper specially made for this from either a small piece of teflon or a soft wooden stick previously soaked in buffer. The cells will come off the substrate in either a single monolayer (if there are tight junctions present, or as a powdery suspension. Quickly take the detached cells from the dish and centrifuge (I use full speed on the benchtop Eppendorf for about 2 seconds to get a pellet). If you do this without protein being present in the fixative (eg serum) the cells will not pellet but will stick to the sides of the tube (smearing). Once the cells have pelleted DO NOT resuspend them. Remove the fixative and medium and replace it with fresh fixative. Leave for the amount of time you want to fix for (eg 1hr). You will find that after the fixation is complete you will be able to remove the pellet from the bottom of the tube and treat it as if it was a piece of tissue. You should be able to cut it into smaller pieces for subsequent embedding. Follow your favorite protocol for this. I am sure that you will have pellets that do not fall apart. The secret is to fix as a pellet a.s.a.p. and then do not disturb until after fixation is complete. I think the fixative cross-links extracellular components of the cells together but this only happens if the cells are in close proximity as fixation occurs.
If you do prefer to process in the tube they were pelleted in, then a neat trick for speeding up all the steps is to centrifuge after each change of solution. You will see how quickly things go into the pellet if you centrifuge in the presence of osmium tetroxide. A 2 min centrifugation at top speed will cause even a large pellet to turn black all the way through. If you continue the processing in this tube then make sure the final infiltration in the resin is sufficient to go allthe way through the pellet. Be careful to remove all dehydrating solutions and propylene oxide from the tube. I tip them upside down for about half an hour before adding the final resin changes. You will soon realize if you did this successfully - a soft pellet will be the result of too much proylene oxide in the resin. I prefer to embed them after cutting into small pieces.
As they are a pellet of individual cells they will be much easier to embed in resin than a solid tissue piece. Any poor morphology you get will probably not be due to what you do them but what happens before you put them in fixative. Trypsin is not good for morphology, neither is transfection. Ccentrifugation or scraping prior to fixation will affect morphology too. It is a good rule to physically handle the cells as little as possible prior to fixation.
The biggest surprize is that they will have poor morphology if they have only been left to grow fror a day or two after passage. It is only after about three days that they really start to recover from this. Unfortunately, they will look really good in the light microscope so poor morphology will be your problem!
I hope this helps.
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
Marilyn Howton 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 new to this group, so if this is question has been covered, please excuse. } I usually do TEM on tissue from biopsies, etc., and now someone wants EM on } cells grown in culture. I have done this for intact monolayers (very tedious), } but these would be on trypsined-off and pelleted cells. I need to know if there } is an easy way to handle them as an entire pellet, so I don't have to cfg. after each } fixation/dehydration step, and so they will be compacted for sectioning. } Thanks in advance for any advice. } Marilyn Howton, Pathology Dept. West Virginia University } } } } } RFC822 header } ----------------------------------- } } Received: from ultra5.microscopy.com [206.69.208.10] by mailhouse.hei.org } with ESMTP } (SMTPD32-6.00) id A73530B01FE; Mon, 11 Dec 2000 21:27:17 -0800 } Received: (from daemon-at-localhost) } by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id OAA00529 } for dist-Microscopy; Mon, 11 Dec 2000 14:40:54 -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 OAA00526 } for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Mon, 11 Dec 2000 } 14:40:24 -0600 (CST) } Received: from gateway.hsc.wvu.edu (gateway.hsc.wvu.edu [157.182.105.18]) } by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id OAA00519 } for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 11 Dec 2000 14:40:13 -0600 (CST) } Received: from HSC-DOM4-Message_Server by gateway.hsc.wvu.edu } with Novell_GroupWise; Mon, 11 Dec 2000 15:39:02 -0500 } Message-Id: {sa34f516.000-at-gateway.hsc.wvu.edu} } X-Mailer: Novell GroupWise Internet Agent 5.5.3.1 } Date: Mon, 11 Dec 2000 15:38:37 -0500 } From: "Marilyn Howton" {mhowton-at-hsc.wvu.edu} } To: {Microscopy-at-sparc5.microscopy.com} } Subject: EM on cell cultures } Mime-Version: 1.0 } Content-Type: text/plain; charset=US-ASCII } Content-Disposition: inline } Content-Transfer-Encoding: 8bit } X-MIME-Autoconverted: from quoted-printable to 8bit by } ultra5.microscopy.com id OAA00520 } Errors-to: Microscopy-request-at-sparc5.microscopy.com } X-RCPT-TO: {pwebster-at-hei.org} } X-UIDL: 273278220 } Status: U }
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Dear Listers,
Can anyone suggest a good reference or share a technique for preparing C-Pt rotary shadowed collagen molecules on mica for TEM?
Thanks.
---------------- Ann Hein Lehman EM Facility Manager Trinity College Hartford, CT 06106 v. 860-297-4289 f. 860-297-2538 e. ann.lehman-at-trincoll.edu
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Dear Marilyn,
I am working with human cell lines in suspension and process the cells in a pellet.
After the usual problems and much help from this list I worked out a procedure which works for me.
It is critical that the pellet is small - in my experience not more than 250 000 cells / pellet. This means you will not or hardly see the pelleted cells until they have been in OsO4. All the dehydration steps and infiltration steps are performed on a rotary shaker.
Here it is:
I transfer the cell suspension into conical BEEM capsules and spin them in home made adapters at 900 x g. (It is essential to use a swing bucket rotor, otherwise your cells smear along the sides of the container and float off during the dehydration steps.)
After carefully removing the supernatant with fine tipped mini pastetts I overlay the pellet with fixative, remove it again with pastetts and so on. As soon as the OSO4 stains the pellet one can see it and that makes the handling easier. I use glass pipetts with OsO4 and propylenoxide and the resin- propylen mixtures.
I usually infiltrate twice with epon-propylenoxide (1+2 and 1+1 respectively) for 45 minutes, then twice with pure resin. The tips of glass Pasteur pipetts have to be cut to widen the opening otherwise it is very difficult to remove the very viscous resin.
For sectioning I carefully flatten the very fine tip of the block with a razor blade. The cells are evenly distributed, there is no need to cut away surplus plastic. Just shape the block face to a trapezoid and it can go straight into the ultramicrotome.
Good luck - if you require more details feel free to ask.
Regards Claudia
Dr. C. Hayward-Costa School of Life Sciences Kingston University Penrhyn Road, Kingston upon Thames Surrey KT1 2EE, UK 44(0)208 547 2000 x 2240 Email: c.hayward-at-kingston.ac.uk
Can anyone tell us where to buy a 12V 50W bulb part number HLWS5-A (by Narva) for Jena scope. Please respond to me directly at the address below.
Thanks
Owen P. Mills Electron Optics Facility Engineer Michigan Technological University Department of Materials Science and Engineering Rm 512 M&M Building Houghton, Mi 49931 Ph: 906-487-2002 FAX: 906-487-2934 Email: opmills-at-mtu.edu URL: http://www.mm.mtu.edu/~opmills/index.html
Here's a summary of the advice I received regarding my falling off section problem. It's long but read the last suggestion, it's my favorite.
Formvar coating the tradition way, or dipping the grid into formvar or 0.01% BSA. Dry by dragging grid across filter paper. Bake the sections on to the grid-use either a 60 or 35 degree C oven overnight. Heat fixing the sections using either a hot plate or slide warmer for 10 minutes. Use Coat-Quick G. Flame the grids with an alcohol lamp until the grid just changes color. When working with Spurr's resin, use Veco grids not Gilder. Make sure the water is utra-clean. Use double distilled, deionized, or microfiltered. Various cleaning techniques: 1. 100% acetone wash. 2. 0.1M Nitric acid dip, rinse with dd water, dry on filter paper. 3. 50% acetic acid, 50% acetic acid, 100% acetone-dip grid into each solution then dry well before use. 4. 40 ml concentrated HCL in 400 ml dd water, add 40 ml acetone, take volume to 500 ml with dd water. Sonicate grids in 50ml beaker for 1-2 minutes, discard solution and rinse 2 X with acetone (30 seconds each) dry on filter paper. Do this every day to prevent oxidation from building up on the grids. 5. 100ml clean beaker, cover grids with 20ml glacial acetic acid, sonicate for 5-10 minutes, rinse with 100% acetone until the smell of acetic acid is gone, rinse with 100% ethanol, pipette off the excess 100% ethanol, invert beaker onto filter paper in a glass petri dish. Place in a 60 degree C oven for about 60 minutes, the grids will fall to the bottom of the petri dish, cover the dish with the glass top, the grids will be good for a while. Glass is used to avoid the static that plastic can have. Put sections on the dull side.
Last but not least......
get a new job or get a technician.
I apologize for this being so lengthy, but I thought others might benefit from this information.
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Dept. of Pathology, Ross Hall rm 505 Electron Microscope Lab 2300 Eye St. Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
Paula Sicurello George Washington Univ. Medical Center Dept. of Pathology, Ross Hall rm 505 Electron Microscope Lab 2300 Eye St. Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
Paula Sicurello George Washington Univ. Medical Center Dept. of Pathology, Ross Hall rm 505 Electron Microscope Lab 2300 Eye St. Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
Can anyone answer this question . Remember Ask-A-Microscopist is a forum that is used by NON-MICROSCOPISTS looking for help so keep try to your technical details at the right level.
Question: Is it possible to identify types of starch glue (rice, wheat, corn, potato, etc)under microscope? Is there any visuals available somewhere for comparisons?
We are trying to do some cryo TEM of organic solvents based samples, and the grids we have used so far were not very satisfactory, some solvents obviously influenced the formvar (and maybe the carbon ?) of the grid making it beam sensitive. We are planning on purchasing another type of grid (pure carbon lacey film, silicon dioxide or maybe silicon nitride) and I would like to know if anybody has tried anything for that kind of application. Any suggestion or comment most welcome !
Thanks to all how ansered my questions about these two FE-SEM. Its a pitty that I heard about MSA Listserver only two weeks ago. It would have been interesting to become these informations a bit sooner. It was very fruitfull to have some return of experience and offline discussions with some. Amazing was that amost only Hitachi owners had something to tell. And they are all pleased about their instrument. True that the Jeol is new, and much different from the former.
We have seen that our conclusions are similar that a lot of others, and so in a few words, what we can say about these two SEMs:
The Jeol 6700F seems to have at low primery energy (1-2 keV) and short working distance (1-3 mm) the best resolution and the easiest way to become nice images. Charging effect are weak, and magnetic samples have no visible propensity to "fly" in the chamber (magnetic nano powder, for exemple) ! The limitation now, is that this good resolution is lost very fast with increasing the working distance (6-8 mm). And the BSE working distance is 6-8 mm, analytical working distance 15 mm. Shure that in BSE or in X ray microanalysis, higher energy will bee used. But...and tilt ? And the vacuum chamber is limited in avaible ports.
On the other hand, the Hitachi may have the capacity to do so good, but it needs more experience of the manipulator and more time. One is shure : the "In Lens" detector of the Hitachi works well (is probably the best) at short and long working distances, gives a realy surface information, and with the last improvement, gives the possibility of composition contrast at low energy (schould be less sensitif to charge effects too, but we didn't be convinced).
So, mesured with a ladle (do you say so in english ? "Mesurer à la louche", in french !) The Jeol seems to be best for ultra high resolution at low energy and short working distance. The Hitachi seems to be best if you want to have the best images possible at analytical distance (12 mm), backscatered detector on place and the two "secondary" dectectors together. And is also capable to reach ultra high resolution. We didn't make detailed tests at high energy (} 15 keV). All instruments seems to be comparable at high energy, and then you don't observe the real surface.
But, and now I'll conclued, the two apparatus are VERY good, and it is VERY difficult to make a choice. If you have monney, buy one of eatch ! We have choiced the Jeol one, but whith some regree for Hitachi's "In Lens" detector, versatility and reliability.
I've said nothing about Leo's 1500 serie and Philips XL30-S-FEG, witch are also interresting. We have test them, but it would be to long to speak about these now. But cold FEG seems to give better high resolutions images,and is to be prefered if you don't need the long term stability of Shottky FE, for intensive XRay microanalysis fot exemple. Gemini's optical concept is very interresting, but more "classical" design give as good results.
I'm still intersted on other opinions or test results, and also avaible for questions, true MSA list server or on my direct mail.
By and thanks to all
J. Faerber IPCMS-GSI 23, rue de Loess 67037 Strasbourg CEDEX
We are trying to do some cryo TEM of organic solvents based samples, and the grids we have used so far were not very satisfactory, some solvents obviously influenced the formvar (and maybe the carbon ?) of the grid making it beam sensitive. We are planning on purchasing another type of grid (pure carbon lacey film, silicon dioxide or maybe silicon nitride) and I would like to know if anybody has tried anything for that kind of application. Any suggestion or comment most welcome !
subject: J. C. Russ, R. T. Dehoff, Practical Stereology, 2nd Edition, Plenum Press, New York, 2000 (!?!), isbn 0-306-46476-4
Some folks on this list have an early draft of my forthcoming (more on that below!) book on stereology, which was distributed on the CD with the Image Processing Tool Kit and also handed out in notes form at the various image analysis workshops that I’ve taught for the past many years. Bob Dehoff (who teaches those workshops with me) and I completed the final version of that book in December, 1999, and sent it in to the publisher. It has taken them a very long time to get around to doing anything with the book, and they are still apparently having a lot of internal technical problems (not worth going into here - but *.doc files and *.tif files on a CD seem to be a little too high tech for these people, and they clearly don’t have any idea what a *.pdf file is good for). At the M&M meeting in Philadelphia, quite a few people asked me about the book and at that time Plenum was promising that it would be available in September. Well, of course, it wasn’t, and still isn’t and frankly I’m getting tired of apologizing for it. None of the delay has anything to do with the authors, it is strictly production difficulties and a high level of technical incompetence at the publisher’s end. Anyway, to make a long story less long, I’ve put a copy of the entire book on the web in the form of a pdf file. In order to keep the download times reasonable (it is still 7.1 MB) the graphics are jpeg compressed, but I think still useful and legible. Anyone who wants to download a copy is welcome to it. When the book eventually (!) comes out the on-line version will evaporate. You may want to purchase a copy to get the index and somewhat better graphics, but I will leave that to your conscience. For the present, you can find a link to it at {http://members.aol.com/ImagProcTK/updates} (look toward the bottom of the page under "Data Analysis") or on my web page {http://members.aol.com/DrJohnRuss} (go to the "links" section). In the on-line file, the contents are linked to the book pages and of course you can use Acrobat Reader to search for any word in the text.
} I am looking for a tissue histology stain for human tongue } epithelium for use with tetramethylrhodamine-labeled probes. The stain } must not } fluoresce under 546nm wavelength light and also must not quench } tetramethylrhodamine fluorescence. Haematoxylin, eosin, and methylene } blue each } fail to meet these requirements. Does anyone have any suggestions? } } Dennis M. Walling, M.D. } Assistant Professor } Division of Infectious Diseases } Department of Internal Medicine } The University of Texas Medical Branch } 301 University Boulevard } Galveston, TX 77555-0835 } } Telephone: (409) 747-2361 } Fax: (409) 772-6527 } E-mail: dwalling-at-utmb.edu }
The Materials Characterization Group in Dow’s Corporate R&D Analytical Science laboratory has a full-time professional level opening for a particle analyst/light microscopist at Dow’s facility in Midland, MI. The primary responsibilities include working with partners to support research and production projects across a broad range of products. 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: * Strong problem solving skills * Operation of particle size characterization instrumentation * Operation of light microscopes and supporting image analysis software packages * Method development * Documentation of work * Compliance with safety and quality programs
Qualifications: * A candidate with a BS or MS degree in material science, chemistry, or physics with experience in light microscopy and particle size characterization. * Experience with common commercially available particle size measurement instrumentation - single-particle, ensemble and fractionation based. * Experience in the general area of polarized light microscopy, digital image acquisition, and quantitative image analysis.
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, Employee Development Center, Workforce Planning, Midland, MI 48674. Email respondents must list Job 00622LUSA/JMK-JB and their last name as the first and second items on the subject line. Only those selected for an interview will be contacted.
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$20 billion. Dow manufactures and supplies chemicals, plastics, and agricultural products for customers in 164 countries and employs approximately 43,000 people worldwide. For more news and information about Dow, visit our web site at www.dow.com.
Qualifications (education, certification, language, etc.) and experience required:
A candidate with a MS or Ph.D. degree in polymer science, material science or chemistry is preferred with prior experience in electron microscopy. Good written and oral communication skills and the ability to work both independently and in a team environment are extremely important.
Job Overview:
The Surface/Microscopy/X-ray (SMX) Group in Dow's Corporate R&D Analytical Science Laboratory has a professional level full time opening for a polymer microscopist. The primary responsibilities include working with partners to support research projects involving new and existing products in Dow's polymer businesses.
Key responsibilities will include:
1. Extensive problem solving. 2. Microscopy preparation technique experience including 3. Ultramicrotomy and cryo-ultramicrotomy. 4. Operation of light microscopes, scanning and transmission electron microscopes. 5. Interpretation of images. 6. Documentation of work. 7. Compliance with safety and quality programs. 8. Active member of project and SMX work teams.
We are an equal opportunity employer and offer a competitive compensation and benefits package including 401k, stock purchase, tuition reimbursement and performance incentives. 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, Employee Development Center, Workforce Planning, Midland, MI 48674. Email respondents must list Job 00623LUSA/JMK-JB and their last name as the first and second items on the subject line.
Only those selected for an interview will be contacted.
Can anyone tell me where I can find information on remote SEM? Is it possible to install remote SEM capabilities on an older existing instrument (JEOL 35C)? Does this technology work for LOM also?
Sam O. Mancuso Special Metals Corporation Research & Development New Hartford, NY 13413 smancuso-at-specialmetals.com
In my opinion, light microscopy is probably most useful in differentiating starch grains, although SEM is sometimes used.
By using a light microscope with two polarizing filters, it is possible to view a "cross" or "x-shaped" pattern within the starch grains. (This has sometimes been referred to as a "Maltese Cross".) The appearance of this cross can be an indicator of starch type. Also, by rotating one of the polarizers you can observe the cross as it comes into view, rotates a bit, then disappears again. Measuring the rotation arc of the polarizer within which the cross is visible gives you another criterion.
Other ways of telling starches apart are, of course, their size and shape. Sizes should be determined by averaging a number of grains, since they can be quite variable---in fact, the amount of variation in size is yet another criterion. Shapes range from round to various types of ovals and irregular ovoids. SEM excels at this, allowing very precise measurements.
If you really want to get involved, it's possible to set up heated water baths and sample a slurry of starch grains as the temperature rises, then count the melted versus unmelted grains in the light microscope. Melted starch grains are amoeba-like blobs, instead of nicely defined shapes. Different starches have different melting characteristics.
There is a two-volume set of work by Edward Tyson Reichert (1913), named "The Differentiation and Specificity of Starches in Relation to Genera, Species, Etc." It is loaded with methods and photos.
Here is a web site you may want to look at: http://www.siu.edu/~ebl/amylose.htm. I used to do starch research in the lab that puts this page out, back in "the old days".
Hope this gets you started. 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/
You will need to carefully define what you mean by "Remote SEM". Do you mean true TelePresence or just TeleObservation.
You should begin by looking at the Microscopy and Microanalysis Proceedings for 1995, 1996, 1998, and 2000 there are a number of full session as well as articles in there by the various groups in the world working in this area.
Viewing images on-line is do-able in virtually any imaging system, with sufficient hardware upgrades, but remote "TeleOperation" can be a much more involved proposition particuliarly for an older instruments.
You might consider getting a copy of the MSA Education Committee Video Tapes made during the Tutorials presented at the Microscopy and Microanalysis 2000 meeting.
I did a presentation therein called Microscopy and Microanalysis over the Internet, which discussed a pretty full range of options. I don't know if 2000 meeting videos are ready yet, but you can contact Greg Erdos for that information at the above WWW page.
There are a number of vendors which can interface older instruments to computer systems for image acquisition which the minimum starting point. This can then, in principle, be adapted to TeleObservation. You should talk to the following companies: 4Pi, Quartz, Gatan , EmiSpec (to name a few there are more so please check the exhibitors list from the M&M 2000 meeting) as well as literally any company that manufacturers EDS equipment (a long list....). Most of the Electron Optical manufacturers also offer or will soon offer this as options on their newer instruments but in all likelihood for your older instrument you will have to go to the accessory market arena.
There is also going to be a session at the Microscopy & Microanalysis Technologist's Forum at the 2001 Meeting which will discuss the issue of TelePresence, you may wish to attend that meeting in Long Beach August 5-9, 2001. http://www.microscopy.com/MSAMeetings/MM01/MMHomePage.html
Nestor Your Friendly Neighborhood SysOp
Disclaimer: I have no commerical interests in the companies mentioned herein, but you might say I have a mild vested interest in "TelePresence" and it's application to Research and Education ;-)
As always my TPM Lab is open to the world.... http://tpm.amc.anl.gov. your welcome to visit anytime...
J. W. Robinson Mechanical, Automotive, and Materials Engineering University of Windsor Windsor, Ontario N9B 3P4
Phone : 519-253-4232, ext 2598
Could anyone send me any information on the L. M. Simard company, maker of the tri-mag sputtering source? I need to order replacement parts and the last phone number I have for them is no longer in service. Any help will be appreciated.
Ooops - guess I need to type more carefully. The correct URL for downloading the Practical Stereology textbook is {http://members.aol.com/ImagProcTK/update.html} . The link there goes to a grad student's computer (someone who had enough storage space that I could use for a while), and from time to time that machine may go off line (when he is doing some "real" work), so if you don't get through at first be patient and try again in a few hours or the next day.
We just installed a Tecnai 12. I am not sure what is a reasonable charge for users to use the TEM. We are at university and a self-supported EM suite. Could you please let me know your opinion or the user fee (hourly rate) charged at your work place? especially at Canadian universities.
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There is a freeware/shareware application WinVNC which basically allows remote control over a network of any other PC and the applications running on that PC. While not a perfect solution, it works well running PC-driven SEMs, such as the JEOL 5500/5600/5900. We have experimented with it in the JEOL UK Application Lab and, considering it is 'free', are happy with it.
Addressing your specific problem, you would first have to convert your JSM-35 to a PC driven SEM. I don't know about the US but in the UK, there are several 3rd party companies offering such conversions. If you don't get any information via the list, contact JEOL Inc.
Once such a conversion has been done, I see no reason why you shouldn't control it over a LAN with WinVNC.
The same applies to any equipment driven by a PC - connect to a LAN and use WinVNC. In fact, I'm told it will work with any platform connected to the LAN - Unix, Mac, etc.
I have a collaborator who wants to do IHC on colon tissue. She just bought some Technovit 8100 but doesn't know how to use it. I use Technovit 7100 for regular histology on plant tissue but I have no protocol experience with 8100, IHC or animal tissue. Does anyone have any suggestions or recommended protocols for doing IHC with Technovit 8100? This is for light microscopy.
w'ere short about 3 plate holders for our Jeol 100cx TEM camera film (3&1/2 by 4"). At $25 apiece new, we were wondering if anyone had some spare? Or if anyone knows which websites would have these sorts of things second-hand?
cheers Liz McKenzie
******************************************************* Geomicrobiology and Electron Microscopy Laboratory Room S9 Cramer Hall 1721 SW Broadway Portland State University Portland OR97201
Post Doctoral Position open in a multiple disciplinary laboratory at NIH We use molecular biology, biochemistry and image analysis (confocal and electron microscopy) techniques for studies on lipid transport in cells.
I am searching for someone who has training in electron microscopy to investigate cholesterol mobilization in cells.
The position is ASAP,
The position is for a person not more than 4 years post PHD.
Those interested should send their CV's as inserts to the E-mail.
Thank You
-------------------------------------------------- Dr. Joan Blanchette-Mackie Chief; Section of Lipid Cell Biology NIDDK / NIH Building 8, Room 427 8, Center Drive Bethesda MD 20892 - 0850
Hello, I am a venture capitalist located in Milwaukee WI and am in need of some basic information on Atom Probe Microscopy because I am evaluating a potential investment in a company that manufactures such a device. What is it and how is it different than TEM and SEM. What are the applications it is used for? Are there companies that sell them? What do they cost? Any references you can direct me to will be appreciated. Thank you.
Daniel Broderick Managing Director Mason Wells Biomedical 770 N. Water St. Milwaukee WI 53202 414 765 7830 fx 414 765 7850
I have EELS spectra from a material contain a mixture of Ce3+ and Ce4+ and wish to determine the relative amounts of each oxidation state. To do this I have collected spectra from a Ce3+ standard and a Ce4+ standard and propose to least squares fit these to the spectrum from my mixed state sample.
I presume this type of thing has been done before and there are probably any number of ways to implement the fitting using Excel or a KaleidaGraph or some purpose written software. I thought I'd ask if anyone would like to make their solution available to me or even just offer suggestions.
The ideal solution would be if the fitting could be done within EL/P or even DigitalMicrograph.
I look forward to any replies. 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.
The answer to this very much depends on what you are charging for. To find an answer, you need to identify (a) the net costs you wish to recover: e.g. capital costs (to be recovered over what period?, at what rate of interest?), running costs, salaries of direct (em technician) and indirect (clerical, admin.) support staff including overheads, any charges for space occupancy and services etc., less any subsidies (b) make a realistic estimate of the maximum number of hours per year that a charge can be recovered from. The answer is a/b. Simple, isn't it? Well, no. In practise you will probably find that b is dependent on a/b since all categories of users are sensitive to price, but you will have to model this differently for different categories of users.
Our charges per hour for self-help access to a Philips CM120 Biotwin are as follows: Departmental £15, Other departments £21, other publicly-funded research organisations £35, industrial £70. Non-attenders incur full charge for the period booked. These numbers were not exactly arrived at using rocket science. We charge extra for all materials consumed (film, processing) and for direct user-support from a technician. We do not include capital costs recovery in our charges.
I would strongly advise imposing a special concessionary charge on users who insist on tipping their specimens and retaining circlips into the guts of the Compustage! An electronically-operated trap door beneath the operator's seat is also an attractive option.
Chris
} From: "Ping Li" {pli-at-is.dal.ca} To: {Microscopy-at-sparc5.microscopy.com}
I've noted no response, so I better offer something. The technique was more commonly used to visualize stretched DNA fibers and you may find more references using the Kleinschmitt technique about 30 years ago.
I have used fixed shadowcasting and negative staining on collagen too and both methods "work". The advantage of rotary shadowcasting seems to be that it makes it easier to follow the continuity of fibers (up-down, round and about). Rotary shadowcasting is usually executed at a shallow angle of 6-12 degrees and so it builds up more metal at the near vertical surfaces of the fibers.
Sticky-tape the very edge of the prepared specimen grid to a flat surface (half of a microscope slide). A bit of tape may be required on two sides to assure that the grid lies flat. Arrange the grid(s) a the rotation center of a rotator within the vacuum chamber and locate the rotator to be between 60 and 100mm from the evaporation source and a little lower than the source to achieve an angle of say 10 degrees. The angle could be checked with a 10 degree card segment.
The source could be C-Pt pellets or 0.1mm dia. Pt wire (try 40mm) wrapped around a 1.5mm carbon cylinder about 5mm long that has been fashioned with a suitable C rod sharpener. The "loaded" C rod is spring tensioned against a second C rod that retains a flat end, but one that is reduced in size to about 3mm dia. using a conventional (but clean, e.g. no pencils) pencil sharpener.
Evacuate the system. Pt/C evaporation gives very fine granularity; about the best short of special equipment, but a good vacuum ( { 5x10-5 torr) is essential. Prior to evaporation start the rotator and adjust the speed to 30 to 60 rev/ minute. Increase the ampere control to achieve red heat, then back off to give the vacuum a moment to recover. Then increase ampere to a white heat and observe using a dark shield until the fashioned carbon tip has evaporated. Hope this helps. This technique is much easier demonstrated than written about. 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 ABN: 99 724 136 560 www.proscitech.com
On Wednesday, December 13, 2000 5:57 AM, Lehman, Ann [SMTP:Ann.Lehman-at-trincoll.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 Listers, } } Can anyone suggest a good reference or share a technique for preparing C-Pt } rotary shadowed collagen molecules on mica for TEM? } } Thanks. } } ---------------- } Ann Hein Lehman } EM Facility Manager } Trinity College } Hartford, CT 06106 } v. 860-297-4289 } f. 860-297-2538 } e. ann.lehman-at-trincoll.edu
If you are a material scientist and have experience in microscopy and the use of computers, then we invite you to apply for this position at the Bridgestone/Firestone Research, Inc. in Akron, Ohio.
Our Microscopy Analysis Laboratories provide research and service in microscopy for Bridgestone/Firestone Research, and, in general, Bridgestone/Firestone (BFS) associates, as well as extend service to non-BFS users. Currently, our laboratories have one 'state-of-the art' TEM/STEM/AIA, one SEM/EDX, as well as one AFM/STM and a number of LOM-s and ancillary equipments.
Duties of the appointee will include sample preparation, instrument operation, analysis, and reporting. The level of appointment will depend on the qualifications, and the experience of the successful candidate.
Please send a letter of application plus your CV (including the names, postal-e-mail addresses, telephone/fax numbers of 3 referees) to: Dr. Pat Sadhukhan Bridgestone/Firestone Research, Inc. Phone: 330-379-7518 1200 Firestone Parkway Fax: 330-379-7530 Akron, OH 44317-0001
Has anyone come across radiation damage to SrTiO3, either knockon or radiolytic?
------------------------------------------------------- Laurence Marks Director, Center for Transportation Nanotechnology Department of Materials Science and Engineering Northwestern University Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -------------------------------------------------------
Workshop May 17-19 2001 "New approaches to the Phase Problem" http://xraysweb.lbl.gov/esg/phasing/index.html
Dear Ping, When we installed our TEM, we calculated the hourly cost of the service contract, based on a 40 hour work week, and used that for a basis for our charge to other users at the University. It is currently 50 dollars per hour. The charge for commercial users is based on what commercial consultant firms charge, in our area about 200 dollars an hour. There are, in my experience, very few commercial users of TEM. At 04:45 PM 12/13/00 -0400, you wrote:
} We just installed a Tecnai 12. I am not sure what is a reasonable charge for } users to use the TEM. We are at university and a self-supported EM suite. } Could you please let me know your opinion or the user fee (hourly rate) } charged at your work place? especially at Canadian universities. } } 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
As a late follow-up to a somewhat recent string on liquid nitrogen safety, I thought I would relate something that happened just this morning. While transferring LN2 from one dewar to another using a funnel, the funnel literally exploded just as the container was filled to the top. The explosion was pretty forceful, sounding like a gunshot and scattering sharp plastic shards all over the room and leaving behind a startled, but otherwise unhurt technician (yup, it was me).
It was a standard-issue, large size plastic lab funnel, nothing special. We had used the same funnel for over a year with no problems whatsoever. I don't have a clue as to why it blew up or how there was enough pressure generated to cause the explosion, but the remainder of the funnel looks a lot like the hat Jughead used to wear in the Archie comics. It is now on our bulletin board with a tastefully done joke notice about my recent brush with eternity, courtesy of the other technician, Cheryl, who shall remain anonymous.
Anyway, WATCH IT if you use the same low-tech LN2 transfer protocol we do (or did).
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
I am looking for comments from individuals who are currently using a PC based program or spreadsheet to treat data from Al or Au ring SAED calibration measurements. I would also like to be able to index minerals such as asbestiform silicates. I am aware of older Fortran based diffraction programs but I need a PC based system that someone is currently using.
On Wed, 13 Dec 2000 DrJohnRuss-at-aol.com-at-sparc5.microscopy.com wrote:
} Ooops - guess I need to type more carefully. The correct URL for downloading } the Practical Stereology textbook is } {http://members.aol.com/ImagProcTK/update.html} . The link there goes to a } grad student's computer (someone who had enough storage space that I could } use for a while), and from time to time that machine may go off line (when he } is doing some "real" work), so if you don't get through at first be patient } and try again in a few hours or the next day.
So far, today, none of the links get one to the download site.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
The fee depends on the purpose of charging.
If you just want to pay the cost of maintenance for the users inside the university, then I think that you just make a calculation based on the costs on liquid N2, N2 gas, films, darkroom expenses, fees for service contract, and the other consumption (everything related to the machine, except the salary of the professional staff). Also the fee could be quite different from university to university. Recently I have been two universities, the same machine, JEOL 2000FX, one charges user USD$35 per hour, the other charges user only USD$20. As I know, when you charge user at high price although it is reasinable, then you will get less users to use, finally you get less "income" to cover the cost. If it happens, you may have to lower the price to encourage users to use. This happened to one university I used to work.
Yes, people always charge those commercial companies at much higher price, 3 - 5 times?
Dear Ping, When we installed our TEM, we calculated the hourly cost of the service contract, based on a 40 hour work week, and used that for a basis for our charge to other users at the University. It is currently 50 dollars per hour. The charge for commercial users is based on what commercial consultant firms charge, in our area about 200 dollars an hour. There are, in my experience, very few commercial users of TEM. At 04:45 PM 12/13/00 -0400, you wrote:
} We just installed a Tecnai 12. I am not sure what is a reasonable charge for } users to use the TEM. We are at university and a self-supported EM suite. } Could you please let me know your opinion or the user fee (hourly rate) } charged at your work place? especially at Canadian universities. } } 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
------------------------------------ Zhenquan Liu (Dr.) Arizona State University CSSS, CHEM Room PSA213, Tempe, AZ 85287 Tel (480) 965 4544 (o) (480) 775 7428 (h) Fax (480) 965 9004 (o) Email zhenquan.liu-at-asu.edu ------------------------------------
Canadian Food Inspection Agency National Centre for Foreign Animal Disease Winnipeg
Salary: $ 45,146 - $ 54,928 per year Tenure: indeterminate
Education: Preference may be given to candidates with a diploma or degree in biology (or a related field) from a recognized technical college or university. Experience: Significant experience in a research/diagnostic laboratory, including performance of electron and light microscopy techniques. Experience in a variety of software applications, including digital imaging Experience in working with infectious agents.
Willingness to work under the rigours of high security biocontainment. Contact with livestock outside of the workplace will be subject to certain restrictions.
Functions:
To run a busy Reserach/Diagnostic EM lab, under supervision of a Principal Investigator:
Prepares samples for diagnostic examination by electron microscopy; operates electron microscope and conducts standard examination procedures for detection, identification and characterization of foreign
animal disease agents by electron microscopy; establishes and confirms primary identification of foreign animal disease agents in emergency disease investigations and/or recruits the required expertise to identify unknown agents; catalogues specimens, micrographs and images and keeps accurate and comprehensive records; develops new technical procedures or modifies existing procedures for sample preparation and examination including observation of frozen hydrated specimens by electron cryomicroscopy. Works on immunohistochemistry techniques to label and localise antigens using both light and electron mciroscopy.
If interested please email application including CV to Dr. Tim Booth, tbooth-at-em.agr.ca Candidates must clearly demonstrate in writing, that they fully meet the
screening criteria. Failure to do so will result in your application being screened out of the selection process with no further consideration being given to your application.
As a late follow-up to a somewhat recent string on liquid nitrogen safety, I thought I would relate something that happened just this morning. While transferring LN2 from one dewar to another using a funnel, the funnel literally exploded just as the container was filled to the top. The explosion was pretty forceful, sounding like a gunshot and scattering sharp plastic shards all over the room and leaving behind a startled, but otherwise unhurt technician (yup, it was me).
It was a standard-issue, large size plastic lab funnel, nothing special. We had used the same funnel for over a year with no problems whatsoever. I don't have a clue as to why it blew up or how there was enough pressure generated to cause the explosion, but the remainder of the funnel looks a lot like the hat Jughead used to wear in the Archie comics. It is now on our bulletin board with a tastefully done joke notice about my recent brush with eternity, courtesy of the other technician, Cheryl, who shall remain anonymous.
Anyway, WATCH IT if you use the same low-tech LN2 transfer protocol we do (or did).
Dear Randy, It is possible that internal stress was present in the funnel, and upon cooling (this time) it built up past the ability of the funnel to adjust. This happens in an altogether different temperature range to quarz high-pressure Hg lamps. Teflon does not become too stiff at 77 K, so this shouldn't happen if you use a teflon funnel. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
FWIW, I no longer use a funnel (would be S.Steel if I did). A number of years ago I experienced the same scenario. My HDPE funnel exploded with great force sending shrapnel ricocheting off the walls...
A bit of spilled nitrogen is not particularly hazardous (don't pour in your shoe :) compared to the high speed funnel shards.
Woody White McDermott Technology
----------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Hi Listers, } } As a late follow-up to a somewhat recent string on liquid } nitrogen safety, I } thought I would relate something that happened just this } morning. While } transferring LN2 from one dewar to another using a funnel, the funnel } literally exploded just as the container was filled to the top. The } explosion was pretty forceful, sounding like a gunshot and } scattering sharp } plastic shards all over the room and leaving behind a startled, but } otherwise unhurt technician (yup, it was me). } } It was a standard-issue, large size plastic lab funnel, } nothing special. We } had used the same funnel for over a year with no problems } whatsoever. I } don't have a clue as to why it blew up or how there was } enough pressure } generated to cause the explosion, but the remainder of the } funnel looks a } lot like the hat Jughead used to wear in the Archie comics. } It is now on } our bulletin board with a tastefully done joke notice about } my recent brush } with eternity, courtesy of the other technician, Cheryl, who } shall remain } anonymous. } } Anyway, WATCH IT if you use the same low-tech LN2 transfer } protocol we do } (or did). } } } 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/ } } } }
Just one point on your comments relating to charge in the Hitachi 4700. In any SEM with an in lens detector the signal will be made up of SE. Should you use the lower detector in the Hitachi you will find an increase in the contribution of BSE and therefore a much lower charge rate.
Training many people in the use of the 4500 and 4700 I find they tend to become upper detector addicts, when the lower detector often has much to offer, particularly with samples that tend towards charging.
Hope this helps?
Steve Chapman
Senior Consultant Protrain Tel & Fax - 44 (0)1280 814774 E-mail- protrain-at-emcourses.com Web Site - www.emcourses.com Protrain for SEM, TEM & EDX Training World Wide with our own full time Professional staff
At 4:45 PM -0400 12/13/00, Ping Li wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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Ping: I run the EM Core facility for our medical college. I had to assure my dean that my fees were in accord with other, similar facilities in the NY-metro area. I called around to many of my colleagues and got their fee schedules. My fees are about average...higher for some things, lower for others. Briefly, our per hour fees for the EM are $50 if the person has been trained and works alone, $75 if they need my assistance. Film is billed at $1 each, which just barely covers the cost...I'll probably have to raise that price this year.
If you'd like to see my whole price list (I have a fee for every service available in the facility), you can go to to the website: http://www.med.cornell.edu/research/cores/index.html Go to the bottom of the page, click on the up-down arrows and scroll to the Electron Microscopy Core.
I am responsible for covering all of the operating costs of the lab, with the exception of my salary (thank heaven!). I am the sole personnel in the facility for now, and handle about 100 clients/year. I have received approval for a part-time tech, and will then offer histology (paraffin embedding/sectioning, cryostat sectioning) as a service rather than just having the equipment here for people to use on their own.
Good luck. Setting up a fee schedule is no easy matter....be prepared for loads of complaints about the cost, and people asking if everyone pays the same rate. Everyone here pays the same fees, from the Sr. Assoc. Dean of Research to the newest Assist. Prof. That way no one gets their nose out of joint!
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 think the point about hazards associated with LN2 that do not directly involve the cold liquid itself is worthy of some note.
Over the 15 years we've been maintaining a LN2 cooled beryllium crystal, there have been 3 incidents where luckily no injuries were sustained. Two involved breaking the 4-L dewer -- once when a student smacked the top edge of the dewer with the LN2 hose nozzle and once when a technician tripped and fell with a full dewer. The first merely resulted in a very loud bang with lots of fog, etc. as described in another post (no aluminum foil, though) and very thankfully, the technician who fell was unhurt by either the fall or the shattered glass. Again, lots of fog and running rivulets of LN2 down the hallway.
The third incident was just a close call and one the was totally unanticipated. Our dewer has a large cork in the top which is replaced after the dewer is filled. After filling, the technician reached down to pick up the nearly full dewer and, in the process, caused the LN2 to slosh and a bit evaporated causing the cork to 'fire' out of the mouth of the dewer (large 'pop gun' effect) and graze her face. She was not seriously hurt beyond a small scrape on her temple...but, had she been hit in the eye at such close range.... Anyway, since that graze incident, we who are responsible for filling the EDAX dewer, wear glasses or goggles and are very careful where the dewer is 'aimed' when we pick it up and carry it.
We have had similar occurrences here. Both old, and new funnels have exploded. It is my guess that it is internal stress in the plastic which causes the event (Or, should I say the rapid release of the stress from being cooled to below minus 200 degrees centigrade?)
What we have done, is to use only stainless steel funnels for LN2.
I'd say the use of a plastic funnel with LN2 is a no no anyway. It probably just shattered from being at that extreme temperature for a certain length of time, which is why the "explosion" coincided with the other dewar being topped off.
-----Original Message----- } From: Tindall, Randy D. [mailto:TindallR-at-missouri.edu] Sent: Thursday, December 14, 2000 10:20 AM To: 'microscopy-at-sparc5.microscopy.com'
Hi Listers,
As a late follow-up to a somewhat recent string on liquid nitrogen safety, I thought I would relate something that happened just this morning. While transferring LN2 from one dewar to another using a funnel, the funnel literally exploded just as the container was filled to the top. The explosion was pretty forceful, sounding like a gunshot and scattering sharp plastic shards all over the room and leaving behind a startled, but otherwise unhurt technician (yup, it was me).
It was a standard-issue, large size plastic lab funnel, nothing special. We had used the same funnel for over a year with no problems whatsoever. I don't have a clue as to why it blew up or how there was enough pressure generated to cause the explosion, but the remainder of the funnel looks a lot like the hat Jughead used to wear in the Archie comics. It is now on our bulletin board with a tastefully done joke notice about my recent brush with eternity, courtesy of the other technician, Cheryl, who shall remain anonymous.
Anyway, WATCH IT if you use the same low-tech LN2 transfer protocol we do (or did).
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/
From root Thu Dec 14 18:01:51 2000 Return-Path: {Microscopy-request-at-sparc5.microscopy.com} Received: (from daemon-at-localhost) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id RAA05412 for dist-Microscopy; Thu, 14 Dec 2000 17:15:12 -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 RAA05407 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Thu, 14 Dec 2000 17:14:42 -0600 (CST) Received: from tmpsmtp102.honeywell.com ([199.64.7.102]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id RAA05400 for {microscopy-at-sparc5.microscopy.com} ; Thu, 14 Dec 2000 17:14:30 -0600 (CST) Received: from 131.127.249.22 by tmpsmtp102.honeywell.com (InterScan E-Mail VirusWall NT); Thu, 14 Dec 2000 16:02:58 -0700 Received: by TMPCN197 with Internet Mail Service (5.5.2650.21) id {YBYJW3NY} ; Thu, 14 Dec 2000 16:13:27 -0700 Message-ID: {3348B4357E7FD4118DAE00508B074950607E0D-at-tmpex177.wins.allied.com}
I'd say the use of a plastic funnel with LN2 is a no no anyway. It probably just shattered from being at that extreme temperature for a certain length of time, which is why the "explosion" coincided with the other dewar being topped off.
-----Original Message----- } From: Tindall, Randy D. [mailto:TindallR-at-missouri.edu] Sent: Thursday, December 14, 2000 10:20 AM To: 'microscopy-at-sparc5.microscopy.com'
Hi Listers,
As a late follow-up to a somewhat recent string on liquid nitrogen safety, I thought I would relate something that happened just this morning. While transferring LN2 from one dewar to another using a funnel, the funnel literally exploded just as the container was filled to the top. The explosion was pretty forceful, sounding like a gunshot and scattering sharp plastic shards all over the room and leaving behind a startled, but otherwise unhurt technician (yup, it was me).
It was a standard-issue, large size plastic lab funnel, nothing special. We had used the same funnel for over a year with no problems whatsoever. I don't have a clue as to why it blew up or how there was enough pressure generated to cause the explosion, but the remainder of the funnel looks a lot like the hat Jughead used to wear in the Archie comics. It is now on our bulletin board with a tastefully done joke notice about my recent brush with eternity, courtesy of the other technician, Cheryl, who shall remain anonymous.
Anyway, WATCH IT if you use the same low-tech LN2 transfer protocol we do (or did).
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
I am currently using Crystal Studio (check google to find the source) to generate diffraction patterns for the amphiboles and others. Although it has some strange characteristics the patterns are reasonable. Be advised that the patterns are based on x-ray diffraction intensities - the actual SADs have different intensity distributions as you might expect.
Email me if you need help indexing the asbestos silicates.
Tom McKee 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 MSA group } } I am looking for comments from individuals who are currently using a PC } based program or spreadsheet to treat data from Al or Au ring SAED } calibration measurements. I would also like to be able to index minerals } such as asbestiform silicates. I am aware of older Fortran based } diffraction programs but I need a PC based system that someone is currently } using. } } Thanks Tom McKee
-- Gordon Nord Small Business Network Design and Construction Macintosh and Windows - Solutions and Conflicts
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I would like just to point that it is not good idea "preheat" carbon rods in the presence of sample. All contaminations from rods will contaminate the sample first and than you will shadow that. Shadowing will develop not only structural details but contamination ether. What I did, I preheat rods without samples, than open the chamber, install the samples and pump down the system again. Even when I done carbon film preparation, I prefer to use the same technique. It takes more time. In general, shadowing quality extremely depends from sample quality and sample preparation procedure. Most air-dried protein samples demonstrate very poor structural integrity preservation. Buffer composition is also important (you will shadow not only the "sample" but salts from buffer as well).
Sergey
} Date: Thu, 14 Dec 2000 20:58:32 +1000 } From: Jim at ProSciTech {jim-at-proscitech.com} } Subject: RE: Rotary shadowing } To: "'Lehman, Ann'" {Ann.Lehman-at-trincoll.edu} , } "Microscopy-at-sparc5.microscopy.com" {Microscopy-at-sparc5.microscopy.com} } Reply-to: "jim-at-proscitech.com" {jim-at-proscitech.com} } 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 Department of Biological Chemistry, School of Medicine University of California, Los Angeles Box 951737 Los Angeles, CA 90095-1737
We have been doing some rotary shadowing to produce replicas on biological samples for examination under the TEM. For shadowing we used V-shaped tungsten wire and pure carbon for support,
They are a few problems we encountered. Every run is a trial and error process. This is especially so with the setting of current to vaporize the metal and the carbon. Every run is a different value although the voltage is constant. At each run we try to follow the prefer parameter but somehow the replica would look different from the previous one. Although, it is better to observe the colour of the glow on the metal and carbon but that is very hurting to the eye. I wonder if anybody has a definite value for the current used to vaporize 0,1 mm pt wire/tungsten and pure carbon rod in the vacuum chamber of 10 -5 torr.
Josephine NUS
-----Original Message----- } From: Jim at ProSciTech [mailto:jim-at-proscitech.com] Sent: Thursday, December 14, 2000 6:59 PM To: 'Lehman, Ann'; Microscopy-at-sparc5.microscopy.com
Josephine This message is only partially related to your topic, but, anyway. This is my comment.
I would say, it's very unlikely to have good reproducibility in your set-up. The easiest and probably cheapest solution is to use thickness monitor. It's relatively cheap (prices vary from $1500 to $5000), precisely measure the thickness starting from a few angstroms, easy to work. You may control the shutter by the thickness monitor in order to close the shutter at some point, say 2 nm.
More "solid" solution is to use electron guns for shadowing. I am using electron guns for decade and they shown outstanding ability to evaporate many substances: W, Pt/C, C in my case. Electron gun evaporated carbon films have outstanding stability (in compare with thermal evaporated) under the beam. I would mention that I forgot the time when I have a problem with drift. Last time it was, probably 15 years ago when I was looking for good technique for carbon film preparation. Additional reason to switch to the Electron Gun - sample thermal damage. Electron guns produces less infra-reds than regular set-up.
Another issue, I would like to comment is a level of vacuum in the evaporators. The level 5x10-5 torr in my point of view is not acceptable for ANY applications related to biology. The abequate range started from 2x10-6 torr I believe. I would point that practically all commercially available evaporators are able to pump system down to the good 10-6 even without LN2 trap. 10-5 is a result of bad evaporator's maintenance. Replace diffusion oil on SANTAVAC 5 (and forget the problems with DP), check the O--rings for cracks, replace old O-rings (do you know how old your system?), keep system clean, run system at least for 24 h a month. I just could not believe that films produced at 10-5 torr would not contaminated by hydrocarbons from oil, from that a problem, many colleagues described: bad hydrophilic properties of the films.
Sergey
} Date: Fri, 15 Dec 2000 12:20:19 +0800 } From: "Howe L. C. Josephine" {michowej-at-nus.edu.sg} } Subject: FW: Rotary shadowing } To: "'microscopy-at-sparc5.microscopy.com'" {microscopy-at-sparc5.microscopy.com} } Cc: "'jim-at-proscitech.com'" {jim-at-proscitech.com} } X-Mailer: Internet Mail Service (5.5.2650.21) } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Pt coating is not as fine as simultaneous Pt - C. Support film (carbon backing) is only needed if you need to dissolve a thick biological specimen; for thin fibres (say unstained biological material under 50nm) a replica is rather cumbersome. I expect that you also have problems with Pt forming an amalgam with W and so varying amounts are evaporated. That is another advantage of using Pt - C. Another problem is that for C evaporation you do best at around 30 Volts and for metal, you achieve much better control with about 10 volts. The volt settings on some evaporators can be changed but the variable that you are using is changing amperes not volts. I expect that the control of one of your electrodes is very touchy. So take a pick, you have several problems and you may be able to eliminate one or two. 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 ABN: 99 724 136 560 www.proscitech.com
On Friday, December 15, 2000 2:20 PM, Howe L. C. Josephine [SMTP:michowej-at-nus.edu.sg] wrote: } We have been doing some rotary shadowing to produce replicas on biological } samples for examination under the TEM. For shadowing we used V-shaped } tungsten wire and pure carbon for support, } } They are a few problems we encountered. Every run is a trial and error } process. This is especially so with the setting of current to vaporize the } metal and the carbon. Every run is a different value although the voltage is } constant. At each run we try to follow the prefer parameter but somehow the } replica would look different from the previous one. Although, it is better } to observe the colour of the glow on the metal and carbon but that is very } hurting to the eye. I wonder if anybody has a definite value for the current } used to vaporize 0,1 mm pt wire/tungsten and pure carbon rod in the vacuum } chamber of 10 -5 torr. } } } Josephine } NUS } } -----Original Message----- } From: Jim at ProSciTech [mailto:jim-at-proscitech.com] } Sent: Thursday, December 14, 2000 6:59 PM } To: 'Lehman, Ann'; Microscopy-at-sparc5.microscopy.com } Subject: RE: Rotary shadowing } } ------------------------------------------------------------------------ } The Microscopy ListServer-Sponsor: The Microscopy Society 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 noted no response, so I better offer something. The technique was more } commonly used to visualize stretched DNA fibers and you may find more } references using the Kleinschmitt technique about 30 years ago. } I have used fixed shadowcasting and negative staining on collagen too and } both methods "work". The advantage of rotary shadowcasting seems to be that } it makes it easier to follow the continuity of fibers (up-down, round and } about). Rotary shadowcasting is usually executed at a shallow angle of 6-12 } degrees and so it builds up more metal at the near vertical surfaces of the } fibers. } Sticky-tape the very edge of the prepared specimen grid to a flat surface } (half of a microscope slide). A bit of tape may be required on two sides to } assure that the grid lies flat. Arrange the grid(s) a the rotation center of } a rotator within the vacuum chamber and locate the rotator to be between 60 } and 100mm from the evaporation source and a little lower than the source to } achieve an angle of say 10 degrees. The angle could be checked with a 10 } degree card segment. } The source could be C-Pt pellets or 0.1mm dia. Pt wire (try 40mm) wrapped } around a 1.5mm carbon cylinder about 5mm long that has been fashioned with a } suitable C rod sharpener. The "loaded" C rod is spring tensioned against a } second C rod that retains a flat end, but one that is reduced in size to } about 3mm dia. using a conventional (but clean, e.g. no pencils) pencil } sharpener. } Evacuate the system. Pt/C evaporation gives very fine granularity; about the } best short of special equipment, but a good vacuum ( { 5x10-5 torr) is } essential. Prior to evaporation start the rotator and adjust the speed to 30 } to 60 rev/ minute. Increase the ampere control to achieve red heat, then } back off to give the vacuum a moment to recover. Then increase ampere to a } white heat and observe using a dark shield until the fashioned carbon tip } has evaporated. Hope this helps. This technique is much easier demonstrated } than written about. } 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 } ABN: 99 724 136 560 www.proscitech.com } } On Wednesday, December 13, 2000 5:57 AM, Lehman, Ann } [SMTP:Ann.Lehman-at-trincoll.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 Listers, } } } } Can anyone suggest a good reference or share a technique for preparing } C-Pt } } rotary shadowed collagen molecules on mica for TEM? } } } } Thanks. } } } } ---------------- } } Ann Hein Lehman } } EM Facility Manager } } Trinity College } } Hartford, CT 06106 } } v. 860-297-4289 } } f. 860-297-2538 } } e. ann.lehman-at-trincoll.edu
OK Folks, I am open to suggestions. The computer with the file on it is still down, and I'm getting tired of all the nagging emails. Who can help me? The file is 7.5 MB. Who has enough space and reasonable access and is willing to host the file? The only conditions are that when the printed version of the book eventually appears, the electronic on-line version will need to be removed. In the meantime, anyone who wants to download the pdf file is welcome. If you are interested in providing host space, please contact me (off the list, please). Also note that there has to be some way for me to get the file to you - many mail systems won't accept such a large attachment.
John Russ John_Russ-at-NCSU.edu or DrJohnRuss-at-AOL.com
Email: William.SHort-at-cpe.amedd.army.mil Name: William Short
Question: One of my tasks is to implement the Verein Deutscher Ingenieur 3492 Method for analyzing asbestos fibers and other indoor pollutants using the Scanning Electron Microscope and an Energy Dispersive X-ray Analyzer. At this point I am nearly completed with the implementation. We have a nice old Hitachi S-450 model SEM and we use the KEVEX EDX to get our spectral data off the unknown samples. I am putting together the paperwork and reports to tell our customers what it is that we analyzed from their sample submissions. I give a Concentration result, the instrument Sensitivity determined by the filter area and air volumes, the raw data, fiber types ( i.e. asbestos, inorganic, calcium sulfate, or other), and a 95% Confidence Interval using the Poisson distribution. Also included are the fiber dimensions, voltages, a picture of the fiber morphology, scale in nanometers, and a copy of a spectral spot return from the EDXA. My problem is that I don't know if I'm producing the correct results for the Confidence Interval calculation using the Poisson function. Aren't there special considerations for the results obtained from getting 0 fibers, 1 fiber, 2, fibers, and 3 fibers ? Can you help me with this area or will I just have to "punt" ? The VDI 3492 Method is being used in Europe for this analysis to comply with the Federal Governing Standards for Host nations.
Any information that you could give me would be greatly
On the subject of metal shadowing, might I recommend the excellent treatise by Willison and Rowe (my ex-head of department!) "Practical Methods in Electron Microscopy - Volume 8" - One of the excellent Glauert edited series.
I am currently in the process of ordering Kodak electron microscope film 4489 (3.25 x 4 in.), however, it seems that I can no longer order the film from Kodak Canada Inc. Kodak is still producing the film but only in the U.S. I guess. This may not seem problematic but the price of the film from the Canadian distributers is at least two times as much (almost three times in one case) as the price paid the last time it was ordered.
I believe that AGFA produces a similar type of film but I do not know the product name or who to contact. If anyone has information on distributers, of either Kodak or the AGFA film, with reasonable prices I would appreciate hearing from you. Alternatively, if anyone has another favorite TEM film that they use and would like to share that information I would be happy to hear from you.
Please send me the information offline at the following address: smithh-at-em.agr.ca
Thank you in advance! Heidi Smith Microscopy Technician Agriculture and Agri-Food Canada Kentville, NS, Canada
I've been running the Core EM Facility at Harvard Medical School in Boston for about 5 years now. When I started the lab, I had absolutely no clue what to charge, so I checked out what other Facilities were charging. After a little research (but still feeling pretty clueless) I started charging 30$/ beam hour for Scope time, which I think was was below average in 1995. There weren't that many EM labs in the area so I took an average of whoever I got hold of across the country..
My idea was to charge a little less to get people interested. Which I think worked well (I had good financial support for a few years, until things took off.. I don't know if your lab there is new or already established..) Now that people are hooked, we have an average use of 50-60 hour/month on 2 scopes. I'm charging 31.50$/hour for Med. School users and 45$/hour for outside, for assisted Scope use I charge 63$/hour for Med. School and 94$/hour for outsiders. With our current use, which has been pretty stable for the last year, the income from scope use covers the service contracts and supplies..
I think my rates are lower than average, but I have a huge user group. I have two half time technicians and the Facility income pretty much covers the operating costs now (..not including my salary..!)
I have a full fee-for serivce pricelist at http://www.hms.harvard.edu/core/em.html
Good luck with the EM lab!
Maria
} At 4:45 PM -0400 12/13/00, Ping Li wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
____________________________
Maria Ericsson Harvard Medical School EM Facility 220 Longwood Avenue Boston, MA 02115 (617) 432 1698 maria_ericsson-at-hms.harvard.edu http://www.hms.harvard.edu/core/em.html
Florida Society for Microscopy-Florida Chapter of the American Vacuum Society & Surface Analysis 2001
Annual Symposium
Technical Meeting March 12-14 - Short Courses March 12-15 University of Central Florida; Student Union Building; Orlando, Florida
Meeting Highlights * Technical Sessions: Microscopy in the Physical and Biological Sciences, Surface Science, Thin Films, Focused Ion Beam Techniques, Biomaterial Interfaces, Environmental Science, Metrology, and Electronic Materials * AVS & Vendor Sponsored Short Courses * Job Fair * 3rd Annual FIB Users Workshop, and Vacuum Educators Workshop * Student Poster Competition (over $5000 in prizes can be awarded) -plus the MSA Traveling Poster Display * Equipment Exhibit with Surface, Analysis, Vacuum, and MicroscopySciences Vendors * No registration fee for Symposium or Equipment Exhibit
Confirmed Invited Speakers to Date * Donald Baer Pacific Northwest National Lab * Alice Dohnalkova Pacific Northwest National Lab * Klaus Edinger University of Maryland * Robert Hull University of Virginia * Bill Lamberti Exxon * Eero Ristolainen Tampere University of Technology * Bruno Schueler Physical Electronics * Catherine Taylor Wright State University * Edgar Voelkl Oak Ridge National Lab
Abstract Submission The abstract deadline is January 5, 2001 for hard copy abstracts and January 12, 2001 for e-mail abstracts. E-mail submissions are strongly encouraged. Contributed papers will be presented as platform presentations or poster presentations. Authors should indicate their session preference and preferred presentation format i.e., poster or platform. For uniformity, all the abstracts must be in the following format: the title of the talk should be in all capital letters followed by the names and addresses of the authors. The presenter's name should be underlined. Abstracts are limited to 200 words. E-mail abstracts should be as text in the body of the e-mail or an attached Microsoft Word file. Send abstracts to Dr. Lucille Giannuzzi, Program Chair, 407 275-4354, lag-at-mail.ucf.edu. Authors will be contacted by their session chair in mid-January 2001 concerning their abstract's status.
Registration You may register on line by clicking on the following link:
General Meeting Information You may find more information by following the "AVS Info; Chapters; Florida Chapter" links at: www.vacuum.org or the "Local Affiliate Societies" link at www.microscopy.org or contact Fred Stevie, 407-371-7626, stevie-at-lucent.com or Jo Ann Moore, jamoore-at-com1.med.usf.edu 1-813-974-9446
In order to receive email updates and future information about the meeting please forward your email address to Pete Ries pjries-at-lucent.com.
******************************************************************* 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."
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Postdoctoral Position in Transmission Electron Microscopy - Rice University
Rice University, Department of Mechanical Engineering and Materials Science, located in Houston, TX, is seeking candidates for a postdoctoral position in transmission electron microscopy of multicomponent oxide thin films. Applicants should have extensive and demonstrated experience in several areas of TEM and a strong background and interest in materials problem solving. Preference will be given to candidates with experience in high-resolution imaging and electron energy-loss spectroscopy as well as conventional diffraction contrast imaging. Facilities at Rice include a JEOL 2010, field-emission SEM and high-resolution X-ray diffractometers, as well as state-of-the-art sample preparation. The project will be carried out in close collaboration with the University of Houston using a state-of-the-art field-emission TEM (JEOL 2010F), with annular dark-field detector, Oxford link EDS, Gatan GIF and STEM capabilities. Ability and interest to take part in an upgrade of existing facilities to high resolution scanning TEM capability is expected. The position is available immediately. Duration about 1-2 years, salary is commensurate with qualifications. Candidates with a Ph.D. in Materials Science or Physics will be given preferred consideration. Interested candidates should send a curriculum vitae, publication list and the names of at least three references with their contact addresses to:
Prof. Susanne Stemmer Rice University Department of Mechanical Engineering and Materials Science MS 321 6100 Main Street Houston, TX 77005-1892 stemmer-at-rice.edu
Applicants must have proof of legal authorization to work in the United States. Rice University is an Affirmative Action/Equal Opportunity Employer.
Unfortunately Agfa have recently stopped supplying their Scienta film that we used in some of our TEMs so we are now changing to Kodak 4489 or SO163 as the Agfa runs out.
Ron
On Fri, 15 Dec 2000 10:21:27 -0500 Heidi Smith {SMITHH-at-em.agr.ca} 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 } -----------------------------------------------------------------------. } } } Seasons greetings to all! } } I am currently in the process of ordering Kodak electron microscope film 4489 (3.25 x 4 in.), however, it seems that I can no longer order the film from Kodak Canada Inc. Kodak is still producing the film but only in the U.S. I guess. This may not seem problematic but the price of the film from the Canadian distributers is at least two times as much (almost three times in one case) as the price paid the last time it was ordered. } } I believe that AGFA produces a similar type of film but I do not know the product name or who to contact. If anyone has information on distributers, of either Kodak or the AGFA film, with reasonable prices I would appreciate hearing from you. Alternatively, if anyone has another favorite TEM film that they use and would like to share that information I would be happy to hear from you. } } Please send me the information offline at the following address: } smithh-at-em.agr.ca } } Thank you in advance! } Heidi Smith } Microscopy Technician } Agriculture and Agri-Food Canada } Kentville, NS, Canada } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
There are now several copies of the book available on different servers, so access should not be a problem. The book is available at:
{http://loner.phys.lsu.edu/Users/brent/Stereology.pdf} (Thanks to Brent Neal at Louisiana State Univ. in Baton Rouge. This was the original host machine, but there was a general power failure just as I was posting the original note - another proof of Murphy's Law - and it took a while to get things back up; it is fine now)
{http://www.abdn.ac.uk/~ort056/stereology.pdf} (Thanks to Jenny Gregory {j.gregory-at-abdn.ac.uk} in Aberdeen, Scotland)
{http://128.101.243.155/Stereology.pdf} (Thanks to Mike Herron {herro001-at-umn.edu} at the University of Minnesota)
{http://www.biotech.ufl.edu/~emcl/Stereology.pdf} (Thanks to Greg Erdos {gwe-at-biotech.ufl.edu} at the University of Florida in Gainesville)
It should be available very soon from {http://www.practical-stereology.org} (Thanks to Greg Strout {gstrout-at-ou.edu} at the University of Oklahoma)
A few more sites are in process, and everything will be linked from the original main pages at {http://members.aol.com/drjohnruss/links.htm} {http://members.aol.com/ImagProcTK/update.htm}
Many thanks to all of the others out there who offered help or advice.
Hi everyone; just before everyone disappears for the weekend.... I will be cutting some cryosections of infected wheat heads (on monday, of course), and wanted to fix briefly, then put into a cryoprotectant to keep the cells from rupturing when I section (for light microscopy). I know I have successfully used sucrose in the past, but I can't seem to find the concentration. (4% rings a very distant bell...). Can anyone out there tell me what works for them? (or if you have other cryoprotectants that you put your plant samples into before sectioning, I'd be interested in those, too!)
thanks, as always, in advance shea
Dr. S.Shea Miller Agriculture & AgriFood Canada Eastern Cereal and Oilseed Research Centre Rm. 2068 Neatby Building Central Experimental Farm Ottawa, Ontario Canada K1A 0C6 Phone: (613) 759-1760 Fax: (613) 759-1701 E-mail: millers-at-em.agr.ca
All this talk about what y'all charge your users is no different than what prompted Congress to pass the Sherman Anti-Trust Act:
to protect Interstate commerce from price-fixing.
The discussion and setting of rates by communication between vendors isn't proper to my legally untrained eye.
Though it's OK to publish your rates, if that's what you really charge.
Now, it also turns out that what you charge users is nowhere near what Amenex Associates as a private, for-profit company has to charge to stay in business:
http://207.103.140.16/webpage/amnxfees.htm
Chuck Garber might want to chime in as to why independent lab owners get steamed up about all this.
Best regards, George Langford amenex-at-amenex.com
I was contacted today by a person who needs to locate some EM images of veterinary specimens. They have an immediate need for images, probably SEM's, of organs of the digestive system.
For further information, please contact them directly, not the listserv, at the following:
Jill Kahn 952-852-7358 kahn-at-collemcvoy.com
I'm just the messenger,
John john.chandler-at-colostate.edu Colorado State University
Thank you to evryone who responded to the free EDS detector. The two that I had are now on their way to the University of North Carolina.
I still have one EDS detector that was removed working from a PHI Auger microprobe. I also have some line voltage stabilizers that were removed from a JEOL 840 SEM. Keep in mind the stabilizers are HEAVY and the shipping costs are proportional.
Please contact me offline for details.
Thank You,
Earl Weltmer Scanservice Corp. SEM Maintenance (714) 573-9158
I can see your point of concern regarding this issue. However, if I understand the context within these folks are discussing fees, there is a different reference basis. They are talking about user fees in an academic environment. These fees are for services to members of their academic community--and in fact, for members of their respective institutions. This is quite different from for-profit enterprises' sphere of interest.
If the universities engage in external fee-based work, your point would tend to have merit. But what these folks are trying to do is to spread their cost center's burden over those who are using it, within their small community of users.
If the situation is other than this, of course, that is a different issue.
gary g.
At 01:29 PM 12/15/00, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I'm somewhat amazed by how many varied responses there are on rotary shadowing technique, and how few seem to agree with what works for us. So, here is yet another response.
We shadow biological molecules from the connective tissue matrix, usually ranging in size from 16 to 300 kd. Many are linear, some are globular. We spray the molecules in solution with 70% glycerol. The other 30% is 100 microgram/ml of protein, preferably in a volatile buffer such as 1% acetic acid or 0.1M ammonium bicarbonate, pH 7.8. Other buffers can be used but salt crystals can be a big problem. Spraying is accomplished using an air brush, and we spray onto freshly cleaved mica discs cut from sheets using a hole punch.
The sample is dried in vacuum, though we often accumulate 20 samples so some drying occurs outside vacuum. We are careful to pre-pump the vacuum chamber and heat the guns thoroughly to out-gas, then vent the jar with nitrogen, introduce the samples, and pump again. Our system uses a turbo-molecular pump. We can vary the angle of the rotary stage from outside the chamber, so we tilt the specimen away from the gun and outgas thoroughly again so that little vacuum loss is seen during evaporation. This is really important. Gas is introduced to the rods whenever the system reaches atmospheric pressure. It is also important to keep the system clean, as outgassing a dirty gun takes much longer than outgassing a clean gun. We try to complete evaporation without entering the 10 -5 range, and we begin in the 10-7 range. We evaporate at a slow rate, usually taking 3 to 5 minutes to complete a run, which also seems to improve resolution. Final film resolution is very proportional to vacuum, the better the vacuum the smaller the grain size. We use a quartz monitor for controlling the amount of Pt-C coming from the electron beam gun, but we also use a folded piece of filter paper placed 90 degrees relative to the source, and monitor the color which should be dark gray (not black, not brown). We evaporate at 6 degrees relative to sample as the sample rotates. Following Pt-C deposition, we then tilt the sample to 90 degrees relative to a resistance carbon source and evaporate a backing film of carbon onto the mica. The thickness of the film is monitored with a piece of folded filter paper placed 90 degrees relative to the carbon source, and the correct amount is just visible tan color (not gray) on the filter paper. Our film thickness monitor is not sensitive enough to monitor carbon deposition. We find this carbon film absolutely necessary for sample stability, perhaps because we use so little PT-C. However, too much carbon will certainly affect final resolution, loosing edge detail. Finally, the replicas are exposed to the vapors of 1% acetic acid in a petri dish for about a half-hour prior to floating in distilled water (the acid is very useful in helping the replica to release from the mica (so that they float off as one intact film ). We use high-transmission 600 mesh grids to support the films.
For many years we evaporated Pt wire from carbon rods using a resistance source. We wound 2.3 cm of wire around a cylindrical jig which was the same diameter as the sharpened carbon rods (about 1mm). Prior to coiling the wire, we passed it through a alcohol burner flame till it was orange, which made it more malleable and perhaps cleaned it a bit. The coil was transferred to the resistance source, spanning the intersection of two rods (therefore the site of most resistance and primary heating) held together with moderate spring tension. The carbon rods with accompanying Pt coil was at 6 degrees and about 11 cm away from the mica discs. Using a welders plate (Fullam #12511), we observed the metal as current was increased through the rods and just after the wire melted, the current was turned up just a bit more and left there until the Pt was seen to evaporate completely. It was necessary to observe this through the welders plate to get a good shadow and know when to turn the current down to avoid too much carbon. We evaporated a backing film of pure carbon from a second source oriented at 90 degrees from a distance of 11 cm so that the rods just started to spark, counted to 1.5 seconds, at which time the amount of carbon was probably about right (as judged by filter paper color and a bit of luck).
Sorry for the novel,
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
Stainless steel funnel poses also some danger - taking it with bare hand after the fill (no such problem with the plastic ones). The funnel I use has a crack which makes it more safe from an internal stress point of view.
Gary Gaugler wrote: ========================================================== I can see your point of concern regarding this issue. However, if I understand the context within these folks are discussing fees, there is a different reference basis. They are talking about user fees in an academic environment. These fees are for services to members of their academic community--and in fact, for members of their respective institutions. This is quite different from for-profit enterprises' sphere of interest.
If the universities engage in external fee-based work, your point would tend to have merit. But what these folks are trying to do is to spread their cost center's burden over those who are using it, within their small community of users. ============================================================= I am obviously not a lawyer, however, one should always keep in mind that Congress never granted any kind of exemptions to the anti-trust laws to tax- exempt organizations. So George Langford is absolutely correct, none of us should be discussing (with competitors) how we set our pricing.
Those of us who follow such things know that the penalties can be quite severe for those who are found to be guilty of violating the anti-trust laws
Gary Gaugler wrote: ========================================================== I can see your point of concern regarding this issue. However, if I understand the context within these folks are discussing fees, there is a different reference basis. They are talking about user fees in an academic environment. These fees are for services to members of their academic community--and in fact, for members of their respective institutions. This is quite different from for-profit enterprises' sphere of interest.
If the universities engage in external fee-based work, your point would tend to have merit. But what these folks are trying to do is to spread their cost center's burden over those who are using it, within their small community of users. ============================================================= I am obviously not a lawyer, however, one should always keep in mind that Congress never granted any kind of exemptions to the anti-trust laws to tax- exempt organizations. So George Langford is absolutely correct, none of us should be discussing (with competitors) how we set our pricing.
Those of us who follow such things know that the penalties can be quite severe for those who are found to be guilty of violating the anti-trust laws
Last Friday I got my first quantitative analytical results out of the JSM-840A which I have been converting to a 1 x eds, 3 x wds JXA version over the past year.
This has been a big fun project, I've learned heaps, and I couldn't have done it without this list and the help of the many listers who have given freely of advice and suggestions, and more.
So thanks, Nestor, and all you good people out there. Please drop in if you ever find yourselves in Auckland, New Zealand, it would be great to be able to replace the mental images which I have of people with factually-based ones!
cheers and happy Christmas
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
I do not agree with your analysis of this situation nor with your conclusion. Drawing on a somewhat fading memory of prior law classes (yes, I too am not a lawyer), I recollect that antitrust is covered by the Sherman Antitrust Act.
The purpose of this Act and those is various states is to prevent trusts from creating restraints on trade or commerce and thus, reducing competition. The Sherman Antitrust Act was designed to maintain economic liberty, and to [try to] eliminate restraints on trade and competition.
The Act applies to all transactions and businesses involved in interstate commerce. If the activities are local, the act applies to transactions affecting interstate commerce. Therefore, as I understand universities, they are not engaged in interstate commerce. Neither are they engaged in commerce, per se. Thus, they do not engage in transactions affecting interstate commerce. Their fundamental basis of action is to recover costs of ownership of university owned equipment and facilities as applied to users of such items at the university. I do not sense a wholesale effort by universities to engage in external commerce. Note also that many of the managers are not recovering the cost of their salaries. These are sunk costs by their respective universities.
It therefore seems to me that antitrust does not apply to universities. They are not engaged in commerce, they do not affect interstate commerce, they do not effect restraints on trade and commerce, and they are not in competition with one another. And I think they would agree that they are not in competition with private industry.
The law works in two ways: it specifies actions which are allowed and may specify actions which are not allowed. Lawyers exist because so much of the law is open to interpretation. I've just provided my interpretation of the law. Since I am not a lawyer, there is no charge for this.
gary g.
At 08:38 AM 12/16/00, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I sincerely thank those of you who provided me with information and support regarding the TEM user fee. I really appreciate it.
Regarding what "Garber, Charles A." wrote:
} .... So George Langford is absolutely correct, none of us } should be discussing (with competitors) how we set our pricing.
I don't think this is relevant to what we have done. What we have discussed is mostly about maintenance cost recovery, not making profit. Furthermore, we are not competitors, and we mostly provide service for research and teaching within our own university or institution. There is no competition among us.
"It therefore seems to me that antitrust does not apply to universities. They are not engaged in commerce, they do not affect interstate commerce, they do not effect restraints on trade and commerce, and they are not in competition with one another. And I think they would agree that they are not in competition with private industry."
Hmmm. This logic is quite interestinng. Then even though Universities provide a service in competition with private enterprise they are not competing. The logic in all of this escapes me. Hmmm.
----- Original Message ----- } From: "Gary Gaugler" {gary-at-gaugler.com} To: "Garber, Charles A." {cgarber-at-2spi.com} Cc: "MSA listserver" {Microscopy-at-sparc5.microscopy.com} Sent: Saturday, December 16, 2000 2:48 PM
I am not friendly with American law ether, but our medical school stated as "non-profit organization". So, to me it mean, that University is not involved in trading and thereafter is not a subject for anti-trust law.
Our hospital, for instance, charge people for the service, but they did not make a profit, same as when we charge people for using our facilities. I don't see any problem here as long as we did not make profit on it.
Merry Christmas! Happy New Year and New Millennium for all ListServer readers! I wish to all of us, that our microscopes will work better in New Millennium and we will have enough users to recover at least 70% of maintaining cost.
Sergey.
} Date: Sat, 16 Dec 2000 14:48:49 -0800 } From: Gary Gaugler {gary-at-gaugler.com} } Subject: Re: User fee discussion } X-Sender: gaugler-at-pop.calweb.com (Unverified) } To: "Garber, Charles A." {cgarber-at-2spi.com} } Cc: MSA listserver {Microscopy-at-sparc5.microscopy.com} } X-Mailer: QUALCOMM Windows Eudora Version 5.0 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Sergey Ryazantsev, Ph.D. Electron Microscopy Department of Biological Chemistry, School of Medicine University of California, Los Angeles Box 951737 Los Angeles, CA 90095-1737
Chuck Garber's reply to the User Fee discussion was truncated, due to a subtle effect of the Email system here. In fairness I am reposting it for him so that you can see the rest of his comments.
For those of you that are curious, the message was truncated because of a line in the original text which started with a "." followed by a blank/empty line. This caused the Email system to believe the message was completed.
============================================================= I am obviously not a lawyer, however, one should always keep in mind that Congress never granted any kind of exemptions to the anti-trust laws to tax- exempt organizations. So George Langford is absolutely correct, none of us should be discussing (with competitors) how we set our pricing.
Those of us who follow such things know that the penalties can be quite severe for those who are found to be guilty of violating the anti-trust laws.
So to my (also) legally untrained eye, for universities to be discussing how they set their pricing is no different than if independent laboratories (like Amenex and Structure Probe) started a parallel discussion on how we should be establishing our pricing. Anyone in academia who has ever written a proposal for funding in recent years knows that universities are in direct competition with each other in ways that are no different from the way independent laboratories compete with each other. George alluded to another issue, that is, the case when universities start offering their services to private companies. I am not aware of anyone having gone to jail for this, but it does raise enormous issues of ethical values, and the impact on students, and their own views of what is right and what is wrong, when the university's facilities are so commercialized. All of us should have some concern about such matters.
Chuck
Disclaimer: Structure Probe, Inc. is an independent for-profit tax-paying laboratory offering electron microscopy services to clients, often times in direct competition with universities who claim they "don't compete with private companies".
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI Structure Probe, Inc. /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: www.2spi.com ############################ ==================================================
Agfa Scientia TEM film was discontinued earlier this year. I do not know of a manufacturer other than Kodak that is selling a "dedicated" TEM in North America.
George Laing National Graphic Supply
Heidi Smith wrote:
} I believe that AGFA produces a similar type of film but I do not } know the product name or who to contact. If anyone has } information on distributers, of either Kodak or the AGFA film, } with reasonable prices I would appreciate hearing from you. } Alternatively, if anyone has another favorite TEM film that they } use and would like to share that information I would be happy to } hear from you. } }
I'd like to add my 0.004$ or R0.02 to the LN2 discussion:
An alternative to both the exploding plastic funnel and freezing finger steel funnel is to make one out of very thick brown paper (held together with staples rather than sticky tape).
I also staple a tissue paper filter in the funnel mouth to try to remove any ice from the LN2.
Regards Alison Tuling
Engineer Industrial Metal and Minerals Research Institute Department Materials science and Metallurgical engineering University of Pretoria South Africa +27 11 420 4556
It is interesting that this discussion comes up every 12 to 18 months or so. I also note a subtle difference in the way the question was posed, and see that the responses have, perhaps without intending, responded to both aspects of the original question. First, it would appear that asking for rates, particularly at potentially competitive facilities, is at least a gray area, if not a violation of the anti-trust laws. Second, asking assistance in how to determine how to set rates seems to be less problematic. It is within that context that anti-trust laws would appear to be less applicable. For individuals who have not had to deal with the issues of making a facility self-supporting, it can seem a daunting task. Thus, getting advice from other facility managers who have done the task aids the requester to determine what costs must be included, how to determine other factors, etc, and not miss any important/essential costs. It is important for all of us to be aware of the differences in the way questions are posed, especially as regards to this type of issue. This has been an enlightening discussion all around.
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc
NB: These are personal opinions only, and in no way reflect any other person or organization.
On Sat, 16 Dec 2000 21:25:04 -0800, Earl Weltmer 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 } -----------------------------------------------------------------------. } } } Gary Gaugler wrote: } } "It therefore seems to me that antitrust does not apply to } universities. They are not engaged in commerce, they do } not affect interstate commerce, they do not effect } restraints on trade and commerce, and they are not } in competition with one another. And I think they would } agree that they are not in competition with private industry." } } Hmmm. This logic is quite interestinng. Then even though Universities } provide a service } in competition with private enterprise they are not competing. The logic in } all of this escapes me. } Hmmm. } } ----- Original Message ----- } } From: "Gary Gaugler" {gary-at-gaugler.com} } To: "Garber, Charles A." {cgarber-at-2spi.com} } Cc: "MSA listserver" {Microscopy-at-sparc5.microscopy.com} } Sent: Saturday, December 16, 2000 2:48 PM } Subject: Re: User fee discussion } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 do not agree with your analysis of this situation nor with } } your conclusion. Drawing on a somewhat fading memory } } of prior law classes (yes, I too am not a lawyer), I recollect } } that antitrust is covered by the Sherman Antitrust Act. } } } } The purpose of this Act and those is various states is to } } prevent trusts from creating restraints on trade or commerce } } and thus, reducing competition. The Sherman Antitrust Act } } was designed to maintain economic liberty, and to [try to] eliminate } } restraints on trade and competition. } } } } The Act applies to all transactions and businesses involved in } } interstate commerce. If the activities are local, the act applies } } to transactions affecting interstate commerce. Therefore, as } } I understand universities, they are not engaged in interstate } } commerce. Neither are they engaged in commerce, per se. } } Thus, they do not engage in transactions affecting interstate } } commerce. Their fundamental basis of action is to recover } } costs of ownership of university owned equipment and facilities } } as applied to users of such items at the university. I do not } } sense a wholesale effort by universities to engage in external } } commerce. Note also that many of the managers are not } } recovering the cost of their salaries. These are sunk costs } } by their respective universities. } } } } It therefore seems to me that antitrust does not apply to } } universities. They are not engaged in commerce, they do } } not affect interstate commerce, they do not effect } } restraints on trade and commerce, and they are not } } in competition with one another. And I think they would } } agree that they are not in competition with private industry. } } } } The law works in two ways: it specifies actions which are } } allowed and may specify actions which are not allowed. } } Lawyers exist because so much of the law is open to } } interpretation. I've just provided my interpretation of the } } law. Since I am not a lawyer, there is no charge for this. } } } } gary g. } } } } } } } } } } } } At 08:38 AM 12/16/00, 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 } } } -----------------------------------------------------------------------. } } } } } } } } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } } } } } Gary Gaugler wrote: } } } ========================================================== } } } I can see your point of concern regarding this issue. However, if I } } } understand the context within these folks are discussing fees, there is a } } } different reference basis. They are talking about user fees in an } academic } } } environment. These fees are for services to members of their academic } } } community--and in fact, for members of their respective institutions. } This } } } is quite different from for-profit enterprises' sphere of interest. } } } } } } If the universities engage in external fee-based work, your point would } tend } } } to have merit. But what these folks are trying to do is to spread their } } } cost center's burden over those who are using it, within their small } } } community of users. } } } ============================================================= } } } I am obviously not a lawyer, however, one should always keep in mind that } } } Congress never granted any kind of exemptions to the anti-trust laws to } tax- } } } exempt organizations. So George Langford is absolutely correct, none of } us } } } should be discussing (with competitors) how we set our pricing. } } } } } } Those of us who follow such things know that the penalties can be quite } } } severe for those who are found to be guilty of violating the anti-trust } laws } } } } } }
_______________________________________________________ Send a cool gift with your E-Card http://www.bluemountain.com/giftcenter/
} Colleagues } } Chuck Garber's reply to the User Fee discussion was } truncated, due to a subtle effect of the Email system here. } In fairness I am reposting it for him so that you can } see the rest of his comments. } -------------------------------------------------- } } ============================================================= } I am obviously not a lawyer, however, one should always keep in mind that } Congress never granted any kind of exemptions to the anti-trust laws to tax- } exempt organizations. So George Langford is absolutely correct, none of us } should be discussing (with competitors) how we set our pricing.
What rubbish!! Of course we can discuss what standards we use to set fees. Businesses do this all the time.
} Those of us who follow such things know that the penalties can be quite } severe for those who are found to be guilty of violating the anti-trust laws.
Everytime someone gets on this list (and other lists) and asks about setting prices for users of lab services, Mr Langford and/or Dr. Garber starts making veiled threats about anti-trust violations. Isn't it interesting that no actual statues are cited, just vague references and generalizations, and that none of the citers are legally trained?
} So to my (also) legally untrained eye, for universities to be discussing how } they set their pricing is no different than if independent laboratories } (like Amenex and Structure Probe) started a parallel discussion on how we } should be establishing our pricing. Anyone in academia who has ever written } a proposal for funding in recent years knows that universities are in direct } competition with each other in ways that are no different from the way } independent laboratories compete with each other.
Not true of NIH funding of individual investigators.
} George alluded to another } issue, that is, the case when universities start offering their services to } private companies.
If you will read the original posting, that is NOT what was being talked about.
} I am not aware of anyone having gone to jail for this,
I am sure that if anyone had even been formally charged, you or Mr. Langford would be sure to let us know!
} } but it does raise enormous issues of ethical values, and the impact on } students, and their own views of what is right and what is wrong, when the } university's facilities are so commercialized. All of us should have some } concern about such matters.
Ethical behavior is a two-way street.
} Chuck } } Disclaimer: Structure Probe, Inc. is an independent for-profit tax-paying } laboratory offering electron microscopy services to clients, often times in } direct competition with universities who claim they "don't compete with } private companies". } } =================================================== } Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 } President 1-(800)-2424-SPI } Structure Probe, Inc. /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: www.2spi.com } ############################ } ==================================================
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 **********************************************
Regarding the contact info that Scott mentioned awhile back, we happen to sell what is probably that film dryer. You can see it at
http://www.tedpella.com/photo_html/photo11.htm
Tom
"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 } -----------------------------------------------------------------------. } } There is a company called California Stainless that made film dryers that } some of the EM supply houses sold. I bought one from them a number of years } ago. Sorry that I don't have the contact info, but I'm sure that this is } just what you want. } } -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: Donald Lovett [mailto:lovett-at-tcnj.edu] } Sent: Monday, November 20, 2000 9:51 AM } To: Microscopy.lst } Subject: Film dryers } } } --------------------------------------------------------------- } --------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserv er/FAQ.html } } } } --------------------------------------------------------------- } --------. } } } } Years ago I worked in a lab that had a small forced (and filtered) film } dryer (with heater). It was just large enough to hold two racks of TEM } negatives. I cannot find one in any of the catalogues. I can } only find } film dryers for 35 mm roll film. Could anyone please } recommend a source } of a similar dryer? (Vendors, please feel free to respond directly to } me). } } 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 } } } }
} It is interesting that this discussion comes up every 12 to 18 months or so.
Yes. Some listmember asks for a little guidance and get several replys. Then two people (the same two people, if my memory serves me) start yelling "anti-trust violation". No facts, no applicable case law, just the suggestion of lawlessness.
} I also note a subtle difference in the way the question was posed, and see } that the responses have, perhaps without intending, responded to both } aspects of the original question. First, it would appear that asking for } rates, particularly at potentially competitive facilities, is at least a } gray area, if not a violation of the anti-trust laws.
How? Please tell us exactly how asking what some other lab charges for XYZ is illegal.
} Second, asking } assistance in how to determine how to set rates seems to be less } problematic. It is within that context that anti-trust laws would appear to } be less applicable. For individuals who have not had to deal with the } issues of making a facility self-supporting, it can seem a daunting task. } Thus, getting advice from other facility managers who have done the task } aids the requester to determine what costs must be included, how to } determine other factors, etc, and not miss any important/essential costs. } It is important for all of us to be aware of the differences in the way } questions are posed, especially as regards to this type of issue. This has } been an enlightening discussion all around. } } Roger Moretz, Ph.D. } Dept of Toxicology } Boehringer Ingelheim Pharmaceuticals, Inc } } NB: These are personal opinions only, and in no way reflect any other person } or organization. } } On Sat, 16 Dec 2000 21:25:04 -0800, Earl Weltmer wrote: } } } Gary Gaugler wrote: } } } } "It therefore seems to me that antitrust does not apply to } } universities. They are not engaged in commerce, they do } } not affect interstate commerce, they do not effect } } restraints on trade and commerce, and they are not } } in competition with one another. And I think they would } } agree that they are not in competition with private industry." } } } } Hmmm. This logic is quite interestinng. Then even though Universities } } provide a service } } in competition with private enterprise they are not competing. The logic } in } } all of this escapes me. } } Hmmm. } } } } ----- Original Message ----- } } } From: "Gary Gaugler" {gary-at-gaugler.com} } } To: "Garber, Charles A." {cgarber-at-2spi.com} } } Cc: "MSA listserver" {Microscopy-at-sparc5.microscopy.com} } } Sent: Saturday, December 16, 2000 2:48 PM } } Subject: Re: User fee discussion } } } } } } } I do not agree with your analysis of this situation nor with } } } your conclusion. Drawing on a somewhat fading memory } } } of prior law classes (yes, I too am not a lawyer), I recollect } } } that antitrust is covered by the Sherman Antitrust Act. } } } } } } The purpose of this Act and those is various states is to } } } prevent trusts from creating restraints on trade or commerce } } } and thus, reducing competition. The Sherman Antitrust Act } } } was designed to maintain economic liberty, and to [try to] eliminate } } } restraints on trade and competition. } } } } } } The Act applies to all transactions and businesses involved in } } } interstate commerce. If the activities are local, the act applies } } } to transactions affecting interstate commerce. Therefore, as } } } I understand universities, they are not engaged in interstate } } } commerce. Neither are they engaged in commerce, per se. } } } Thus, they do not engage in transactions affecting interstate } } } commerce. Their fundamental basis of action is to recover } } } costs of ownership of university owned equipment and facilities } } } as applied to users of such items at the university. I do not } } } sense a wholesale effort by universities to engage in external } } } commerce. Note also that many of the managers are not } } } recovering the cost of their salaries. These are sunk costs } } } by their respective universities. } } } } } } It therefore seems to me that antitrust does not apply to } } } universities. They are not engaged in commerce, they do } } } not affect interstate commerce, they do not effect } } } restraints on trade and commerce, and they are not } } } in competition with one another. And I think they would } } } agree that they are not in competition with private industry. } } } } } } The law works in two ways: it specifies actions which are } } } allowed and may specify actions which are not allowed. } } } Lawyers exist because so much of the law is open to } } } interpretation. I've just provided my interpretation of the } } } law. Since I am not a lawyer, there is no charge for this. } } } } } } gary g. } } } } } } } } } } } } } } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } } } } } } } Gary Gaugler wrote: } } } } ========================================================== } } } } I can see your point of concern regarding this issue. However, if I } } } } understand the context within these folks are discussing fees, there } is a } } } } different reference basis. They are talking about user fees in an } } academic } } } } environment. These fees are for services to members of their academic } } } } community--and in fact, for members of their respective institutions. } } This } } } } is quite different from for-profit enterprises' sphere of interest. } } } } } } } } If the universities engage in external fee-based work, your point } would } } tend } } } } to have merit. But what these folks are trying to do is to spread } their } } } } cost center's burden over those who are using it, within their small } } } } community of users. } } } } ============================================================= } } } } I am obviously not a lawyer, however, one should always keep in mind } that } } } } Congress never granted any kind of exemptions to the anti-trust laws } to } } tax-exempt organizations. So George Langford is absolutely correct, none } of us } } } } should be discussing (with competitors) how we set our pricing. } } } } } } } } Those of us who follow such things know that the penalties can be } quite } } } } severe for those who are found to be guilty of violating the } anti-trust } } laws } } _______________________________________________________ } Send a cool gift with your E-Card } http://www.bluemountain.com/giftcenter/
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 **********************************************
Ah yes, this reminds me of a similar Close Encounter with liq N2 that I had: I removed a stainless steel rack from a Dewar, the type used for holding 5"X5" boxes as a cell freezer. Heavy, and dripping liq N2, I placed it down on one of those gray round rubber-topped step stools found in most labs. It was quite exciting when the rubber exploded with a CRACK and sent shards of frozen rubber in every direction. Luckily, I was alone and never 'fessed up to the missing rubber top on the step stool.............. Marilyn
Best wishes for an environmentally conscious, socially responsible, low stress, non-addictive, gender neutral, winter solstice holiday, practiced within the most joyous traditions of the religious persuasion of your choice, but with due respect for the religious persuasion of others who choose to practice their own religion, as well as those who choose not to practice a religion at all.
Additionally, (topical bit here!) a fiscally successful, personally fulfilling, and medically uncomplicated recognition of the generally accepted calendar year 2001, but not without due respect for the alternative calendars of choice of other cultures whose contributions have helped make our society great, without regard to the race, creed, color, religious, or sexual preferences of the wishes.
May the deity of our choice bless us each and every one but should you not be a believer (as above) then please accept our collective wish as fellow sentient (?) beings for the full development of your inner self.
Yours, Chris ==================================================
(Disclaimer: This greeting is subject to confirmation, clarification or withdrawal. It implies no promise by the sendeer actually to implement any of the expressed wishes for her/himself or others and no responsibility for any unintended emotional stress these greetings may or may not bring to those not caught up in the holiday spirit.)
Hope y'all manage to get a life this Christmas ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 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
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Reply to: RE: Fwd: Re: MRS follow-up
Dear Experts of Quasicrystals,
Recently I received two e-mails from the MRS QC meeting organizer to this entire group of recipients I am addressing, concerning the binary QCs. Since the binary QCs are interesting, I would like to make some more discussions.
In the literature, many binary QCs have been observed, including the Al-, Cd-, Ti-, and Cr-based binary systems (to see Pr. Guyot's review, Rep. Prog. Phys., 1991, 54, p. 1373, Table 1). The Ta- and Ga-based systems found later are not listed there. Some of these constituents may be substituted by other elements, not only for the Cd-Cu system (Bendersky et al., Scripta metall., 1987, 21, 531) but also for other alloy systems. Since the substitution seems to be relatively easier in binary alloys, probably there are some more chances to explore new QC alloys by the substitution method (little changes in composition maybe needed), Among those QCs observed so far, some may still remain unclear if they can form from the liquid directly or not.
The interesting thing to us is that why some QCs can form from the liquid directly. However, even if a QC forms directly from the liquid, it doesn't mean that it must be a stable phase (for example, we recently observed that the Mg-Zn-RE IQCs are not thermodynamically stable). Therefore, if a QC is not thermodynamically stable, it should be more accurately renamed as an "as-cast" QC instead of a "stable" QC.
by ultra5.microscopy.com (8.9.3+Sun/8.9.1) id QAA02924 for dist-Microscopy; Mon, 18 Dec 2000 16:51:47 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with SMTP id QAA02921 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Mon, 18 Dec 2000 16:51:17 -0600 (CST) Received: from pdx.edu (tuttle.oit.pdx.edu [131.252.120.29]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with ESMTP id QAA02914 for {Microscopy-at-sparc5.microscopy.com} ; Mon, 18 Dec 2000 16:51:02 -0600 (CST) Received: from [131.252.192.235] (host-192-235.dhcp.pdx.edu [131.252.192.235]) by pdx.edu (8.11.1/8.9.3) with ESMTP id eBIMnnu22753 for {Microscopy-at-sparc5.microscopy.com} ; Mon, 18 Dec 2000 14:49:49 -0800 (PST) Message-Id: {200012182249.eBIMnnu22753-at-pdx.edu} X-Mailer: Microsoft Outlook Express Macintosh Edition - 4.5 (0410)
Hi everyone,
thanks for the deluge of TEM film holders - we have plenty now thanks.
Yet another plea - last week we noticed we were suddenly getting nasty streaks on our SEM photos from light getting to our film holder. Close examination revealed that the plastic casing has cracked through.
In light of the fact that lots of people are going digital with their SEM images, does anyone have an old 550 Polaroid film holder that they are willing to sell to us? If not, we are willing to sell off our 30 packs of polaplan film that goes in this type of holder.
cheers Liz McKenzie
******************************************************* Geomicrobiology and Electron Microscopy Laboratory Room S9 Cramer Hall 1721 SW Broadway Portland State University Portland OR97201
I must protest the blatant and insensitive hemispherism implicit in this posting.
Merry er....just be happy, OK?
Sally Stowe (Canberra, Australia, where its 30 degrees and rising).
Chris Jeffree wrote:
---
Dear All
Best wishes for an environmentally conscious, socially responsible, low stress, non-addictive, gender neutral, winter solstice holiday, practiced within the most joyous traditions of the religious persuasion of your choice, but with due respect for the religious persuasion of others who choose to practice their own religion, as well as those who choose not to practice a religion at all. ........... etc
Folks, this is the year 2000. Anti trust laws dont mean, nor do diddly squat. It is all rhetoric spun off to make it look like things are peachy keen. In reality it is pure B_ _ _S_ _T. Same for when the gurus talk about how great the economy is. Look into your own wallet and assess if you are REALLY doing better than you were 10-20 year ago. The answer is NOT. Lou Solebello
From root Mon Dec 18 20:28:14 2000 Return-Path: {Microscopy-request-at-sparc5.microscopy.com} Received: (from daemon-at-localhost) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) id TAC00250 for dist-Microscopy; Mon, 18 Dec 2000 19:38:18 -0600 (CST) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with SMTP id SAA03116 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Mon, 18 Dec 2000 18:52:01 -0600 (CST) Received: from [206.69.208.21] (mac21.zaluzec.com [206.69.208.21]) by ultra5.microscopy.com (8.9.3+Sun/8.9.1) with ESMTP id SAA03109 for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 18 Dec 2000 18:51:51 -0600 (CST) X-Sender: zaluzec-at-ultra5.microscopy.com Message-Id: {v03130301b66461281365-at-[206.69.208.21]} Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Folks, this is the year 2000. Anti trust laws dont mean, nor do diddly squat. It is all rhetoric spun off to make it look like things are peachy keen. In reality it is pure B_ _ _S_ _T. Same for when the gurus talk about how great the economy is. Look into your own wallet and assess if you are REALLY doing better than you were 10-20 year ago. The answer is NOT. Lou Solebello
To anyone who has experience in attempting to view and determine emulsions bead size using a SEM and determining average particle size of a past wax/paint sealant.
Please respond to Thomas Van Doozer tvd-at-gvtc.com Thank you.
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Zaluzec-at-sparc5.microscopy.com wrote:
} Colleagues } } Chuck Garber's reply to the User Fee discussion was } truncated, due to a subtle effect of the Email system here. } In fairness I am reposting it for him so that you can } see the rest of his comments. } -------------------------------------------------- } } ============================================================= } I am obviously not a lawyer, however, one should always keep in mind that } Congress never granted any kind of exemptions to the anti-trust laws to tax- } exempt organizations. So George Langford is absolutely correct, none of us } should be discussing (with competitors) how we set our pricing.
What rubbish!! Of course we can discuss what standards we use to set fees. Businesses do this all the time.
} Those of us who follow such things know that the penalties can be quite } severe for those who are found to be guilty of violating the anti-trust laws.
Everytime someone gets on this list (and other lists) and asks about setting prices for users of lab services, Mr Langford and/or Dr. Garber starts making veiled threats about anti-trust violations. Isn't it interesting that no actual statues are cited, just vague references and generalizations, and that none of the citers are legally trained?
} So to my (also) legally untrained eye, for universities to be discussing how } they set their pricing is no different than if independent laboratories } (like Amenex and Structure Probe) started a parallel discussion on how we } should be establishing our pricing. Anyone in academia who has ever written } a proposal for funding in recent years knows that universities are in direct } competition with each other in ways that are no different from the way } independent laboratories compete with each other.
Not true of NIH funding of individual investigators.
} George alluded to another } issue, that is, the case when universities start offering their services to } private companies.
If you will read the original posting, that is NOT what was being talked about.
} I am not aware of anyone having gone to jail for this,
I am sure that if anyone had even been formally charged, you or Mr. Langford would be sure to let us know!
} } but it does raise enormous issues of ethical values, and the impact on } students, and their own views of what is right and what is wrong, when the } university's facilities are so commercialized. All of us should have some } concern about such matters.
Ethical behavior is a two-way street.
} Chuck } } Disclaimer: Structure Probe, Inc. is an independent for-profit tax-paying } laboratory offering electron microscopy services to clients, often times in } direct competition with universities who claim they "don't compete with } private companies". } } =================================================== } Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 } President 1-(800)-2424-SPI } Structure Probe, Inc. /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: www.2spi.com } ############################ } ==================================================
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 **********************************************
Hi guys, Anybody who can give me tips on interpretation of thin film micrographs. Or where I can get info on the same subject on the net. Thanks for all your help.
Mr. O. O. ILORI DEPARTMENT OF ELECTRONIC/ELECTRICAL ENGINEERING OBAFEMI AWOLOWO UNIVERSITY, ILE-IFE, OSUN STATE NIGERIA.
We are trying to take the low-power photomicrographs for negative images to identify the connections past the interstitial subnucleus in the Nucleus tractus solitarius by neuronal tracer. Any guidance on this would be very much appreciated Thank you in advance.
Hyung-Tae Kim
Postdoctoral Fellow Laryngeal and Speech Section, MNB, NINDS Bldg. 10 Rm 5D38 10 Center Drive, MSC 1416 Bethesda, MD 20892-1416
At 3:27 PM -0500 12/15/00, Shea Miller wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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
Best wishes for an environmentally conscious, socially responsible, low stress, non-addictive, gender neutral, winter solstice holiday, practiced within the most joyous traditions of the religious persuasion of your choice, but with due respect for the religious persuasion of others who choose to practice their own religion, as well as those who choose not to practice a religion at all.
Additionally, (topical bit here!) a fiscally successful, personally fulfilling, and medically uncomplicated recognition of the generally accepted calendar year 2001, but not without due respect for the alternative calendars of choice of other cultures whose contributions have helped make our society great, without regard to the race, creed, color, religious, or sexual preferences of the wishes.
May the deity of our choice bless us each and every one but should you not be a believer (as above) then please accept our collective wish as fellow sentient (?) beings for the full development of your inner self.
Yours, Chris ==================================================
(Disclaimer: This greeting is subject to confirmation, clarification or withdrawal. It implies no promise by the sendeer actually to implement any of the expressed wishes for her/himself or others and no responsibility for any unintended emotional stress these greetings may or may not bring to those not caught up in the holiday spirit.)
Hope y'all manage to get a life this Christmas ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 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
Thanks for your all your replies on this topic. Some liked it, and some didn't but there was a substantial number of spoiled ballots, so the jury is still out. Best wishes to you all anyway! 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
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ University of Oregon EPMA/SEM Technician ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
The Department of Geological Sciences at the University of Oregon invites applications for an electron beam specialist in a modern microanalytical facility. The facility houses a four-spectrometer Cameca SX-50 microprobe and a JEOL JSM-6300 SEM. It serves faculty and students in the Department of Geological Sciences, Chemistry, Physics and the Materials Science Institute. Primary responsibilities include instrument maintenance, instruction and research collaboration. Additional responsibilities include maintenance and operation of support and sample preparation equipment as well as optical microscopes and a network of computers used for file archival and data/image analysis.
We seek an individual with expertise and experience in WDS and EDS analytical techniques, SEM imaging and instrument maintenance. Additional desired skills include experience with network computers (UNIX/Windows), applied computer-aided microscopy and instruction in instrument use. An advanced degree in petrology, mineralogy or analytical chemistry is desirable.
This is a full-time, annually renewable position. Salary and title will be commensurate with experience and education. Applicants should send curriculum vitae, a statement of experience and interests, and the names, addresses, phone numbers and email addresses of at least three referees to the EPMA/SEM Search Committee, Department of Geological Sciences, 1272 University of Oregon, Eugene OR 97403-1272. We will begin reviewing completed applications February 1, 2001, and will continue until the position is filled.
The University of Oregon is an equal opportunity/affirmative action institution committed to cultural diversity and compliance with the Americans with Disabilities Act.
A PDF of the text is available ... we would genuinely appreciate this announcement's distribution. http://epmalab.uoregon.edu/epma_position.pdf
Feel free to visit the facility's wwwsite for more information regarding the Dep't of Geological Sciences, the University of Oregon, and the community of Eugene. http://epmalab.uoregon.edu/ ...
to ask any specific questions ... about the facility: mailto:epmalab-at-darkwing.uoregon.edu about the position: Jack Rice mailto:jrice-at-darkwing.uoregon.edu
... and a special thanx to my peers ... http://epmalab.uoregon.edu/images/zircon-xmas.jpg
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/
We at university-based facilities are not competing with commercial labs, nor are we out to steal any business away from you. Believe me, most of us have our hands full trying to keep up with the needs/demands of our in-house clients. We are here to provide service at reasonable cost to the research faculty, students, and staff. I know that my facility is expected to cover its non-personnel operating costs (service contracts, supplies, maintenance contracts) and NOT turn a profit. That is against NIH guidelines (take a look at the tapes/transcripts of the M&M sessions devoted to facility management). Since most if not all of my clients are federally funded, I am not allowed to charge them more than a fee calculated to cover my expenses for the procedure involved. My institution maintains a separate EM facility for clinical applications, in part because of the billing differences. I think that the original question posted was a plea for help in establishing de novo an approximation of what is a reasonable price....high enough to cover operating costs and keep from operating too far "in the red", but not so high as to be unreasonable or out of the range for people operating within grant budgets.
It is not unreasonable for people to want to be able to get work done without having to send it out or schlepp it half-way across town. It has become too costly for individuals to maintain this type of equipment in their own labs, and many people need access only on a sporadic basis. That is why central core facilities get set up.
Chill out guys.
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 Colleagues: } } We are trying to take the low-power photomicrographs for negative images to } identify the connections past the interstitial subnucleus in the Nucleus } tractus solitarius by neuronal tracer. } Any guidance on this would be very much appreciated } Thank you in advance. } } } Hyung-Tae Kim } } Postdoctoral Fellow } Laryngeal and Speech Section, MNB, NINDS } Bldg. 10 Rm 5D38 } 10 Center Drive, MSC 1416 } Bethesda, MD 20892-1416 } } email : kimth-at-ninds.nih.gov } Phone : 301-402-0157 } Fax : 301-480-0803
Could you be more specific about the problems you are encountering? What is keeping you from getting useful images?
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 am looking for a used control unit for Reichert Ultracut Ultramicrotome. This is a separate box sits right next to the main microtome and some key functions of the microtome can be controlled from it. If anyone has one for sale, I will be very interested in buying it. Please contact me directly, not to the listserve.
Thank you,
Soumitra
************************************************************* Soumitra Ghoshroy Ph.D. Electron Microscopy Lab and Fluorescence Imaging Facility Department of Biology Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3600 Fax: 505-646-5665 e-mail:sghoshro-at-nmsu.edu http://confocal.nmsu.edu/eml
Help! I'm looking for objective lenses for a Zeiss STEMI SV8 stereomicroscope.
The Zeiss part numbers include: 47 50 73 S 50 mm. 47 50 61 S 100 mm. 47 50 62 PLS 100 mm. 47 50 63 DS 100 mm. 47 50 72 Photo S 100 mm. 47 50 74 S 150 mm. and 47 50 71 S 200 mm.
Please respond to me directly at nessonm-at-ucs.orst.edu with prices and condition. DON'T just hit the REPLY button and bother everyone else in the newsgroup.
TIA, M. Nesson _______________________________________________________________________ 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
How might I find out if there is anything we can do with our old RCA EMU3G TEM other than simply scrapping it? How could I list it where someone who might need it for parts would see it? It was working until the day we stopped teaching the course here in 1990, and with the arrival of our new SEM in January the old TEM has to go to make room for it.
Thanks for any assistance.
Sincerely, Lee Hadden Professor of Biology Department of Biology Wingate University Wingate, NC 28174
Can anybody tell me how to prepare TEM samples for biomaterials such as coral? Thanks. Feng ************************************************** Dr. Feng Wu Dept. of Materials Engineering Ben-Gurion University of the Negev Beer-Sheva 84105, Israel
Philips delivered their first commercial electron microcope in October 1949 (bought by 'Statens Serum Institute' in Denmark for research into the polio virus)
Source: FEI Company - Scope 11 - December 1999 - page 3
Decraen Erwin
Belgian Building Research Institute (BBRI) Division of Materials Av. P. Holoffe 21 B - 1342 Limelette
-----Original Message----- From: Ekstrom, Harry [SMTP:harry.ekstrom-at-honeywell.com] Sent: Wednesday, December 20, 2000 4:57 AM To: '"Geinfam-at-aol.com"-at-sparc5.microscopy.com'; Microscopy-at-sparc5.microscopy.com Subject: RE: Electron microscope first demonstration
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-----Original Message----- } From: "Geinfam-at-aol.com"-at-sparc5.microscopy.com [mailto:"Geinfam-at-aol.com"-at-sparc5.microscopy.com] Sent: Tuesday, December 19, 2000 4:36 PM To: Microscopy-at-sparc5.microscopy.com Subject: Electron microscope first demonstration
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I have a date of Tues 07 April 1931 for the first two-stage (magnification of an electron image further magnified by a second lens) image of a platinum grid being obtained by Ruska.
Ref. T. Mulvey, Dept. Physics, The University of Aston, Birmingham. - An article based upon the opening address of the fifth European congress on electron microscopy, EMCON 72 held in Manchester 5-12 September 1972.
S.C. Hyman Chief Technician The Electron Microscope Laboratory Faculty of Medicine and Biological Sciences Adrian Building University of Leicester University Road Leicester LE1 7RH
Tel. (0116) 252 3370
-----Original Message----- } From: Ekstrom, Harry [mailto:harry.ekstrom-at-honeywell.com] Sent: 20 December 2000 03:57 To: '"Geinfam-at-aol.com"-at-sparc5.microscopy.com'; Microscopy-at-sparc5.microscopy.com
My source quotes M.Knoll first demonstrated the SEM in 1935 but no specific date.
} Can anyone tell me the exact date in the "30's when the electron microscope } was first demonstrated?
I have a little brochure in front of me called "The First North American Electron Microscope", prepared by U.M. Franklin, G.C. Weatherly and G.T. Simon, in 1978. In a section about the early European design efforts, they state that Ruska had "constructed the pioneer two-stage transmission model in 1933", but don't give any further information on it.
Frank Thomas Geological Survey of Canada Atlantic Dartmouth, Nova Scotia Canada
Hello! This is to inform members and other interested individuals that the newsletter for the Southeastern Microscopy Society is now online at http://www.biotech.ufl.edu/sems/ . This issue contains information about our annual meeting to be held May 23-25, 2001 at Clemson University. The issue also includes information and forms for joining the Society. Thanks John P. Shields, PhD Center for Ultrastructural Research 151 Barrow Hall University of Georgia Athens, GA 30602 706-542-4080 FAX 706-542-4271 jshields-at-cb.uga.edu
Eragonite or polyp? Hard or soft? 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 ABN: 99 724 136 560 www.proscitech.com
On Wednesday, December 20, 2000 6:35 PM, Feng Wu [SMTP:fwu-at-gbumail.bgu.ac.il] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Can anybody tell me how to prepare TEM samples for biomaterials such as } coral? } Thanks. } Feng } ************************************************** } Dr. Feng Wu } Dept. of Materials Engineering } Ben-Gurion University of the Negev } Beer-Sheva 84105, Israel } } ************************************************ }
In 1935? That was a TEM, not an SEM. Knoll was Ernst Ruska's PhD supervisor and later collaborator. They designed and built the first TEMs. The very first model had no specimen chamber and just proved that the lens system could work. So the first model, which I seem to remember was before 1935 was really not a microscope. Von Ardenne described the operating principle of an SEM in the late 30th, but I believe none was build until the early 60th at Cambridge University, UK. 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 ABN: 99 724 136 560 www.proscitech.com
On Wednesday, December 20, 2000 1:57 PM, Ekstrom, Harry [SMTP:harry.ekstrom-at-honeywell.com] wrote: } } } My source quotes M.Knoll first demonstrated the SEM in 1935 but no specific } date. } } Harry Ekstrom } Materials Laboratory } } (602) 231-2744 } e-mail: harry.ekstrom-at-honeywell.com } } } -----Original Message----- } } From: "Geinfam-at-aol.com"-at-sparc5.microscopy.com } [mailto:"Geinfam-at-aol.com"-at-sparc5.microscopy.com] } Sent: Tuesday, December 19, 2000 4:36 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Electron microscope first demonstration } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Can anyone tell me the exact date in the "30's when the electron microscope } was first demonstrated? } } geinfam-at-aol.com }
I think Ernst Ruska is generally credited for building the protootype of today's microscope as his PhD project. He and Max Knoll published a description in 1932 (Knoll M, Ruska E. The electron microscope. Z Physik 78:318-39, 1932.).
However, others contributed to the design and knowledge of electron behavior and lenses, including de Broglie on wave theory (Phil Mag 47:446-58, 1924) and Busch on lenses (Ann Physik 15 (Ser 5):145-66, 1932). Also Rudenberg received patents describing and EM in 1932, and Bruche and Johanson built the first electrostatic EM that could enlarge images of self-luminous objects in 1932.
The first commercial EM was developed by von Borries and Ruska and produced in Germany by Siemens and Halske in 1939, and the first commercial EM in the US was the RCA developed by Hillier, Vance, and some others.
This is probably more than you wanted to know, but "the" electron microscope was really a series of developments, and today's microscopes benefitted from additional contributions and refinements of others.
Hope this helps. Sara Miller
On Tue, 19 Dec 2000, Ekstrom, Harry wrote:
} Date: Tue, 19 Dec 2000 20:56:52 -0700 } From: Ekstrom, Harry {harry.ekstrom-at-honeywell.com} } To: "'\"Geinfam-at-aol.com\"-at-sparc5.microscopy.com'" {"Geinfam-at-aol.com"-at-sparc5.microscopy.com} , } Microscopy-at-sparc5.microscopy.com } Subject: RE: Electron microscope first demonstration } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 source quotes M.Knoll first demonstrated the SEM in 1935 but no specific } date. } } Harry Ekstrom } Materials Laboratory } } (602) 231-2744 } e-mail: harry.ekstrom-at-honeywell.com } } } -----Original Message----- } } From: "Geinfam-at-aol.com"-at-sparc5.microscopy.com } [mailto:"Geinfam-at-aol.com"-at-sparc5.microscopy.com] } Sent: Tuesday, December 19, 2000 4:36 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Electron microscope first demonstration } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Can anyone tell me the exact date in the "30's when the electron microscope } was first demonstrated? } } geinfam-at-aol.com } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
An excellent, brief history of EM development, written by Ernst Ruska himself (as the text of his Nobel Prize address) appears in Reviews of Modern Physics Vol. 59 No. 3 Part I p.p. 627-638 (1987). It includes some interesting photos, micrographs, and drawings
To answer the question, in this article Ruska states that he and Max Knoll first demonstrated magnified electron images of a specimen on 7 April 1931. ........................................................................... ...................................................... Jeffrey A. Fortner Argonne National Laboratory CMT/205 9700 S. Cass Avenue Argonne, IL 60439-4837
(630) 252-5594 (voice) (630) 252-4771 (fax)
} ---------- } From: Jim at ProSciTech } Reply To: jim-at-proscitech.com } Sent: Wednesday, December 20, 2000 7:53 AM } To: 'Ekstrom, Harry'; '"Geinfam-at-aol.com"-at-sparc5.microscopy.com'; } Microscopy-at-sparc5.microscopy.com } Subject: RE: Electron microscope first demonstration } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 1935? That was a TEM, not an SEM. Knoll was Ernst Ruska's PhD } supervisor and } later collaborator. They designed and built the first TEMs. } The very first model had no specimen chamber and just proved that the lens } } system could work. So the first model, which I seem to remember was before } 1935 } was really not a microscope. } Von Ardenne described the operating principle of an SEM in the late 30th, } but I } believe none was build until the early 60th at Cambridge University, UK. } 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 } ABN: 99 724 136 560 www.proscitech.com } } On Wednesday, December 20, 2000 1:57 PM, Ekstrom, Harry } [SMTP:harry.ekstrom-at-honeywell.com] wrote: } } } } } } My source quotes M.Knoll first demonstrated the SEM in 1935 but no } specific } } date. } } } } Harry Ekstrom } } Materials Laboratory } } } } (602) 231-2744 } } e-mail: harry.ekstrom-at-honeywell.com } } } } } } -----Original Message----- } } } From: "Geinfam-at-aol.com"-at-sparc5.microscopy.com } } [mailto:"Geinfam-at-aol.com"-at-sparc5.microscopy.com] } } Sent: Tuesday, December 19, 2000 4:36 PM } } To: Microscopy-at-sparc5.microscopy.com } } Subject: Electron microscope first demonstration } } } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Can anyone tell me the exact date in the "30's when the electron } microscope } } was first demonstrated? } } } } geinfam-at-aol.com } } } }
I could not find an exact date ether but Burton and Kohl "The Electron Microscope," Reinhold publishing corp., 1942 in a chapter on History of the Electron Microscope says the following (p149) " In 1932 both types of electron microscopes were described and the first images were shown. E. Bruche and H. Hohannson produced the first electron images of an oxide cathode with an aperture lens system utilizing 300-volt electron beams. M. Knoll and E. Ruska developed the first magnetic electron microscope utilizing 60,000-volt electron beams. The theory of electron lenses was largely developed by J. Picht, O. Shcherzer and W. Henneberg." In the Acknowledgments at the start of the book Dr. Kohl (one of the authors) is said to have given a demonstration of the oxide cathodes experiments: "Such an electron microscope was demonstrated before the seminar of the McLennan Laboratory in April, 1934; this probably was the first such demonstration in Canada." according to Burton. This puts the non-scanning types back a few years before the SEM date given below. Burton and Kohl don't provide any better dates than these, that I could find. Hope this might help you in your search. Jim.
James L. Roberts Firearm and Toolmark Examiner Ventura Co. Sheriff's Lab (805) 654-2308 James.Roberts-at-mail.co.ventura.ca.us
} } } "Ekstrom, Harry" {harry.ekstrom-at-honeywell.com} 12/19/00 07:56PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
My source quotes M.Knoll first demonstrated the SEM in 1935 but no specific date.
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Good luckn and Happy Holidays
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
Can anybody tell me how to prepare TEM samples for Single crystal? Thanks for your interested
Dear Erdem, It depends on the crystal and other parameters. Are you doing selected-area (SAED), convergent beam (CBED), or reflection (RHEED) diffraction? Is your specimen radiation resistant--such as a mineral--or labile--such as an organic? Will it stay solid in the vacuum, or will it need to be kept at LN2 temperature (as does anthracene)? What do you need to measure, intensities or lattice constants? What voltage will you use? The preparation can be very simple--e.g., MgO, which is made by burning Mg and collecting the smoke on a formvar-coated grid--or complicated--e.g., growing copper perchlorophthalocyanine epitaxially on freshly-cleaved alkali halide from a vapor or isolating a protein and finding crystallization conditions that produce well-ordered platy single crystals. Let me know some details, and I'll see if I can help. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
Many thanks for the awful warnings, confessions and suggestions. We use two ordinary 9cm plastic funnels which have cracked and one 20cm funnel labelled KENUTUF, supplied with the mic to top up the Xray analysis dewar, which still seems as good as new after at least six years.
I love the Origami solution but to replace the little funnels I'll go with the stainless type for durability and add an insulated handle.
On the bung popping dewar front, the answer is not tin hats and a shout of 'incoming' but to provide a small vent. Even pressurised dewars need a relief valve.
Try Dr. Joseph L. Scott at The College of William and Mary (Williamsburg, VA)... jlscot-at-facstaff.wm.edu His research includes TEM of coralline algae.
Patty Clow
~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Patricia A. Clow, Ph.D. Staff Scientist I / Microscopy Genzyme Corporation One Mountain Road PO Box 9322 Framingham, MA 01701-9322 phone: 508-270-2053 fax: 508-872-9080 email: patricia.clow-at-genzyme.com
-----Original Message----- } From: Feng Wu [mailto:fwu-at-gbumail.bgu.ac.il] Sent: Wednesday, December 20, 2000 3:35 AM To: Microscopy-at-sparc5.microscopy.com
Can anybody tell me how to prepare TEM samples for biomaterials such as coral? Thanks. Feng ************************************************** Dr. Feng Wu Dept. of Materials Engineering Ben-Gurion University of the Negev Beer-Sheva 84105, Israel
The current discussion of user fees appears to have strong, contrary opinions, so it seems worthwhile to take a look at what is posted at the US Department of Justice web site.
The following is taken "http://www.usdoj.gov/atr/foia/divisionmanual/ch2.htm".
Trusts, etc., in restraint of trade illegal; penalty
Every contract, combination in the form of trust or otherwise, or conspiracy, in restraint of trade or commerce among the several States, or with foreign nations, is declared to be illegal. Every person who shall make any contract or engage in any combination or conspiracy hereby declared to be illegal shall be deemed guilty of a felony, and, on conviction thereof, shall be punished by fine not exceeding $10,000,000 if a corporation, or, if any other person, $350,000, or by imprisonment not exceeding three years, or by both said punishments, in the discretion of the court.
§ 2 Sherman Act, 15 U.S.C. § 2
Monopolizing trade a felony; penalty
Every person who shall monopolize, or attempt to monopolize, or combine or conspire with any other person or persons, to monopolize any part of the trade or commerce among the several States, or with foreign nations, shall be deemed guilty of a felony, and, on conviction thereof, shall be punished by fine not exceeding $10,000,000 if a corporation, or, if any other person, $350,000, or by imprisonment not exceeding three years, or by both said punishments, in the discretion of the court.
(There are many more sections, but I believe these two summarize the main issues of the Act.) { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { { {
Based on a common sense reading of the above, it seems to me that "restraint of trade" and the "attempt to monopolize" are the key factors that that Sherman Antitrust Act is designed to prevent, however, the issue of "intent" is not obvious.
I would think that a discussion by EM facility managers about how they charge for the use of their facilities and services, where the main emphasis seems to be how to cover the expenses of running the facilities, does not appear to be an intentional "restraint of trade" or an "attempt to monopolize" their trade.
I would agree that any suggestion of price fixing or how to undercut the rates of private companies would certainly violate the Sherman Antitrust Act, but I hardly think anyone can argue that any such comments have been made in the current discussion.
However, the question remains that if the well intentioned discussions between managers of nonprofit and/or university supported facilities on how to charge to cover their operating costs results in wide spread rates that actually do undercut private companies, is that illegal? I will refrain from presenting my opinion, since I believe that only someone with the appropriate legal background can give an authoritative answer to this question. And considering that every lawsuit has lawyers representing the opposing sides, the answer may only be clear if there is already a precedent on record where a similar case has actually gone through the courts.
Joe _________________________________________ Joe Nabity, Ph.D. JC Nabity Lithography Systems E-Beam Lithography using Commercial SEMs & STEMs PO Box 5354, Bozeman, MT 59717 USA Voice: (406) 587-0848 FAX: (406) 586-9514 E-mail: info-at-jcnabity.com Web: www.jcnabity.com
Personnel salaries ARE part of the operating costs of running any type of laboratory facility. In addition, the cost of an electron microscope is also part of the operating expense of operating a lab. To suggest that only the non personnel costs such as service contracts, supplies etc. should be counted as operating costs really distorts the true picture.
I came from a university lab that originally was small and able to meet all of its operating expenses including salaries. Eventually visionaries saw the need for a larger more grand facility that would include some instruments with an annual service contract around 35K, other contract costs were in the range of 9K to 20K per year. The facility was built and the rates were kept low for academic users (about $25/hr.) and rates for non university users were scaled depending upon association with the facility. It didn't matter that it required about 1400 hours of beam time on one instrument just to pay the service contract because the facility received 8-10 million dollars per year in support from NSF. Principle instruments were obtained in grants and other subsidies not related to operating expenses. Eventually the NSF contract expired and now with aging equipment technology is passing them by.
The point of this is that nearly all university facilities will eventually have to be accountable for all of their expenses and adjust their rates to cover the costs. But universities still will have an advantage in that they usually don't have to purchase the principle equipment from operating expenses. How NSF granted instrumentation can be used is described in NSF Important Rule 91.
When university labs have to account for all of their operating expenses, they will become competitors of private non-subsidized labs. If they offer instruments rates less than private lab, they are cheating.
John Humenansky/Staff Scientist Physical Electronics, Inc. (PHI) 6509 Flying Cloud Drive Eden Prairie, MN 55344 952-828-6387
I get a rare opportunity to renovate some space to move our EM suite and have to turn in the ideal lab concept very soon. I know there is a book on this very thing. Anyone recall the title and author??
I am now in charge of a Phillips XL30 ESEM and have a couple of questions. What are you doing to prevent contamination of the specimen chamber. I had a user wanting to view human tissue today unfixed and am reluctant to do so. Anyone have a solution. Mine was to fix and process for standard SEM but that may not be an option for some of the in-situ studies or preserved museum specimens they don't want to wash or damage. Thanks
} Small stainless steel funnels are used to fill many LN2 } anticontamination devices on electron microscopes. I have found that a } Java jacket (designed for holding hot coffee cups) makes a great } reuseable jacket for picking up these funnels when they are cold.
I want to cheaply mount an inexpensive video camera onto the optical microscope of, yes, you guessed it, a JEOL 840. Economically, you might say.
Somebody been there, or near there?
I'm not very knowledgeable about optics, do I need to get
1 a lensless video camera and take off the microscope eyepiece,
2 a lensless video camera and retain the microcope eyepiece,
3 a video camera with a lens and take off the microscope eyepiece,
4 a video camera with a lens and retain the microscope eyepiece,
or
5 none of the above?
I'm sure I can devise the neccessary spacer/adaptor
tia
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
One volume is "Design of the Electron Microscope Laboratory", by Ronald Alderson. It is part of the Glauert series, "Practical Methods in Electron Microscopy". The edition we have is from 1975 and I don't know if there is a newer one.
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: Scott Whittaker [mailto:Whittaker.scott-at-nmnh.si.edu] Sent: Wednesday, December 20, 2000 3:08 PM To: Microscopy-at-sparc5.microscopy.com
I get a rare opportunity to renovate some space to move our EM suite and have to turn in the ideal lab concept very soon. I know there is a book on this very thing. Anyone recall the title and author??
I think the point we are missing here is geinfam-at-aol.com asked when the first EM microscope was first "demonstrated". My source states the theoretical "description" of the SEM was brought to us by H. Stintzing in 1929. However the first "demonstration" of an SEM was in 1935 by M. Knoll. In 1932, the first TEM was "constructed" by Knoll and Ruska. I wonder how they all felt about the electoral college.
----- Original Message ----- } From: Fortner, Jeffrey A. {fortner-at-cmt.anl.gov} To: 'Microscopy Listserver' {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, December 21, 2000 3:32 AM
I worked out the real, full-costs-recovery cost of our emlab a few years ago, and came to the conclusion that we would have to charge around £1250 a day to our users for access to a TEM. The charge for basic access to a top-grade analytical FEG TEM at a commercial rate could easily be in the order of £3000 per day. The task for most of us, as you indicate, is to keep the running costs down to a level which the University can tolerate. Most University facilities I know of in the UK are free of the requirement to earn the replacement cost of the equipment, and have to earn only limited staff costs. I am aware that there is ever-increasing pressure on Universities to earn more of the costs of their operations. The latest rumour is that electricity and water charges are just around the corner, but that one is at least a decade old now. I'll believe it when they install the meters. Maybe it'll come, maybe it won't. But in the past 15 years of operating a facility I swear that the user attitudes to charging don't reflect any recognition of this. Internal users still lobby for access to be free, or at least to appear free at the point of use. We therefore make just a few k per annum towards our costs by selling a little slack time to any local industries that have a need for our services. Who exactly are we cheating by doing this? An alternative way of looking at it is that we are offering the public some access to a facility they paid for in the first place, at a rate which is no more than is necessary to keep the machines running. I personally don't know of any commercial operation anywhere within hundreds of miles of Edinburgh that we are in competition with. Our operation is strictly self-limiting. once we have made the few k required to cover our costs the focus is back on internal user support. If there is any real industrial EM business out there any competent commercial operation would have no difficulty whatever in surviving whatever level of pseudo-commercial activity Universities are involved in. If there is any real evidence to the contrary I would like to hear it (and would be amazed) We're no threat, so please don't keep shooting us! Chris
} The point of this is that nearly all university facilities will eventually
} have to be accountable for all of their expenses and adjust their rates to } cover the costs. But universities still will have an advantage in that } they usually don't have to purchase the principle equipment from operating } expenses. How NSF granted instrumentation can be used is described in NSF } Important Rule 91. } } When university labs have to account for all of their operating expenses, } they will become competitors of private non-subsidized labs. If they offer } instruments rates less than private lab, they are cheating. } } John Humenansky/Staff Scientist } Physical Electronics, Inc. (PHI) } 6509 Flying Cloud Drive } Eden Prairie, MN 55344 } 952-828-6387 } }
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 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
Can anybody suggest how to measure thickness of the conductive coating for EM specimen? If it is transparent, ellipsometer could be used. Since metallic or carbon coating is not transparent, I could not use that method. Then do I have only way to measure directly, like measuring thickness on the cross-section or on the surface by using EM or SPM? Is there any other way?
I have the following problem during my TEM observation. I am looking at titania (TiO2), rutile, particles which are placed on top of a carbon coated cooper grid. Those particles are ca. 90 nm long and ca. 15 nm wide. At 200 kV the particles are very unstable under the electron beam. I think charging occurs. The attempt to coat the sample with carbon did show some success, but even after very long sputtering times (15 min. at 25 A) most particles twinkle.
If anybody has an idea for me how to avoid the twinkling, preferentially without sputtering any carbon on top of the sample, I would be very thankful.
Sincerely, Joon
************************************************************ Joon Hwan CHOI, Ph.D. Post-doctoral Researcher Materials Department University of California at Santa Barbara (UCSB) Santa Barbara, CA 93106
Ernst Ruska received in 1986 the Nobel Prize in Physics "for his fundamental work on electron optics and for the design of the first electon microscope".
You can read more on the Official Web Site of the Nobel Foundation: http://www.nobel.se/physics/laureates/1986/index.html
Cheers,
Paul ====================== Paul Baggethun Alcoa Technical Center Alcoa Center, PA 15069 USA ======================
-----Original Message----- } From: "Geinfam-at-aol.com"-at-sparc5.microscopy.com [mailto:"Geinfam-at-aol.com"-at-sparc5.microscopy.com] Sent: Tuesday, December 19, 2000 6:36 PM To: Microscopy-at-sparc5.microscopy.com
I think the twinkling problem is more electron beam heating of your sample as opposed to charging. If the samples were charging, they would be moving on the carbon support film. Carbon coating the samples will help. Are you using holey carbon grids or continuous support films. The latter should help somewhat, but you have about 200A of carbon that adds to the thickness. You could also try a cold stage to see if that could help. You could also try a higher voltage machine if one is available to you. Higher voltages allow the electrons to pass through your sample with less interaction and therefore less heating occurs.
-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: Joon Hwan CHOI [mailto:jchoi-at-engineering.ucsb.edu] Sent: Wednesday, December 20, 2000 9:32 PM To: Microscopy-at-sparc5.microscopy.com Subject: TEM specimen preparation of titania particles
--------------------------------------------------------------- --------- The Microscopy ListServer -- Sponsor: The Microscopy Society 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 the following problem during my TEM observation. I am looking at titania (TiO2), rutile, particles which are placed on top of a carbon coated cooper grid. Those particles are ca. 90 nm long and ca. 15 nm wide. At 200 kV the particles are very unstable under the electron beam. I think charging occurs. The attempt to coat the sample with carbon did show some success, but even after very long sputtering times (15 min. at 25 A) most particles twinkle.
If anybody has an idea for me how to avoid the twinkling, preferentially without sputtering any carbon on top of the sample, I would be very thankful.
Sincerely, Joon
************************************************************ Joon Hwan CHOI, Ph.D. Post-doctoral Researcher Materials Department University of California at Santa Barbara (UCSB) Santa Barbara, CA 93106
About the firts TEM, the question is answered. But about the SEM, a few precisions.
Some interresting papers gives the essential informations about its history. But we must keep in mind that the early SEM was a "reflexion microscope" designed in analogy with the optical microscope. And the development were made in paralell with the TEM. So the design looks like a modified TEM, and not like what we know today as a SEM. It was also the case with the first Stereoscan.
The papers :
M. Von Ardenne Zur Geschichte der Rasterelektronenmikroskopie und der Elektronenmikrosonde. Optik 50 (1978) No 3, 177-188
C.W. Oatley The early history of the scanning electron microscope J. Appl. Phys. 53(2) Feb 1982
The first papers of Von Ardenne were from 1937 and 39 (Zeitschrift fur Physik), and he had built an instrument and published some photography. The intrument was destroyed by the bombardement at the end of WW II.
An other interresting paper is :
V.K. Zworykin, J. Hillier and R.L.Snder A Scanning Electron Microscope ASTM bulletin August 1942
After a lot of nice experiments (with FE guns!) they conclued that it was too expensive, with a poor signal and to less resolution.
J. Faerber IPCMS-GSI 23, rue de Loess 67037 Strasbourg CEDEX France
We fix human cells for safety reasons but sometimes look at them wet (hard work!). Chris Gilpin argues that fixation helps specimens resist beam damage and that most distortion occurs during drying rather than fixation.
Dave
On Wed, 20 Dec 2000 16:15:13 -0500 Scott Whittaker {Whittaker.scott-at-nmnh.si.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 am now in charge of a Phillips XL30 ESEM and have a couple of questions. What are you doing to prevent contamination of the specimen chamber. I had a user wanting to view human tissue today unfixed and am reluctant to do so. Anyone have a solution. Mine was to fix and process for standard SEM but that may not be an option for some of the in-situ studies or preserved museum specimens they don't want to wash or damage. Thanks } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
At 4:08 PM -0500 12/20/00, Scott Whittaker wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
******************* Hi Scott, I just pulled the volume off my shelf. It is part of the series edited by Audrey Glauert: Practical Mehods in Electron Microscopy Volume 4 "Design of the Electron Microscope Laboratory" written by Ronald H. Alderson original copyright 1975. There may be a more recent edition, but this one certainly covers the main considerations.
I envy your opportunity to "start from scratch" many of us end up retrofitting spaces designed for other purposes.Have fun.
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
By analogy, any combination (even among nonprofits) for the purposes of setting rates must be illegal if the well publicized divvying up of applicants to graduate school was an illegal restraint of trade.
Private for-profit industry was not involved in that particular conspiracy in any way.
On the other hand, the mere listing of categories of expense and the discussion of how best to allocate those costs between users and providers seem appropriate. That's essentially the same as a discussion of the principles of accounting or the geometry of the Ewald sphere.
When specific rates are mentioned, that's what looks like price fixing.
Best regards, George Langford, Sc.D. amenex-at-amenex.com
I disagree that posting rates implies core facilities are attempting price-fixing. Surely commercial entities look at the published costs of their competitors all the time. When one airline drops its rates, the others usually follow within 24 hrs. Price-fixing only results if you make your decisions in concert with your competitors. Knowing how much other core facilities charge has several benefits. First, the core knows what level of fees allow investigators to afford to use a technology. If I charged enough to fully reimburse the true expenses of my core, most of my researchers would simply switch to alternative techniques since they have limited grant dollars. Secondly, it allows core directors to demonstrate to administrators what portion of operating expenses other cores attempt to recover from user fees. I always tell my administrators that I am to make a profit that rivals our university library! Lots of cores say they try to recover operating expenses but unlike industry, we never account for building costs, electricity and heat, maintenance, etc. Researchers pay for those types of things with their indirect costs that accompany virtually all grants. This makes a comparison of commercial vs university fees impossible to judge. Indirect costs also help many universities pay for the "matching portion" that most federal grant agencies require as a part of matching equipment grants. Whether I use the flow cytometer on my campus or not, my indirect costs help pay for it. In addition, as a tax-paying citizen of the state of Missouri, my tax dollars help subsidize our University. We pay these taxes, in part, because we want our students to have access to the cutting edge technologies that will make them competitive citizens. What good would having all these fun toys be if they were too expensive to allow our undergrads and grads some time on them. These subsidies have no equivalent with private industry.
Anyone wanting to see what MU charges for confocal and other types of LM, should surf over to http://www.biotech.missouri.edu/mcc/prices.html . The costs for our EM core are at http://www.biotech.missouri.edu/mbp/cores/ . I am responsible for posting the LM core prices but Randy Tindall is responsible for the EM core prices being posted so please direct all lawsuits regarding them at him (Sorry, Randy). See you in court.
Merry Christmas and Happy Holidays to all. 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)
Dear Scott I had the same opportunity to design for an EM lab when I moved to the University of Nebraska -- new facility, new space. Fortunately the lab spaces offered included one that I checked to be virtually ideal. This was in the basement of a building that had 3 floors above ground plus the basement and that was intended to be suitable for adding another 3 floors on top. The footings and the (1 foot thick reinforced concrete) wall surrounding the lab were really massive and confer the best stability I've heard of -- floor vibration velocities { 0.5micrometers per second (at frequencies 1 to } 50 Hz) and amplitudes { 0.5 micrometers (peak to peak) and AC magnetic fields {0.5 mG (before EM equipment installation). I got the architectural and construction team to provide several important provisions for use of the space for EM: 1. laminar air inlets to rooms (positioned so as not to affect the TEM columns!), 2. near-floor level return air vent in the TEM rooms (to satisfy needs to remove SF6 since it is denser than air), 3. acoustic lining of air inlet ducts, 4. temperature regulation to +/- 1 C with a maximum rate 1 C / hour, 5. humidity regulation in the range 40 to 60% relative humidity year round 6. water chillers in a mechanical facilities room adjacent to, but outside, the EM facility.
The above aspects overlap and go beyond the necessarily somewhat outdated recommendations in Anderson's book.
There were still issues that required care in order for the TEM to meet the image drift specifications I required of the manufacturers. These measures included adjustment of the air flow in the HREM room and reduction of the temperature difference between the TEM water temperature and the room air temperature. The TEM water chiller also required to be equipped with the precision electronic temperature regulation option. Under these conditions, there is no need to install curtains or otherwise to curtail air circulation during high resolution work.
I am very happy that our facilities design and management staff, construction and maintenance people, and microscope manufacturer's representatives could all be persuaded to achieve the desired end result -- continued, essentially trouble-free, stable operations of our lab in all conditions of weather and use of the rest of the building. It is definitely worth taking great care over the major and minor details before you start!
If I can provide other information. please let me know. Best wishes Brian Robertson
*********************************************************** Assoc. Prof. Brian W. Robertson Department of Mechanical Engineering and Center for Materials Research and Analysis University of Nebraska-Lincoln, N124 WSEC, 17th & Vine Sts., Lincoln, NE 68588-0656, USA ** Office 402 472 8308; FAX 472 1465; Labs 472 8762 and 472 5498 **
HI Ritchie, here is my cheap fix ( {1K$US) As you mention in 1-4 the eye piece is the trick. If you have std 23 mm eye pieces (For larger slip in eye pieces you can make a bushing) you can buy a relay lens from Edmund Scientific & others (23 mm x c-mount). Then buy a color security camera w/c-mount & connect to your video recording device. I use a Snappy TM interface to my laptop. I can throw all this in my laptop tote sack & aquire pretty decent images at almost any optical u-scope I walk up to. ball park on economics, optics $500, camera $200-300, Snappy {$100US For higher res. images there has been a lot of buzz on this list server about interfacing the Cool Pic TM digital camera. ck the archives. Don't forget to recalibrate you magnification.
If you have thread on eye pieces... I'd like to know what you find out.
disclaimer: no financial interest in companies or products mentioned.
Happy holidays, 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 } -----------------------------------------------------------------------. } } Hi again } } I want to cheaply mount an inexpensive video camera onto the optical } microscope of, yes, you guessed it, a JEOL 840. Economically, you } might say. } } Somebody been there, or near there? } } I'm not very knowledgeable about optics, do I need to get } } 1 a lensless video camera and take off the microscope eyepiece, } } 2 a lensless video camera and retain the microcope eyepiece, } } 3 a video camera with a lens and take off the microscope eyepiece, } } 4 a video camera with a lens and retain the microscope eyepiece, } } or } } 5 none of the above? } } I'm sure I can devise the neccessary spacer/adaptor } } tia } } 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
Tell you what, folks. Next time we marauding academic pirates are in need of guidance on fees, let's just ask for links to web pages where other labs already have their rates published. Maybe that will make everybody happy and serve the same purpose as our usual blatant conspiracy does. That way we can take over the whole world legally!
Yeah, that's the ticket.....
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/
Has some a recept to clean the ceramic holder from a LaB6 filament. It is shorted (300 kohm at rohm temp., 0 at working temp.) with the filament. The tip is OK. Is it a way to remove the evaported coating (LaB6, decomposed LaB6 or carbon ?) on the ceramic, without dammaging the tip. Mechanically seems to bee inpracticable, it's too small (10 mm diameter, 5 mm heigth between the ceramic holder and the graphite blocs), and I have no "microbillage" (I don't know in english). But chemicaly ? I don't know the manufacturer of the tip, it comes from a LEED optic.
Thanks
J. Faerber IPCMS-GSI 23, rue de Loess 67037 Strasbourg CEDEX France
Your reply proves my point. If you were a business your personnel salaries and instrument costs would have to be factored into the ACTUAL operating costs. The instrument rates that you charge for outside clients are based on YOUR operating costs which do not include salaries or instruments and can therefore be much lower than what a business MUST charge to recover their costs and make a profit. Many universities are not in competition to private business because they are funded internally and exist primarily for education and research. Too often however the economics of operating a university facility collide with the reduction in institutional funding and then outside users are sought which is the unfair competition aspect unless the university rates for outside users are the same as what the businesses are charging for similar equipment.
John Humenansky/Staff Scientist Physical Electronics, Inc. (PHI) 6509 Flying Cloud Drive Eden Prairie, MN 55344 952-828-6387
} We pay these taxes, in part, because we want our } students to have access to the cutting edge technologies that will } make them competitive citizens. What good would having all these fun } toys be if they were too expensive to allow our undergrads and grads } some time on them.
I would also argue that students who never have a chance to see or use an electron microscope during their training are less likely to seek out those services later when they go to work in the private sector. I strongly suspect that the availability of EM in Universities is an important factor in creating a market for private sector EM providers.
Marie
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology, U-2131 University of Connecticut Storrs, CT 06269-2131 Phone: 860-486-3588 Fax: 860-486-6369
} } } One point that seems to have been missed is that the various } university labs do not compete with each other but rather need to recover } costs within their institution. This is a cost recovery and } redistribution process from within a single fiscal entity. (All grants } etc. are university funds and no longer belong to the granting agency or } the PI.) So I don't get this restraint of trade nonsense. We do not } sell anything within our institution. We provide a service that must be } financially supported in one way or another and it is most appropriate to } have the users of our facility provide all or part of those costs. So I } figure we can discuss anything we damn well please among professionals } who provide these services. } } At 10:01 AM 12/21/2000 -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 think all opinions have been aired and nothing new has been said for awhile. I'm going to suggest that the thread be left to ferment for another year when I'm sure the question will get asked once again.
It's a good bet that it will work. If it doesn't, try changing the distance between the microscope and the camera. Also, if you cannot fill the video chip completely with the image, or if it is over filled or distorted, try using the video lens with an eyepiece preceding it before you consider purchasing any additional optics. The alignment of the eyepiece and video lens from side to side, as well as the distance separation them are critical. A small deviation can lead to large changes in the image appearance and quality. There is a sweet spot, so patience prevails. Again, this is not the optimal way to interface a video camera and a microscope, but it is cost effective and performs well in many non-critical applications. If the video lens has an adjustable focus, it may be possible to use it without the eyepiece, and it is worth a try if all else fails.
Best of luck
John
John Turner, Ph.D. Director, Advanced Chemical Imaging Facility Cleveland State University Cleveland, Ohio 44115
I beleive the first high resolution SEM was built by Oliver Wells as a PhD project in 1959. It was enabled by the Everhart-Thornley secondary electron detector. My understanding is that the theory of how an SEM should work was developed in the '30s but specimen current and other available signals couldn't be amplified sufficiently or cleanly enough to be useable. Most of them still can't be used for high resolution work.
Ken Converse owner Quality Images third party SEM service Delta, PA
Jim at ProSciTech 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 } -----------------------------------------------------------------------. } } In 1935? That was a TEM, not an SEM. Knoll was Ernst Ruska's PhD supervisor and } later collaborator. They designed and built the first TEMs. } The very first model had no specimen chamber and just proved that the lens } system could work. So the first model, which I seem to remember was before 1935 } was really not a microscope. } Von Ardenne described the operating principle of an SEM in the late 30th, but I } believe none was build until the early 60th at Cambridge University, UK. } 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 } ABN: 99 724 136 560 www.proscitech.com } } On Wednesday, December 20, 2000 1:57 PM, Ekstrom, Harry } [SMTP:harry.ekstrom-at-honeywell.com] wrote: } } } } } } My source quotes M.Knoll first demonstrated the SEM in 1935 but no specific } } date. } } } } Harry Ekstrom } } Materials Laboratory } } } } (602) 231-2744 } } e-mail: harry.ekstrom-at-honeywell.com } } } } } } -----Original Message----- } } } From: "Geinfam-at-aol.com"-at-sparc5.microscopy.com } } [mailto:"Geinfam-at-aol.com"-at-sparc5.microscopy.com] } } Sent: Tuesday, December 19, 2000 4:36 PM } } To: Microscopy-at-sparc5.microscopy.com } } Subject: Electron microscope first demonstration } } } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Can anyone tell me the exact date in the "30's when the electron microscope } } was first demonstrated? } } } } geinfam-at-aol.com } }
Is there somebody who know how to calculate fine structures near L23 edges by using ICXANES, while this code is restricted to spectra involving core levels of s symmetry, i.e. K, L1...?
} Are you going to fill us in on the answer, or do you even know it? } ... } Inquiring minds want to know.
I thought I had ... it is a mystery to me how some posts, for example this facility supervisor vacancy's announcement, get cross-posted everywhere, and why the subsequent discussion of this image missed this list :o)
http://epmalab.uoregon.edu/images/zircon-xmas.jpg
It is an image of e-beam induced CL as acquired via RGB filters. The blue is somewhat typical of igneous (or primary) quartz ... the red halo is evidence of radiation damage from a nearby zircon inclusion. Other images from the same rock actually show us the "smoking gun". We investigated a number of sandstones, and otherwize quartz bearing rocks from this environment, and another type of radiation induced CL we observed was red indistinct rims on qtz grains in sandstone. We imagined radioactive fluids percolating through the rocks.
My turn for True Confessions or surviving liquid nitrogen incidents.
Many years ago in another time warp (70's) a "green" technician (no, not environmental green) was dutifully following his supervisor's orders to fill EDX detector Dewars with liquid nitrogen. The staff had used a heavy-duty plastic Nalgene pitcher, wrapped with lab tape, to accomplish this chore. Thus, the technician followed protocol and used the same pitcher to collect the cryogenic fluid. And it worked well for many, many weeks. However, one day as he was filling the wrapped container, there was an enormous explosion (or implosion?). The technician was left holding the handle of the pitcher with the rest of the pitcher vanished into hundreds of small plastic pieces and liquid nitrogen splattered everywhere. Holy mackerel! But for the grace of God, not one piece of plastic or drop of liquid nitrogen hit the SURPRISED technician!
However, to this day that technician remembers his mother's admonitions about wearing clean underwear, you never know when an eighteen wheeler will hit you.
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-commserver.srrc.usda.gov
=================================================== Fernando D. Balducci Laboratorio de Microscopia Electrónica Facultad de Ingeniería - Bioingenieria Universidad Nacional de Entre Rios Argentina e-mail: microsc-at-fi.uner.edu.ar tel: 54 43 975100 fax: 54 43 975077
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Hi, all Thanks for your attention to my question on coral TEM specimen. The coral is hard, but not dense. A lot of platelets arranged in radiation way were observed in optical microscope. Can anybody give me some suggestion to prepare it? Thanks again. ************************************************** Dr. Feng Wu Dept. of Materials Engineering Ben-Gurion University of the Negev Beer-Sheva 84105, Israel
fax 972-7-6472944 tel 972-7-6461473(o) -6281432(h) email: fwu-at-bgumail.bgu.ac.il ************************************************ ----- Original Message ----- } From: Feng Wu {fwu-at-bgumail.bgu.ac.il} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, December 20, 2000 10:34 AM
Dear Dr. Feng Wu,
I have used the following procedure to fix specimen of Leptogorgia virgulata, an octocoral (sea whip) found off the eastern coast of the USA. When fixed according to this procedure, whole tissue specimen, as well as the harvested sclerites by themselves, can be prepared for viewing in either SEM or TEM. I hope you find this information helpful.
Regards,
Elizabeth Bray Plant Chemist SCE&G Central Laboratory 2102 N. Lake Dr. Columbia, SC 29212 USA
Fixing and Embedding Protocol for Octocorals and Their Sclerites ================================================================
Reagent Preparation: 1. Purchase the following: 16% Paraformaldehyde (PFA) 50% Gluteraldehyde (GA) NaCacodylate, reagent grade [Sodium cacodylate (CH3)2AsO2Na · 3H2O, MW = 214.02] LR White acrylic embedding medium
2. 0.1 M NaCacodylate (CH3)2AsO2Na · 3H2O, MW = 214.02 21.4 g NaCacodylate diluted to 1000 mL with DW. Adjust the pH to 7.4 with 4% NaOH or 1:10 HCl.
3. Fixative: 2% PFA, 0.15% GA, 0.05 M NaCacodylate in filtered Sea Water: 6.3 mL of 16% PFA 0.15 mL of 50% GA 25.0 mL of 0.1 M NaCacodylate, pH = 7.4 18.55 mL of Millipore filtered Sea Water Total Volume of fixative = 50 mL
Procedure: 1. Fix the specimen in PFA/GA/NaCaco fixative for 1 hour. 2. Rinse in 0.05 M NaCaco Buffer 3 x 10 minutes. 3. Dehydrate in a series of EtOH (ethyl alcohol) washes of ever increasing strength: 50% EtOH 10 minutes. 75% EtOH 10 minutes. 95% EtOH 15 minutes. 100% EtOH 15 minutes x 2, done under vacuum.
4. Three (3) changes of LR White, all done under vacuum. The first two changes for at least 30 minutes, and the last change overnight. The last change should be done in the BEEM capsules in which the specimen will be cured.
NOTE: Be sure that when you fill the capsules with the LR White for the final time, there are no air bubbles in them. This will cause the level of embedding medium to become lower in the capsule as it sits overnight under vacuum, and when you cap them for curing in the oven, the air inside will lead to incomplete curing making it difficult, if not impossible, to properly section the hard coral material. To ensure that there are no bubbles present, allow the capsules to sit under vacuum for 30 minutes to 1 hour, go back and top off any capsules that need it, then reestablish the vacuum and allow them to sit overnight. Before capping, ensure that the level of embedding medium is at the top of the capsule. If it is not, carefully add sufficient medium to restore the proper level and proceed with capping.
5. Cap to ensure the exclusion of air and cure overnight in a 60o C oven.
A position as a TEM Engineer is open at Infineon Technologies, Richmond.
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Job Responsibilities: To coordinate, optimize and perform TEM analysis in support of DRAM fabrication.
The candidate will perform: 1: Coordination of TEM analysis jobs with lab customers and TEM technicians 2: Compilation, documentation and presentation of analysis results. 3: Coordination of tool maintenance and development. 4: TEM sample preparation using mechanical polishing and focused ion beam (FIB) techniques. 5: TEM inspections of CMOS semiconductor interfaces, dislocations, films and defects.
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You may be referring to Vol. 4 of Practical Methods in Electron Microscopy, A. Glauert, ed., North Holland Pub., 1975. The book is a monograph by Ronald H Alderson on "Design of the Electron Microscope Laboratory". It has a lot of good information, but may be getting a bit dated for the ultimate level of environment for today's most sophisticated and sensitive instruments.
hope this helps...
Larry
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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
Hi, I am looking for a protocol for embedding plant tissues in LR White resin. I am working with endosperm and plant embryo of seeds of Ilex paraguariensis (maté). The endosperm contains a large amount of proteins and lipids. Thank's.
Dr. Rinaldo Pires dos Santos Dept. of Botany - UFRGS Brazil E-mail: rinaldop-at-uol.com.br Tel: + 55 51 4964053 (home) + 55 51 3167634 (UFRGS)
Thank you for your kind replies. The equipment that you mentioned seems too expensive for me to use, especially when I need just a couple of times thickness measurement for the calibration of my sputter coater. Is there other way to measure the thickness? Tobias suggested to use ellipsometry technique because the metallic or carbon coating transmits light when it is very thin. Have any you ever tried this before?
Best wishes,
Jondo Yun Kyungnam University, Korea jdyun-at-hanma.kyungnam.ac.kr
There are special machines called "thickness monitor" on the market. You have to go to your preferable EM supplier and ask them about that. It's very usual piece of equipment. It measures directly the thickness of the deposited material on the sensor. Sensor is mounted inside the vacuum chamber close to your sample. It is pretty sensitive (down to the 0.1 nm). The cost - couple of thousand backs. If you have further questions - I'll be happy to help you.
Have a good holiday season.
Sergey.
Sergey Ryazantsev, Ph.D. Electron Microscopy Department of Biological Chemistry, School of Medicine University of California, Los Angeles Box 951737 Los Angeles, CA 90095-1737
=============================================================== Jondo, Keep in mind that unless you deposit a really thick coating, carbon or metal coats do transmit light so ellipsometry could work. I have no personal experience with this, I just wanted to make the point. Good luck. TB
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I am currently involved with a cryo substitution project with human red blood cells. I have replaced the non crystalline ice with 2% Osmium in 100% acetone and have OK morphology, but the contrast is a bit low. I am thinking about trying to follow the Osmium step with uranyl acetate, before embedding in plastic. Does any one out there know if 1%uranly acetate will be OK in 100% acetone? Thanks, Tim
Timothy G. Schneider Director of Electron Microscopy Department of Pathology Room 229 Jefferson Hall Thomas Jefferson University 1020 Locust St. Philadelphia Pa. 19107 215-503-4798 work 610-613-8170 cellular timothy.schneider-at-mail.tju.edu
Email: msoulek-at-foodprorecruiters.com Name: Michael Soulek
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I am looking for a person to fill an available Sr. Microscopist position in the Midwest. Salary of $60-75k and a 5% bonus potential. Desire PHD or equivalent experience in Histo and or Micro Chemistry area of knowledge. Selected candidate will work in a million dollar + lab in a service group on problems that are brought to the group to solve. Solid communication and management of priorities is strongly sought in this person.
Food or plant background is sought after experience. Company does work on a World Wide Basis
Some of the projects are to work in coloring,marking,compound ID,looking at the different types to identify the various types of proteins,carbohydrates,lipids or elements of the various tissues. The last hired person came from USDA.
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Dear Timothy, We also worked with acetone and I have found that if one uses 4% OsO4 in acetone the contrast should be rather good.
Happy New Year!
Yours, Sasha
On Thu, 28 Dec 2000, Timothy Schneider 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 Out There: } } I am currently involved with a cryo substitution project with human red } blood cells. I have replaced the non crystalline ice with 2% Osmium in 100% } acetone and have OK morphology, but the contrast is a bit low. I am } thinking about trying to follow the Osmium step with uranyl acetate, before } embedding in plastic. Does any one out there know if 1%uranly acetate will } be OK in 100% acetone? Thanks, Tim } } Timothy G. Schneider } Director of Electron Microscopy } Department of Pathology } Room 229 Jefferson Hall } Thomas Jefferson University } 1020 Locust St. } Philadelphia Pa. 19107 } 215-503-4798 work } 610-613-8170 cellular } timothy.schneider-at-mail.tju.edu } } }
Can you deposit your coating onto a sample of polymerized TEM embedding media, then cut the sample at 90 degrees on a microtome, then measure the thickness using a TEM?
Good luck,
Doug
On Thu, 28 Dec 2000 15:38:21 +0900 =?ks_c_5601-1987?B?Sm9uZG8gWXVuIMCxwbi1tQ==?= {jdyun-at-kyungnam.ac.kr} 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 Sergey, Tobias and other listers: } } Thank you for your kind replies. } The equipment that you mentioned seems too expensive for } me to use, especially when I need just a couple of times } thickness measurement for the calibration of my sputter } coater. Is there other way to measure the thickness? } Tobias suggested to use ellipsometry technique because the } metallic or carbon coating transmits light when it is very } thin. Have any you ever tried this before? } } Best wishes, } } Jondo Yun } Kyungnam University, Korea } jdyun-at-hanma.kyungnam.ac.kr } } ==================================================================== } Hello Jondo } } It looks like nobody answer your question. } } There are special machines called "thickness monitor" on } the market. You have to go to your preferable EM supplier } and ask them about that. It's very usual piece of } equipment. It measures directly the thickness of the } deposited material on the sensor. Sensor is mounted inside } the vacuum chamber close to your sample. It is pretty } sensitive (down to the 0.1 nm). The cost - couple of } thousand backs. If you have further questions - I'll be } happy to help you. } } Have a good holiday season. } } Sergey. } } Sergey Ryazantsev, Ph.D. } Electron Microscopy } Department of Biological Chemistry, School of Medicine } University of California, Los Angeles } Box 951737 } Los Angeles, CA 90095-1737 } } (310) 825-1144 (office) } Pager: (310) 845-0248 } FAX: (310) 206-5272 (departmental) } mailto:sryazant-at-ucla.edu } } =============================================================== } Jondo, } Keep in mind that unless you deposit a really thick } coating, carbon or metal coats do transmit light so } ellipsometry could work. I have no personal experience with } this, I just wanted to make the point. Good luck. TB } } } } Jondo,?------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy } Society 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: } } } Can anybody suggest how to measure thickness of the } conductive } coating for EM specimen? If it is transparent, } ellipsometer could be } used. Since metallic or carbon } coating is not transparent, I could } not use that method. } Then do I have only way to measure directly, } like } measuring thickness on the cross-section or on the surface } by } using EM or SPM? Is there any other way? } } } Jondo Yun } } jdyun-at-kyungnam.ac.kr } } -- } _ ____ __ ____ Tobias I. Baskin } / \ / / \ / \ \ 109 } Tucker Hall / / / / \ \ \ } Biological Sciences /_ / __ /__ \ \ \__ } University of Missouri / / / \ \ } \ Columbia, MO USA / / / } \ \ \ 65211-7400 / / ___ / \ } \__/ \ ____ voice: 573-882-0173 fax: } 573-882-0123 } } }
---------------------- 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
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