Hello all, Would anyone be able to tell me why corn oil is often recommended as the correct oil to use to neutralise osmium tetroxide waste? Would any vegetable oil do just as well, or does corn oil have special properties? I have been unable to find it in supermarkets, and wonder if another kind would be just as good. Thanks. Lyn Waterhouse CEMMSA Centre for Electron Microscopy and Microstructure Analysis University of Adelaide Adelaide SA 5005 Ph: (08) 8303 4074 Fax: (08) 8303 4356 Website: http://www.adelaide.edu.au/CEMMSA
We have a LaB6 equipped ESEM installed in early 2000. It is still using the original filament which has now logged over 1000 hours. Although we have had various problems with the instrument vacuum levels have not been part of it. Gun vacuum is in the range 2.0 - 2.6 x10-7.
We bake it out overnight every time air is admitted to the gun for cleaning or repair. We are in the process of installing a high vac valve between the ion pump and the gun to save on bake outs at pressured times.
Sounds like you have a 'microleak' - get the agents onto it while you are still under warranty ! Baking out every two weeks might appeal to the purists who want to ensure 'ultimate vacuum' but it seems like overkill - unless it is compensating for a latent problem.
Another point that may be of interest - we have found that the instrument (with mixed lo and hivac usage) will last about three months before it starts getting wobbly ie gun instability and uncorrectable astigmatism. This is an indicator to clean the Wehnelt - itself no mean task since you are cleaning off an entirely transparent but totally nonconductive layer of Lanthanum (?) from the inside of the cap - checking for continuity with a multimeter as you clean. Once baked out after this process it performs like a star once again ! Thus far we are cleaning with polish, but are investigating less exhausting chemical cleaning methods !
The local service agents for FEI/Philips Anaspec-at-icon.co.za have assisted us a great deal in coming to terms with the idiosyncrasies of a LaB6 equipped ESEM - a concept which has given us some interesting moments.
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/department/default.asp?dept=cemunp Email:bruton-at-nu.ac.za postal address; Private Bag X01, Scottsville, 3209 KwaZulu-Natal South Africa
} } } "FABBRI" {fabbri-at-cigssrv1.unimo.it} 05/31/01 09:55AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We are using a quite new FEI XL-30 LaB6 SEM. It's a long time the we are trying to understand the normal working conditions of the vacuum system. Looking at what is written in the operating manual, our vacuum system is not able to work in safety conditions for the Lab6 filament ( { 2x10-7 mbar ). To do this we need to bakeout the gun frequently ( weekly )and are able to work in that safety ( { 2x10-7 )condition only for few hours. FEI says that the info on the manual are too restrictive (!!) and that we can work safetly with vacuum levels {6 x10-7 baking out every two weeks.
Any thoughts would be very welcome. Thank you very much in advance.
Best regards P.L. Fabbri
--------------------------------------------- Dr. Pier Luigi Fabbri C.I.G.S. - Centro Interdip. Grandi Strumenti Università di Modena via Campi 213/A - 41100 Modena, Italy Tel +39-059-2055231 / +39-059-370551 Fax +39-059-370551 E-mail: fabbri-at-mail.cigs.unimo.it ---------------------------------------------
Access to cryoSEM or ESEM would save a lot of work.
(Without any real experience of clays) I would be inclined to just air dry the sample at 40 deg. C in an oven.
The meniscus is required when replacing the intermediate fluid, to prevent exposing the sample, since you would not have one I guess it is irrelevant.
Dave
On Fri, 01 Jun 2001 10:06:22 +1000 John V Nailon {J.Nailon-at-mailbox.uq.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 } -----------------------------------------------------------------------. } } } G'day Bruce, } What is wrong in going the other way and simply dry your sample using a } small vacuum chamber?? } High temperature and high pressure spell trouble to me. } Regards } JVN } } Bruce Girrell wrote: } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 here familiar with any attempts to achieve critical point drying } } of a water saturated sample without the use of an intermediate fluid, i.e., } } by using the critical point of water itself at 374 oC/3212 PSI? } } } } I'm interested in looking at clay samples. I am concerned that acetone will } } cause structural rearrangement of the clay particles because of dissimilar } } physical and electrical properties between acetone and water. Unlike } } biological specimens, clay would not care the least about a temperature of } } 374 degrees, as it does not begin its dehydroxylization process until over } } 500 degrees. I work in an oil field related industry where we have ready } } access to equipment that would consider 5000 PSI to be "light duty" so an } } appropriate pressure bomb would not be difficult to construct. } } } } Some questions: } } } } 1) All CPD devices that I have seen have a window that allows you to observe } } the state of the meniscus. Is this essential, or can I assume that the } } process will go as physics says it should without actually observing the } } meniscus? } } } } 2) I do not want to place the clay samples in water and allow the water to } } provide pressure as it is heated, as any water in contact with the clay will } } start to decompose the sample. Can I use an external pressure source (high } } pressure air, for example) to keep the pressure inside the bomb high enough } } to avoid evaporation of any water until the critical point is reached? Would } } it suffice to simply build a little platform that would keep the clay sample } } above the water level? } } } } 3) Am I nuts for even considering this? What am I missing? } } } } Thanks for your help. } } } } Bruce Girrell } } Microline Technology Corp. } } 2397 Traversefield Dr. } } Traverse City, MI 49686 } } http://www.microlinetc.com } } } } (231) 935-1585 (Voice) } } (231) 922-5099 (FAX) } } bigirrell-at-microlinetc.com } } -- } John Nailon } Operations Manager } The Centre for Microscopy and Microanlaysis } The University of Queensland } St Lucia QLD 4072 } Tel: +61-7-33654214 } Fax: +61-7-33654422 } WWW: http://www.uq.edu.au/nanoworld }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
Johathan, I have the same Sony camera match on my Axiophot and Axiovert . I have couple the cameras using optical couplers with a 0.6X magnification. This overcomes some of the magnification factor and allows a much larger section of the field to be captured. The couplers are made by Diagnostic Instruments. If you want more info let me know.
-----Original Message----- } From: jmkrupp-at-cats.ucsc.edu [mailto:jmkrupp-at-cats.ucsc.edu] Sent: Thursday, May 31, 2001 4:00:PM To: Microscopy-at-sparc5.microscopy.com
Hi:
A colleague wishes to get guidance and information about coupling his video camera to his compound microscope.
He has a Sony DXC-970MD video camera. The camera has an ENG 1/2" bayonet lens mount. He wants to use it on a Zeiss Axiophot.
I am used to simple C-mount adapters, the bayonet mount is the ringer for me.
According to his research Sony offers an adapter, HRT045ENG12, that they say is used to mount this camera to a microscope. His question is 'Does the Sony adapter substitute for those offered by Zeiss for this purpose, or is the Sony adapter used to convert the bayonet mount to something compatable with the adapters offered by Zeiss?'
Finally, back in the old days, I remember learning that one could use a simple T-mount to attach a 35 mm camera to a microscope, but that it was not as good as having an eyepiece camera because it lacked the ocular lens. Is the same true of mounting a video camera? I have always just used simple, lenless C-mounts and it has seemed fine. If we are looking for ultimate image quality, should we be using a more sophisticated adapter?
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
Hi Jonathon, Remember that the image projected by the ocular of an LM on the CCD array is REAL ("Please draw a diagram of image formation in your light microscope", he asked the students in the audience!). The lens in the 'C' mounts are usually reduction lenses. [I haven't seen them all, so I am depending on my long-held view that if one wanted more resolution, one would use better objectives and 4x5" film(plates.] One may go without a reduction lens on camera and adapter, but then the video image will only be a subset [whose size (area) is dependent on the length of the 'C' adapter] of the area that one 'sees' in the 'virtual' image space. I have a Nikon bayonet-'C' adapter for a cooled CCD camera with a built-in reduction lens. My purpose was to extend the camera for some macro work using Nikon macro lenses that I already had for my Nikon 35mm camera. I also have two different types of 'C' mount adapter. The lensLESS adapter is used with a CCD camera that already has a reduction lens fitted to it, while the lensED adapter is used with a vidicon that lacks a reduction lens. I mismatched the adapters once and used the lensLESS adapter on the lensLESS vidicon (did use the ocular!). Only a subset, and no intensity! The only way to make a decision is to try both Nikon and Zeiss bayonet adapters, because one cannot determine - except empirically - what design criteria the engineers used, though my suspicion would be that they would not differ by much if both are used with an ocular. Also, watch the length of the adapter. A little long, and the camera (I am not familiar with its form.) may have to be connected more securely to avoid vibration effects.
READ ON AT YOUR OWN RISK (OF WASTING TIME!)
Film Photomicrography? How many listers have ever used a bellows camera setup for photomicrography? Your age is determined by the thickness of the catalog you received from the manufacturer of your microscope. What ARE we going to do with all that stuff in the darkroom?
Leitz ORTHOLUX? Catalogs? Speaking of that, I still have catalogs of Leitz (now Leica) from the OrthoLUX epoch (1950's-1970+). If anyone needs something, a picture or a number, don't hesitate to ask.
Video on TEM in 80's? I remember an old TEM that I once used had a video (vidicon) camera mounted under the viewing plate. The computer resided against half of a 9 foot wall, and was ROBUST. A user wanted to count mitochondria in the cytoplasm of cells. The only way I could get a single mitochondrion in the video camera view was to reduce the TEM mag almost to scan. So, while I could see a large part of the grid on the EM view screen the camera 'grabbed' only the center of a grid square and didn't show a bar! To program the computer, one had to remove boards and change jumpers. AH! The good OLD days!
Regards to all,
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
} ---------- } From: jmkrupp-at-cats.ucsc.edu } Sent: Thursday, May 31, 2001 3:59 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: LM- Video camera/microscope coupler } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 colleague wishes to get guidance and information about coupling his } video } camera to his compound microscope. } } He has a Sony DXC-970MD video camera. The camera has an ENG 1/2" bayonet } lens mount. He wants to use it on a Zeiss Axiophot. } } I am used to simple C-mount adapters, the bayonet mount is the ringer for } me. } } According to his research Sony offers an adapter, HRT045ENG12, that they } say is used to mount this camera to a microscope. His question is 'Does } the } Sony adapter substitute for those offered by Zeiss for this purpose, or is } the Sony adapter used to convert the bayonet mount to something compatable } with the adapters offered by Zeiss?' } } Finally, back in the old days, I remember learning that one could use a } simple T-mount to attach a 35 mm camera to a microscope, but that it was } not as good as having an eyepiece camera because it lacked the ocular } lens. } Is the same true of mounting a video camera? I have always just used } simple, lenless C-mounts and it has seemed fine. If we are looking for } ultimate image quality, should we be using a more sophisticated adapter? } } Thanks } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } } } }
This reply is a bit off the Microscopy theme of the post, but here goes ---
Adding high pressure air to a "pressure bomb" will not prevent the evaporation of the water (though it will slow it down). The vapor pressure of any gas in equilibrium with its liquid phase is independent of the pressure of other gases present. In Bruce's example, if he added air to give a pressure of 3212psi at the critical temperature, the pressure in the container would actually rise (assuming enough water was present) to 6424psi. This is why, when using a pressure cooker, you have to allow the steam to vent for a few moments before closing the valve and allowing the pressure to rise - the boiling point of the water depends only upon the pressure of the water vapor above the liquid, not the total pressure.
Tony Garratt-Reed.
At 05:07 PM 5/31/2001 -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: (+) 1-617-258-6478 **
I don't have a LaB6 XL30, but I used to have its predecessor, a very early ElectroScan E3. We ran that with LaB6 in a routine vacuum of about 5 - 7x10-7 Torr. It did not have the capability of being baked. The LaB6 ran fine - we almost never were able to run filaments to their "natural" life, as the manual control allowed students to abuse the current and emission ratings - a facility they took advantage of regularly, destroying the filaments in the process! However, we certainly had filaments run for up to 1000 hrs.
As it aged (it was 10 years old when we replaced it, and still with its original ion pump) we did not experience ultimate vacuum changes, although the ion pump did become noticably harder to start after a gun vent.
As a general comment, I can't believe that a well-designed vacuum system (as I assume the thermionic XL30 is), pumped by an ion pump and baked, can't maintain a vacuum in the low -7's, unless there is a leak or some other problem.
Tony G-R
} -----Original Message----- } } From: FABBRI {fabbri-at-cigssrv1.unimo.it} } To: microscopy-at-sparc5.microscopy.com {microscopy-at-sparc5.microscopy.com} } Date: Thursday, May 31, 2001 6:39 AM } Subject: XL30 - LaB6 Vacuum system } } } ------------------------------------------------------------------------ } 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 **
I'd like to echo the comments of Scott and Roger, who replied earlier. Their wisdom is worth repeating. I wrote SOPs for a commercial analytical service (not SEM specifically) in a past position........including the SOP on "writing SOPs". Meet the requirements, but do not go overboard with too many restrictions.
When considering a quality program it is so important to generate documentation that you can adhere to and follow. It is possible to create an incredibly "thorough" document which appears to cover every possible eventuality......but is also impossible to execute. Quality is then not achieved. Those who solicit feedback and allow their quality program to evolve over time will have more success. Having a strict internal review program that documents and corrects existing problems is actually a benefit in terms of passing external reviews. The internal documentation is typically the first thing examined by external examiners and it places you in a better light if it is clear you are making an effort to improve your program.
I would offer only a few suggestions for your specific analytical document:
If you have protocols for specific types of analyses that require more detailed instruction incorporate these as separate work instructions that are distinct from your SOP. The advantage is you can format your SOP in such a manner that the WIs can be amended or additional WIs can be added, without having to formerly generate an entire new version of your SOP. I have omitted the details, but consult one of the many manuals offered by the host of officials registrars. Referencing manuals is also fine for broader scope descriptions and procedures.
Maintanance Records and Statistical Process Control (SPC) procedures (i.e., calibration/sensitivity of EDS for example) are important components of your quality documentation.
A certain amount (not all) of the quality program information for commercial analytical service companies is available to customers (public). You may find some useful information by requesting information from their quality manager and/or asking specific departments or individuals about their quality procedures.
Very old book, perhaps on someone's shelf (Old instructor of EM class) Kay, D.H.,1965), Techniques for Electron Microscopy(2nd ed.), Chapter 8, "Preparation of Thin Sections" (Glauert, A.M. and Phillips, R.), Section: "Metallurgical and Crystallographic Applications of Thin Sectioning", pp. 248-253 (includes references), F.A. Davis, Philadelphia, PA, USA[Reprinted by Blackwell Scientific Pubs, Great Britain., 1967].
Kay, et al. promise that Zn is in there somewhere.
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
} ---------- } From: Dujin Wang } Sent: Thursday, May 31, 2001 3:54 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: TEM need help on embedding and cutting of ZnO crsytals } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 am trying to do ultrathin sectioning work of ZnO crystals, which size } is about 2-3microns(L) and 1-2microns(W). I am really a freshman } on TEM, so I hope to get help on this work. Any suggestions for the } choosing of embedding media and following procedures will be greatly } appreciated. } Thank you in advance. } } Dujin Wang } Max-Planck-Institute for Polymer Research } Ackermannweg 10, Mainz D-55128 } Germany } } Tel: 0049-6131-379226 } Fax: 0049-6131-379100 } Email: wangd-at-mpip-mainz.mpg.de } } }
Corn oil is used because it has lots of double bonds for the osmium to react with, so any highly unsaturated vegetable oil would do, or even be better than corn oil. Hey! Maybe there is a use for Vegemite ...
Phil
} Hello all, } Would anyone be able to tell me why corn oil is often recommended as the } correct oil to use to neutralise osmium tetroxide waste? Would any } vegetable oil do just as well, or does corn oil have special properties? } I have been unable to find it in supermarkets, and wonder if another } kind would be just as good. } Thanks. } Lyn Waterhouse } CEMMSA } Centre for Electron Microscopy } and Microstructure Analysis } University of Adelaide } Adelaide SA 5005 } Ph: (08) 8303 4074 } Fax: (08) 8303 4356 } Website: http://www.adelaide.edu.au/CEMMSA
-- }}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{ 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)
This post was a few days ago - apologies for the delayed response, but I've not seen others make quite the same points.
SOP's are merely recipes for achieving a goal. In Colin's case, he is needing to achieve quality control, others may be needing safe operation, ISO standard certification, or whatever. The point is that the SOP will vary depending on the goal. There is no single "Standard Operating Procedure" that covers every operating mode (unless it is so full of qualifications, conditions, and alternatives as to be practically useless.) For EDX, the manufacturer's instructions may be a good guide, and at least a useful reference.
After stating the goal, the SOP will put down, in writing, all the essential steps that must be followed to achieve this goal. This will include such items as checking the performance of the system (mag. and beam voltage calibration of the microscope, energy calibration and resolution of the EDX, for example), as well as things like specimen preparation and mounting (for quant. EDX, the specimen must be polished, and mounted in an accurately known orientation, etc., etc.).
Hope this helps.
Tony Garratt-Reed
} -----Original Message----- } Sent: Tuesday, May 29, 2001 1:20 AM } To: MSA Listserver } Subject: Writing SOP's for SEM/X-Ray Analysis } } } ------------------------------------------------------------------------ } 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 **
You could freeze-dry specimens and that would be a rather more obvious approach. The cost of building a high temperature/ pressure bomb aside the question is: what would be the effect of the high temperature on the structure of the clay. I think what is missing from your consideration is that water is intrinsic to clay and its nature is changed irreversible when all water is removed from its chemical structure. I know that quite a few people have worked on the fine structure of clay and it appears that this seemingly simple task is completely elusive. I don't know, but I am doubtful that much useful could be gleaned from modest SEM magnifications of dried clay. 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 01, 2001 7:07 AM, Bruce Girrell [SMTP:bigirrell-at-microlinetc.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 } -----------------------------------------------------------------------. } } } Has anyone here familiar with any attempts to achieve critical point drying } of a water saturated sample without the use of an intermediate fluid, i.e., } by using the critical point of water itself at 374 oC/3212 PSI? } } I'm interested in looking at clay samples. I am concerned that acetone will } cause structural rearrangement of the clay particles because of dissimilar } physical and electrical properties between acetone and water. Unlike } biological specimens, clay would not care the least about a temperature of } 374 degrees, as it does not begin its dehydroxylization process until over } 500 degrees. I work in an oil field related industry where we have ready } access to equipment that would consider 5000 PSI to be "light duty" so an } appropriate pressure bomb would not be difficult to construct. } } Some questions: } } 1) All CPD devices that I have seen have a window that allows you to observe } the state of the meniscus. Is this essential, or can I assume that the } process will go as physics says it should without actually observing the } meniscus? } } 2) I do not want to place the clay samples in water and allow the water to } provide pressure as it is heated, as any water in contact with the clay will } start to decompose the sample. Can I use an external pressure source (high } pressure air, for example) to keep the pressure inside the bomb high enough } to avoid evaporation of any water until the critical point is reached? Would } it suffice to simply build a little platform that would keep the clay sample } above the water level? } } 3) Am I nuts for even considering this? What am I missing? } } Thanks for your help. } } Bruce Girrell } Microline Technology Corp. } 2397 Traversefield Dr. } Traverse City, MI 49686 } http://www.microlinetc.com } } (231) 935-1585 (Voice) } (231) 922-5099 (FAX) } bigirrell-at-microlinetc.com
Would you consider (freeze-fracturing, freeze-drying) or (freeze-drying, dry-fracturing) techniques? How about cryo-SEM and ESEM? If you decide to build and use a super CPD, please let us know the results.
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
} } } "Bruce Girrell" {bigirrell-at-microlinetc.com} 05/31 5:07 PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Has anyone here familiar with any attempts to achieve critical point drying of a water saturated sample without the use of an intermediate fluid, i.e., by using the critical point of water itself at 374 oC/3212 PSI?
I'm interested in looking at clay samples. I am concerned that acetone will cause structural rearrangement of the clay particles because of dissimilar physical and electrical properties between acetone and water. Unlike biological specimens, clay would not care the least about a temperature of 374 degrees, as it does not begin its dehydroxylization process until over 500 degrees. I work in an oil field related industry where we have ready access to equipment that would consider 5000 PSI to be "light duty" so an appropriate pressure bomb would not be difficult to construct.
Some questions:
1) All CPD devices that I have seen have a window that allows you to observe the state of the meniscus. Is this essential, or can I assume that the process will go as physics says it should without actually observing the meniscus?
2) I do not want to place the clay samples in water and allow the water to provide pressure as it is heated, as any water in contact with the clay will start to decompose the sample. Can I use an external pressure source (high pressure air, for example) to keep the pressure inside the bomb high enough to avoid evaporation of any water until the critical point is reached? Would it suffice to simply build a little platform that would keep the clay sample above the water level?
3) Am I nuts for even considering this? What am I missing?
Thanks for your help.
Bruce Girrell Microline Technology Corp. 2397 Traversefield Dr. Traverse City, MI 49686 http://www.microlinetc.com
} ****************************************************************************** } ************************************************* } } International SEMATECH, a consortium of worldwide semiconductor manufacturing } companies, has an opening for a Materials Analysis Laboratory Manager. We } offer our employees an attractive package of benefits, including medical and } dental coverage, group legal insurance, college tuition reimbursement, 12 paid } holidays plus vacation. Additionally, we offer a generous retirement and } shared savings plan, in which you are fully vested after two years of } employment, PLUS a bonus program. } } To learn more about International SEMATECH, visit our website at } www.sematech.org. To apply for the Materials Analysis Laboratory Manager } position, send your resume to: staffing.hr-at-sematech.org. Principals only } please. } } } ****************************************************************************** } ***************** } } Materials Analysis Laboratory Manager Job Description } Job Summary: } Management of the daily operation in the (Advanced Tool Development Facility) } ATDF's Materials Analysis Laboratory in a cost effective method, including all } logistical planning and scheduling activities. Continual interactions with the } customer on the prioritizing and dispositioning of data and samples is a major } component of this job. } Job Responsibilities: } * Maintain a safe working environment: be mindful of any potential } hazards, report and/or correct anything that needs to be made safe, } participate in monthly group self-inspections. Work with safety to update } policies and procedures and ensure they are appropriate. } * Maintain group specific safety training records. Manage technical } leadership of group to "realize the roadmap" } * Maintain a quality-staffing plan through guidance and feedback, } encourage appropriate training, professional interaction, presentations and } publications, retain (attract if needed) and develop quality staff. } * Maintains & Reports on the MA lab's budget to both management and } customers. } * Ownership of MA indices and develops plans for continuous improvement of } those indices } * Promotes team interactions to optimize resources with sister FA lab } * Provides the highest level of Customer satisfaction and Member Company } value. } * Maintain leading edge tool set through creative means, i.e. partnering, } reduced cost of purchase, etc. } Qualifications: } * Minimum 4 years experience in management of an analytical laboratory in } semiconductor industry } * Current and comprehensive working knowledge of semiconductor } manufacturing technology and associated characterization and metrology } equipment and techniques } * Direct semiconductor industry experience with all or most of the } following techniques: Auger, SIMS-ICP-MS, TEM, SEM, FIB/SIM, AFM, TXRF, and } VPD-ICP-MS } * Degree in: Material Sciences, Surface Chemistry, Physics or Chemical } Engineering preferred, PhD, MS, EE and/or BS with commensurate experience. } * Currently Published articles in related field(s) } Special Conditions: } * Travel Required {10% of the time. } * Shift/Hours worked:1st }
Lyn Waterhouse wrote: ================================================= Would anyone be able to tell me why corn oil is often recommended as the correct oil to use to neutralise osmium tetroxide waste? Would any vegetable oil do just as well, or does corn oil have special properties? I have been unable to find it in supermarkets, and wonder if another kind would be just as good. ============================================= Whether one kind of oil works better vs. another is a function of the unsaturation present. However, from an environmental and also a recycling standpoint, none of these "oils" is very environmentally friendly because once the osmium is put into this state, the economics of recovery become so terrible, that the concept of recovery and recycling becomes impossible (at least at current market prices). The only fate is incineration and burial in a land fill.
SPI Supplies is now beta testing our own SPI Osmium Recyling Kit. It consists of a 4 liter bottle with some formulated "potion" inside. When the bottle is filled up, the idea is for it to be returned to SPI Supplies for recycling. The chemistry is not rocket scientist logic, but navigating the various regulatory challenges, especially since they tend to vary from country to country, not to mention the rules being constantly changing as well, is what presents the real challenges. We believe the economics are such that we could even "give back" some new osmium tetroxide in amoules upon the return of such a bottle.
We would be happy to send you, with our compliments, a beta test kit. Just let us know if you would like us to do that. We would give you instructions for returning it to SPI for recycling. We are doing this on a carefully controlled basis for now, because every time we think we have all the bases covered, we encounter some other surprise. However, we plan to make some kind of more formal announcement about it on our website later this year, if not earlier at the M&M meeting, when the kit is officially introduced as a regular product of SPI Supplies.
But in the mean time, rather than doing the neutralization in an "oil", use 1N KOH solution. You will get the final reduction of the tetroxide to the dioxide, and the resulting system could at some future time, literally be "added" to the SPI Osmium Recycling Kit when it is more widely available. Now this suggestion might not be right for everyone but it is an alternative to rendering the material to a fate outside of the stream of commerce from which it is then lost for the benefit of future generations.
Disclaimer: SPI Supplies would like to promote the concept of recycling instead of incineration and burial of this non-renewable resource. This is called "passionate commercialism", something I hope is permitted on this listserver......!
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 ############################ ==================================================
What volume ratio of 0.1 M KOH to OsO4 (say, 1% in buffer) do you recommend?
Marie
You wrote:
} But in the mean time, rather than doing the neutralization in an "oil", use } 1N KOH solution.
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
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Hello all, Would anyone be able to tell me why corn oil is often recommended as the correct oil to use to neutralise osmium tetroxide waste? Would any vegetable oil do just as well, or does corn oil have special properties? I have been unable to find it in supermarkets, and wonder if another kind would be just as good. Thanks. Lyn Waterhouse CEMMSA Centre for Electron Microscopy and Microstructure Analysis University of Adelaide Adelaide SA 5005 Ph: (08) 8303 4074 Fax: (08) 8303 4356 Website: http://www.adelaide.edu.au/CEMMSA
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This reply is a bit off the Microscopy theme of the post, but here goes ---
Adding high pressure air to a "pressure bomb" will not prevent the evaporation of the water (though it will slow it down). The vapor pressure of any gas in equilibrium with its liquid phase is independent of the pressure of other gases present. In Bruce's example, if he added air to give a pressure of 3212psi at the critical temperature, the pressure in the container would actually rise (assuming enough water was present) to 6424psi. This is why, when using a pressure cooker, you have to allow the steam to vent for a few moments before closing the valve and allowing the pressure to rise - the boiling point of the water depends only upon the pressure of the water vapor above the liquid, not the total pressure.
Tony Garratt-Reed.
At 05:07 PM 5/31/2001 -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: (+) 1-617-258-6478 **
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Dear XL30 LaB6 users
We have a LaB6 equipped ESEM installed in early 2000. It is still using the original filament which has now logged over 1000 hours. Although we have had various problems with the instrument vacuum levels have not been part of it. Gun vacuum is in the range 2.0 - 2.6 x10-7.
We bake it out overnight every time air is admitted to the gun for cleaning or repair. We are in the process of installing a high vac valve between the ion pump and the gun to save on bake outs at pressured times.
Sounds like you have a 'microleak' - get the agents onto it while you are still under warranty ! Baking out every two weeks might appeal to the purists who want to ensure 'ultimate vacuum' but it seems like overkill - unless it is compensating for a latent problem.
Another point that may be of interest - we have found that the instrument (with mixed lo and hivac usage) will last about three months before it starts getting wobbly ie gun instability and uncorrectable astigmatism. This is an indicator to clean the Wehnelt - itself no mean task since you are cleaning off an entirely transparent but totally nonconductive layer of Lanthanum (?) from the inside of the cap - checking for continuity with a multimeter as you clean. Once baked out after this process it performs like a star once again ! Thus far we are cleaning with polish, but are investigating less exhausting chemical cleaning methods !
The local service agents for FEI/Philips Anaspec-at-icon.co.za have assisted us a great deal in coming to terms with the idiosyncrasies of a LaB6 equipped ESEM - a concept which has given us some interesting moments.
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/department/default.asp?dept=cemunp Email:bruton-at-nu.ac.za postal address; Private Bag X01, Scottsville, 3209 KwaZulu-Natal South Africa
} } } "FABBRI" {fabbri-at-cigssrv1.unimo.it} 05/31/01 09:55AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We are using a quite new FEI XL-30 LaB6 SEM. It's a long time the we are trying to understand the normal working conditions of the vacuum system. Looking at what is written in the operating manual, our vacuum system is not able to work in safety conditions for the Lab6 filament ( { 2x10-7 mbar ). To do this we need to bakeout the gun frequently ( weekly )and are able to work in that safety ( { 2x10-7 )condition only for few hours. FEI says that the info on the manual are too restrictive (!!) and that we can work safetly with vacuum levels {6 x10-7 baking out every two weeks.
Any thoughts would be very welcome. Thank you very much in advance.
Best regards P.L. Fabbri
--------------------------------------------- Dr. Pier Luigi Fabbri C.I.G.S. - Centro Interdip. Grandi Strumenti Università di Modena via Campi 213/A - 41100 Modena, Italy Tel +39-059-2055231 / +39-059-370551 Fax +39-059-370551 E-mail: fabbri-at-mail.cigs.unimo.it ---------------------------------------------
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Access to cryoSEM or ESEM would save a lot of work.
(Without any real experience of clays) I would be inclined to just air dry the sample at 40 deg. C in an oven.
The meniscus is required when replacing the intermediate fluid, to prevent exposing the sample, since you would not have one I guess it is irrelevant.
Dave
On Fri, 01 Jun 2001 10:06:22 +1000 John V Nailon {J.Nailon-at-mailbox.uq.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 } -----------------------------------------------------------------------. } } } G'day Bruce, } What is wrong in going the other way and simply dry your sample using a } small vacuum chamber?? } High temperature and high pressure spell trouble to me. } Regards } JVN } } Bruce Girrell wrote: } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 here familiar with any attempts to achieve critical point drying } } of a water saturated sample without the use of an intermediate fluid, i.e., } } by using the critical point of water itself at 374 oC/3212 PSI? } } } } I'm interested in looking at clay samples. I am concerned that acetone will } } cause structural rearrangement of the clay particles because of dissimilar } } physical and electrical properties between acetone and water. Unlike } } biological specimens, clay would not care the least about a temperature of } } 374 degrees, as it does not begin its dehydroxylization process until over } } 500 degrees. I work in an oil field related industry where we have ready } } access to equipment that would consider 5000 PSI to be "light duty" so an } } appropriate pressure bomb would not be difficult to construct. } } } } Some questions: } } } } 1) All CPD devices that I have seen have a window that allows you to observe } } the state of the meniscus. Is this essential, or can I assume that the } } process will go as physics says it should without actually observing the } } meniscus? } } } } 2) I do not want to place the clay samples in water and allow the water to } } provide pressure as it is heated, as any water in contact with the clay will } } start to decompose the sample. Can I use an external pressure source (high } } pressure air, for example) to keep the pressure inside the bomb high enough } } to avoid evaporation of any water until the critical point is reached? Would } } it suffice to simply build a little platform that would keep the clay sample } } above the water level? } } } } 3) Am I nuts for even considering this? What am I missing? } } } } Thanks for your help. } } } } Bruce Girrell } } Microline Technology Corp. } } 2397 Traversefield Dr. } } Traverse City, MI 49686 } } http://www.microlinetc.com } } } } (231) 935-1585 (Voice) } } (231) 922-5099 (FAX) } } bigirrell-at-microlinetc.com } } -- } John Nailon } Operations Manager } The Centre for Microscopy and Microanlaysis } The University of Queensland } St Lucia QLD 4072 } Tel: +61-7-33654214 } Fax: +61-7-33654422 } WWW: http://www.uq.edu.au/nanoworld }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
Hi !!! I was doing EDAX on a sample to determine the surface constituents. Can you let me know answers to the following questions: 1. When I vary the accelerating voltage the atomic % (concentration of = } elements) varies. Why is that? 2. What is the depth of penetration of EDAX? Does this depend on = } accelerating voltage? Please send copies of your reply to bala-at-sc.edu. Thanks in advance Sincerely, Bala Haran
Hi !!! I was doing EDAX on a sample to determine the surface constituents. Can you let me know answers to the following questions: 1. When I vary the accelerating voltage the atomic % (concentration of = } elements) varies. Why is that? 2. What is the depth of penetration of EDAX? Does this depend on = } accelerating voltage? Please send copies of your reply to bala-at-sc.edu. Thanks in advance Sincerely, Bala Haran
Optem and Diagnostic Instrument make adapters for this camera to most any microscope.
gary g.
At 12:59 PM 5/31/2001, you wrote:
} Hi: } } A colleague wishes to get guidance and information about coupling his video } camera to his compound microscope. } } He has a Sony DXC-970MD video camera. The camera has an ENG 1/2" bayonet } lens mount. He wants to use it on a Zeiss Axiophot. } } I am used to simple C-mount adapters, the bayonet mount is the ringer for me. } } According to his research Sony offers an adapter, HRT045ENG12, that they } say is used to mount this camera to a microscope. His question is 'Does the } Sony adapter substitute for those offered by Zeiss for this purpose, or is } the Sony adapter used to convert the bayonet mount to something compatable } with the adapters offered by Zeiss?' } } Finally, back in the old days, I remember learning that one could use a } simple T-mount to attach a 35 mm camera to a microscope, but that it was } not as good as having an eyepiece camera because it lacked the ocular lens. } Is the same true of mounting a video camera? I have always just used } simple, lenless C-mounts and it has seemed fine. If we are looking for } ultimate image quality, should we be using a more sophisticated adapter? } } 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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Hi Jonathon, Remember that the image projected by the ocular of an LM on the CCD array is REAL ("Please draw a diagram of image formation in your light microscope", he asked the students in the audience!). The lens in the 'C' mounts are usually reduction lenses. [I haven't seen them all, so I am depending on my long-held view that if one wanted more resolution, one would use better objectives and 4x5" film(plates.] One may go without a reduction lens on camera and adapter, but then the video image will only be a subset [whose size (area) is dependent on the length of the 'C' adapter] of the area that one 'sees' in the 'virtual' image space. I have a Nikon bayonet-'C' adapter for a cooled CCD camera with a built-in reduction lens. My purpose was to extend the camera for some macro work using Nikon macro lenses that I already had for my Nikon 35mm camera. I also have two different types of 'C' mount adapter. The lensLESS adapter is used with a CCD camera that already has a reduction lens fitted to it, while the lensED adapter is used with a vidicon that lacks a reduction lens. I mismatched the adapters once and used the lensLESS adapter on the lensLESS vidicon (did use the ocular!). Only a subset, and no intensity! The only way to make a decision is to try both Nikon and Zeiss bayonet adapters, because one cannot determine - except empirically - what design criteria the engineers used, though my suspicion would be that they would not differ by much if both are used with an ocular. Also, watch the length of the adapter. A little long, and the camera (I am not familiar with its form.) may have to be connected more securely to avoid vibration effects.
READ ON AT YOUR OWN RISK (OF WASTING TIME!)
Film Photomicrography? How many listers have ever used a bellows camera setup for photomicrography? Your age is determined by the thickness of the catalog you received from the manufacturer of your microscope. What ARE we going to do with all that stuff in the darkroom?
Leitz ORTHOLUX? Catalogs? Speaking of that, I still have catalogs of Leitz (now Leica) from the OrthoLUX epoch (1950's-1970+). If anyone needs something, a picture or a number, don't hesitate to ask.
Video on TEM in 80's? I remember an old TEM that I once used had a video (vidicon) camera mounted under the viewing plate. The computer resided against half of a 9 foot wall, and was ROBUST. A user wanted to count mitochondria in the cytoplasm of cells. The only way I could get a single mitochondrion in the video camera view was to reduce the TEM mag almost to scan. So, while I could see a large part of the grid on the EM view screen the camera 'grabbed' only the center of a grid square and didn't show a bar! To program the computer, one had to remove boards and change jumpers. AH! The good OLD days!
Regards to all,
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
} ---------- } From: jmkrupp-at-cats.ucsc.edu } Sent: Thursday, May 31, 2001 3:59 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: LM- Video camera/microscope coupler } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 colleague wishes to get guidance and information about coupling his } video } camera to his compound microscope. } } He has a Sony DXC-970MD video camera. The camera has an ENG 1/2" bayonet } lens mount. He wants to use it on a Zeiss Axiophot. } } I am used to simple C-mount adapters, the bayonet mount is the ringer for } me. } } According to his research Sony offers an adapter, HRT045ENG12, that they } say is used to mount this camera to a microscope. His question is 'Does } the } Sony adapter substitute for those offered by Zeiss for this purpose, or is } the Sony adapter used to convert the bayonet mount to something compatable } with the adapters offered by Zeiss?' } } Finally, back in the old days, I remember learning that one could use a } simple T-mount to attach a 35 mm camera to a microscope, but that it was } not as good as having an eyepiece camera because it lacked the ocular } lens. } Is the same true of mounting a video camera? I have always just used } simple, lenless C-mounts and it has seemed fine. If we are looking for } ultimate image quality, should we be using a more sophisticated adapter? } } 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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Hello all, Would anyone be able to tell me why corn oil is often recommended as the correct oil to use to neutralise osmium tetroxide waste? Would any vegetable oil do just as well, or does corn oil have special properties? I have been unable to find it in supermarkets, and wonder if another kind would be just as good. Thanks. Lyn Waterhouse CEMMSA Centre for Electron Microscopy and Microstructure Analysis University of Adelaide Adelaide SA 5005 Ph: (08) 8303 4074 Fax: (08) 8303 4356 Website: http://www.adelaide.edu.au/CEMMSA
Staining cardiac vessel (e.g. aorta) wall is accompanied by a lot of autofluorescence, probably from elastin and perhaps other extracellualr matrix material.
Should attempts to reduce autofluorescence concentrate on the source of the problem, in other words, are there specific treatments for different types, such as lipofuscin versus elastin.
I have tried Sudan black B, sodium borohydride, toluidine blue. The first 2 reduced the red wavelength contribution only while toluidine blue actually increased the red autofluorescence. The green signal was reduced by only a small amount, all compared to just a PBS wash.
Any suggestions would be highly appreciated.
Judy Trogadis Bio-Imaging Coordinator St. Michael's Hospital 30 Bond St. Toronto, ON M5B 1W8
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Access to cryoSEM or ESEM would save a lot of work.
(Without any real experience of clays) I would be inclined to just air dry the sample at 40 deg. C in an oven.
The meniscus is required when replacing the intermediate fluid, to prevent exposing the sample, since you would not have one I guess it is irrelevant.
Dave
On Fri, 01 Jun 2001 10:06:22 +1000 John V Nailon {J.Nailon-at-mailbox.uq.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 } -----------------------------------------------------------------------. } } } G'day Bruce, } What is wrong in going the other way and simply dry your sample using a } small vacuum chamber?? } High temperature and high pressure spell trouble to me. } Regards } JVN } } Bruce Girrell wrote: } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 here familiar with any attempts to achieve critical point drying } } of a water saturated sample without the use of an intermediate fluid, i.e., } } by using the critical point of water itself at 374 oC/3212 PSI? } } } } I'm interested in looking at clay samples. I am concerned that acetone will } } cause structural rearrangement of the clay particles because of dissimilar } } physical and electrical properties between acetone and water. Unlike } } biological specimens, clay would not care the least about a temperature of } } 374 degrees, as it does not begin its dehydroxylization process until over } } 500 degrees. I work in an oil field related industry where we have ready } } access to equipment that would consider 5000 PSI to be "light duty" so an } } appropriate pressure bomb would not be difficult to construct. } } } } Some questions: } } } } 1) All CPD devices that I have seen have a window that allows you to observe } } the state of the meniscus. Is this essential, or can I assume that the } } process will go as physics says it should without actually observing the } } meniscus? } } } } 2) I do not want to place the clay samples in water and allow the water to } } provide pressure as it is heated, as any water in contact with the clay will } } start to decompose the sample. Can I use an external pressure source (high } } pressure air, for example) to keep the pressure inside the bomb high enough } } to avoid evaporation of any water until the critical point is reached? Would } } it suffice to simply build a little platform that would keep the clay sample } } above the water level? } } } } 3) Am I nuts for even considering this? What am I missing? } } } } Thanks for your help. } } } } Bruce Girrell } } Microline Technology Corp. } } 2397 Traversefield Dr. } } Traverse City, MI 49686 } } http://www.microlinetc.com } } } } (231) 935-1585 (Voice) } } (231) 922-5099 (FAX) } } bigirrell-at-microlinetc.com } } -- } John Nailon } Operations Manager } The Centre for Microscopy and Microanlaysis } The University of Queensland } St Lucia QLD 4072 } Tel: +61-7-33654214 } Fax: +61-7-33654422 } WWW: http://www.uq.edu.au/nanoworld }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
Dear Listers! Thank you very much for your interest in our problems. I got yours recommendation for HT check. Thank you very much again. Next week I will be in Moscow and I will be back on Friday June 8, 2001. Happy week end. Best regards Anton mailto:gut-at-thermo.isp.nsc.ru
Below is the result of your feedback form. It was submitted by (alsamszw-at-aol.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Saturday, June 2, 2001 at 15:48:55 ---------------------------------------------------------------------------
Email: alsamszw-at-aol.com Name: Dr S Z AL-SAM
Organization: Mid Essex Hospitals, Chelmsford, UK
Education: Graduate College
Location: Chelmsford, United Kingdom
Question: Please tell me more about 3D microscopy and has this been used to study cytological smears before. Who supply these microscopes and how much do they cost? Many thanks.
Please tell me more about 3D images and 3D microscopes. Have these been used to study cytological smears and histological sections? How much do they cost and are there catalogues available? thank you AL-SAM
Research Technician (part-time) Electron Microscopy and Biochemistry.
We are seeking a technician preferably with experience in biochemistry and/or electron microscopy to join this BBSRC funded project “Trans-cellular Ca2+ transport and Ca2+ homeostasis in calcifying microalgae”. The post holder will be responsible for preparation and analysis of algal cells using electron microscopy in addition to purification of calcium binding proteins using gel electrophoresis.
The post is offered as a 50% fixed term appointment for 18 months with starting salary £13,717 (pro-rata), although other suitable working arrangements will be considered.
For further details of this project you can visit the following website: http://www.mba.ac.uk
Informal enquires may be addressed to Dr Alison Taylor (tel: +44 (0)1752 633290, e-mail: arta-at-mba.ac.uk) or Professor Colin Brownlee (tel: +44 (0)1752 633246, e-mail: cbr-at-wpo.nerc.ac.uk).
Closing Date: open until appointed but start by November 2001
Electron Microscopy(full time, permanent) Technician —Health Research Inc., Wadsworth Center, Albany, NY. We are looking for a motivated, mature and responsible individual to join a state-of-the-art Federal funded biomedical research laboratory. Flexible job hours in a challenging and stimulating research environment. Will train for specialized tasks.
Minimum qualification: B.S. in biology or related filed.
Preferred qualification: B.S. with experience in specimen sectioning and electron microscopy.
Responsibilities: 1. Embedding various biological specimens for transmission electron microscopy analyses. 2. Thin and thick (serially) sectioning of specimens. 3. Scanning and photographing sections with the EM 4. Conducting 3D analyses.
Contact: Richard Cole Research Scientist III Director of the Laser Microsurgery and Advanced Light Microscopy Core Unit Wadsworth Center P.O. Box 509 Albany N.Y. 12201-0509 rcole-at-wadsworth.org 518-474-7048 Phone 518-486-4901 Fax
We do alot of Immunofluorescence on sections of human skin. The main source of atuofluorescence is the elastin in the dermis and concentrated around the larger vessels. The best success we have had is using a CY5 label. The autofluorescence for elastin goes down dramaticly in the longer wavelengths. Or if we are using a single antiboby, we cature an image of the autofluorescence in a different channel and digitally subtract it out of the final image using image math. Then actually, most of the time we just leave it in and acknowledge that it is elastin (it is actually a good structural landmark and quite beautiful).
Bob Dermatology Research Center U of Washington, Seattle
On Fri, 1 Jun 2001, Judy Trogadis wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Staining cardiac vessel (e.g. aorta) wall is accompanied by a lot of autofluorescence, probably from elastin and perhaps other extracellualr matrix material. } } Should attempts to reduce autofluorescence concentrate on the source of the problem, in other words, are there specific treatments for different types, such as lipofuscin versus elastin. } } I have tried Sudan black B, sodium borohydride, toluidine blue. The first 2 reduced the red wavelength contribution only while toluidine blue actually increased the red autofluorescence. The green signal was reduced by only a small amount, all compared to just a PBS wash. } } Any suggestions would be highly appreciated. } } } } Judy Trogadis } Bio-Imaging Coordinator } St. Michael's Hospital } 30 Bond St. } Toronto, ON M5B 1W8 } } ph: 416-864-6060 x6337 } fax: 416-864-6043 } trogadisj-at-smh.toronto.on.ca } } } }
We just acquired a new high resolution SEM (FEG) and want to keep it "clean". How can I find out about out-gassing rates and the relative "cleanliness" of various sample preparation supplies:
We just acquired a new high resolution SEM (FEG) and want to keep it "clean". How can I find out about out-gassing rates and the relative "cleanliness" of various sample preparation supplies:
DIRECTOR, CELLULAR IMAGING CENTER: The Department of Pathology and Anatomy seeks Ph.D. applicants for a faculty position in fall 2001 as director of our cellular imaging core facility. The facility is equipped with transmission and scanning electron microscopes, a new laser scanning confocal microscope, image analysis computer, and conventional fluorescence microscopes.. Strong credentials in state of the art light and/or electron microscopy (e.g., confocal applications, FRET) are desired. In addition to overseeing the imaging core facility, the director will be expected to develop independent and/or collaborative research in one of five research areas: Cancer, Cardiovascular/Renal, Neuroscience, Reproductive Endocrinology, and Virology. Please send curriculum vitae, statement of research interests, and names, addresses, telephone numbers and email addresses of three references to:
William F. Glass II, M.D. Ph.D., Chair Department of Pathology and Anatomy Eastern Virginia Medical School P.O. Box 1980 Norfolk, VA 23501
Eastern Virginia Medical School is an Affirmative Action/Equal Opportunity Employer.
Questions may be directed to Dr. Glass or Dr. Earl Godfrey (godfreew-at-borg.evms.edu).
Please inform qualified individuals of this opportunity. Thank you very much.
Earl W. Godfrey, Ph.D. Dept. of Pathology and Anatomy Eastern Virginia Medical School P.O. Box 1980 Norfolk, VA 23501
street address: 700 W. Olney Rd. Lewis Hall Rm. 3077a Norfolk, VA 23507
We just acquired a new high resolution SEM (FEG) and want to keep it "clean". How can I find out about out-gassing rates and the relative "cleanliness" of various sample preparation supplies:
We just acquired a new high resolution SEM (FEG) and want to keep it "clean". How can I find out about out-gassing rates and the relative "cleanliness" of various sample preparation supplies:
DIRECTOR, CELLULAR IMAGING CENTER: The Department of Pathology and Anatomy seeks Ph.D. applicants for a faculty position in fall 2001 as director of our cellular imaging core facility. The facility is equipped with transmission and scanning electron microscopes, a new laser scanning confocal microscope, image analysis computer, and conventional fluorescence microscopes.. Strong credentials in state of the art light and/or electron microscopy (e.g., confocal applications, FRET) are desired. In addition to overseeing the imaging core facility, the director will be expected to develop independent and/or collaborative research in one of five research areas: Cancer, Cardiovascular/Renal, Neuroscience, Reproductive Endocrinology, and Virology. Please send curriculum vitae, statement of research interests, and names, addresses, telephone numbers and email addresses of three references to:
William F. Glass II, M.D. Ph.D., Chair Department of Pathology and Anatomy Eastern Virginia Medical School P.O. Box 1980 Norfolk, VA 23501
Eastern Virginia Medical School is an Affirmative Action/Equal Opportunity Employer.
Questions may be directed to Dr. Glass or Dr. Earl Godfrey (godfreew-at-borg.evms.edu).
Please inform qualified individuals of this opportunity. Thank you very much.
Earl W. Godfrey, Ph.D. Dept. of Pathology and Anatomy Eastern Virginia Medical School P.O. Box 1980 Norfolk, VA 23501
street address: 700 W. Olney Rd. Lewis Hall Rm. 3077a Norfolk, VA 23507
Below is the result of your feedback form. It was submitted by (davidbock-at-mindspring.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Sunday, June 3, 2001 at 20:06:47 ---------------------------------------------------------------------------
Email: davidbock-at-mindspring.com Name: David B.
Organization: --
Education: Graduate College
Location: LA, CA
Question: I would like to create a very large image of human blood at ~4000 times magnification. Ideally, it would be a single image that I would print at 10 feet x 30 feet. I imagine each red blood cell would be about 3" across in the final printed version.
Is it possible to take such a picture? Would I have to take a series of shots and then join them with Photoshop? What about resolution?
Below is the result of your feedback form. It was submitted by (bubidabub-at-worldnet.att.net) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, June 4, 2001 at 01:18:52 ---------------------------------------------------------------------------
Email: bubidabub-at-worldnet.att.net Name: Robin Olsen
Organization: Northampton Community College
Education: Undergraduate College
Location: Easton, PA Northampton County
Question: Could you please tell me what type of microorganisms one can expect to find in environmental dust taken from outdoor playground equipment? I am a college student taking microbiology at NCC and I am having a difficult time finding resources for research regarding organisms in dust and dirt. If you could tell me some things I could expect to culture from a sample taken from a playground, or tell me some resources where I could find this information I would greatly appreciate it! Thanks!!
Below is the result of your feedback form. It was submitted by (bubidabub-at-worldnet.att.net) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, June 4, 2001 at 01:18:52 ---------------------------------------------------------------------------
Email: bubidabub-at-worldnet.att.net Name: Robin Olsen
Organization: Northampton Community College
Education: Undergraduate College
Location: Easton, PA Northampton County
Question: Could you please tell me what type of microorganisms one can expect to find in environmental dust taken from outdoor playground equipment? I am a college student taking microbiology at NCC and I am having a difficult time finding resources for research regarding organisms in dust and dirt. If you could tell me some things I could expect to culture from a sample taken from a playground, or tell me some resources where I could find this information I would greatly appreciate it! Thanks!!
I have an old, but brilliant, book 'Leybold Vacuum Handbook' by K. Diels and R. Jaeckel (Translated by H. Adam and J. Edwards) Pergamon Press, 1966. It contains explainations of these problems, descriptions of experiments to measure outgassing and desorption, and rates for most elements and compounds that may be used in vacuum systems.
If you want a simple way to test the various materials find a pumping rig and pump it down to it's ultimate pressure at least overnight, ensure that the ultimate pressure is of the same order as your microscope. Then vent it to dry nitrogen and pump down again noting the pressure change with time to it's ultimate pressure (say for 1 hour). Then vent it and introduce the unknown component and pump down again noting the time and pressure. The change in pumping speed (rate of decrease in pressure) will be due to the outgassing. No fancy units but a direct comparison for your problem. Remember that to make a real comparison of outgassing rates you need to have similar amounts of material in similar forms, ie. same surface areas and volumes. However, I guess you are more concerned with the comparison between different mounting techniques so similar specimens mounted by the various techniques would be OK.
To save you a lot of time in experimenting I suggest that you use the minimum amount of mounting material, you want the smallest possible surface area to see the vacuum and you want the lowest desorption (outgassing) rate and lowest vapour pressure.
Of your 4 options I would not use sticky pads or waxes. I would only use epoxy or conductive inks and pastes fully underneath the specimen and ensure that the small amount used was fully cured and dried by pumping it out in a vacuum rig before loading it into the microscope. I would also store it under vacuum before loading.
Good luck, Ron
} } We just acquired a new high resolution SEM (FEG) and want to keep it "clean". How can I find out about out-gassing rates and the relative "cleanliness" of various sample preparation supplies: } } 1. sticky dots } 2. conductive inks and pastes } 3. epoxies } 4. waxes } } Is there an accepted method for rating these materials? } }
---------------------- 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
Below is the result of your feedback form. It was submitted by (kellyburns-at-cfl.rr.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html#form on Monday, June 4, 2001 at 22:29:41 ---------------------------------------------------------------------------
Email: kellyburns-at-cfl.rr.com Name: Kelly Burns
Organization: Valencia Community College
Education: Undergraduate College
Location: Ocoee, FL, US
Question: When preparing a smear, why is it important to use tap water versus distilled water?
Karen Dye wrote the following: ========================================================== We just acquired a new high resolution SEM (FEG) and want to keep it "clean" . How can I find out about out-gassing rates and the relative "cleanliness" of various sample preparation supplies:
Is there an accepted method for rating these materials? ========================================================== We at SPI Supplies have been dealing with this particular issue for more than thirty years. I don't think your question has the kind of absolute answer you might have been seeking, for the following reasons:
a) There are at least several different sources of "sticky dots", and they do not all behave the same. Some off-gas more than others and some are more sensitive to the beam than others. Also other factors such as age and storage conditions can effect the behavior of some of these adhesive materials in vacuum.
b) The conductive inks and pastes similarly represent different compositions, they are not all the same, and do not all come from the same place, but in addition to that, the "off-gassing characteristics" also depend on how thick of a layer has been applied and whether there is a "skin" that forms on the top, resulting in a lingering "wet" area underneath the skin. This effect can be even more problematic for a paste. The details of the formulation can influence the tendency of the composition to form such a skin, which acts as a barrier to diffusion.
c) The typical epoxy can be cured using more or less hardener, and it can be cured at room temperature or at higher temperatures. A room temperature cure with less rather than more hardener increases the chances for there being more unpolymerized material being present to off-gas and cause problems. The use of more hardener and some residence time at a higher temperature tends to minimize the off-gassing effect.
d) Waxes tend to be nonconductive and beam sensitive. Some have plasticizers that migrate to the surface and off-gas. We try to avoid waxes as a mounting medium for this kind of application in any kind of SEM. I have waxes like dental waxes in mind in the making of that comment.
We have adopted our own "test" for UHV compatibility. Simply put, it is a measure of the deterioration of vacuum when a test specimen is inserted into a UHV system. We found that the SPI double sided conductive carbon sheet material, when a 10 mm square was used, was inserted into the UHV system, there was no visible "meter deflection" on the LED display for the vacuum. Since the adhesive is the same as what is used in the SPI carbon tape and double sided discs, we would expect the same result would be found for them as well. We know that some similar appearing materials offered by others will perform as well on such a test. But not in all instances. We know this might not be the perfect test, but over the years it has been quite effective in terms of validating those materials that are better for this kind of application than others. Obviously, the worst performing material, if just a tiny bit is used, could end up "passing" the test, just as an inordinately large piece of the SPI double sided conductive sheets could flunk on such a test. So it is important to standardize on the area to be exposed to the UHV conditions if one is doing such a comparative test.
Having said all of that, we believe that the "dry adhesives" are far better than wet paints. But we also believe that the best laboratory practice would involve the use of the smallest amount of adhesive possible, and for that reason, we usually recommend the use of the sheets, which are cut out to the size desired for each sample, rather than the discs for which the smallest is still probably bigger than what is needed for many samples. Of course my other factors, such as personal preference and experience come into play so I am sure there could be selection based on factors other than purely that of off-gassing.
Disclaimer: SPI Supplies offers all four types of mountants for use for the mounting of SEM samples.
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 ############################ ==================================================
I provide a copy of the following URL to a site with rich detail on imaging for those of us in the mood to learn.
http://www.tasi.ac.uk/welcome.html
This site is TASI: "Technical Advisory Service for Images". Suggest further, to find tree of info, click on "FRAMEWORK"
Regards to all,
Fred Monson
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
I am posting this for a college, Lauri Wyner, in the Pathology Core. I have no idea how to section this stuff...!
Thanks! Maria Ericsson
My question is: I have a carbon coated porous tantalum matrix approximately 1 cm in circumference and 2 mm thick. This matrix has fixed cells adhered to the surface and throughout. I would like to embed this, make slides and stain for H&E to confirm the presences of cells as well as immunohistochemistry to characterize them. I am looking for suggestions on how I can section this matrix while maintaining its overall structure. Any information would be greatly appreciated.
Lauri Wyner DF/HCC Central Pathology Cores Coordinator Harvard Medical School G1-126, Goldenson Building 220 Longwood Avenue Boston, MA 02115 Tele: (617) 432-4947 Fas: (617) 432-6474 Lauri_wyner-at-hms.harvard.edu
This is a test for being able to send from our institution to the MSA List server. MS Outlook97 is annoying in being somewhat inscrutable for sending html unless you know how to find it's hidden secrets. This we have done. : ) ------------------------------------- Name: Charles Gilbert VOC: (704) 355-0604 Carolinas Medical Center FAX: (704) 355-8424 Dept of Pediatric Research digPager: (704) 355-4088 : 2058 PO Box 32861 Charlotte, NC 28232-2861
DISCLAIMER: {"The opinions are my own and not necessarily shared by my employer."}
Sent by Outlook under the 60 Hz electron recycling project ------------------------------------- "I know of no safe depository of the ultimate powers of the society but the people themselves; and if we think them not enlightened enough to exercise their control with wholesome discretion, the remedy is not to take it from them, but to inform their discretion by education." (Thomas Jefferson; Letter to Wm. C. Jarvis, 1820)
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-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Karen Dye wrote the following: ========================================================== We just acquired a new high resolution SEM (FEG) and want to keep it "clean" How can I find out about out-gassing rates and the relative "cleanliness" of various sample preparation supplies:
Is there an accepted method for rating these materials? ========================================================== We at SPI Supplies have been dealing with this particular issue for more than thirty years. I don't think your question has the kind of absolute answer you might have been seeking, for the following reasons:
a) There are at least several different sources of "sticky dots", and they do not all behave the same. Some off-gas more than others and some are more sensitive to the beam than others. Also other factors such as age and storage conditions can effect the behavior of some of these adhesive materials in vacuum.
b) The conductive inks and pastes similarly represent different compositions, they are not all the same, and do not all come from the same place, but in addition to that, the "off-gassing characteristics" also depend on how thick of a layer has been applied and whether there is a "skin" that forms on the top, resulting in a lingering "wet" area underneath the skin. This effect can be even more problematic for a paste. The details of the formulation can influence the tendency of the composition to form such a skin, which acts as a barrier to diffusion.
c) The typical epoxy can be cured using more or less hardener, and it can be cured at room temperature or at higher temperatures. A room temperature cure with less rather than more hardener increases the chances for there being more unpolymerized material being present to off-gas and cause problems. The use of more hardener and some residence time at a higher temperature tends to minimize the off-gassing effect.
d) Waxes tend to be nonconductive and beam sensitive. Some have plasticizers that migrate to the surface and off-gas. We try to avoid waxes as a mounting medium for this kind of application in any kind of SEM. I have waxes like dental waxes in mind in the making of that comment.
We have adopted our own "test" for UHV compatibility. Simply put, it is a measure of the deterioration of vacuum when a test specimen is inserted into a UHV system. We found that the SPI double sided conductive carbon sheet material, when a 10 mm square was used, was inserted into the UHV system, there was no visible "meter deflection" on the LED display for the vacuum. Since the adhesive is the same as what is used in the SPI carbon tape and double sided discs, we would expect the same result would be found for them as well. We know that some similar appearing materials offered by others will perform as well on such a test. But not in all instances. We know this might not be the perfect test, but over the years it has been quite effective in terms of validating those materials that are better for this kind of application than others. Obviously, the worst performing material, if just a tiny bit is used, could end up "passing" the test, just as an inordinately large piece of the SPI double sided conductive sheets could flunk on such a test. So it is important to standardize on the area to be exposed to the UHV conditions if one is doing such a comparative test.
Having said all of that, we believe that the "dry adhesives" are far better than wet paints. But we also believe that the best laboratory practice would involve the use of the smallest amount of adhesive possible, and for that reason, we usually recommend the use of the sheets, which are cut out to the size desired for each sample, rather than the discs for which the smallest is still probably bigger than what is needed for many samples. Of course my other factors, such as personal preference and experience come into play so I am sure there could be selection based on factors other than purely that of off-gassing.
Disclaimer: SPI Supplies offers all four types of mountants for use for the mounting of SEM samples.
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 ############################ ==================================================
Listers, Can anyone refer me to a company or lab that can do sub-micron particle size and distribution studies, especially but not necessarily, in the southeast US? It involves a study injecting small wear debris particles into a skeletal joint. Thanks for any help. ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Winston W. Wiggins, Supervisor June 4, 2001 2:44 PM Cannon Electron Microscopy Lab Ofc: 704-355-1267 Carolinas Medical Center Lab: 704-355-7220 P.O. Box 32861 (Ship to: 1000 Blythe Blvd ) Fax: 704-355-0589 Charlotte, NC 28232-2861 (Ship to: 28203 ) Winston.Wiggins-at-CarolinasHealthCare.org {mailto:WWiggins-at-Carolinas.org} ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
*********************************************************************** This electronic message may contain information that is confidential and/or legally privileged. It is intended only for the use of the individual(s) and entity named as recipients in the message. If you are not an intended recipient of this message, please notify the sender immediately and delete the material from any computer. Do not deliver, distribute or copy this message, and do not disclose its contents or take any action in reliance on the information it contains. Thank you.
The general question of outgassing and its effect on the performance of vacuum systems is discussed in some detail in Section 2.9 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 details).
There is one interesting way in which unexpectedly large amounts of solvents and gases can get introduced into an SEM when carbon or silver paint is used to mount a specimen. This can occur if the paint is smeared on a mounting stub in such a way that the entire area underneath the specimen is covered, and then only a relatively short time is allowed for the paint to dry. Then only the paint around the edges of the specimen will dry, while that underneath the specimen can remain wet. When inserted into the SEM the solvent from the wet paint underneath the specimen can slowly diffuse outward through the 'dry' paint around the edges, thereby providing a strong source of outgassing for a long time.
If you really must use a paing to mount specimens, I think it is best to only put a few small dabs of it at several spots around the periphery of the specimen. Then there will be a better opportunity for the solvent to evaporate out from each small dab, and if sufficient time is allowed for the drying process, no wet paint will be left trapped underneath the specimen.
Better yet, make some simple clips or clamps that will hold the specimen in place mechanically, without the use of solvents or adhesives. With a little ingenuity and some pliers you can fashion a clip to hold almost any specimen from a piece of heavy copper wire or a paperclip. -- Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan 3062 Dow Bldg.; 2300 Hayward St. Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
My Oxford EDS x-ray detector needs to be returned to Oxford for repair, and unfortunately, I cannot locate the original cover for the EDS port on the microscope. Neither Oxford not JEOL can supply a cover. I do not want to lose the use of the SEM for imaging while the detector is repaired. The microscope is a JEOL JXA-840. As far as I know, the column is identical to the JSM-840. Is there anyone out there able and willing to help me with a sale, rent, or loan of a suitable cover until my x-ray detector is repaired.
Thank you.
Dave Wilbur
-- __________________________________ David J. Wilbur, Ph.D. Instrumentation Specialist Department of Chemistry Tufts University voice: 617-627-2163 Fax: 617-627-3443 email: dwilbu01-at-tufts.edu __________________________________
I can agree with Ron about the waxes. I've used a JEOL 6600F and Hitachi S-4700s and neither of these instruments likes wax. The wax will outgas so badly that the loadlocks won't pump down, so you never get it into the main chamber. Superglue-type glues work good (fully cured) and I've had no trouble with black carbon double-sided tape. I also use carbon (graphite) paint after a 15-minute bake at about 40-50 degrees C. A vacuum dryer could also be used if your specimen was heat-sensitive. I don't much care for silver paint; it takes too long to dry and is hard to remove if necessary. For the ultimate in cleanliness, remember to handle your mounts, stages, etc. (anything that goes into the main chamber) with gloves. This will prevent finger oils from being dispersed in the system.
Ron Doole wrote: ----------- {snip} --------------
} To save you a lot of time in experimenting I suggest that } you use the minimum amount of mounting material, you want } the smallest possible surface area to see the vacuum and } you want the lowest desorption (outgassing) rate and lowest } vapour pressure. } } Of your 4 options I would not use sticky pads or waxes. I } would only use epoxy or conductive inks and pastes fully } underneath the specimen and ensure that the small amount } used was fully cured and dried by pumping it out in a } vacuum rig before loading it into the microscope. I would } also store it under vacuum before loading. } } Good luck, } Ron } } } } } We just acquired a new high resolution SEM (FEG) and want to keep it "clean". How can I find out about out-gassing rates and the relative "cleanliness" of various sample preparation supplies: } } } } 1. sticky dots } } 2. conductive inks and pastes } } 3. epoxies } } 4. waxes } } } } Is there an accepted method for rating these materials? } } } } } } ---------------------- } 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
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-598-1291 (pager) DSPS Packaging Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Position available in an independent testing lab in a suburb of Detroit, MI. Knowledge of ISI SS 40 required. Materials testing experience a plus. For information contact 734-668-3309 and leave message.
Several questions about safety.. What do you use for gloves when handling both resins and fixatives? What procedures are out there for precipitating lead? Is anyone aware of any suspected instances of EM chemical exposure (resins, osmium), documented cases and/or references about such? Our institution would appreciate any information to help them answer questions about this unique part of the lab.
Rons advice is all very sound, but you have overlooked the effect of temperature. Even a modest rise in temperature increases vapour pressure and therefore outgassing markedly. Unless a cold-trap is used the objective area is rather warmer than room temperature. To cure samples so that later outgassing is lessened, store specimens at a suitably increased temperature and under a modest vacuum. 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, June 05, 2001 6:13 PM, Ron Doole [SMTP:ron.doole-at-materials.oxford.ac.uk] wrote: } } } HI Karen, } } I have an old, but brilliant, book 'Leybold Vacuum } Handbook' by K. Diels and R. Jaeckel (Translated by H. Adam } and J. Edwards) Pergamon Press, 1966. It contains } explainations of these problems, descriptions of } experiments to measure outgassing and desorption, and rates } for most elements and compounds that may be used in vacuum } systems. } } If you want a simple way to test the various materials find } a pumping rig and pump it down to it's ultimate pressure at } least overnight, ensure that the ultimate pressure is of } the same order as your microscope. Then vent it to dry } nitrogen and pump down again noting the pressure change } with time to it's ultimate pressure (say for 1 hour). Then } vent it and introduce the unknown component and pump down } again noting the time and pressure. The change in pumping } speed (rate of decrease in pressure) will be due to the } outgassing. No fancy units but a direct comparison for } your problem. Remember that to make a real comparison of } outgassing rates you need to have similar amounts of } material in similar forms, ie. same surface areas and } volumes. However, I guess you are more concerned with the } comparison between different mounting techniques so similar } specimens mounted by the various techniques would be OK. } } To save you a lot of time in experimenting I suggest that } you use the minimum amount of mounting material, you want } the smallest possible surface area to see the vacuum and } you want the lowest desorption (outgassing) rate and lowest } vapour pressure. } } Of your 4 options I would not use sticky pads or waxes. I } would only use epoxy or conductive inks and pastes fully } underneath the specimen and ensure that the small amount } used was fully cured and dried by pumping it out in a } vacuum rig before loading it into the microscope. I would } also store it under vacuum before loading. } } Good luck, } Ron } } } } } We just acquired a new high resolution SEM (FEG) and want to keep it } } "clean". How can I find out about out-gassing rates and the relative } } "cleanliness" of various sample preparation supplies: } } } } 1. sticky dots } } 2. conductive inks and pastes } } 3. epoxies } } 4. waxes } } } } Is there an accepted method for rating these materials? } } } } } } ---------------------- } 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
Dave, Unless you have been running an exceptionally clean and good vacuum, a rubber stopper of the correct size will do nicely as a temporary substitute. In particular, make sure that it's not too small so that it doesn't get sucked through the port at 15 psi. Otherwise, after a couple of hours of pumping, the outgassing should be minimal (better after overnight).
Clean the sides of the stopper with IPA or some similar solvent and use a very small amount of your favorite vacuum grease around the sides.
Ken Converse owner Quality Images third party SEM service Delta, PA
David Wilbur wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com {mailto:ListServer-at-MSA.Microscopy.Com} } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } My Oxford EDS x-ray detector needs to be returned to Oxford for repair, } and unfortunately, I cannot locate the original cover for the EDS port } on the microscope. Neither Oxford not JEOL can supply a cover. I do } not want to lose the use of the SEM for imaging while the detector is } repaired. The microscope is a JEOL JXA-840. As far as I know, the } column is identical to the JSM-840. Is there anyone out there able and } willing to help me with a sale, rent, or loan of a suitable cover until } my x-ray detector is repaired. } } Thank you. } } Dave Wilbur } } -- } __________________________________ } David J. Wilbur, Ph.D. } Instrumentation Specialist } Department of Chemistry } Tufts University } voice: 617-627-2163 } Fax: 617-627-3443 } email: dwilbu01-at-tufts.edu {mailto:dwilbu01-at-tufts.edu} } __________________________________ } } } } }
We currently use either Denka or Fei filaments in our FESEM which have generally given similar results over similar lifetimes . However for various reasons we are mow experiencing occasional stray magnetic field interference with corresponding deterioration in image quality . I have never really thought it through but what's actually happening at the tip ?..
1. Apart from occasional irritating beam 'sway' is physical damage being done to the tungsten tip with this interference and
2. Assuming a fluid zirconia film/ball exists at the tip of the tungsten filament is this being consumed more quickly influencing lifetime ?
Best regards
Martyn
From M Harris Device Engineering ESM Ltd , Cardiff Rd . Newport , South Wales NP10 8YJ .
A comprehensive microscopy salary survey was published in the old EMSA Bulletin in the early 80's. It is too old to be useful. More recently, there was a smaller survey in "Microscopy Today." Check with Don Grimes at: microtoday-at-mindspring.com
Perhaps it's time for a new comprehensive survey.
Ron Anderson, IBM, Hopewell Jct., New York, USA. anderron-at-us.ibm.com
IBM Analytical Services; http://www.chips.ibm.com/services/asg
"Rajesh Patel" {rpatel-at-umdnj.edu} on 06/05/2001 01:27:32 PM
To: {microscopy-at-sparc5.microscopy.com} cc:
I beleive that some time a go a salray survey was done for electron microscopy field. How and where can I access that info.
Rajesh Patel Robert Wood Johnson Medical School Dept. of Pathology Electron Microscopy Lab 675 Hoes Lane Piscataway, NJ 08854
Dear Dave, While the exact fit would be nice, the dimensions of the requirement would be nicer. Also, if you haven't called the JEOL service center near you, you might try that. I have parts galore, but I have to know more about the item.
Fred Monson
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
} ---------- } From: David Wilbur } Sent: Tuesday, June 5, 2001 3:37 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: SEM/EDS Need port cover for JEOL 840 microscope } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 Oxford EDS x-ray detector needs to be returned to Oxford for repair, } and unfortunately, I cannot locate the original cover for the EDS port } on the microscope. Neither Oxford not JEOL can supply a cover. I do } not want to lose the use of the SEM for imaging while the detector is } repaired. The microscope is a JEOL JXA-840. As far as I know, the } column is identical to the JSM-840. Is there anyone out there able and } willing to help me with a sale, rent, or loan of a suitable cover until } my x-ray detector is repaired. } } Thank you. } } Dave Wilbur } } -- } __________________________________ } David J. Wilbur, Ph.D. } Instrumentation Specialist } Department of Chemistry } Tufts University } voice: 617-627-2163 } Fax: 617-627-3443 } email: dwilbu01-at-tufts.edu } __________________________________ } } } }
I would suggest glycol methacrylate embedment (as a start) and thin sectioning with a carbide knife (expensive, but well worth the cost when materials are 'hard')
Fred Monson
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
} ---------- } From: Maria Ericsson } Sent: Tuesday, June 5, 2001 11:53 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: sectioning tantalum matrix } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 posting this for a college, Lauri Wyner, in the Pathology Core. } I have no idea how to section this stuff...! } } Thanks! } Maria Ericsson } } My question is: } I have a carbon coated porous tantalum matrix approximately 1 cm in } circumference and 2 mm thick. This matrix has fixed cells adhered to the } surface and throughout. I would like to embed this, make slides and stain } for H&E to confirm the presences of cells as well as immunohistochemistry } to characterize them. I am looking for suggestions on how I can section } this matrix while maintaining its overall structure. Any information would } } be greatly appreciated. } } } } Lauri Wyner } DF/HCC Central Pathology Cores Coordinator } Harvard Medical School } G1-126, Goldenson Building } 220 Longwood Avenue } Boston, MA 02115 } Tele: (617) 432-4947 } Fas: (617) 432-6474 } Lauri_wyner-at-hms.harvard.edu } } }
I keep running into users who believe that High magnification for an SEM, say 20,000X and greater is a benefit to EDS analysis. I have always been under the impression that high magnification is not a benefit to EDS due to the large interaction volume of X-rays. Comments please.
The keyword here is analysis. You are absolutely correct for this. The interaction volume will definitely be much larger than the particular feature that the beam can fully resolve. I preached this for many years and still do to a certain extent. However, it was shown to me by a colleague here after we had been arguing about it that in fact you can get meaningful qualitative information from areas much smaller than the interaction volume. He showed me that putting the beam on a surface feature on a thin film on glass that had a lateral spatial dimension less than 0.1 um and comparing it to an area adjacent to this area gave a significant difference in the spectra. Enough of a difference that it enabled him to pinpoint the problem. We do quite a lot of qualitative analysis and very little quantitative analysis. From that standpoint, higher magnifications can be quite useful, but not for quantitative analysis.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Ingram, Mike [mailto:MIngram-at-rodel.com] Sent: Wednesday, June 06, 2001 11:06 AM To: 'Microscopy-at-MSA.Microscopy.Com'
I keep running into users who believe that High magnification for an SEM, say 20,000X and greater is a benefit to EDS analysis. I have always been under the impression that high magnification is not a benefit to EDS due to the large interaction volume of X-rays. Comments please.
I agree that good vacuum practice is the first line of defense in keeping your new FEG SEM clean. The use of most of these materials may dirty your microscope.
XEI Scientific does offer a system to remove outgassed hydrocarbons from the walls and atmosphere of your microscope. The details about our EVACTRON SEM-CLEAN system may be found at www.SEMCLEAN.com This system was specifically developed for keeping FEG SEMs clean.
Ron Vane XEI Scientific 3124 Wessex Way Redwood City, CA 94061 (650)-369-0133
-----Original Message----- } From: Karen Dye {karen.dye-at-medtronic.com} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
} My Oxford EDS x-ray detector needs to be returned to Oxford for repair, } and unfortunately, I cannot locate the original cover for the EDS port } on the microscope. Neither Oxford not JEOL can supply a cover. I do } not want to lose the use of the SEM for imaging while the detector is } repaired. The microscope is a JEOL JXA-840. As far as I know, the } column is identical to the JSM-840. Is there anyone out there able and } willing to help me with a sale, rent, or loan of a suitable cover until } my x-ray detector is repaired. } David,
There are actually at least two different ports that could be used for mounting of the xray detector or, in this case, the blanking plate for the removed detector.
Is it the high take off angle port or the side mount, circular port?
P.S. I would advise against using a rubber stopper. This technique could leave you needing more than a blanking plate.
Bob Roberts EM Lab Services, Inc. 2409 S. Rural Rd Suite C Tempe, Arizona 85282 (480) 967-3946 "www.emlabservices.com"
I would like to hear from users out there about experience with the Sutter Lambda DG4 fluorescence source/filter changer. It looks very nice on paper. How is brightness/field uniformity/flexibility for general use. We want ratio capability which it appears good at, but we also want versatility for normal single fluor imaging at various wavelengths. Any comments about using this as opposed to a standard mercury, standard xenon, or monochromator for a general fluorescence microscopy facility are appreciated. Thanks- Dave --
************************************************************ 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 ************************************************************
Generally, nothing is happening to the tip by the magnetic fields as the only components that affect filament life are: pressure, overdriving, & poor filament construction.
Earl
----- Original Message ----- } From: {"HARRISm-at-esm-semi.co.uk"-at-sparc5.microscopy.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, June 06, 2001 7:19 AM
A rubber stopper, well-greased with appropriate grease, worked fine on my 840 in a similar situation, no degradation of vacuum. Just make absolutely sure that it's big enough that there's no chance of its being sucked right in, and that nobody knocks into it!
cheers
rtch
} } } } My Oxford EDS x-ray detector needs to be returned to Oxford for repair, } } and unfortunately, I cannot locate the original cover for the EDS port } } on the microscope. Neither Oxford not JEOL can supply a cover. I do } } not want to lose the use of the SEM for imaging while the detector is } } repaired. The microscope is a JEOL JXA-840. As far as I know, the } } column is identical to the JSM-840. Is there anyone out there able and } } willing to help me with a sale, rent, or loan of a suitable cover until } } my x-ray detector is repaired. } } } David, } } There are actually at least two different ports that could } be used for mounting of the xray detector or, in this case, } the blanking plate for the removed detector. } } Is it the high take off angle port or the side mount, } circular port? } } P.S. I would advise against using a rubber stopper. This } technique could leave you needing more than a blanking plate. Dr 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 recently performed an x-ray map and line scan on a cross sectioned Ag on Cu sample at 20,000X (15kV) and was able to show little to no migration of the Ag into the Cu. I was very surprised at how sharp the demarcations between the layers were at 20,000 in the x-ray maps and line scans. I would be happy to share the data w/ any interested parties. It is my opinion that high magnification EDS analysis can be useful. Of' course one can lower the kV or work with very thin samples to push the limits.
Ric
-----Original Message----- } From: Ingram, Mike [mailto:MIngram-at-rodel.com] Sent: Wednesday, June 06, 2001 11:06 AM To: 'Microscopy-at-MSA.Microscopy.Com'
I keep running into users who believe that High magnification for an SEM, say 20,000X and greater is a benefit to EDS analysis. I have always been under the impression that high magnification is not a benefit to EDS due to the large interaction volume of X-rays. Comments please.
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EMF does not generally affect SEMs in the electron column, sufficient shielding being provided by the normal design of these areas. Once the beam leaves the final pole piece and enters the sample chamber, though, it is a different matter. You can tell if the major influence is in the chamber by doing a simple qualitative measure of the interference at different working distances. If the problem is greater at the longer working distance, then the EMF is influencing the beam during its traverse through the sample chamber.
The same is basically true for vibrational effects. Within the column, physical movement of the instrument laterally is somewhat cancelled by the changing proximity to the applied fields of the electron optic system. Once free of those fields, though, movement of the instrument (and thus the sample) change the apparent position of the beam. Once again, the effect is reduced with a reduction in the working distance.
The first step has to be to determine whether the problem is EMF or vibration - quite possibly both. Since many vibrational sources are linked to the electrical line frequency, this can sometimes be tough. However, most modern SEMs should be quite capable of damping 60Hz. vibrations, but if EMF of any low frequency can come in then all low frequencies will be allowed in - EMF shielding is far less specific than vibrational.
This leads to the usual trick of trying to determine the offensive frequency by counting the number of 'saw-tooth' edges in a portion of a slow image sweep. If the frequency is close to 60Hz then it is probably EMF.
If the appearance of a problem like this is rather sudden, then the natural tendency is to find a temporal link to some change in the system. Taking this approach, though, you have to be sure to include all possible changes that may have taken place. For both problems, you have to consider if any changes have been made in the environment around the SEM. Has any equipment been added or moved in the area that may affect the instrument? Can the effect be due to a seasonal change in the HVAC equipment in use? Have there been any increases in the current load on any power distribution lines near the instrument?
More intimate causes also have to be accessed. Have any electrical cables, air, water or vacuum lines on the SEM been repositioned? EMF isolation generally involves putting a distance between any sources and any receptors. Vibrational isolation generally involves that as well as several levels of damping that require isolation between them. A simple cable or tube that is moved and bridges the isolation of one or more stages can destroy their effectiveness. Moving a computer monitor or external vacuum gauge controller a few inches can make profound changes in received EMF.
Finally, one has to consider time. Time, along with other influences, can produce changes such as corrosion of electrical connections that can cause high resistance in ground connections (in this case a 'high' resistance of only a few ohms can cause ground loop currents in instruments that can become a real problem). Another problem with aging instruments is in the filter capacitors of their power supplies. Capacitors age, and in doing so, their values often change. If a circuit is very tightly designed, this change can result in an increase in a 60 Hz ripple seen in power supplies. If circuit designs rely heavily on bypass capacitors to reduce ripple to individual circuits from power supply circuits, a failure of one or more of those capacitors can also result in increased supply ripple to circuits. Such failures will have the appearance of an induced EMF effect, but without the dependence on working distance.
Seems like I've said a lot, but I haven't even scratched the surface. SEMs, and particularly FESEMs are extremely sensitive instruments. Their design, implementation and continued operation are a continual balance of many factors. However, the problems you describe are probably not related to the use of third-party emitters.
You didn't mention the make and model of the instrument you are using, or the particular orientation of the emitters you are using. Could you elaborate on these?
On Wednesday, June 06, 2001 9:19 AM, "HARRISm-at-esm-semi.co.uk"-at-sparc5.microscopy.com [SMTP:"HARRISm-at-esm-semi.co.uk"-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 } -----------------------------------------------------------------------. } } } } } Dear Colleagues , } } We currently use either Denka or Fei filaments in our } FESEM which have generally given similar results over similar } lifetimes . } However for various reasons we are mow experiencing occasional stray } magnetic field interference with corresponding deterioration in image } quality . } I have never really thought it through but what's actually happening } at the tip ?.. } } 1. Apart from occasional irritating beam 'sway' is physical damage } being done to the tungsten tip with this interference and } } 2. Assuming a fluid zirconia film/ball exists at the tip of the } tungsten filament is this being consumed more quickly influencing } lifetime ? } } Best regards } } Martyn } } } From M Harris } Device Engineering } ESM Ltd , Cardiff Rd . } Newport , South Wales } NP10 8YJ . } } Email harrism-at-esm-semi.co.uk } } }
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
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Interesting question. I generally let users know that the interactions of electrons and x-ray generation and flourescence limit the analytical volume to a sphere of 50 - 100 microns in diameter. However, that assumes that you want to quantify that specific volume. There are many times when you want different criteria. For example, if one were to do a line scan across the interface of two differing materials. In this case, we can artificially determine the boundary as a function of the constantly varying intensities of their different components even though the compositional change takes place in an area much smaller than we can reasonable measure.
Such measurement, however, has to be made with the caviat of the experimental conditions and the 'thresholds' used for determination. Ideally, these are determined using similar matrix standards - i.e., similar materials whose dimensional composition can be verified by other means.
Thin film or small particle determinations are made using assumptions that are probably quite accurate, but have to be taken with consideration of the experimental conditions and their effects on known compositions. One has to be careful to ensure that the assumptions made by a particular routine are applicable to their particular circumstances.
Case in point - you currently have the beam located on the exact center of the interface of two widely different materials. In this case, you would expect an equal contribution from both materials. However, you have to consider many factors of x-ray generation by incident electron radiation, the flourescence and cross-flourescence of the individual materials as well as the absorption of each for varying wavelengths. Now add into the mix the morpholgy of your specific samples. We don't currently have all the answers to these complex problems and rely, instead, on empirical data. The closer the empirical data to the materials you're looking at, the better the analysis.
In brief, very minor differences between extremely close materials is possible, but what you can make of those differences depends on the effort you put into it.
On Wednesday, June 06, 2001 10:06 AM, Ingram, Mike [SMTP:MIngram-at-rodel.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 keep running into users who believe that High magnification for an SEM, } say 20,000X and greater is a benefit to EDS analysis. I have always been } under the impression that high magnification is not a benefit to EDS due } to the large interaction volume of X-rays. Comments please. } } } Mike Ingram } Rodel Inc. }
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
Tantalum is very hard and tough, so I doubt that it will section with either diamond or tungsten carbide knives. Preparing the material like a geological section by grinding is a possibility, but not a good one. Chances are that the grinding material would fill the voids and the tissue would be ground away first (although ProSciTech and others supply diamond grit embedded in plastic disks, which are much better in that regard) Proper vacuum infiltration with a low viscosity resin, cutting with a diamond blade, grinding with a diamond disc would be my preparation trial. After that either stain the surface and try your luck with a high power reflection (metallurgical) microscope, or more likely digest the plastic and view the specimen in SEM. If the tantalum could be fractured open, an ESEM my offer some "insights". There is no simple solution to the preparation of very hard and very soft materials in one specimen. Cheers Jim Darley Disclaimer: ProSciTech supplies both diamond and TC knives. I advised against the use of either in this particular case. 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, June 06, 2001 1:54 AM, Maria Ericsson [SMTP:maria_ericsson-at-hms.harvard.edu] wrote: } } } } I am posting this for a college, Lauri Wyner, in the Pathology Core. } I have no idea how to section this stuff...! } } Thanks! } Maria Ericsson } } My question is: } I have a carbon coated porous tantalum matrix approximately 1 cm in } circumference and 2 mm thick. This matrix has fixed cells adhered to the } surface and throughout. I would like to embed this, make slides and stain } for H&E to confirm the presences of cells as well as immunohistochemistry } to characterize them. I am looking for suggestions on how I can section } this matrix while maintaining its overall structure. Any information would } be greatly appreciated. } } } } Lauri Wyner } DF/HCC Central Pathology Cores Coordinator } Harvard Medical School } G1-126, Goldenson Building } 220 Longwood Avenue } Boston, MA 02115 } Tele: (617) 432-4947 } Fas: (617) 432-6474 } Lauri_wyner-at-hms.harvard.edu } }
we are in the process of cost our EM service, that is a cost breakdown for doing EM. i know the actual costs are very small. i was wondering if anyone has done it recently. we are a diagnostic lab. john hoffpauir cooper hospital
Just out of curiousity. . . how much X-radiation penetrates the rubber stopper?
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
I vote with Ritchie. Rubber stoppers are great for temporary problems, particularly for finding vacuum leaks. One stopper can eliminate a more complex assembly. I somehow prefer the silicone rubber stoppers. However, do consider X-ray penetration. Particularly on instruments above 20kV. 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, June 07, 2001 10:44 PM, Ritchie Sims [SMTP:r.sims-at-auckland.ac.nz] wrote:
} } } A rubber stopper, well-greased with appropriate grease, worked fine } on my 840 in a similar situation, no degradation of vacuum. } Just make absolutely sure that it's big enough that there's no chance } of its being sucked right in, and that nobody knocks into it! } } cheers } } rtch } } } } } } } } } } } } My Oxford EDS x-ray detector needs to be returned to Oxford for repair, } } } and unfortunately, I cannot locate the original cover for the EDS port } } } on the microscope. Neither Oxford not JEOL can supply a cover. I do } } } not want to lose the use of the SEM for imaging while the detector is } } } repaired. The microscope is a JEOL JXA-840. As far as I know, the } } } column is identical to the JSM-840. Is there anyone out there able and } } } willing to help me with a sale, rent, or loan of a suitable cover until } } } my x-ray detector is repaired. } } } } } David, } } } } There are actually at least two different ports that could } } be used for mounting of the xray detector or, in this case, } } the blanking plate for the removed detector. } } } } Is it the high take off angle port or the side mount, } } circular port? } } } } P.S. I would advise against using a rubber stopper. This } } technique could leave you needing more than a blanking plate. } Dr 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 order to re-send this email, I had to change the subject line to stop a rejection because of the r word.
-----Original Message----- } From: Beauregard, Paul A. Sent: Thursday, June 07, 2001 11:39 AM To: Microscopy-at-sparc5.microscopy.com
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Jim Darley wrote:
==================================================== Tantalum is very hard and tough, so I doubt that it will section with either
diamond or tungsten carbide knives. Preparing the material like a geological
section by grinding is a possibility, but not a good one. Chances are that the grinding material would fill the voids and the tissue would be ground away first (although ProSciTech and others supply diamond grit embedded in plastic disks, which are much better in that regard) Proper vacuum infiltration with a low viscosity resin, cutting with a diamond blade, grinding with a diamond disc would be my preparation trial. After that either stain the surface and try your luck with a high power reflection (metallurgical) microscope, or more likely digest the plastic and
view the specimen in SEM. If the tantalum could be fractured open, an ESEM my offer some "insights". There is no simple solution to the preparation of very hard and very soft materials in one specimen. ===================================================== Actually Ta **can** be thin sectioned if it is done by someone with the proper experience and who is using the right kind of diamond knife. We have offered our SPI #02888-AB sectioned Ta foil on a grid for AEM studies for some number of years; see URL http://www.2spi.com/catalog/standards/aem.html
The Ta system that prompted the original posting was thicker, however, it is also porous, and if properly infiltrated with resin, and assuming the porosity is above some point, in principle, at least, there is no reason why it could not be thin sectioned for TEM. No method is really artifact free, but some artifacts are more easy to recognize than others. Knife induced artifacts are anisotripic (e.g. directional) in nature and can be more easily recognized (as artifacts) than artifacts caused by say, ion milling, which are isotropic in nature.
When I say "right kind of diamond knife", I am not suggesting that the SPI diamond knife could do something above and beyond what other diamond knives could do. What I am saying is that there is a process of selection of the optimum knife angle, because one might have to be prepared to use a knife with a fairly low (for materials science work) knife angle, rather than a more blunt angle, but with the downside is that it will wear out more quickly. That might make for great business for a diamond knife supplier, but it does tend to get expensive for the user who is not so experienced with this kind of sectioning.
Disclaimer: SPI Supplies offers diamond knives, both materials and life science, and we have done this kind of sectioning, as a service, for commercial clients for over thirty years.
Chuck
PS: Please remember that we are nearly 100% paperless and we would ask that any reply to this message be by way of the "reply" feature on your software, so that the entire string of correspondence can come back to us and all be in one place.
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 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 ############################ ==================================================
Following up on Jim's comments about using mechanical cutting and grinding techniques, I have some insight that may help. Jim mentioned that "There is no simple solution to the preparation of very hard and very soft materials in one specimen". This is true to some degree, but there are solutions. For example, the use of a wire saw is actually ideal for cutting through materials containing both hard and soft phases. The wire saw can be used either with an abrasive slurry or with a diamond impregnated wire. Use with an abrasive slurry is ideal for cutting materials of various hardness without damage or the smearing effect that you would see with a diamond wheel saw.
To be honest, I have no idea if this saw would do what you want it to do in this application, however if the difficulty is the hard/soft combination, the wire saw is one potentially viable solution. Also, I'm not sure if you need to embed the sample, but I would suggest forgoing tat step, if possible, if oyu are going to try the wire saw technique.
DISCLAIMER: South Bay Technology pioneered the development of the wire saw in the early 60s and continues to produce wire saws for many applications today.
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} } Tantalum is very hard and tough, so I doubt that it will section with either } diamond or tungsten carbide knives. Preparing the material like a geological } section by grinding is a possibility, but not a good one. Chances are that the } grinding material would fill the voids and the tissue would be ground away } first (although ProSciTech and others supply diamond grit embedded in plastic } disks, which are much better in that regard) } Proper vacuum infiltration with a low viscosity resin, cutting with a diamond } blade, grinding with a diamond disc would be my preparation trial. } After that either stain the surface and try your luck with a high power } reflection (metallurgical) microscope, or more likely digest the plastic and } view the specimen in SEM. } If the tantalum could be fractured open, an ESEM my offer some "insights". } There is no simple solution to the preparation of very hard and very soft } materials in one specimen. } Cheers } Jim Darley } Disclaimer: ProSciTech supplies both diamond and TC knives. I advised against } the use of either in this particular case. } 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, June 06, 2001 1:54 AM, Maria Ericsson } [SMTP:maria_ericsson-at-hms.harvard.edu] wrote: } } } } } } } } I am posting this for a college, Lauri Wyner, in the Pathology Core.
} } I have no idea how to section this stuff...! } } } } Thanks! } } Maria Ericsson } } } } My question is: } } I have a carbon coated porous tantalum matrix approximately 1 cm in } } circumference and 2 mm thick. This matrix has fixed cells adhered to the } } surface and throughout. I would like to embed this, make slides and stain } } for H&E to confirm the presences of cells as well as immunohistochemistry } } to characterize them. I am looking for suggestions on how I can section } } this matrix while maintaining its overall structure. Any information would } } be greatly appreciated. } } } } } } } } Lauri Wyner } } DF/HCC Central Pathology Cores Coordinator } } Harvard Medical School } } G1-126, Goldenson Building } } 220 Longwood Avenue } } Boston, MA 02115 } } Tele: (617) 432-4947 } } Fas: (617) 432-6474 } } Lauri_wyner-at-hms.harvard.edu } } } }
-- David Henriks TEL: +1-949-492-2600 South Bay Technology, Inc. FAX: +1-949-492-1499 1120 Via Callejon San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
*** www.southbaytech.com ***
-- David Henriks TEL: +1-949-492-2600 South Bay Technology, Inc. FAX: +1-949-492-1499 1120 Via Callejon San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
While infiltrating a set of specimens (insect abdomens and thoraxes) in LR White, something occurred that has so far defied my attempts to figure out. I was doing side by side microwave and conventional fixations, dehydrations, and embeddings. The microwave specimens were dehydrated in acetone, behaved normally, and we have polymerized blocks. However I dehydrated the conventional set in an ethanol series, as we have done many times before in LR White. The first infiltration step was 2 parts ethanol to one part LR White Medium Grade at room temp, and upon going later to change into 1:1, I found that the resin/ETOH mix had partially polymerized into a spongy, granular mass. Uh-oh, bad resin, I said, along with a few other choice words.
After the initial panic, I ended up putting the chunks into pure LR White from a newly opened bottle and the partially polymerized resin seemed to go back into solution. I ran them through a couple more changes of pure resin, then left them on a rocker overnight at room temperature. Checking them this morning, I found two of the samples were fine, and the third had polymerized into a rubbery mass. Same bottle of resin, same identically processed samples, same everything.
An additional tube with a sample of the "bad" resin was also happily unpolymerized, as was the remainder of the "bad" resin in the bottle, which I had left in the fume hood overnight at room temperature. The "bad" resin was slightly more than a year old and has been refrigerated at 4 C since we got it. The second resin is about 10 months old and has also been constantly refrigerated. Neither had any accelerator in them (at least none added by us).
I have my share of problems with LR White, but this one really has us puzzled. Could there have been something in the samples that triggered a polymerization? Can LR White react with some plastics in this way? (We were using a relatively new batch of Eppendorf tubes that seem more hydrophobic than our previous ones.) Could it have been the full moon?
Regards, 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 http://biotech.missouri.edu/emc
Does anyone have a reference for or some transmission electron micrographs that illustrate the effects of osmolarity of buffers on animal cells? I would like to show students on tour of our facility shrinkage and swelling of cells and organelles. I have other wonderful artifacts to show, including one of my first sections with chatter so bad it looks like mini-blinds...
Mahalo! 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* ****************************************************************************
Biomedical Electron Microscopy by Maunsbach A B & Afzelius B A (1999) has several pages of illustrations of shrinkage etc.
Dave
On Thu, 7 Jun 2001 16:29:12 -1000 (HST) Tina Carvalho {tina-at-pbrc.hawaii.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- } } Does anyone have a reference for or some transmission electron micrographs } that illustrate the effects of osmolarity of buffers on animal cells? I } would like to show students on tour of our facility shrinkage and swelling } of cells and organelles. I have other wonderful artifacts to show, } including one of my first sections with chatter so bad it looks like } mini-blinds... } } Mahalo! } 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* } **************************************************************************** } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
Chuck - the point of the original request was not to section Ta - which is hard enough, but do show the cells attached to the Ta. If you think that you can section Ta without the cells being ripped away, then do it. I will praise your skills when I can see useful results. 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 08, 2001 8:26 AM, Garber, Charles A. [SMTP:cgarber-at-2spi.com] wrote: } } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } Jim Darley wrote: } } ==================================================== } Tantalum is very hard and tough, so I doubt that it will section with either } } diamond or tungsten carbide knives. Preparing the material like a geological } } section by grinding is a possibility, but not a good one. Chances are that } the } grinding material would fill the voids and the tissue would be ground away } first (although ProSciTech and others supply diamond grit embedded in } plastic } disks, which are much better in that regard) } Proper vacuum infiltration with a low viscosity resin, cutting with a } diamond } blade, grinding with a diamond disc would be my preparation trial. } After that either stain the surface and try your luck with a high power } reflection (metallurgical) microscope, or more likely digest the plastic and } } view the specimen in SEM. } If the tantalum could be fractured open, an ESEM my offer some "insights". } There is no simple solution to the preparation of very hard and very soft } materials in one specimen. } ===================================================== } Actually Ta **can** be thin sectioned if it is done by someone with the } proper experience and who is using the right kind of diamond knife. We have } offered our SPI #02888-AB sectioned Ta foil on a grid for AEM studies for } some number of years; see URL } http://www.2spi.com/catalog/standards/aem.html } } The Ta system that prompted the original posting was thicker, however, it is } also porous, and if properly infiltrated with resin, and assuming the } porosity is above some point, in principle, at least, there is no reason why } it could not be thin sectioned for TEM. No method is really artifact free, } but some artifacts are more easy to recognize than others. Knife induced } artifacts are anisotripic (e.g. directional) in nature and can be more } easily recognized (as artifacts) than artifacts caused by say, ion milling, } which are isotropic in nature. } } When I say "right kind of diamond knife", I am not suggesting that the SPI } diamond knife could do something above and beyond what other diamond knives } could do. What I am saying is that there is a process of selection of the } optimum knife angle, because one might have to be prepared to use a knife } with a fairly low (for materials science work) knife angle, rather than a } more blunt angle, but with the downside is that it will wear out more } quickly. That might make for great business for a diamond knife supplier, } but it does tend to get expensive for the user who is not so experienced } with this kind of sectioning. } } Disclaimer: SPI Supplies offers diamond knives, both materials and life } science, and we have done this kind of sectioning, as a service, for } commercial clients for over thirty years. } } Chuck } } PS: Please remember that we are nearly 100% paperless and we would ask that } any reply to this message be by way of the "reply" feature on your software, } so that the entire string of correspondence can come back to us and all be } in one place. }
} Hi, All- } } Does anyone have a reference for or some transmission electron micrographs } that illustrate the effects of osmolarity of buffers on animal cells? I } would like to show students on tour of our facility shrinkage and swelling } of cells and organelles. I have other wonderful artifacts to show, } including one of my first sections with chatter so bad it looks like } mini-blinds... } } Mahalo! } Tina
Arborgh et al. J. Ultrastruc. Res. 56:339-350, 1976. A classic study.
} } } 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* } ****************************************************************************
-- ********************************************** 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 utility of rubber stoppers to seal a SEM has been doubted. 1 Somebody may chose too smaller stopper size and the stopper may be sucked into the column. Good point, just like noting that the stopper must be inserted from the outside of the SEM. Yes!
2 The X-ray objection is one which needs to be addressed. I had looked up that data some years ago, when in need of temporary blanks. Because of the questions I again looked for the data, now on the marvelous Internet . The common Monte Carlo programs would give similar results, but the show the bulk of the X-ray interaction, whereas we are interested in maximum penetration. I recommend http://www-cxro.lbl.gov/optical_constants/atten2.html This site allows some data entry and then calculates the attenuation length at which the X-ray intensity falls of to 1eV at the surface. Assuming that Teflon is close to rubber in absorption, then 20keV X-ray photons are attenuated in 5mm of rubber. Whereas 30keV photons are only attenuated in about 12mm of rubber. Clearly, it would be unwise to run the kV of a TEM, when blanked with a rubber stopper. It is also clear that a SEM/ Probe when operated at 20 kV would not produce X-rays capable of penetrating a normal rubber stopper. Incidentally, a 20kV beam would produce somewhat lower KeV X-ray photons. X-ray generation depends on the interaction with electron shells. Only the heavier metals can produce powerful X-rays and to reach full fluorescence (maximum production) the kV needs to be about 2x that of the X-ray KeV. Cheers Jim Darley PS Red stoppers contain Iron oxide pigment, whereas black stoppers (I presume) have negligible metal content. Question: Considering that iron has greater X-ray stopping power than carbon, should we assume that it is smarter to use red stoppers??? The opposing argument would be that the iron could generate more high energy X-rays.
I don't know the answer (would be amused to learn though), but I prefer silicone rubber stoppers because they form a better vacuum seal.
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, June 07, 2001 10:14 PM, Marie E. Cantino [SMTP:cantino-at-uconnvm.uconn.edu] wrote: } } } Just out of curiousity. . . how much X-radiation penetrates the rubber } stopper? } } Marie }
} I vote with Ritchie. Rubber stoppers are great for temporary problems, } particularly for finding vacuum leaks. One stopper can eliminate a more complex } assembly. I somehow prefer the silicone rubber stoppers. } However, do consider X-ray penetration. Particularly on instruments above } 20kV. } 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
} } } } } A rubber stopper, well-greased with appropriate grease, worked fine } } on my 840 in a similar situation, no degradation of vacuum. } } Just make absolutely sure that it's big enough that there's no chance } } of its being sucked right in, and that nobody knocks into it! } } } } cheers } } } } rtch } } } } 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 }
I usually wrap several layers of aluminum foil over the stopper if I need to use it. This will stop most of the X-rays. Fortunately, the EDS port is to the rear of the column and pointing upward.
Earl
----- Original Message ----- } From: "Arthur Day" {ard-at-ansto.gov.au} To: {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, June 07, 2001 9:53 PM
-------- Original Message --------
Pehaps someone could provide a better analysis, but I have a couple of anecdotes:
I had a Canadian customer (20+ years ago) who specialized in very long EDS x-ray sweeps in either dot map form or a combined YZ modulated form. As I recall, some of these scans were up to 16 hours long. He had a Plexiglass port cover so that he could easily see his chamber geometry. This had a metal cover that was velvet lined for light leakage. Once he put his radiation badge inside the metal cover for a month, then turned it in. There apparently was nothing out of the ordinary, as his rad peo;le never said "boo". Perhaps it's a commentary on the rad people, but a full month exposure?
Second, I had another customer get me some rad figures on an SEM gun operating at 30kV and 150uA (1.5x10E-4). The bottom line was that the x-ray output from the gun was very serious, but all the guns are enclosed in steel. No problem. At the specimen for EDS work we're usually talking about 200-600pA (2-6x10E-10), or roughly 6 orders of magnitude less current and the stopper is going to absorb some of the x-rays generated. Is this really a problem?
I'm not talking about TEMs, as they can be very dangerous. The current at the screen is much higher, the excitation voltage is much higher and there is this large expanse of glass that MUST be leaded. Perhaps the currents used for WDS would also present a problem (10-100 nA or 10E-8 to 10E-7) at 2 to 3 orders of magnitude more than EDS.
Don't forget, those nice color displays on the four computers surrounding you in your lab also generate x-rays and their beam currents are in the mA range (10E-3) and an accelerating voltage of about 25kV. There may be a decelerating grid and leaded glass on the front, but I think you will find the the shielding on the rear of the monitor is not so rigorous, Your dentist's x-rays are about 70kV and only one order of magnitude greater current.
I'd love some feed-back from someone who knows radiation because I've felt that applying TEM rules to SEMs is gross over-kill, especially with so many color monitors around. How often do you have your decelerating grid checked for proper operation? If it doesn't operate correctly, your exposure could be very dangerous, given the time that one sits in front of these things and their distance from your face.
If I'm way off base, I'd like to know and know why.
Ken Converse owner Quality Images third party SEM service Delta, PA
Arthur Day wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 } } -----------------------------------------------------------------------. } } } } } } Just out of curiousity. . . how much X-radiation penetrates the rubber } } stopper? } } } } Marie } } } } Doesn't anybody supply X-ray proof rubber bungs for microscope ports? } } No worries. Just wrap a few sheets of lead foil around the column and } make sure pregnant operators stay below 5kV ;-( } } Surely something somewhat less desperate could be cobbled together out } of metal without too much extra work? } } } }
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Jim Darley wrote:
==================================================== Tantalum is very hard and tough, so I doubt that it will section with either
diamond or tungsten carbide knives. Preparing the material like a geological
section by grinding is a possibility, but not a good one. Chances are that the grinding material would fill the voids and the tissue would be ground away first (although ProSciTech and others supply diamond grit embedded in plastic disks, which are much better in that regard) Proper vacuum infiltration with a low viscosity resin, cutting with a diamond blade, grinding with a diamond disc would be my preparation trial. After that either stain the surface and try your luck with a high power reflection (metallurgical) microscope, or more likely digest the plastic and
view the specimen in SEM. If the tantalum could be fractured open, an ESEM my offer some "insights". There is no simple solution to the preparation of very hard and very soft materials in one specimen. ===================================================== Actually Ta **can** be thin sectioned if it is done by someone with the proper experience and who is using the right kind of diamond knife. We have offered our SPI #02888-AB sectioned Ta foil on a grid for AEM studies for some number of years; see URL http://www.2spi.com/catalog/standards/aem.html
The Ta system that prompted the original posting was thicker, however, it is also porous, and if properly infiltrated with resin, and assuming the porosity is above some point, in principle, at least, there is no reason why it could not be thin sectioned for TEM. No method is really artifact free, but some artifacts are more easy to recognize than others. Knife induced artifacts are anisotripic (e.g. directional) in nature and can be more easily recognized (as artifacts) than artifacts caused by say, ion milling, which are isotropic in nature.
When I say "right kind of diamond knife", I am not suggesting that the SPI diamond knife could do something above and beyond what other diamond knives could do. What I am saying is that there is a process of selection of the optimum knife angle, because one might have to be prepared to use a knife with a fairly low (for materials science work) knife angle, rather than a more blunt angle, but with the downside is that it will wear out more quickly. That might make for great business for a diamond knife supplier, but it does tend to get expensive for the user who is not so experienced with this kind of sectioning.
Disclaimer: SPI Supplies offers diamond knives, both materials and life science, and we have done this kind of sectioning, as a service, for commercial clients for over thirty years.
Chuck
PS: Please remember that we are nearly 100% paperless and we would ask that any reply to this message be by way of the "reply" feature on your software, so that the entire string of correspondence can come back to us and all be in one place.
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 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 ############################ ==================================================
Randy: This has happened a bunch of times to me. It only happens with osmicated tissues. I have run up the same tisse +/- osmication and premature polymerization only occurs in the osmicated ones - not every time or at the same rate. I guess there is something that doesn't get rinsed out that can trigger it - perhaps more so when the LRW is aging. I am starting a LRW infiltration with osmicated tissue using a brand new bottle of LRW and I will let you know what happens. my tissues were in glass vials so it isn't the plastic tubes. it has happened at both 4 C and room temp. i have a vague recollection posting this on the microscopy listserver and not getting much of a response. maddening problem, isn't it?
} ------------------------------------------------------------. } } } Dear Listers, } } While infiltrating a set of specimens (insect abdomens and thoraxes) in LR } White, something occurred that has so far defied my attempts to figure out. } I was doing } side by side microwave and conventional fixations, dehydrations, and } embeddings. The microwave specimens were dehydrated in acetone, behaved } normally, and we } have polymerized blocks. However I dehydrated the conventional set in an } ethanol series, as we have done many times before in LR White. The first } infiltration step } was 2 parts ethanol to one part LR White Medium Grade at room temp, and } upon going later to change into 1:1, I found that the resin/ETOH mix had } partially } polymerized into a spongy, granular mass. Uh-oh, bad resin, I said, along } with a few other choice words. } } After the initial panic, I ended up putting the chunks into pure LR White } from a newly opened bottle and the partially polymerized resin seemed to go } back into solution. I } ran them through a couple more changes of pure resin, then left them on a } rocker overnight at room temperature. Checking them this morning, I found } two of the } samples were fine, and the third had polymerized into a rubbery mass. } Same bottle of resin, same identically processed samples, same everything. } } An additional tube with a sample of the "bad" resin was also happily } unpolymerized, as was the remainder of the "bad" resin in the bottle, which } I had left in the fume } hood overnight at room temperature. The "bad" resin was slightly more } than a year old and has been refrigerated at 4 C since we got it. The } second resin is about 10 } months old and has also been constantly refrigerated. Neither had any } accelerator in them (at least none added by us). } } I have my share of problems with LR White, but this one really has us } puzzled. Could there have been something in the samples that triggered a } polymerization? Can } LR White react with some plastics in this way? (We were using a } relatively new batch of Eppendorf tubes that seem more hydrophobic than our } previous ones.) Could } it have been the full moon? } } Regards, } 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 } http://biotech.missouri.edu/emc }
-- 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)
You might have nailed it. These were osmicated tissues, since they weren't intended for immuno. Our client prefers LR White for making thick sections for light microscopy, so these specimens were prepared with osmium and UA post-fixations. Given the large size and difficult nature of insect parts, it's very possible that something remained in the tissue despite several lengthy washes. At least we have back-up specimens!
Thanks for the response. At least it's something to work with.
Randy
-----Original Message----- } From: Tom Phillips [mailto:PhillipsT-at-missouri.edu] Sent: Friday, June 08, 2001 9:24 AM To: Tindall, Randy D. Cc: Microscopy-at-msa.microscopy.com
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Colleagues....
I hate to say this, but alot of mail got rejected today by accident.
I was updating the junk mail filters last night and unintentionally created a new filter which rejected the majority of the mail posted today.
If you received a rejection message indicating that you had a subject line of \b$ please repost your message. It was not your fault, but mine.
Listers, Thanks to all who responded to my particle size analysis posting recently. Reasons, including a heavy workload, would not allow us to accept the project at this time. We now have several resources from which to choose. I appreciate the time and effort from those who offered constructive answers. From the one who offered questionable musings... contact me off-line and I'll be more
specific about what can be done with those. Winston ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Winston W. Wiggins, Supervisor June 8, 2001 10:07 AM Cannon Electron Microscopy Lab Ofc: 704-355-1267 Carolinas Medical Center Lab: 704-355-7220 P.O. Box 32861 (Ship to: 1000 Blythe Blvd ) Fax: 704-355-0589 Charlotte, NC 28232-2861 (Ship to: 28203 ) WWiggins-at-Carolinas.org {mailto:WWiggins-at-Carolinas.org} ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
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I also have a number of old Leitz "Technical Information Bulletins" that will be listed later.
Hope there is a use.
FCM
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
I concluded from the original information given that the tantalum matrix the young lady was speaking about was Cytomatrix (http://www.cytomatrix.com). The matrix is reportedly a tantalum coated graphite core material that was first use as a support for bone marrow/bone cell regeneration. That's why I suggested the glycol methacrylate, though you may still be correct about it's 'hardness'.
Fred Monson
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
} ---------- } From: Jim at Proscitech } Reply To: jim-at-proscitech.com } Sent: Thursday, June 7, 2001 6:25 AM } To: 'Maria Ericsson'; Microscopy-at-sparc5.microscopy.com } Subject: RE: sectioning tantalum matrix } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Tantalum is very hard and tough, so I doubt that it will section with } either } diamond or tungsten carbide knives. Preparing the material like a } geological } section by grinding is a possibility, but not a good one. Chances are that } the } grinding material would fill the voids and the tissue would be ground away } } first (although ProSciTech and others supply diamond grit embedded in } plastic } disks, which are much better in that regard) } Proper vacuum infiltration with a low viscosity resin, cutting with a } diamond } blade, grinding with a diamond disc would be my preparation trial. } After that either stain the surface and try your luck with a high power } reflection (metallurgical) microscope, or more likely digest the plastic } and } view the specimen in SEM. } If the tantalum could be fractured open, an ESEM my offer some "insights". } There is no simple solution to the preparation of very hard and very soft } materials in one specimen. } Cheers } Jim Darley } Disclaimer: ProSciTech supplies both diamond and TC knives. I advised } against } the use of either in this particular case. } 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, June 06, 2001 1:54 AM, Maria Ericsson } [SMTP:maria_ericsson-at-hms.harvard.edu] wrote: } } } } } } } } I am posting this for a college, Lauri Wyner, in the Pathology Core. } } I have no idea how to section this stuff...! } } } } Thanks! } } Maria Ericsson } } } } My question is: } } I have a carbon coated porous tantalum matrix approximately 1 cm in } } circumference and 2 mm thick. This matrix has fixed cells adhered to the } } surface and throughout. I would like to embed this, make slides and } stain } } for H&E to confirm the presences of cells as well as } immunohistochemistry } } to characterize them. I am looking for suggestions on how I can section } } this matrix while maintaining its overall structure. Any information } would } } be greatly appreciated. } } } } } } } } Lauri Wyner } } DF/HCC Central Pathology Cores Coordinator } } Harvard Medical School } } G1-126, Goldenson Building } } 220 Longwood Avenue } } Boston, MA 02115 } } Tele: (617) 432-4947 } } Fas: (617) 432-6474 } } Lauri_wyner-at-hms.harvard.edu } } } } } } }
I agree with David, there are several ways to cut and grind samples with multiple materials of varying hardnesses. The EXAKT Cutting/Grinding Systems were developed just for this purpose. One of the major issues in cutting or grinding these types of materials is the force required to cut the hard material vs. the force required to cut the softer material. Like the diamond wire saw a diamond embedded band system designed to section with a minimum of force can also achieve excellent results. The reduced force technology (reduction of the active cutting or grinding surface) is not new but has new applications in material research. And the technology can be applied to the grinding process as well to achieve flat surfaces at material interfaces.
Many of these techniques have been developed for LM in the biomaterials industry where the materials can vary from cobalt chrome implanted in bone or bone cement to memory metal stents implanted in soft arteries. The technology works equally well for rubber bonded to metal or solders layered on ceramics or carbon fibers embedded in unpolymerized resin. One of the key features of reducing the force and changing the cutting or grinding direction is also to prevent smearing of one material over an interface. Biomaterials researchers have been the driving force in developing this technology for the past 15 years and for LM it is a viable alternative to standard metallographic preparations.
Disclaimer: EXAKT Technologies, Inc. is the North American Distributor for EXAKT Apparatebau the manufacturer of various cutting devices and grinders for multiple material applications.
Linda Durbin
-----Original Message----- } From: David Henriks [mailto:henriks-at-southbaytech.com] Sent: Thursday, June 07, 2001 7:48 PM To: Microscopy Listerver
Following up on Jim's comments about using mechanical cutting and grinding techniques, I have some insight that may help. Jim mentioned that "There is no simple solution to the preparation of very hard and very soft materials in one specimen". This is true to some degree, but there are solutions. For example, the use of a wire saw is actually ideal for cutting through materials containing both hard and soft phases. The wire saw can be used either with an abrasive slurry or with a diamond impregnated wire. Use with an abrasive slurry is ideal for cutting materials of various hardness without damage or the smearing effect that you would see with a diamond wheel saw.
To be honest, I have no idea if this saw would do what you want it to do in this application, however if the difficulty is the hard/soft combination, the wire saw is one potentially viable solution. Also, I'm not sure if you need to embed the sample, but I would suggest forgoing tat step, if possible, if oyu are going to try the wire saw technique.
DISCLAIMER: South Bay Technology pioneered the development of the wire saw in the early 60s and continues to produce wire saws for many applications today.
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------.
} } Tantalum is very hard and tough, so I doubt that it will section with either } diamond or tungsten carbide knives. Preparing the material like a geological } section by grinding is a possibility, but not a good one. Chances are that the } grinding material would fill the voids and the tissue would be ground away } first (although ProSciTech and others supply diamond grit embedded in plastic } disks, which are much better in that regard) } Proper vacuum infiltration with a low viscosity resin, cutting with a diamond } blade, grinding with a diamond disc would be my preparation trial. } After that either stain the surface and try your luck with a high power } reflection (metallurgical) microscope, or more likely digest the plastic and } view the specimen in SEM. } If the tantalum could be fractured open, an ESEM my offer some "insights". } There is no simple solution to the preparation of very hard and very soft } materials in one specimen. } Cheers } Jim Darley } Disclaimer: ProSciTech supplies both diamond and TC knives. I advised against } the use of either in this particular case. } 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, June 06, 2001 1:54 AM, Maria Ericsson } [SMTP:maria_ericsson-at-hms.harvard.edu] wrote: } } } } } } } } I am posting this for a college, Lauri Wyner, in the Pathology Core.
} } I have no idea how to section this stuff...! } } } } Thanks! } } Maria Ericsson } } } } My question is: } } I have a carbon coated porous tantalum matrix approximately 1 cm in } } circumference and 2 mm thick. This matrix has fixed cells adhered to the } } surface and throughout. I would like to embed this, make slides and stain } } for H&E to confirm the presences of cells as well as immunohistochemistry } } to characterize them. I am looking for suggestions on how I can section } } this matrix while maintaining its overall structure. Any information would } } be greatly appreciated. } } } } } } } } Lauri Wyner } } DF/HCC Central Pathology Cores Coordinator } } Harvard Medical School } } G1-126, Goldenson Building } } 220 Longwood Avenue } } Boston, MA 02115 } } Tele: (617) 432-4947 } } Fas: (617) 432-6474 } } Lauri_wyner-at-hms.harvard.edu } } } }
-- David Henriks TEL: +1-949-492-2600 South Bay Technology, Inc. FAX: +1-949-492-1499 1120 Via Callejon San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
*** www.southbaytech.com ***
-- David Henriks TEL: +1-949-492-2600 South Bay Technology, Inc. FAX: +1-949-492-1499 1120 Via Callejon San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
I have have a task of quantifying the actin distribution in cultured endothelial cells and subsequently comparing different culture conditions. So far I have managed to isolate the filaments and threshold them. Now...I wonder what is the best way to put a number on the distribution within each cell. One possibility I was tinking of, is using a sobel operator to give the direction of the lines representing the filaments and then display the data as histograms of filament orientations.
Has anyone done this or know of a good wayto go about this type of analysis?
We have been using an older Balzer's sputter coater to coat SEM samples with Pt. Lately the grain structure of the Pt coating has become much more obvious (at around 100,000x.) Any experts have a thought on why this is occurring and how to improve the process? Thanks for any guidelines and help. Regards, Mike ******************************************************************** Michael L. Boucher Sr. mboucher-at-tc.umn.edu Lab Manager Rm 55 Office Ph 612-624-6590 I.T. Characterization Facility Rm 12 Main Off Ph 612-626-7594 University of MN Fax 612-625-5368 12 Shepherd Labs 100 Union Street S.E. Minneapolis, MN 55455 http://resolution.umn.edu ********************************************************************
I did some looking into costs/hr and finally came to the conclusion that if I made an assumption that well funded institutions with shared facilities had set prices for instrument use, it probably reflected what the market - i.e. the granting agencies - would bear. What I discovered was prices that varied but stayed in the range of $25 to $35. $29 seemed to be popular, though I wondered why not the infinitely more popular prince of $29.95.
Fred Monson
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
} ---------- } From: "JHoffpa464-at-aol.com"-at-sparc5.microscopy.com } Sent: Thursday, June 7, 2001 8:03 AM } To: microscopy-at-sparc5.microscopy.com } Subject: EM COSTING } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 in the process of cost our EM service, that is a cost breakdown } for } doing EM. i know the actual costs are very small. i was wondering if } anyone } has done it recently. we are a diagnostic lab. } john hoffpauir } cooper hospital } }
A little more insight on Xray absorption of materials: Twenty years ago when I was doing EDS XRF, I did a lot of work on Xray filters on the primary Xray beam from Xray tubes. I designed both edge filters (the absorption edges of each element can be used to strongly remove X-rays above certain energies} and "white" filters to remove all Xrays below certain energies. My white filters were usually layers of filter paper (no Ti pigment). My experience was that at below 30KV on the Xray tube almost nothing would get through 1/2 inch of organic materials. Above 30 KeV we had worry about X-ray leakage. The Xrays tubes (and Electron microscopes) give off both primary lines and bremsstrahlung xrays. The Bremsstrahlung peak intensity energy in KeV by rule of thumb is about 2/3 the energy of the electron beam. You may observe in your SEM by EDS the X-ray energy spectrum. It is that background spectrum. Since I had a spectrometer I could ready see what energies passed or started leaking through my filters. You too can do the Xray absorption spectrum test inside your chamber by placing the stopper or other material in front of your EDS detector window and see what comes through. The EDS detector is much more sensitive than a Geiger counter or film.
Therefore, if the SEM is used below 30 kV, then a thick rubber stopper will absorb the X-rays. Still worried? Stick a radiation badge on the outside of the stopper to check.
Ronald Vane XEI Scientific
-----Original Message----- } From: Ken Converse {qualityimages-at-netrax.net} To: Arthur Day {ard-at-ansto.gov.au} ; MSA, listserver {Microscopy-at-sparc5.microscopy.com}
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Tina Carvalho wrote:
} Hi, All- } } Does anyone have a reference for or some transmission electron micrographs } that illustrate the effects of osmolarity of buffers on animal cells? I } would like to show students on tour of our facility shrinkage and swelling } of cells and organelles. I have other wonderful artifacts to show, } including one of my first sections with chatter so bad it looks like } mini-blinds... } } Mahalo! } Tina
Arborgh et al. J. Ultrastruc. Res. 56:339-350, 1976. A classic study.
} } } 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* } ****************************************************************************
-- ********************************************** 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 **********************************************
You can try JB-4 embedding media instead of LR White. It works pretty well for thick sectioning and maitains excellent structure preservation.
Soumitra
************************************************************* Soumitra Ghoshroy Ph.D. Director, Electron Microscopy Lab Graduate Faculty, Department of Biology Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3600 Fax: 505-646-5665 e-mail:sghoshro-at-nmsu.edu URL:http://confocal.nmsu.edu/eml
On Fri, 8 Jun 2001, 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 } -----------------------------------------------------------------------. } } } Tom, } } You might have nailed it. These were osmicated tissues, since they weren't } intended for immuno. Our client prefers LR White for making thick sections } for light microscopy, so these specimens were prepared with osmium and UA } post-fixations. Given the large size and difficult nature of insect parts, } it's very possible that something remained in the tissue despite several } lengthy washes. At least we have back-up specimens! } } Thanks for the response. At least it's something to work with. } } Randy } } -----Original Message----- } } From: Tom Phillips [mailto:PhillipsT-at-missouri.edu] } Sent: Friday, June 08, 2001 9:24 AM } To: Tindall, Randy D. } Cc: Microscopy-at-msa.microscopy.com } Subject: Re: TEM: LR White weirdness } } } Randy: This has happened a bunch of times to me. It only happens } with osmicated tissues. I have run up the same tisse +/- osmication } and premature polymerization only occurs in the osmicated ones - not } every time or at the same rate. I guess there is something that } doesn't get rinsed out that can trigger it - perhaps more so when the } LRW is aging. I am starting a LRW infiltration with osmicated tissue } using a brand new bottle of LRW and I will let you know what happens. } my tissues were in glass vials so it isn't the plastic tubes. it has } happened at both 4 C and room temp. i have a vague recollection } posting this on the microscopy listserver and not getting much of a } response. maddening problem, isn't it? } } } ------------------------------------------------------------. } } } } } } Dear Listers, } } } } While infiltrating a set of specimens (insect abdomens and thoraxes) in } LR } } White, something occurred that has so far defied my attempts to figure out. } } I was doing } } side by side microwave and conventional fixations, dehydrations, and } } embeddings. The microwave specimens were dehydrated in acetone, behaved } } normally, and we } } have polymerized blocks. However I dehydrated the conventional set in } an } } ethanol series, as we have done many times before in LR White. The first } } infiltration step } } was 2 parts ethanol to one part LR White Medium Grade at room temp, and } } upon going later to change into 1:1, I found that the resin/ETOH mix had } } partially } } polymerized into a spongy, granular mass. Uh-oh, bad resin, I said, } along } } with a few other choice words. } } } } After the initial panic, I ended up putting the chunks into pure LR } White } } from a newly opened bottle and the partially polymerized resin seemed to go } } back into solution. I } } ran them through a couple more changes of pure resin, then left them on } a } } rocker overnight at room temperature. Checking them this morning, I found } } two of the } } samples were fine, and the third had polymerized into a rubbery mass. } } Same bottle of resin, same identically processed samples, same everything. } } } } An additional tube with a sample of the "bad" resin was also happily } } unpolymerized, as was the remainder of the "bad" resin in the bottle, which } } I had left in the fume } } hood overnight at room temperature. The "bad" resin was slightly more } } than a year old and has been refrigerated at 4 C since we got it. The } } second resin is about 10 } } months old and has also been constantly refrigerated. Neither had any } } accelerator in them (at least none added by us). } } } } I have my share of problems with LR White, but this one really has us } } puzzled. Could there have been something in the samples that triggered a } } polymerization? Can } } LR White react with some plastics in this way? (We were using a } } relatively new batch of Eppendorf tubes that seem more hydrophobic than our } } previous ones.) Could } } it have been the full moon? } } } } Regards, } } 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 } } http://biotech.missouri.edu/emc } } } } -- } 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) } }
At 12:55 PM 6/8/01 -0400, you wrote: } ------------------------------------------------------------------------ } 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 am considering acquiring a cryostat for the laboratory. Does anyone have an experience, positive or negative, with the Bright OTF cryostat? Please respond to me offline.
I am in need of a reference EELS spectum of ZnO (or probably any other Zn[+2] compound ) to help me interpret some data. I've already checked the CEMES EELS database to no avail, and I've done some web-searching with lots of hits but mostly to conference abstracts but no hard data. I'm attempting to distinguish between Zn metal, for which I have a reference EELS spectrum, and Zn[+2}.
Tia, Mike 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
You are right in the comparison of SEMs and TEMs. The accelerating voltages typically used in TEMs creates a much greater concern as do the glass viewing ports directly open to the electron beam - sample interactions. In fact, the majority of problems in TEMs have been inadequate lead content of the glass in those ports. SEMs are inherently safe from external emissions in that their design results in a generous metallic shielding surrounding all portions that can generate x-rays.
However, even in SEMs, there can be exceptions. Remember the small aluminum plug in the columns of ETEC SEMs, the one opposite where the scan coil cables enter? If that plug is not in place, then there is a potential for external radiation. While the designs may be inherently safe, there is still room for error.
Many states require a regular scan for external radiation on any x-ray generating equipment. While I have heard of quite a few measurable responses on TEMs (including one just two days ago at a DOE site), I have never heard of any detectable radiation from an SEM.
Regarding computer monitors, don't worry. While early TV sets did produce considerable x-rays externally, modern TVs and computer monitors really don't. Unless you plan on spending 24 hours a day hugging the back side of a monitor, I wouldn't worry.
Finally, regarding the temporary use of a rubber stopper - It shouldn't pose a problem. Most of the ports on an SEM, and particularly those for EDS systems, are on the rear of the chamber. SEMs, given their usual operating conditions and limitations, are not prodigious x-ray producers. Most would be absorbed by the thickness of a stopper capable of withstanding the vacuum pressures, and what little could escape would be directed away from anyone using the instrument. Since any resulting radiation would be attenuated by the square of the distance from the source (in this case the x-rays penetrating the stopper) there would normally be no detectable exposure except, perhaps, at the surface of the stopper.
I do have a problem with permanent plastic viewports, however. These are generally placed on the sides of the sample chamber where a more direct path to the operator is possible. Whenever I find a customer with a plastic viewport, I always strongly suggest that they keep a suitable cover over it when they aren't being used to view the sample presentation. Surprisingly thin ports can be made of materials like polycarbonate that are capable of withstanding atmospheric pressures.
The real questions should be, if this is a situation that may occur from time to time, should a proper port cover be fabricated and kept on hand so that there are no such questions, now or in the future?
On Friday, June 08, 2001 9:16 AM, Ken Converse [SMTP:qualityimages-at-netrax.net] wrote: } } Pehaps someone could provide a better analysis, but I have a couple of } anecdotes: } } I had a Canadian customer (20+ years ago) who specialized in very long } EDS x-ray sweeps in either dot map form or a combined YZ modulated } form. As I recall, some of these scans were up to 16 hours long. He } had a Plexiglass port cover so that he could easily see his chamber } geometry. This had a metal cover that was velvet lined for light } leakage. Once he put his radiation badge inside the metal cover for a } month, then turned it in. There apparently was nothing out of the } ordinary, as his rad peo;le never said "boo". Perhaps it's a commentary } on the rad people, but a full month exposure? } } Second, I had another customer get me some rad figures on an SEM gun } operating at 30kV and 150uA (1.5x10E-4). The bottom line was that the } x-ray output from the gun was very serious, but all the guns are } enclosed in steel. No problem. At the specimen for EDS work we're } usually talking about 200-600pA (2-6x10E-10), or roughly 6 orders of } magnitude less current and the stopper is going to absorb some of the } x-rays generated. Is this really a problem? } } I'm not talking about TEMs, as they can be very dangerous. The current } at the screen is much higher, the excitation voltage is much higher and } there is this large expanse of glass that MUST be leaded. Perhaps the } currents used for WDS would also present a problem (10-100 nA or 10E-8 } to 10E-7) at 2 to 3 orders of magnitude more than EDS. } } Don't forget, those nice color displays on the four computers } surrounding you in your lab also generate x-rays and their beam currents } are in the mA range (10E-3) and an accelerating voltage of about 25kV. } There may be a decelerating grid and leaded glass on the front, but I } think you will find the the shielding on the rear of the monitor is not } so rigorous, Your dentist's x-rays are about 70kV and only one order of } magnitude greater current. } } I'd love some feed-back from someone who knows radiation because I've } felt that applying TEM rules to SEMs is gross over-kill, especially with } so many color monitors around. How often do you have your decelerating } grid checked for proper operation? If it doesn't operate correctly, } your exposure could be very dangerous, given the time that one sits in } front of these things and their distance from your face. } } If I'm way off base, I'd like to know and know why. } } Ken Converse } owner } Quality Images } third party SEM service } Delta, PA } } Arthur Day wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 } } } -----------------------------------------------------------------------. } } } } } } } } } Just out of curiousity. . . how much X-radiation penetrates the rubber } } } stopper? } } } } } } Marie } } } } } } } Doesn't anybody supply X-ray proof rubber bungs for microscope ports? } } } } No worries. Just wrap a few sheets of lead foil around the column and } } make sure pregnant operators stay below 5kV ;-( } } } } Surely something somewhat less desperate could be cobbled together out } } of metal without too much extra work? } } } } } } } } } }
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
} I have in my possession the following [Old Microscope Catalogs]: } I also have a number of old Leitz "Technical Information Bulletins" that } will be listed later. } Hope there is a use. } Fred -
If you don't get takers for all of these, there's a microscopy book dealer in England who can probably find them a home: http://www.savonabooks.free-online.co.uk It would be a pity to just trash them...
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 know someone makeing a CDROM of Leitz information that is not copyrighted. It is going to be a very reasonably priced commerical project.
I would post his name but I am in the middle of changeing computers and I don't have his email address at hand.
Gordon Couger
----- Original Message ----- } From: "Caroline Schooley" {schooley-at-mcn.org} To: "Monson, Frederick C." {fmonson-at-wcupa.edu} Cc: {Microscopy-at-sparc5.microscopy.com} Sent: Saturday, June 09, 2001 10:32 AM
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The most important advertising dollar being spent should be for search engine TRAFFIC.
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You Can't Get Your Companies Web Site Indexed by the Search Engines?
Unfortunately, this is all too common of a Problem. You're not the only one frustrated about the length of time it takes to be listed, or all the pitfalls involved. It takes anywhere from 2 days to as much as 6 months to be listed on all the search engines. Waiting several weeks to months can be extremely frustrating.
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Engines can at any time delay their indexing for maintenance and many other reasons.
So do you feel lucky? Do YOU?
Lucky if you submit A PAGE just days before an engine does a complete refresh of their many indexes/databases.
WE Know exactly how each search engine works, and we know when to submit and what to submit. Search engines are changing Daily and we study them each day. Your competitors ARE -at- the mercy of OUR Marketing Departments. 6 Plus Years Now in the search engine wars, We are Masters of words like: MP3 - BOOKS - WEB SITE HOSTING - MARKETING - - FREE WEB SITES - CASINO - - CASINO REVIEWS - BALLS - - LOGOS - ART - ATTORNEY'S - NEW CAR PARTS - OLD CAR PART - - NETWORK MARKETING - WATER FILTERS - - SCALES - AND THE LIST GOES ON - -
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Contact:
THE CYBER TRAFFIC TECHNICIANS.
You need is us in your corner, we will take control of the submission cycle of your domain/s. We Provide you with: a real report that shows you what's been done and when completed, we offer free rank reporting.
The cost is 79.00 US Dollars month to month - 69.00 US Dollars a month when paying for 3 months at a time - 59.00 US Dollars a month when paying for 12 months. This Price is good for the next 5 orders only. DON'T delay in ordering.
If You fax a check, there is no need for you to mail the original. We will draft a new check, with the exact information from your original check. All checks will be held for bank clearance. (7-10 days) Make payable to: "CYBERCONTROL"
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-------------------------------------------------------------------------- --------------------------------------- To be removed from future mailings!!!! All REMOVE requests AUTOMATICALLY honored upon receipt. href="mailto: notraffic1-at-excite.com?subject=NOBIZ"mailto: notraffic1-at-excite.com?subject=NOBIZ PLEASE understand that any effort to disrupt, close or block this REMOVE account can only result in difficulties for others wanting to be removed from our mailing list as it will be impossible to take anyone off the list if the remove instruction is not received.
Unwanted polymerisation of LR White resin during infiltration is an occasional problem with the resin. I have never experienced it with fresh batches, indeed the only times I have had this problem was with batches over 1 year old (the recommended use by time). An possible explanation was suggested to me by Roy Gillett of London Resin a number of years ago:
Quote "The reason this pre-polymerisation occurs only with tissue must be something to do with a tissue constituent catalysing polymerisation. Older resin is much more susceptibe to this that fresh monomer becaue of the significant polymer growth that will inevitably have occurred in the monomer. The most likely 'endogenousd catalyst' from previous experience is likely to be an amine or peroxide moiety in the tissue"
We have used LR White resin extensively for a long time - perhaps the reason we don't have a problem is that our resin never gets anything like a year old before we use it/
Best Wishes
Ian
} } Dear Listers, } } While infiltrating a set of specimens (insect abdomens and thoraxes) in LR } White, something occurred that has so far defied my attempts to figure out. } I was doing } side by side microwave and conventional fixations, dehydrations, and } embeddings. The microwave specimens were dehydrated in acetone, behaved } normally, and we } have polymerized blocks. However I dehydrated the conventional set in an } ethanol series, as we have done many times before in LR White. The first } infiltration step } was 2 parts ethanol to one part LR White Medium Grade at room temp, and } upon going later to change into 1:1, I found that the resin/ETOH mix had } partially } polymerized into a spongy, granular mass. Uh-oh, bad resin, I said, along } with a few other choice words. } } After the initial panic, I ended up putting the chunks into pure LR White } from a newly opened bottle and the partially polymerized resin seemed to go } back into solution. I } ran them through a couple more changes of pure resin, then left them on a } rocker overnight at room temperature. Checking them this morning, I found } two of the } samples were fine, and the third had polymerized into a rubbery mass. } Same bottle of resin, same identically processed samples, same everything. } } An additional tube with a sample of the "bad" resin was also happily } unpolymerized, as was the remainder of the "bad" resin in the bottle, which } I had left in the fume } hood overnight at room temperature. The "bad" resin was slightly more } than a year old and has been refrigerated at 4 C since we got it. The } second resin is about 10 } months old and has also been constantly refrigerated. Neither had any } accelerator in them (at least none added by us). } } I have my share of problems with LR White, but this one really has us } puzzled. Could there have been something in the samples that triggered a } polymerization? Can } LR White react with some plastics in this way? (We were using a } relatively new batch of Eppendorf tubes that seem more hydrophobic than our } previous ones.) Could } it have been the full moon? } } Regards, } 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 } http://biotech.missouri.edu/emc } }
Ian Hallett HortResearch Mt Albert Research Centre Private Bag 92 169 Auckland, New Zealand Fax 64-9-815 4201 Telephone 64-9-815 4200 EMail ihallett-at-hortresearch.co.nz
______________________________________________________ The contents of this e-mail are privileged and/or confidential to the named recipient and are not to be used by any other person and/or organisation. If you have received this e-mail in error, please notify the sender and delete all material pertaining to this e-mail. ______________________________________________________
SEARCH ENGINES DELIVER TARGETED UNIQUE TRAFFIC TO WEB SITE
If you don't find what you're looking for in the first 10 to 40 matches ON A SEARCH ENGINE, what do you do? If you are like most people, you simply go to the next search engine and try again.
If your site is not coming up in the TOP 40 matches On a search engine, you're losing MONEY!
IT 'S THAT SIMPLE!
Improving search engine TRAFFIC means:
MORE HITS MORE BUSINESS MORE SUCCESS
The most important advertising dollar being spent should be for search engine TRAFFIC.
1. Approximately 95% of all Internet users start with a search engine query.
2. Anyone who comes to your site from a major search engine is 100 times more likely to become a customer because they were specifically looking for your product, goods or services.
3. Search engine traffic gives you substantially more for your advertising dollar than banners. Moreover Banners are done on impressions. An impression is just someone that went to a web page with your Banner ad on the page. It does not mean your Ad was seen.
You Can't Get Your Companies Web Site Indexed by the Search Engines?
Unfortunately, this is all too common of a Problem. You're not the only one frustrated about the length of time it takes to be listed, or all the pitfalls involved. It takes anywhere from 2 days to as much as 6 months to be listed on all the search engines. Waiting several weeks to months can be extremely frustrating.
WHEN TO DO YOUR SUBMISSION?
Engines can at any time delay their indexing for maintenance and many other reasons.
So do you feel lucky? Do YOU?
Lucky if you submit A PAGE just days before an engine does a complete refresh of their many indexes/databases.
WE Know exactly how each search engine works, and we know when to submit and what to submit. Search engines are changing Daily and we study them each day. Your competitors ARE -at- the mercy of OUR Marketing Departments. 6 Plus Years Now in the search engine wars, We are Masters of words like: MP3 - BOOKS - WEB SITE HOSTING - MARKETING - - FREE WEB SITES - CASINO - - CASINO REVIEWS - BALLS - - LOGOS - ART - ATTORNEY'S - NEW CAR PARTS - OLD CAR PART - - NETWORK MARKETING - WATER FILTERS - - SCALES - AND THE LIST GOES ON - -
If you've submitted your site and come to find no listing, what do you do now?
Contact:
THE CYBER TRAFFIC TECHNICIANS.
You need is us in your corner, we will take control of the submission cycle of your domain/s. We Provide you with: a real report that shows you what's been done and when completed, we offer free rank reporting.
The cost is 79.00 US Dollars month to month - 69.00 US Dollars a month when paying for 3 months at a time - 59.00 US Dollars a month when paying for 12 months. This Price is good for the next 5 orders only. DON'T delay in ordering.
If You fax a check, there is no need for you to mail the original. We will draft a new check, with the exact information from your original check. All checks will be held for bank clearance. (7-10 days) Make payable to: "CYBERCONTROL"
Address:
State: City: Zip:
Contact Name:
Contact Telephone w/best time to call:
Contact Fax:
Contact E-mail Address:
Web site Address:
__ Yes, I want you to start the one time domain submission for 19.95 US Dollars!
__ Yes, I want to start the monthly submission program And my info is completely filled out! I want to pay for: You must check one
___ONE MONTH -79.00 US Dollars
___3 MONTH - 207.00 US Dollars
___12 MONTH 708.00 US Dollars
A few of your top Keywords:
Questions:
-------------------------------------------------------------------------- --------------------------------------- To be removed from future mailings!!!! All REMOVE requests AUTOMATICALLY honored upon receipt. href="mailto: notraffic1-at-excite.com?subject=NOBIZ"mailto: notraffic1-at-excite.com?subject=NOBIZ PLEASE understand that any effort to disrupt, close or block this REMOVE account can only result in difficulties for others wanting to be removed from our mailing list as it will be impossible to take anyone off the list if the remove instruction is not received.
I have a question about measuring small cellular structures at the light microscope level using Argus 10 enhancement and measurement software.
I am measuring vesicle diameters in living cells of the fungus Neurospora. They measure about 350-400 nm in diameter using the Argus 10 software. With cryofixed TEMs of the same cell type, the only vesicles with the same distribution as at the LM level measure 150 nm diameter and there are no structures at 350-400 nm.
All my calibrations of both the LM and TEM have been checked and double-checked.
It is my understanding that video-enhanced LM can amplify signal (or sizes) so that structures below limit of LM resolution can be detected (eg, imaging of individual microtubules with DIC/VELM). I am wondering if this is the source of this discrepancy that it see and if there are references that describe this.
Thanks so much,,,,,, Robby Roberson
*************************************** Robert W. Roberson, PhD Department of Plant Biology Molecular and Cellular Biology Program Arizona State University Tempe, AZ 85282 Office/Lab 480-965-8618
At 7:53 AM -0700 6/8/01, Eric wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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
We have notice a strange effect of colored Microcentrifuge tubes. The colored tubes will cause hardening of the LR White at room temperature. Something in the coloring dyein the tubes must be acting as a catalyst.
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: IAN HALLETT [mailto:ihallett-at-hortresearch.co.nz] Sent: Sunday, June 10, 2001 3:35 PM To: microscopy-at-sparc5.microscopy.com
Randy and others:
Unwanted polymerisation of LR White resin during infiltration is an occasional problem with the resin. I have never experienced it with fresh batches, indeed the only times I have had this problem was with batches over 1 year old (the recommended use by time). An possible explanation was suggested to me by Roy Gillett of London Resin a number of years ago:
Quote "The reason this pre-polymerisation occurs only with tissue must be something to do with a tissue constituent catalysing polymerisation. Older resin is much more susceptibe to this that fresh monomer becaue of the significant polymer growth that will inevitably have occurred in the monomer. The most likely 'endogenousd catalyst' from previous experience is likely to be an amine or peroxide moiety in the tissue"
We have used LR White resin extensively for a long time - perhaps the reason we don't have a problem is that our resin never gets anything like a year old before we use it/
Best Wishes
Ian
} } Dear Listers, } } While infiltrating a set of specimens (insect abdomens and thoraxes) in LR } White, something occurred that has so far defied my attempts to figure out. } I was doing } side by side microwave and conventional fixations, dehydrations, and } embeddings. The microwave specimens were dehydrated in acetone, behaved } normally, and we } have polymerized blocks. However I dehydrated the conventional set in an } ethanol series, as we have done many times before in LR White. The first } infiltration step } was 2 parts ethanol to one part LR White Medium Grade at room temp, and } upon going later to change into 1:1, I found that the resin/ETOH mix had } partially } polymerized into a spongy, granular mass. Uh-oh, bad resin, I said, along } with a few other choice words. } } After the initial panic, I ended up putting the chunks into pure LR White } from a newly opened bottle and the partially polymerized resin seemed to go } back into solution. I } ran them through a couple more changes of pure resin, then left them on a } rocker overnight at room temperature. Checking them this morning, I found } two of the } samples were fine, and the third had polymerized into a rubbery mass. } Same bottle of resin, same identically processed samples, same everything. } } An additional tube with a sample of the "bad" resin was also happily } unpolymerized, as was the remainder of the "bad" resin in the bottle, which } I had left in the fume } hood overnight at room temperature. The "bad" resin was slightly more } than a year old and has been refrigerated at 4 C since we got it. The } second resin is about 10 } months old and has also been constantly refrigerated. Neither had any } accelerator in them (at least none added by us). } } I have my share of problems with LR White, but this one really has us } puzzled. Could there have been something in the samples that triggered a } polymerization? Can } LR White react with some plastics in this way? (We were using a } relatively new batch of Eppendorf tubes that seem more hydrophobic than our } previous ones.) Could } it have been the full moon? } } Regards, } 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 } http://biotech.missouri.edu/emc } }
Ian Hallett HortResearch Mt Albert Research Centre Private Bag 92 169 Auckland, New Zealand Fax 64-9-815 4201 Telephone 64-9-815 4200 EMail ihallett-at-hortresearch.co.nz
______________________________________________________ The contents of this e-mail are privileged and/or confidential to the named recipient and are not to be used by any other person and/or organisation. If you have received this e-mail in error, please notify the sender and delete all material pertaining to this e-mail. ______________________________________________________
I would like to be of some assistance on the subject of a solution to:-'There is no simple solution to the preparation of very hard and very soft materials in one specimen.'
As our Bright/Hacker OTF/AS/D/LT cryostat is able to section a hard and soft specimen incorporated into one specimen. If this is of interest I would be pleased to receive sample and return our results to those who have a problem.
Best Regards
Alan Bright
Bright Instrument Co.Ltd. St Margaret's Way Huntingdon Cambridgeshire PE29 6EU 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: Garber, Charles A. [mailto:cgarber-at-2spi.com] Sent: 07 June 2001 23:26 To: MICROSCOPY BB
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Jim Darley wrote:
==================================================== Tantalum is very hard and tough, so I doubt that it will section with either
diamond or tungsten carbide knives. Preparing the material like a geological
section by grinding is a possibility, but not a good one. Chances are that the grinding material would fill the voids and the tissue would be ground away first (although ProSciTech and others supply diamond grit embedded in plastic disks, which are much better in that regard) Proper vacuum infiltration with a low viscosity resin, cutting with a diamond blade, grinding with a diamond disc would be my preparation trial. After that either stain the surface and try your luck with a high power reflection (metallurgical) microscope, or more likely digest the plastic and
view the specimen in SEM. If the tantalum could be fractured open, an ESEM my offer some "insights". There is no simple solution to the preparation of very hard and very soft materials in one specimen. ===================================================== Actually Ta **can** be thin sectioned if it is done by someone with the proper experience and who is using the right kind of diamond knife. We have offered our SPI #02888-AB sectioned Ta foil on a grid for AEM studies for some number of years; see URL http://www.2spi.com/catalog/standards/aem.html
The Ta system that prompted the original posting was thicker, however, it is also porous, and if properly infiltrated with resin, and assuming the porosity is above some point, in principle, at least, there is no reason why it could not be thin sectioned for TEM. No method is really artifact free, but some artifacts are more easy to recognize than others. Knife induced artifacts are anisotripic (e.g. directional) in nature and can be more easily recognized (as artifacts) than artifacts caused by say, ion milling, which are isotropic in nature.
When I say "right kind of diamond knife", I am not suggesting that the SPI diamond knife could do something above and beyond what other diamond knives could do. What I am saying is that there is a process of selection of the optimum knife angle, because one might have to be prepared to use a knife with a fairly low (for materials science work) knife angle, rather than a more blunt angle, but with the downside is that it will wear out more quickly. That might make for great business for a diamond knife supplier, but it does tend to get expensive for the user who is not so experienced with this kind of sectioning.
Disclaimer: SPI Supplies offers diamond knives, both materials and life science, and we have done this kind of sectioning, as a service, for commercial clients for over thirty years.
Chuck
PS: Please remember that we are nearly 100% paperless and we would ask that any reply to this message be by way of the "reply" feature on your software, so that the entire string of correspondence can come back to us and all be in one place.
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 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 ############################ ==================================================
Hi, The optics of forming an image is constrained by diffraction so that when the light comes together in a point it actually has a finite breadth. This is given by the numerical aperture of your objective and condenser (and the wavelength of light) and has a theoretical min of around 250 nm for green light. If you are using a high NA lens (say 1.3) and an equally good condenser, which must be oiled to the slide to get its full resolution, then you should be able to reach this limit. To the extent that your lenses are not that high NA, the limit will be larger. Vesicles smaller than the limit may be imaged (indeed, single microtubules can be imaged) but the diameter will read out on the image as being whatever your diffraction limit is. You can find a good discussion of this in Inoué's book on Video Microscopy, to name just one source.
As ever, Tobias
} } Hello all, } } I have a question about measuring small cellular structures at the light } microscope level using Argus 10 enhancement and measurement software. } } I am measuring vesicle diameters in living cells of the fungus Neurospora. } They measure about 350-400 nm in diameter using the Argus 10 software. } With cryofixed TEMs of the same cell type, the only vesicles with the same } distribution as at the LM level measure 150 nm diameter and there are no } structures at 350-400 nm. } } All my calibrations of both the LM and TEM have been checked and } double-checked. } } It is my understanding that video-enhanced LM can amplify signal (or sizes) } so that structures below limit of LM resolution can be detected (eg, } imaging of individual microtubules with DIC/VELM). I am wondering if this } is the source of this discrepancy that it see and if there are references } that describe this. } } Thanks so much,,,,,, } Robby Roberson } } } *************************************** } Robert W. Roberson, PhD } Department of Plant Biology } Molecular and Cellular Biology Program } Arizona State University } Tempe, AZ 85282 } Office/Lab 480-965-8618
Frederick C. Monson, PhD West Chester University of Pennsylvania Center for Advanced Scientific Imaging (CASI) Schmucker II Science Center (Room: SS024(Basement)) South Church Street West Chester, PA, 19383 MailDrop: Department of Geology/Astronomy Phone: 610-738-0437 Fax: 610-436-3036 email: fmonson-at-wcupa.edu Please call before visiting.
} ---------- } From: Alan J. Kruger } Sent: Friday, June 8, 2001 1:05 PM } To: Microscopy } Subject: LM: Formula for calculating dept of focus } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 lead me to a reference or the formula to calculate dept of } focus on light microscopes? } } Al Kruger } USDA Meat Animal Research Center } } }
A few days ago someone asked about sources for used and renovated microscopy equipment. Just today I received a leaflet from Microscopy Laboratories, (P.O. Box 338, Red Bank, NJ 07701, email:micro-at-mail.superlink.net; Tel: 732-747-6228) that lists a wide variety of items of reconditioned and guaranteed equipment they are offering for sale. -- Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan 3062 Dow Bldg.; 2300 Hayward St. Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
Dear All, I have been contacted by someone wishing urgently to obtain an image of skin with psoriasis for a science exhibition aimed at children. They specified an SEM image but possibly TEM or LM would do. We have nothing along these lines in our lab - is anyone out there in the wide world of microscopy able and willing to provide such an image?
Best regards,
Richard
Richard Easingwood South Campus Electron Microscope Unit School of Medical Sciences, University of Otago PO Box 913, Dunedin NEW ZEALAND
} Can anyone lead me to a reference or the formula to calculate dept of } focus on light microscopes?
Al,
"Photomicrography. A comprehensive treatise" by Roger P. Loveland John Wiley & Sons, 1970, contains an appendix on calculating depth of field. For compound microscopes Loveland gives several formulae, which produce similar but not identical results in his worked examples.
I can fax it to you if you can't find a copy locally.
Regards
Stephen Edgar
Pathology Department Faculty of Medicine & Health Sciences University of Auckland Private Bag 92019 Auckland New Zealand
This thread may never finish. Fred Monson wrote to me making the point that he thought that the original request (which I have appended last) related to a carbon sponge coated with Ta. Fred also had and so he appended an article in pdf format, which deals with such material. There also is what appears to be an SEM image of a thick section of this very open material. It seems that it had been sectioned, presumably with a diamond or TC knife. It does however, not show any cell inclusions, though the article says that purpose of the matrix is the growths of T-cells. Plastic infiltrated Ta coated C sponge with cell inclusions "may" section. The original correspondence referred to a Ta sponge coated with carbon with cell inclusions and that would be a whole lot more difficult. The publication is: Contact: Mark J. Pykett, Cytomatrix, (781) 939-0995, mpykett-at-cytomatrix.com I think that we could have a delightful time with a hundred protagonist writing to the unsuspecting "Contact" 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, June 12, 2001 12:38 AM, Alan Bright [SMTP:bright-at-dial.pipex.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 would like to be of some assistance on the subject of a solution } to:-'There is no simple solution to the preparation of very hard and very } soft materials in one specimen.' } } As our Bright/Hacker OTF/AS/D/LT cryostat is able to section a hard and soft } specimen incorporated into one specimen. If this is of interest I would be } pleased to receive sample and return our results to those who have a } problem. } } Best Regards } } Alan Bright } } Bright Instrument Co.Ltd. } St Margaret's Way } Huntingdon } Cambridgeshire } PE29 6EU } 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: Garber, Charles A. [mailto:cgarber-at-2spi.com] } Sent: 07 June 2001 23:26 } To: MICROSCOPY BB } Subject: Sectioning Ta } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 } -----------------------------------------------------------------------. } } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } Jim Darley wrote: } } ==================================================== } Tantalum is very hard and tough, so I doubt that it will section with either } } diamond or tungsten carbide knives. Preparing the material like a geological } } section by grinding is a possibility, but not a good one. Chances are that } the } grinding material would fill the voids and the tissue would be ground away } first (although ProSciTech and others supply diamond grit embedded in } plastic } disks, which are much better in that regard) } Proper vacuum infiltration with a low viscosity resin, cutting with a } diamond } blade, grinding with a diamond disc would be my preparation trial. } After that either stain the surface and try your luck with a high power } reflection (metallurgical) microscope, or more likely digest the plastic and } } view the specimen in SEM. } If the tantalum could be fractured open, an ESEM my offer some "insights". } There is no simple solution to the preparation of very hard and very soft } materials in one specimen. } ===================================================== } Actually Ta **can** be thin sectioned if it is done by someone with the } proper experience and who is using the right kind of diamond knife. We have } offered our SPI #02888-AB sectioned Ta foil on a grid for AEM studies for } some number of years; see URL } http://www.2spi.com/catalog/standards/aem.html } } The Ta system that prompted the original posting was thicker, however, it is } also porous, and if properly infiltrated with resin, and assuming the } porosity is above some point, in principle, at least, there is no reason why } it could not be thin sectioned for TEM. No method is really artifact free, } but some artifacts are more easy to recognize than others. Knife induced } artifacts are anisotripic (e.g. directional) in nature and can be more } easily recognized (as artifacts) than artifacts caused by say, ion milling, } which are isotropic in nature. } } When I say "right kind of diamond knife", I am not suggesting that the SPI } diamond knife could do something above and beyond what other diamond knives } could do. What I am saying is that there is a process of selection of the } optimum knife angle, because one might have to be prepared to use a knife } with a fairly low (for materials science work) knife angle, rather than a } more blunt angle, but with the downside is that it will wear out more } quickly. That might make for great business for a diamond knife supplier, } but it does tend to get expensive for the user who is not so experienced } with this kind of sectioning. } } Disclaimer: SPI Supplies offers diamond knives, both materials and life } science, and we have done this kind of sectioning, as a service, for } commercial clients for over thirty years. } } Chuck } original request was posted on 6 June 2001
I am posting this for a college, Lauri Wyner, in the Pathology Core. I have no idea how to section this stuff...!
Thanks! Maria Ericsson
My question is: I have a carbon coated porous tantalum matrix approximately 1 cm in circumference and 2 mm thick. This matrix has fixed cells adhered to the surface and throughout. I would like to embed this, make slides and stain for H&E to confirm the presences of cells as well as immunohistochemistry to characterize them. I am looking for suggestions on how I can section this matrix while maintaining its overall structure. Any information would be greatly appreciated.
POSTDOCTORAL POSITIONS IN NANOTECHNOLOGY AND BIOPHYSICS AT THE DEPARTMENT OF PHYSICS, UCLA
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Current Research Areas: Carbon Nanotubes Molecular biophysics of DNA and proteins Structure and function of macromolecular assemblies
The compensation package is competitive, and medical benefits are included. Interested person should send cv and three letters of recommendation to: Professor Ching-Hwa Kiang Department of Physics & Astronomy 6-130 Knudsen Hall University of California Los Angeles, CA 90095-1547 chkiang-at-physics.ucla.edu
----------------------------------------------------------------- Dr. Ching-Hwa Kiang 6-130 Knudsen Department of Physics Phone: (310) 206-0563 University of California Fax: (310) 825-5734 Los Angeles, CA 90095-1547 chkiang-at-physics.ucla.edu http://www.physics.ucla.edu/people/faculty_members/kiang -----------------------------------------------------------------
A fellow here in the department has the interest in knowing by which mean an FEG W-crystal needle is cut. I only know that in the case of Schottky emitter, the submicron needle is cut in {100} direction and coated with ZrO2 to reduce the work function. But I have no knowledge in the manufacturing process of the tip (FIB?). Anybody who knows please shed some lights. Thanks in advance.
********************************* Chaoying Ni Electron Microscopy Laboratory Materials Science and Engineering College of Engineering University of Delaware Newark, DE 19716 *********************************
I have a question as to technique or method regarding the quantification of Au/Sn in a "solder bump" application for an InP microcircuit. Essentially what I've been asked to do is provide the weight % of each element after this "solder" has been melted and re-flowed. The samples are cylindrical in shape with an x-section of 65 uM and a thickness of 34 uM. The Au/Sn portion sits atop this structure and is a thickness of ~ 2 uM.
My question is, how can I quantify, in Wt. %, the ratios of the two metals? I have EDX at my disposal but I am uncertain that I will be able to accurately quantify this alloy without a standard. One other issue is that the underlying metallization that primarily constructs this "bump" is also Au for the next 32 uM.
Any insight into this problem would be greatly appreciated by myself and my colleagues.
Peter Tomic Analytical Services Group Anadigics, Inc. 35 Technology Drive Warren, New Jersey U.S.A. 07090
An excellent Philips CM-10 transmission electron microscope is being offered for sale by Michigan State University. It is 1987 vintage and has been on service contract since new. We can demo this instrument at any time. Please contact Dr. Xudong Fan with questions, fanx-at-msu.edu, 517-353-4525.
Stanley L. Flegler, Acting Director Center for Advanced Microscopy
Chaoying Ni , I don't know about the current crop of FE cathodes, but in the past an electrolytic cell was set up with the W needle as part of the circuit. The needle was slowly dipped and removed several times from the NaOH electrolyte. Essentially, it was the same technique as was (and is) used to produce micomanipulator needles. I think the set-up was shown and described well in a Microscopy Today article within the past 12 months.
Ken Converse owner Quality Images Delta, PA
Chaoying Ni wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 lister, } } A fellow here in the department has the interest in knowing by which mean } an FEG W-crystal needle is cut. I only know that in the case of Schottky } emitter, the submicron needle is cut in {100} direction and coated with } ZrO2 to reduce the work function. But I have no knowledge in the } manufacturing process of the tip (FIB?). Anybody who knows please shed } some lights. Thanks in advance. } } ********************************* } Chaoying Ni } Electron Microscopy Laboratory } Materials Science and Engineering } College of Engineering } University of Delaware } Newark, DE 19716 } ********************************* } } } }
I was cleaning today and I have a complete ready-to-plug wehnelt assembly for a JEOL JSM-5200 SEM to give away. We inherited this piece and it is just a bit different from the unit in our 5400, so please only ask if you really have a 5200 as it probably won't fit other models. Claim goes to the first school or non-profit that requests it, otherwise others.
Contact me if interested. Be willing to pay postage. ($10???)
Dale Callaham +++++++++++++++++ Dale A. Callaham Senior Microscopist, Stuck Jar Lid Opener, Odd Job Bear Central Microscopy Facility Morrill 4 North, Room 1 639 North Pleasant Street The University of Massachusetts Amherst, MA 01003-9278 ------------------------- Phone 413-545-3751 FAX 413-545-3243 email dac-at-bio.umass.edu http://www.bio.umass.edu/microscopy
After more than 20 years of faithful service at IBM Research we will be retiring our Cameca SX50 electron microprobe. The machine has three wavelength dispersive detectors as well as one energy dispersive detector. The machine is operational except for the computer system, which is a PDP-11. The machine has been dormant, but under vacuum for roughly 3 years. If anyone is interested in purchasing the machine, please send your offers to :
Dr Andrew J. Kellock Ion Beams Lab Almaden Research Center (408) 927 2353 kellock-at-almaden.ibm.com
We have an old Zeiss Photomicroscope II at our lab, mainly in wery good condition, but there are electronic problems with the automatic camera system. There are some faults in the large electronic box placed in the microscope table. We have possibilities to repair the microscope here at the university, but unfortunately we have only been able to get a copy of the "Gesamtschaltplan zum Photomicroscope III" (the electronic diagram), but that is entirely different. In order to repair it we need the original electronic diagram (or a copy of it) for the Photomicroscope II. The Carl Zeiss Factory could not help us. If someone could send us a copy, we would highly appreciate it.
Sincerely:
Morten Motzfeldt Laane Professor of Biology (electron-microscopy) Biological Institute P.O.Box 1066, 0316 Blindern, Oslo, Norway
Dear Sir or Madame, I have been asked by my employer to investigate setting my laboratory up to perform what was called "hot-stage microscopy". I understand this to be the ability to observe the melting of (in this case) a polymer. I was hoping that your organization could point me in the right direction. Any information you could give me would be greatly appreciated.
I am looking for a colouring agent for oil (emollients) which needs to applied to human skin. If we apply the oil as such we do not have enough contrast for measurements. This colouring agent needs to be skin friendly, leave no stain on the skin and it must not change the viscosity of the oil.
Any information would be very helpful and appreciated.
Thanks,
Nancy Meijer
IMPORTANT NOTICE: This email may be confidential, may be legally privileged, and is for the intended recipient only. Access, disclosure, copying, distribution, or reliance on any of it by anyone else is prohibited and may be a criminal offence. Please delete if obtained in error and email confirmation to the sender.
Below is the result of your feedback form. It was submitted by (lilaeloise-at-cs.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, June 12, 2001 at 16:14:22 ---------------------------------------------------------------------------
Email: lilaeloise-at-cs.com Name: Lila Allen
Organization: Sunrise Hospital
Education: Graduate College
Location: Nevada
Question: I've been looking for information regarding the differences between laser confocal microscope and digital confocal microscopes, including the advantages/disadvantages and uses of each. Thank you, Lila
Thanks to all who replied to my request for references to images of osmolarity problems. The book below is a wonderful resource for images of all kinds of treatments ranging from the effects of various fixation protocols to analysis of digital images. I don't know how this book escaped my notice when it came out, but I'm buying it, and heartily recommend it!
} Biomedical Electron Microscopy by Maunsbach A B & Afzelius } B A (1999) has several pages of illustrations of shrinkage } etc. }
} } Hi, All- } } } } Does anyone have a reference for or some transmission electron micrographs } } that illustrate the effects of osmolarity of buffers on animal cells? I } } would like to show students on tour of our facility shrinkage and swelling } } of cells and organelles. I have other wonderful artifacts to show, } } including one of my first sections with chatter so bad it looks like } } mini-blinds... } } } } Mahalo! } } 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* ****************************************************************************
Hello, } From time to time I need to scan some negatives with flat bed scanner (Umax PowerLook II). Sometimes there is a problem with Newton rings in the scanned image. Does anybody know some tricks how to solve this problem? Best regards Oldrich +-----------------------------------+ 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
Emispec Systems, Inc., a rapidly growing company that creates software for electron microscopy, is looking for a domestic technical sales engineer. We are looking for someone with strong interpersonal skills and experience with customer interactions.
Some of the job requirements will include:
-Strong technical knowledge of products. -Ability to give product demonstrations at tradeshows and at customer sites. -Providing product information and quotations. -Defining potential customers. -Qualifying leads and determining customer needs. -Adding/updating contact information in the sales database. -Strong customer support throughout the purchasing cycle and after. -Handling initial customer service requests, and coordinating with the service department to see that they are handled appropriately. -Extensive travel.
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Below is the result of your feedback form. It was submitted by (ggauss-at-dynacocorp.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, June 12, 2001 at 18:12:49 ---------------------------------------------------------------------------
Email: ggauss-at-dynacocorp.com Name: Gordon Gauss
Organization: Dynaco Corporation
Education: Undergraduate College
Location: Tempe, Arizona
Question: We recently received a microscope with no manual. It is an aus JENA Jenavert. It is in excellent condition. We believe it was East German made. Is anyone familiar with this microscope and how can we obtain a manual in either german or english. Thank you...
If Newton rings are the same as Moire patterns (constructive/destructive interference lines)....
Many scanner drivers have a "de-screen" option or the like. My Microtek system seems to do a decent job of removing the lines. Also, you might try scanning at a much higher dpi (dots per inch ...or cm), then lightly average pixels and reduce in array size to meet needs.
Woody White McDermott Technology, 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 } -------------------------------------------------------------- } ---------. } } } Hello, } } From time to time I need to scan some negatives with flat } bed scanner } (Umax PowerLook II). Sometimes there is a problem with Newton } rings in } the scanned image. Does anybody know some tricks how to solve this } problem? } Best regards Oldrich } +-----------------------------------+ } 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 } } }
I run a research based service lab at Ohio State University and I've been asked to find out what other universities charge their internal customers for the use of TEM, SEM, confocal microscopy and technical assistance. I'm particularly interested in biological labs at large state universities, big 10 universities, and Ohio universities. Please respond directly to me. Thanks. Kathleen S. Wolken Senior Electron Microscopist Campus Microscopy and Imaging Facility (CMIF) 4029 Graves Hall 333 West 10th Avenue Columbus, OH 43210-1239
Phone 614-292-9786 FAX 614-688-8742 WEB http://www.med.ohio-state.edu/cmif
The Newton rings are interference fringes due to the small air gap when the film not quite touches the glass. The way to get around the problem is to raise the film off of the glass just slightly. Most of the better quality scanners provide holders, although not TEM sized one. Make your own from two pieces of a file folder cut to the appropriate size and held together at one edge with tape. There is no problem with a defocus of the image.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Oldrich Benada [mailto:benada-at-biomed.cas.cz] Sent: Wednesday, June 13, 2001 2:45 AM To: microscopy-at-sparc5.microscopy.com
Hello, } From time to time I need to scan some negatives with flat bed scanner (Umax PowerLook II). Sometimes there is a problem with Newton rings in the scanned image. Does anybody know some tricks how to solve this problem? Best regards Oldrich +-----------------------------------+ 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
Try speaking to Hitachi in Gaithersburg, MD. Their Applications Lab. should have that info. at their fingertips.
Peter Tomic Anadigics
-----Original Message----- } From: Ken Converse [mailto:qualityimages-at-netrax.net] Sent: Tuesday, June 12, 2001 4:39 PM To: Chaoying Ni; MSA, listserver
Chaoying Ni , I don't know about the current crop of FE cathodes, but in the past an electrolytic cell was set up with the W needle as part of the circuit. The needle was slowly dipped and removed several times from the NaOH electrolyte. Essentially, it was the same technique as was (and is) used to produce micomanipulator needles. I think the set-up was shown and described well in a Microscopy Today article within the past 12 months.
Ken Converse owner Quality Images Delta, PA
Chaoying Ni wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 lister, } } A fellow here in the department has the interest in knowing by which mean } an FEG W-crystal needle is cut. I only know that in the case of Schottky } emitter, the submicron needle is cut in {100} direction and coated with } ZrO2 to reduce the work function. But I have no knowledge in the } manufacturing process of the tip (FIB?). Anybody who knows please shed } some lights. Thanks in advance. } } ********************************* } Chaoying Ni } Electron Microscopy Laboratory } Materials Science and Engineering } College of Engineering } University of Delaware } Newark, DE 19716 } ********************************* } } } }
I am looking for a LKB Nova ultramicrotome. I presently have a Reichert Ultracut E which works absolutely fine, its just that I'm really a fan of LKB ultramicrotomes (the last one was a LKB III which is not usable anymore). I have used LKB's for over 20 yrs and I would prefer to go back to that brand. Does anyone out there want to swap or sell one?
Karen Jensen, M.S. Associate Scientist and Project Manager University of Rochester Medical Center Electron Microscope Research Core Pathology and Lab. Med. Rochester, NY 14642 716-275-1954
We have been looking at melting and crystallization of polymers under hot stages for twenty years. We have always found Mettler-Toledo Hot Stages very good, and have bought three of them.
For information, go to:
http://www.mt.com
then click on "Products" in the bar at top;
in the window 2.Product Search type "stage";
in the next window, under the text "Thermal Data - FP82HT/FP84HT"
click on the little button "English"
which will take you to a screen where you can request more information;
(that's all one line, in case the e-mail server has chopped it in two.)
** Disclaimer ** we have no commercial interest in M-T, simply we've found them very good and helpful.
+------------------------------------------------------------------------+ | 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 Tue, 12 Jun 2001 Dalton.Pierson-at-sealedair.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 } -----------------------------------------------------------------------. } } } Dear Sir or Madame, } I have been asked by my employer to investigate setting my laboratory up to } perform what was called "hot-stage microscopy". I understand this to be the } ability to observe the melting of (in this case) a polymer. I was hoping } that your organization could point me in the right direction. Any } information you could give me would be greatly appreciated. } } Thank you, } Dalton Pierson } }
You do not indicate whether the objective is to measure the total amount of the two metals in the assembly as a whole, or whether you want to measure their proportions in each of the different areas: the tin solder, the intermetallic layers and so on. You also don't indicate the purpose to which the results are to be put. To some extent the answer depends on knowing this, as a biased but highly repeatable measure of something is often useful for internal purposes.
Someone with specific experience in this industry might have a more specific reply but in general, the problem is that the same elements are shared by very different phases in close proximity to one another. For the Au/Sn intermetallic compounds which have quite specific compositions, the absorption /enhancement effects will be different than for a low-concentration solid solution of either of the metals in the other. Without a standard comprised of a known intermetallic phase, the results could be consistent (that is, you might be able to use the measure as a means of process control) but not very accurate (as compared with the absolute amounts present). Another frequently overlooked problem with quantifying the results of e-probe analysis of a multiphase alloy is that the size distribution of imiscible but intimately mixed phases may change due to process variables (e.g. thermal history) even though the total proportion of the metals remains the same. The micro-environment (matrix effect) experienced by an emitted x-ray will depend on its probability of passing through the same material or a different phase on its way to reaching the detector. Obviously this can be affected by the grainsize of a eutectic mixture.
You haven't indicated whether the sample is to be sectioned into a planar surface for analysis or examined as is. Another frequently overlooked problem is that of x-rays emitted through secondary interactions with structures that are not illuminated by the beam but which are "visible" within the detector collimator's view. A planar cross section presents the least chance for interference of this kind. On the other hand, a gold-metallized surface projection that resides next to your solder structure could be a source of enhanced Au-m x-rays produced through secondary fluorescence by the Sn-l x-rays under the beam. (even though secondary emmission is not a very efficient process, the number of Sn x-rays fluorescing the adjacent gold is much higher than the number that happen to be emitted in the direction of the X-ray detector. Meanwhile the X-ray detector is looking at the whole scene indiscriminately.
John Twilley
Peter Tomic wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Folks; } } I have a question as to technique or method regarding the quantification of } Au/Sn in a "solder bump" application for an InP microcircuit. Essentially } what I've been asked to do is provide the weight % of each element after } this "solder" has been melted and re-flowed. The samples are cylindrical in } shape with an x-section of 65 uM and a thickness of 34 uM. The Au/Sn } portion sits atop this structure and is a thickness of ~ 2 uM. } } My question is, how can I quantify, in Wt. %, the ratios of the two metals? } I have EDX at my disposal but I am uncertain that I will be able to } accurately quantify this alloy without a standard. One other issue is that } the underlying metallization that primarily constructs this "bump" is also } Au for the next 32 uM. } } Any insight into this problem would be greatly appreciated by myself and my } colleagues. } } Peter Tomic } Analytical Services Group } Anadigics, Inc. } 35 Technology Drive } Warren, New Jersey } U.S.A. 07090 } } } }
Hi, I've found two things that helped eliminate this problem. If you place the negative directly onto the scanner bed, use an anti-static brush to wipe both the bed and the negative first. The second solution was to modify a negative holder. I work mostly with TEM sized negatives and we just cut out the back (solid) part of a TEM negative holder, then we place the negative into the holder which keeps it from contacting the scanner bed directly. If your working with a different size negative you could fashion a holder from anything, even a piece of poster board should work. I think the fringes are caused by a build up of static between the scanner bed and the negative. I have also found that if you go to adjust contrast in print shop and adjust the contrast through the entire range from too light to too dark you can pick up a fringe before you take the negative off the scanner. That eliminates having to go back and re-do a picture because the fringe wasn't real apparent when you scanned it. Hope this works for you...it has made my life a lot easier. Dorrance
} ---------- } From: Oldrich Benada } Sent: Tuesday, June 12, 2001 11:45 PM } To: microscopy-at-sparc5.microscopy.com } Subject: Unwanted Newton rings } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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, } } From time to time I need to scan some negatives with flat bed scanner } (Umax PowerLook II). Sometimes there is a problem with Newton rings in } the scanned image. Does anybody know some tricks how to solve this } problem? } Best regards Oldrich } +-----------------------------------+ } 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 } } } } }
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2. SALES ORDER. Once a quotation is accepted, the final quotation information can be transformed into a Sales Order for your client's signature on a "point and click" basis. The Sales Order can be modified and re issued if necessary. A history if kept of all Sales Orders for future reference, or modification for other clients. All sales orders and revisions are "auto numbered," including versions. Inventory status can be accessed from this section for reference.
3. CUSTOMER LETTERS can be created from the Quotation and Sales Order sections.
4. SHOP TRAVELER/WORK ORDER. Once a Sales Order is accepted, the sales order information can be transformed into a shop traveler/work order on a "point and click" basis. Each item on the Sales Order becomes a shop traveler/work order, with each step of production of the item then listed on the traveler/work order. Each such traveler/work order is tied back into the Sales Order. The shop traveler/work order allows for the entry of line items, and notes on each line item. The shop traveler/work order contains a "notes" section. The Shop traveler/work order allows for the storing or attachment of drawings to the traveler/work order. The shop traveler/work order also contains a "drop down," from which standard processes can be selected for inclusion on the shop traveler/work order. The shop traveler/work order numbers progress in order of production sequence, and re numbers them if new steps are added. The shop traveler/work order allows for change orders or revisions, and numbers changes in sequence of he original shop traveler/work order number; i.e., 100, 100-1, 100-2, etc. All shop traveler/work orders and related revisions are retained in memory for future reference. The shop traveler/work order is bar coded for tracking of production step by step, and production of ongoing client status reports. Bar coding includes the ability for an employee to "swipe" their own ID bar code for recording in the system as to who upgraded what step. The shop traveler/work order function also allows for manual update of production status. The shop traveler/work order allows for quality control sign off, and the final production of certifications, either from a "canned" list, or hand typed in on a case by case basis.
5. INVENTORY. The application includes an inventory section, which allows operations to check materials inventory in and out. The inventory section allows for the comparison of inventory received against a P.O., and produces an "overage/underage" report of inventory received as compared against the P.O. The inventory section allows for the setting of minimum (re-order now!) and maximum inventory amounts, and produces reports showing what inventory needs to be ordered, as well as inventory that is at or above the maximum set to have in house. The inventory section also tracks "partially shipped" orders, which are tied in to the shipping function. This section shows how much completed product under a particular order has been actually shipped to a client, and how much remains to be shipped. The balance is adjusted as shipments are made.
6. REQUEST FOR PURCHASE. The application allows operators to produce a Request For Purchase for accounting for any inventory items, which need to be ordered. Inventory items have a drop down of approved vendors for each item.
7. REQUEST FOR BID. The application allows operators to produce a Request For Bid for accounting to send to Vendors for any inventory items, which need to be ordered. Inventory items have a drop down of approved vendors for each item to which Requests For Bid can be sent.
8. INVOICE. The application produces an invoice/invoice detail for all completed items ready to be billed/shipped to clients.
9. PRODUCTION OUTPUT STATUS. The application produces a date range selectable report on how much product, and the value of the product, which was completed during a selected date range. The application also produces a report on how many orders, and the value of those orders, which remain to be completed during a selected date range.
10. The application produces SHIPPING DOCUMENTS as per selected shippers, and produces a PACKING SLIP.
11. The application has a "FIND" FUNCTION in selected sections, allowing for searches by customer name, work order number, etc.
12. The application has "AUTO FILL;" i.e., when an operator starts to type in a name, number, etc. all related information auto fills after the first few letters or numbers are typed in.
Job Master is currently being sold in the marketplace for $2,495.00 per package. However, if we receive your order by June 22th, your total price will be $1,495.00.
Again, if you have any questions at all, or would like to place your order, please call me on my direct line, (661) 254-9926. Thank you!
You have received this newsletter because you signed up for updates on our tracking software. If you want to unsubscribe from this newsletter, please send a reply email with "REMOVE" in the subject line.
Listers: Here's info about another source of used instruments.
- - - - - - - - - - - -
} From: "A. Greene" {ablue-at-io.com} } To: {bigelow-at-umich.edu} } Subject: Source } Date: Mon, 11 Jun 2001 20:05:13 -0500 } X-Priority: 3 } Status: } } Hello Professor Bigelow, } } Here is another source, as well. I have been in the instrument } business for over 30 years, used to be with Philips and then at the } University of Illinois for 10 years. Now, for the past eight years } have had my own little business in Texas. I probably have more } parts for older Philips microscopes than Philips. Also, I sell } reconditioned, used electron microscopes of most brands. } } 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 } } Sustaining Member - MICROSCOPY SOCIETY OF AMERICA
-- Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan 3062 Dow Bldg.; 2300 Hayward St. Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
We have a graduate student who is using a fluorescent antibody labelling technique on a neuronal cell culture plate. Briefly, she fixes the cells in 4% paraformaldehyde, permeabilizes them with Triton-X and continues with the immunostaining. The cells stay on the plate (they can only grow on plates, not slides or coverslips). The question we have is how to coverslip them and then observe them with a fluorescent microscope. Will glycerol work? Can we simply put a drop of glycerol on the plate and add a coverslip or do we need a fluorescent mounting medium? I have limited experience with fluorescent microscopy, so I would appreciate any help that you can provide.
Thank you very much, Sandy Hancock
Laboratory for Neurotoxicity Studies VMRCVM VA Tech
I am using transparency for Xerox machine with "windows" cut in it in such manner, that it's slightly smaller than actual negative's size. In my particular case I find that rings happens between negative and scanners bed. So, I put transparency on the bed and than negatives (4 per transparency sheet) in the way, when negatives situated across the "windows". In my particular case, transparency presence does not affect scanned image quality. You may put second sheet over the negatives if necessary.
Good luck!
Sergey.
At 10:00 AM 6/13/01 -0600, 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
Recently, there was a question about TEM and osmolarity. I have now a question about SEM and osmolarity. Has anyone have some references about this topic ? We are studying the canine uterus. We have some problems concerning (we think !) the osmolarity & the epithelial cells of the uterus. Any comment would be appreciated !
Photobleaching, or fading, causes loss of fluorescent signal in part due to photodynamic events which involve the interaction of the fluorophore with light energy and oxygen. Fluorescent labels have a characteristic life span, for instance, an average fluorescein molecule will emit 30k to 40k photons during its photo-chemical life span. Higher intensity light excites the fluorophor to emit a greater yield of photons and the service life of the fluorescent label is proportionally reduced. For this reason, it is generally a good idea to add some form of bleaching protection (anti-fading reagent) to a glycerol-based mounting media for fluorescent microscopy. P-phenylenediamine (Sigma Chemical Co.) used in a concentration of 0.1% in [10% phosphate buffered saline/90% glycerol] is good. Diazabicyclo-octane or n-propyl-gallate (~0.1-0.2% in 10% PBS/Glycerol are decent alternatives. The disadvantage to this approach is that the coverslip must be sealed (nail varnish) in order to fix the coverslip permanently. Vectrashield (Vector Co.) is a propriety product for fluorescent microscopy which seems to work quite well with the added advantages of convenience and greater permanence. One other consideration is the degree to which your fixation protocol affects the antigenicity of your target-a short (10 min/10% buffered formalin) is a good place to start-some antigens seem to be very sensitive to the cross-linking effects of aldehyde fixation, acetone or ethanol/methanol fixation may prove helpful if this is the case. If the target is a membrane protein one wants to be cautious with the Triton-X. Residual Triton-X could also interfere with the antigen-antibody interaction in a sensitive system. Acetone also permeablizes cell membranes, and if I recall correctly is a favorable fixative for immunofluoresent studies of the cytoskeleton. Culturing cells on slides or coverslips may not be an issue as long as you can get the culure plates into the scope. However, I believe theromonox (sp?) coverslips are designed to allow cell adhesion. Some workers coat slides and/or cover slips with poly-l-lysine for cell adhesion. Coverslipping is important to allow the optical system of the microscope to function at full potential. This is less of an issue with inverted microscopes designed to accomodate tissue culture vessels. Regards, Karl Garsha
You can de-stain both thick and thin section w/ 0.2 M EDTA. Only the time differs. One caveat, if the section has been irradiated, i.e. in the TEM then this will much less effective.
Good luck
Richard Cole Research Scientist III Director: Advanced Light Microscopy Core Unit Wadsworth Center P.O. Box 509 Albany N.Y. 12201-0509 518-474-7048 Phone 518-486-4901 Fax
-----Original Message----- } From: "tbargar-at-unmc.edu"-at-sparc5.microscopy.com [mailto:"tbargar-at-unmc.edu"-at-sparc5.microscopy.com] Sent: Wednesday, June 13, 2001 4:10 PM To: Microscopy-at-sparc5.microscopy.com
Hello;
Is it possible to destain a thin section? After having stained with uranyl acetate and lead citrate. All help appreciated, thanks.
At the University of Rochester Electron Microscope Research Core, we charge $75.00/hour with an Electron Microscopist running & photographing using our Hitachi 7100 TEM, without the EM person, a trained researcher or grad student must be trained on its use and the rate becomes $50.00/hour.
Our processing and preparation of tissue is at $75.00/hour and that only counts the hands-on times. For instance, usually it could take 30 minutes to do 1 micron sections and thin section the same block. We bill for .5hr=$37.50 to cut one block, .25 hr=$18.75 if no screening 1 micron sections are needed.
Most investigators have us embedd 2-6 blocks of tissue, etc. We cut a representative block, review the 1 micron with them if necessary and then proceed with the thin-sectioning. Staining is usually .25hr...etc, If the investigator wanted more than one block cut and thin sectioned(3-5 grids), they would all be stained together using a Hiroka staining kit(holds up to 40 grids) and therefore would only be charged the .25hr rate. However, the methods and materials you use and speed in which they are performed can vary from one lab to another.
We only billed $35,000 last year and that doesn't begin to cover salaries. So the University of Rochester has to subsidize the EM Core. However, the Core(s) is a big marketing tool for those researchers which are actively being recruited. Many Molecular Science researchers utilize the EM Core for ImmunoEM procedures. In fact, the majority of my work is performing pre- and postembedding immunolabeling for EM. The average bill for this varies from $00-1,000.
Hope this helps you.
Karen Jensen, M.S. Associate Scientist and Project Manager University of Rochester Medical Center Electron Microscope Research Core Pathology and Lab. Med. Rochester, NY 14642
-----Original Message----- } From: Kathy Wolken [mailto:wolken.1-at-osu.edu] Sent: Wednesday, June 13, 2001 9:15 AM To: microscopy-at-sparc5.microscopy.com
I run a research based service lab at Ohio State University and I've been asked to find out what other universities charge their internal customers for the use of TEM, SEM, confocal microscopy and technical assistance. I'm particularly interested in biological labs at large state universities, big 10 universities, and Ohio universities. Please respond directly to me. Thanks. Kathleen S. Wolken Senior Electron Microscopist Campus Microscopy and Imaging Facility (CMIF) 4029 Graves Hall 333 West 10th Avenue Columbus, OH 43210-1239
Phone 614-292-9786 FAX 614-688-8742 WEB http://www.med.ohio-state.edu/cmif
As a general rule, I am less than thrilled about using anti-fade agents for fixed cells unless absolutely necessary for the reasons explained below. Live cell work may require anti-oxidants but I am less familiar with this application. I think it is better to use more stable fluorochromes (e.g., Alexa 488 instead of FITC)
PPD is a skin sensitizer, photosensitive, and Giloh & Sedat (1982) Science 217:1252 says it has no effect on rhodamine fading. I believe they actually showed storage of rhodamine specimens in 5% n-propyl gallate (NPG) in glycerol for 2-3 days decreased fluorescence (suggest rinsing slides for storage (what a pain!). Krienk et al., 1989 J. Immuno. Methods 117:91 compared DABCO, PPD and NPG and concluded DABCO the least effect. Valnes & Brandtzaeg (1985) J Histochem Cytochem 33:755 also said DABCO not as effective as PPD or NPG but also say NPG and PPD only good if slides made recently and examined promptly - recommended re-mounting in PVA without quencher if you were going to store. I have tried Vectashield and other commercial anti-fade compounds with ok results but it is hard to say exactly how much they helped. I don't think there are any systematic published studies examining if they work equally well with all fluorochromes or if they hurt the sample with storage.
Be cautious about using FITC in any PBS buffered mounting medium since it is much more fluorescent at alkaline (pH 8.6) than neutral (pH 7.0) pH's.
} ------------------------------------------------------------------------ } 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)
I'm trying to embed and section mouse cochleas that have been immunoreacted en bloc for BrdU with DAB as the chromagen. The soft tissues of the cochlea were separated from the bony structures, leaving a soft tissue spiral containing the organ of Corti.
During processing for Epon-Araldite for 5 micron sections for light microscopy (without osmium and using increasing concentrations of aqueous acetone as the dehydrant), it appears as if the chromagen was lost (no labeled cells were seen) and a purple/grey background was left behind. Whole mounts immunoreacted at the same time retained an amber coloring with dark brown-labeled nuclei.
Does anyone have any hints as to what happened here or what we can do in the future to retain the labeling throughout plastic processing. I've done this embedding in the past with the same type of tissue (but still in the bony labyrinth) and had no problems.
Thank you,
Jaclynn M. Lett, Research Assistant jlett-at-cid.wustl.edu
Faye and Carl Simons Center for Biology of Hearing and Deafness Central Institute for the Deaf 4560 Clayton Avenue St. Louis, MO 63110
Just an FYI- we have found thermanox to be autoflourescent.
Lara Muffley Dermatology Dept University of Washington Seattle, WA muffley-at-u.washington.edu
On Thu, 14 Jun 2001, Karl Garsha wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 Sandy, } } Photobleaching, or fading, causes loss of fluorescent signal in part due to } photodynamic events which involve the interaction of the fluorophore with } light energy and oxygen. } Fluorescent labels have a characteristic life span, for instance, an average } fluorescein molecule will emit 30k to 40k photons during its photo-chemical } life span. Higher intensity light excites the fluorophor to emit a greater } yield of photons and the service life of the fluorescent label is } proportionally reduced. } For this reason, it is generally a good idea to add some form of } bleaching protection (anti-fading reagent) to a glycerol-based mounting } media for fluorescent microscopy. P-phenylenediamine (Sigma Chemical Co.) } used in a concentration of 0.1% in [10% phosphate buffered saline/90% } glycerol] is good. Diazabicyclo-octane or n-propyl-gallate (~0.1-0.2% in } 10% PBS/Glycerol are decent alternatives. The disadvantage to this approach } is that the coverslip must be sealed (nail varnish) in order to fix the } coverslip permanently. } Vectrashield (Vector Co.) is a propriety product for fluorescent } microscopy which seems to work quite well with the added advantages of } convenience and greater permanence. } One other consideration is the degree to which your fixation protocol } affects the antigenicity of your target-a short (10 min/10% buffered } formalin) is a good place to start-some antigens seem to be very sensitive } to the cross-linking effects of aldehyde fixation, acetone or } ethanol/methanol fixation may prove helpful if this is the case. If the } target is a membrane protein one wants to be cautious with the Triton-X. } Residual Triton-X could also interfere with the antigen-antibody interaction } in a sensitive system. Acetone also permeablizes cell membranes, and if I } recall correctly is a favorable fixative for immunofluoresent studies of the } cytoskeleton. } Culturing cells on slides or coverslips may not be an issue as long as } you can get the culure plates into the scope. However, I believe theromonox } (sp?) coverslips are designed to allow cell adhesion. Some workers coat } slides and/or cover slips with poly-l-lysine for cell adhesion. } Coverslipping is important to allow the optical system of the microscope to } function at full potential. This is less of an issue with inverted } microscopes designed to accomodate tissue culture vessels. } Regards, } Karl Garsha } } keg-at-uwm.edu } www.uwm.edu/~keg } } } } } } } } } } } }
I am in the market to purchase a new ultramicrotome. I am aware of the Leica and RMC ultramicrotomes. Are there other companies manufacturing ultramicrotomes today?
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-umn.edu
I have some problems with our EM chillers. The water in EM cooling circulation is bad contaminated by algae, bacteria, maybe more microorganisms. Does somebody here know some water cleaner for chillers of EM? Appreciate for the help.
Haixin Xu
Biological Sciences University of Maryland, Baltimore County Baltimore, Maryland
Do you have filters somewhere in the recirculation lines? A lot of places have 2 barrel-type filters (I don't know the correct term - they look like tubing, not flat membranes) in-line to clean up anything that might grow. Change them every 6 months or when they start to look like they might become sentient.
I've been told that you should not put algicides in the water; the filters are probably the best way to go.
Tamara
On Thu, 14 Jun 2001, Haixin Xu wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 lister, } } I have some problems with our EM chillers. The water in EM cooling } circulation is bad contaminated by algae, bacteria, maybe more } microorganisms. Does somebody here know some water cleaner for chillers of } EM? Appreciate for the help. } } Haixin Xu } } Biological Sciences } University of Maryland, Baltimore County } Baltimore, Maryland } } }
|--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward-at-unm.edu |--------------------------------------------------|
Below is the result of your feedback form. It was submitted by (Whunter-at-ushrl.ars.usda.gov) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Thursday, June 14, 2001 at 07:58:06 ---------------------------------------------------------------------------
Email: Whunter-at-ushrl.ars.usda.gov Name: Wayne Hunter
Organization: USDA { ARS
Education: Graduate College
Location: Ft. Pierce, FL, USA
Question: I am looking for a protocol to embed insects in Paraplast, and doing thick sections, Thank you, Sincerely, Wayne Hunter REsearch Entomologist
Growth of algae, bacteria etc may also be prevented/reduced by using black, intransparant tubes instead of white transparant ones. They were installed on my chiller for an ESEM 2020, after considerable problems. With best regards, Emond de Roever, Ondeo Nalco Europe Technical Centre, Oegstgeest, the Netherlands
"I have some problems with our EM chillers. The water in EM cooling circulation is bad contaminated by algae, bacteria, maybe more microorganisms. Does somebody here know some water cleaner for chillers of EM? Appreciate for the help."
Haixin Xu
Biological Sciences University of Maryland, Baltimore County Baltimore, Maryland
Haixin, I've found that using distilled or de-ionised water in the chiller in addition to the measures already suggested will also greatly reduce the growth of microorganisms by denying them nutrients.
Regards, Eric lachowski
------------ Dr Eric Lachowski University of Aberdeen Department of Chemistry Meston Walk Old Aberdeen AB24 3UE Scotland Tel. +44 (0)1224 272934 Fax +44(0)1224 272921 e.lachowski-at-abdn.ac.uk
I am using an Epson 980. I find it satisfactory for both B&W and color prints and it is reasonably fast. It is also relatively inexpensive. Choice of paper grade/type is very important in determining available print quality.
Woody White McDermott technology, Inc
} -----Original Message----- } From: treese [mailto:treese-at-marinebio.mbl.edu] } Sent: Friday, June 15, 2001 1:00 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: printers } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } 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 } } ------------------------------------------------------------- } ----------. } } } } } We are going to replace our old Epson InkJet printer which produces } nice B+W prints of micrographs, when it works. I haven't kept up } with Epson vs HP for electron microscopists and wondered if a clear } preference for one of these, or maybe another mfg or technology has } emerged. We want B+W quality and reliability, and are willing to pay } a bit more,and sacrifice speed and color quality...Thanks...Tom Reese }
I have seen a retrofit for Epson printers that looks quite interesting at
http://www.piezography.com/
I haven't actually seen it in action but it looks like it should be good for B/W prints. The advertising samples on the web site look very nice. Basically they replace the color cartridges with different densities of black ink. By allowing gray inks, they reduce the amount of dithering required. The product consists of the replacement cartridges and the revised printer driver software.
Has anyone actually tried this approach?
Cheers, Henk Colijn
At 12:00 AM 6/15/01 -0500, treese wrote:
} We are going to replace our old Epson InkJet printer which produces nice } B+W prints of micrographs, when it works. I haven't kept up with Epson vs } HP for electron microscopists and wondered if a clear preference for one } of these, or maybe another mfg or technology has emerged. We want B+W } quality and reliability, and are willing to pay a bit more,and sacrifice } speed and color quality...Thanks...Tom Reese
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://web.ceof.ohio-state.edu Fools are pleased when they discover error. The wise are pleased when they discover truth.
Does anyone happen to know in what issue of microscopy Today the following information on sharpening W needles was found? (or how to make them) If so please send me a note at, MRoberson-at-dow,com
I don't know about the current crop of FE cathodes, but in the past an electrolytic cell was set up with the W needle as part of the circuit. The needle was slowly dipped and removed several times from the NaOH electrolyte. Essentially, it was the same technique as was (and is) used to produce micomanipulator needles. I think the set-up was shown and described well in a Microscopy Today article within the past 12 months.
Michael Roberson (517)636-8656 SMX Analytical Sciences Midland, MI 48667
The question of avoiding algal growth in water chillers is discussed at length on p. 216 of my book 'Vacuum Methods in Electron Microscopy' (see http://www.2spi.com/catalog/books/book48.html and http://www.pup.princeton.edu/titles/6484.html)
One important way of reducing algal growth is to exclude light from the system by keeping the reservior tank covered and by using opaque tubing. If this is not sufficient then we have had great success over many years by adding the algacide known as 'Chloramine-T' to the water in the amount of about 0.25 grams per liter. This stuff is available from most speciality chemical companies (Aldrich, Sigma, Polysciences) under the chemical name of N-chloro-p-toluensulphonamide. It is not highly soluble, and so is not likely to produce deposits in the system. We have not had any trouble with damage to any of our systems in which this material has been used. -- Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan 3062 Dow Bldg.; 2300 Hayward St. Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
-----Original Message----- } From: Jensen, Karen Sent: Thursday, June 14, 2001 10:23 AM To: 'Kathy Wolken' Cc: 'microscopy-at-msa.microscopy.com'
Dear Kathy:
At the University of Rochester Electron Microscope Research Core, we charge $75.00/hour with an Electron Microscopist running & photographing using our Hitachi 7100 TEM, without the EM person, a trained researcher or grad student must be trained on its use and the rate becomes $50.00/hour.
Our processing and preparation of tissue is at $75.00/hour and that only counts the hands-on times. For instance, usually it could take 30 minutes to do 1 micron sections and thin section the same block. We bill for .5hr=$37.50 to cut one block, .25 hr=$18.75 if no screening 1 micron sections are needed.
Most investigators have us embedd 2-6 blocks of tissue, etc. We cut a representative block, review the 1 micron with them if necessary and then proceed with the thin-sectioning. Staining is usually .25hr...etc, If the investigator wanted more than one block cut and thin sectioned(3-5 grids), they would all be stained together using a Hiroka staining kit(holds up to 40 grids) and therefore would only be charged the .25hr rate. However, the methods and materials you use and speed in which they are performed can vary from one lab to another.
We only billed $35,000 last year and that doesn't begin to cover salaries. So the University of Rochester has to subsidize the EM Core. However, the Core(s) is a big marketing tool for those researchers which are actively being recruited. Many Molecular Science researchers utilize the EM Core for ImmunoEM procedures. In fact, the majority of my work is performing pre- and postembedding immunolabeling for EM. The average bill for this varies from $300-1,000.
Hope this helps you.
Karen Jensen, M.S. Associate Scientist and Project Manager University of Rochester Medical Center Electron Microscope Research Core Pathology and Lab. Med. Rochester, NY 14642
-----Original Message----- } From: Kathy Wolken [mailto:wolken.1-at-osu.edu] Sent: Wednesday, June 13, 2001 9:15 AM To: microscopy-at-sparc5.microscopy.com
I run a research based service lab at Ohio State University and I've been asked to find out what other universities charge their internal customers for the use of TEM, SEM, confocal microscopy and technical assistance. I'm particularly interested in biological labs at large state universities, big 10 universities, and Ohio universities. Please respond directly to me. Thanks. Kathleen S. Wolken Senior Electron Microscopist Campus Microscopy and Imaging Facility (CMIF) 4029 Graves Hall 333 West 10th Avenue Columbus, OH 43210-1239
Phone 614-292-9786 FAX 614-688-8742 WEB http://www.med.ohio-state.edu/cmif
you can purchase algicide from Fisher, VWR or other lab suppliers.
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
} } } Haixin Xu {xu-at-gl.umbc.edu} 06/14 5:22 PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi lister,
I have some problems with our EM chillers. The water in EM cooling circulation is bad contaminated by algae, bacteria, maybe more microorganisms. Does somebody here know some water cleaner for chillers of EM? Appreciate for the help.
Haixin Xu
Biological Sciences University of Maryland, Baltimore County Baltimore, Maryland
I don't remember which issue of MT, but the following two paragraphs are three items that I sent to the Listserver recently describing sample preparation of emitters.
1)(Ni) I'm trying to remember back to grad school, but I'm pretty sure that it was a 10%HCl solution in water. I used it for Ni FIM samples. I'll look it up in my dissertation and see if I have it. Hren's book or Bowkett and Smith's book on FIM had the reference for Ni. I do remember that the solution worked better after it was used some. It took on a greenish tinge. So what I started doing with a fresh batch, was taking a little crystalline NiCl (green powder) a putting just a bit into solution to get the color just right. I can't remember if I used ac of dc.
2)(W) 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.
3)(Fe & floating layer method) You can use Field Ion Microscopy sample prep techniques to prepare very sharp needles of the wires. Basically, you can dip them in a beaker and electropolish them. The liquid/air interface will preferentially polish them. You need to move the interface up and down the sample by moving either the sample or liquid. If the wires are long enough, you can float electrolyte on top of CCl4 layer and you can get two samples when the bottom part drops off. Look up some of the standard electropolishing solutions and conditions. One is based on percholoric/butylcelsolve and the other on chromic acid/acetic acid -I think.
FEG note: Someone asked recently about FEG tips. One problem with these tips that anyone off the streets would have is getting material that is oriented. Normal W wires have a [011] texture. The desired orientation for emitters in order for them to have a low work function and high brightness is either [112] or [113] (I can't remember). Other than that, I think anyone with experience making FIM samples could probably duplicate their construction within a short time, even Schottky design.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Roberson, Michael (M) [mailto:MRoberson-at-dow.com] Sent: Friday, June 15, 2001 9:07 AM To: 'Microscopy-at-MSA.Microscopy.Com'
Does anyone happen to know in what issue of microscopy Today the following information on sharpening W needles was found? (or how to make them) If so please send me a note at, MRoberson-at-dow,com
I don't know about the current crop of FE cathodes, but in the past an electrolytic cell was set up with the W needle as part of the circuit. The needle was slowly dipped and removed several times from the NaOH electrolyte. Essentially, it was the same technique as was (and is) used to produce micomanipulator needles. I think the set-up was shown and described well in a Microscopy Today article within the past 12 months.
Michael Roberson (517)636-8656 SMX Analytical Sciences Midland, MI 48667
I installed the Lyson grayscale inks in our Epson 850N. The results are sometimes amazing, but always at least good.
Optimal results is greatly affected by paper type and quality, it seems more so than printing color. You will also need to fiddle with the colorspace gamma. Sorry I can't give more details, but I haven't had time to work with it too much since most people here are printing color. I think it has a great deal of promise and is worth checking out. The Lyson website has a number of tips and print comparisons for optimizing results.
As far as choosing printers, find people who have used any particular model. Epsons are slow to start printing. Out of the 4 epsons I work with (3 850N, 1 3000) and another 3 in a colleague's lab. at least 1, often 2, are misfeeding paper, engaging in randoms acts of stubborness, dropping off the network, etc. The 3000 is extremely touchy about how you load the paper, backing sheets, etc., and regularly soils itself with ink. Epson drivers have improved - sometimes, they even allow use to automatically scale to fit the page at print time. However, 2 of the 850N printers hasve never failed, always ready to go, so these problems probably reflect Epson QC issues..
Regards, Glen
"Hendrik O. Colijn" wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Tom, } } I have seen a retrofit for Epson printers that looks quite interesting at } } http://www.piezography.com/ } } I haven't actually seen it in action but it looks like it should be good } for B/W prints. The advertising samples on the web site look very } nice. Basically they replace the color cartridges with different densities } of black ink. By allowing gray inks, they reduce the amount of dithering } required. The product consists of the replacement cartridges and the } revised printer driver software. } } Has anyone actually tried this approach? } } Cheers, } Henk Colijn } } At 12:00 AM 6/15/01 -0500, treese wrote: } } } We are going to replace our old Epson InkJet printer which produces nice } } B+W prints of micrographs, when it works. I haven't kept up with Epson vs } } HP for electron microscopists and wondered if a clear preference for one } } of these, or maybe another mfg or technology has emerged. We want B+W } } quality and reliability, and are willing to pay a bit more,and sacrifice } } speed and color quality...Thanks...Tom Reese } } Hendrik O. Colijn colijn.1-at-osu.edu } Campus Electron Optics Facility Ohio State University } (614) 292-0674 http://web.ceof.ohio-state.edu } Fools are pleased when they discover error. } The wise are pleased when they discover truth.
-- Glen MacDonald Microscopy and Imaging Facility University of Washington Core for Communication Research 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 have always used Epson and have never been disappointed with its best performance, but I do have an occasional problem with banding. I am on my third model, an Epson stylus 900, over the last 6 years. It is of the 1440 DPI Varity. Now they have 2880, although I am usually happy with 720DPI. I get the impression that HP's asset is reliability. HP has not focused on photo image quality as much as Epson, but both printers have surpassed the photo-realistic hurdle a few years ago, not to say that they are as good as film of' course. It would be interesting to buy both, do a side by side comparison and send the one that fails back. Let me know what you find.
On a side note I have been playing around with wide format prints. There is some good in printing larger hard copies and scanning at lower magnification to get a better representation of the surface, but where the limits are I don't know.
Don't write off speed too much. Speed usually only comes at the cost of dollars, not quality with these inkjets. I like USB as well, may be faster.
Ric
-----Original Message----- } From: treese [mailto:treese-at-marinebio.mbl.edu] Sent: Friday, June 15, 2001 1:00 AM To: Microscopy-at-sparc5.microscopy.com
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Tom -
I've gotten quite passionate about marine invertebrate macrophotography as a retirement project (see the URL below if you like such things). I have a lot of prints on display in various local educational venues, so I've been very concerned with print longevity - at a reasonable cost, since I buy the equipment with my own $$$. I suggest that you look at http://www.luminous-landscape.com/1280.htm for a critical review of the Epson 1280. Epson's 6-color process is supposed to give 25-year color prints & 100-year B&W (see http://www.wilhelm-research.com/). I've just bought a closeout 1270, for $250; 1440 dpi rather than 2880, but what a deal!
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
Thank you for all your replies to my question regarding oil red O. Here is my attempt to summarize those replies (and resulting discussions) starting with the earliest reply. Please forgive the haphazard manner of organization.
Jaclynn Lett, Research Assistant jlett-at-cid.wustl.edu
Faye and Carl Simons Center for Biology of Hearing and Deafness Central Institute for the Deaf 4560 Clayton Avenue St. Louis, MO 63110
voice: 314.977.0257 fax: 314.977-0030 Jaclynn M. Lett, Research Assistant jlett-at-cid.wustl.edu
Faye and Carl Simons Center for Biology of Hearing and Deafness Central Institute for the Deaf 4560 Clayton Avenue St. Louis, MO 63110
voice: 314.977.0257 fax: 314.977.0030
Ms. Allan-Wojtas,
In response to your request regarding oil red O lipid staining, here are the replies I received (along with some discussions as well). I have not had time to thoroughly digest everything myself, so please forgive the delay with which I replied and the lenghthy and unorganized manner of compiling the replies (listed in the order in which they were received).
Jaclynn M. Lett, Research Assistant jlett-at-cid.wustl.edu
Faye and Carl Simons Center for Biology of Hearing and Deafness Central Institute for the Deaf 4560 Clayton Avenue St. Louis, MO 63110
voice: 314.977.0257 fax: 314.977.0030
---------------------------------------------------------------------------- --------------------------------------- } Has anyone used Oil Red O to stain lipids in tissues embedded in plastics } (Epon or Epon-Araldite)? If so, has this been done by staining en bloc or } by staining the sections. Sections would range from 1-4 microns in } thickness.
After the tissue is embedded, the solvents used (alcohol, propylene oxide) will have dissolved most or all of the lipids so there will not be much to stain. If you stain before embedding, dehydration will remove the lipid and the stain.
} We would also consider tissues embedded in glycol methacrylate.
Forget it if you are using alcohol (or acetone) for dehydration. I don't know of any dehydrating agents that will not remove lipids. Perhaps freeze drying would work, but you still need an embedding media that won't dissolve lipids. Even after osmium, some lipid is lost in dehydration.
} We'd like } to avoid frozen sections because we'd prefer the higher level of detail } possible with plastic.
So would a lot of us! This is why frozen sections are used for lipid staining.
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 lipids will be removed by the solvents used in processing. In tissues post fixed with osmium tetroxide BEFORE embedding in these plastics, the osmium will stain lipids black.
Gayle Callis MT,HT,HTL(ASCP) Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman Bozeman MT 59717-3610
406 994-6367 404 994-4303 (FAX) ---------------------------------------------------------------------------- ----------------------------------- The problem with trying to do lipid staining in epon or plastic embedded tissue is that inorder to embed in epon or gma you usually have to process in solvents which will dissolve the lipids not to mention that the plastic itself is a solvent, at least GMA is somewhat.
Patsy Ruegg ---------------------------------------------------------------------------- ------------------------------------ I don't know anyone who has been able to "successfully" stain enbloc or sections from resins with oil red O, Sudan black or any of the other common lipid stains because the solvents used in the processing are all designed to remove fats and lipids. The only way I could get true accurate staining was with cryo sections. I have tried so many methods/ resin schedules/ types of resin and the same problems always occur, if there is enough lipid remaining to stain, it doesn't give a true representation of location and quantity. I'm sorry if this doesn't help, other than to perhaps save you some time and effort. As always, if you do happen to find someone who swears by their method, I would love you to email me a copy of it. Cheers, Kerrie
Kerrie Holmes Histology, Microscopy Research Research Division, Peter MacCallum Cancer Institute Locked Bag #1 A'Beckett St. East Melbourne 8006 Phone: 9656 1244 / 1242 Fax: 9656 1411 Email: k.holmes-at-pmci.unimelb.edu.au ---------------------------------------------------------------------------- ----------------------------------- I do alot of samples in GMA and have tried oil red O on a couple of occasions with no luck. I would be very interested to know if anyone has done this and using what method. Good luck. Regards Liz
Elizabeth Cox Fisheries Biologist Queensland Department of Primary Industries Northern Fisheries Centre PO Box 5396 Cairns, Queensland, Australia 4870 Fax: 07 4035 1401 Ph: 07 4035 0158 ---------------------------------------------------------------------------- --------------------------------------- Has anyone used Oil Red O to stain lipids in tissues embedded in plastics (Epon or Epon-Araldite)? If so, has this been done by staining en bloc or by staining the sections. Sections would range from 1-4 microns in thickness.
A couple of people so far have commented on how hard this would be, and our experience agrees with theirs. HOWEVER Jaclynn also said:
We would also consider tissues embedded in glycol methacrylate. We'd like to avoid frozen sections because we'd prefer the higher level of detail possible with plastic.
In THAT case things look better! Would you settle for Sudan black, rather than Oil red staining of the lipid? If 'yes' then there is a method - which does indeed show even tiny droplets of lipid very clearly. This was worked out by the one-time king of GMA staining Peter Gerrits, and can be found in J Neurosci Methods, as follows:
Gerrits PO, Brekelmans-Bartels M, Mast L, 's-GrAavenmade EJ, Horobin RW and Holstege G. (1992).. Staining myelin and myelin-like degradation products in the spinal cords of chronic experimental allergic encephalomyelitis (Cr-EAE) rats using Sudan Black B staining of glycol methacrylate-embedded material. J. Neuroscience Methods. 45, 99-105
Bye - Richard Horobin
Institute of Biomedical & Life Sciences, University of Glasgow T direct 01796-474 480 --- E RichardWHorobin-at-aol.com "What should we expect? Everything." ---------------------------------------------------------------------------- ------------------------------------ Richard-
Whenever I've worked with those plastics, there has always been a clearing stage through acetone. Since acetone would remove all non-bound fat, Oil Red O would have nothing to go into.
When I've worked with these plastics, I also did a post-fixation in osmium tetroxide, which does a very good job of staining fats and lipid (it turns them black). Perhaps this would work for your purposes.
Joe
Joseph A. Saby, BA, HT(ASCP) Drug Safety Evaluation Pfizer Global Research and Development 2800 Plymouth Road Ann Arbor, MI 48105 Phone: (734)-622-3631 FAX: (734)-622-3866 E-mail: joseph.saby-at-pfizer.com ---------------------------------------------------------------------------- -------------------------------------- Certain stains for light microscopy are not usable if you are trying to stain for lipid and increase resolution of the sectioned tissue. One has to adjust one's thinking and look only at how lipid can be preserved AND processed into a plastic, which involves solvents such as alcohol during the dehydration and infiltration steps. If you are an experienced EM person, you have probably noticed that lipid has been seen as a green color in your semithin secions. If you look at Toluidine Blue stained semithin sections, lipid remains green, and the nuclei and cytoplasm are blue. The resolution of epoxy and the green color allows for subcellular identification of lipid.
HOWEVER, the preservation of lipid by osmium can be greatly enhanced by p-phenylenediamine (use carefully, it can cause contact dermatitis and asthma). Osmicate normal time, then start you dehyration procedure 25%, 50%,then put tissue in a 1.0% p-phenylenediamine in 70% ethanol for 15-25 minutes, then finish dehydratation procedure as normal, up to 100% ethanol, into ethanol/propylene oxide, etc....Semithin (1-4u)sections without any additional staining will show lipid by light microscopy. If needed, counterstain nuclei with methylene blue stain or Toluidine Blue.
Glycol methacry. would not work, because none of the lipid will remain, due to the alcohols used in processing. Only osmication, and especially post- osmication treatment of tissue with p-phenylenediamine perserves and stablizes lipid so it doesn't wash out during dehyration steps.
Good Luck!
Karen Jensen, M.S. Associate Scientist and Project Manager University of Rochester Medical Center Electron Microscope Research Core Pathology and Lab. Med. Rochester, NY 14642 ---------------------------------------------------------------------------- ---------------------------------------------- Years ago I prepared a demonstration slide of testes for the Leydig cells that came from a conventional EM block, Glut/Cacod and Osmium. 1 micron section stained with Sudan black then Toluidine blue. Beautiful result but it was only a one-off, although something I'll try again if needed. Ian.
Dr. Ian Montgomery, West Medical Building, University of Glasgow, Glasgow, G12 8QQ. Tel: 0141 339 8855. Extn:6602. Fax: 0141 330 2923 e-mail: ian.montgomery-at-bio.gla.ac.uk ---------------------------------------------------------------------------- ---------------------------------- A question: is this really the case? Osmium binds across double-bonds of lipids, which is why it shows and preserves membranes, but it binds poorly or not all at to saturated lipids. So I wouldn't expect OsO4 to show fatty deposits if the fats are saturated. Oil Red O and Sudan Black, which more mix into the lipids and don't react with them, would show fat deposits that OsO4 doesn't, and are better fat stains.
Phil
} Agreed, there would be no need to do special fat stains if the tissue is } processed with osmium. In fact, this can be done for paraffin embedding as } well to show fat distribtion in some diseases with vastly better morphology } and localization than frozen sections. } } Tim Morken } CDC, Atlanta ---------------------------------------------------------------------------- --------------------------------------- Do you have any fixed, unprocessed tissues to work with?? If so, do frozens on them instead of your blocks. This is what all the ORO protocols that I have say to do, which makes sense when you consider that all processing will subject the tissues to lipid solvents, thus all or most of the lipids will have been removed and plastics are especially good lipid solvents.
Connie McManus ---------------------------------------------------------------------------- ---------------------------------------- The block was post fixed with osmium tetroxide/cacodylate then stained with Sudan black. It was only a wee trial just to see what happened. At the time I naively thought that the Sudan black must be binding to the osmium fixed fat. It was only a once of on a single slide. I stained another slide with Toluidine blue/Pyronine Y and it was just as lovely. Goodness knows what the answer was. At the time I used to try all sorts of staining combinations on semi-thin Glut/Osmium resin sections. Some were awful but others showed promise, unfortunately time didn't permit further investigation. The best staining for Glut/Osmium resin sections, in our hands was Toluidine Blue/Pyronine Y (Ito & Winchester 1963 J. Cell Biol) and a Polychrome technique (Pasyk, Bartok & Fabry 1989 Stain Technol 64 (3) 149. Ian.
X-Sender: uvsgc-at-trex2.oscs.montana.edu X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.6 (32)
MicroListers,
This is a timely topic for me. We are getting a new chiller for our two EM's and I'm reconsidering what to use in it. For over 20 years I've run them with tap water and a dusting of dichlorophene fungicide on the surface of the water in the tank, which powder slowly dissolves into the water. Have had no problems at all with corrosion, plugged up lines (well, only ONCE, when I'd failed to add fungicide after water change), so I'm inclined to keep doing that, pending manufacturer's recommendations when I get the new chiller.
However, other voices have advised using 10% propylene glycol (pure stuff, NOT as in automobile anti-freeze) in de-ionized water.
Would like to revisit the pro's and con's of each method.
Thanks for your advice, in advance,
Gib Ahlstrand
} Haixin, } I've found that using distilled or de-ionised water in the chiller in } addition to the measures already suggested will also greatly reduce the } growth of microorganisms by denying them nutrients. } } Regards, } Eric lachowski
-- Gib Ahlstrand Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)625-5754 FAX, giba-at-puccini.cdl.umn.edu http://www.cbs.umn.edu/ic/
Just out of curiousity. . . how much X-radiation penetrates the rubber stopper?
Marie
Dear Marie, The short answer is, "Nearly all of it." The flux of x-rays depends on such parameters as the location of the stopper with respect to things in the column which can scatter the incident beam, produce brehmsstrahlung, etc. The easiest thing to do is to take a counter and make the measurement. For x-rays, the appropriate detector is an ionization chamber, such as a hand-held QT-Pi; however, a Geiger counter can also be used to give a relative measurement between the original set-up and that with the stopper. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
Luminos http://www.lumijet.com/mono.htm also has quadtone inksets for many Epson printers. With any of the quadtone type ink systems, you will really need to dedicate the printer to printing only black and white. It is impractical to switch between color and monochrome ink sets as the printer needs to be flushed out before each switch.
I have seen some "fine art" photo samples printed with this ink system and the results are stunning.
George
George Laing National Graphic Supply v:(518) 438-8411 X3109 f:(518) 438-0940 email: scisales-at-ngscorp.com } } Tom, } } I have seen a retrofit for Epson printers that looks quite interesting at } } http://www.piezography.com/ } } I haven't actually seen it in action but it looks like it should be good } for B/W prints. The advertising samples on the web site look very } nice. Basically they replace the color cartridges with different } densities of black ink. By allowing gray inks, they reduce the amount of } dithering required. The product consists of the replacement cartridges and the } revised printer driver software. } } Has anyone actually tried this approach? } } Cheers, } Henk Colijn
I too have heard this tick from a company in long Island NY. I called, got sample prints, looked good, but I could tell better with my own images. It also might be cheaper since you can do continuous feed on some Epson products and buy ink in bulk.
-----Original Message----- } From: Hendrik O. Colijn [mailto:colijn.1-at-osu.edu] Sent: Friday, June 15, 2001 8:09 AM To: treese; Microscopy-at-sparc5.microscopy.com
Tom,
I have seen a retrofit for Epson printers that looks quite interesting at
http://www.piezography.com/
I haven't actually seen it in action but it looks like it should be good for B/W prints. The advertising samples on the web site look very nice. Basically they replace the color cartridges with different densities of black ink. By allowing gray inks, they reduce the amount of dithering required. The product consists of the replacement cartridges and the revised printer driver software.
Has anyone actually tried this approach?
Cheers, Henk Colijn
At 12:00 AM 6/15/01 -0500, treese wrote:
} We are going to replace our old Epson InkJet printer which produces nice } B+W prints of micrographs, when it works. I haven't kept up with Epson vs } HP for electron microscopists and wondered if a clear preference for one } of these, or maybe another mfg or technology has emerged. We want B+W } quality and reliability, and are willing to pay a bit more,and sacrifice } speed and color quality...Thanks...Tom Reese
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://web.ceof.ohio-state.edu Fools are pleased when they discover error. The wise are pleased when they discover truth.
Fortunately, the EDS port is to the rear of the column and pointing upward.
Dear Earl, Fortunate for the user; however, an often-neglected aspect of radiation shielding is that sufficiently energetic x-rays will penetrate floors and ceilings as well as walls, so check with the folks upstairs if you have a TEM--especially an IVEM or HVEM. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
I'd love some feed-back from someone who knows radiation because I've felt that applying TEM rules to SEMs is gross over-kill, especially with so many color monitors around. How often do you have your decelerating grid checked for proper operation? If it doesn't operate correctly, your exposure could be very dangerous, given the time that one sits in front of these things and their distance from your face.
If I'm way off base, I'd like to know and know why.
Dear Ken, I think I qualify as someone who knows radiation, so here goes. Yes, TEMs and SEMs are different, but the standards for stray radiation should be the same. It is a lot easier to meet the standards with a low-voltage, low-current machine, so the necessary shielding for a SEM would be less difficult than for a TEM, but there should still be less than the specified x-ray flux at the surface of the column. The Electron Microscopy Safety Handbook (2nd Ed., 1994), p 49 gives a standard set by the Radiation Control for Health and Safety Act of 1968 as 0.5 mR/hr at 5 cm from the surface of the column. I am not aware of any update of this standard, but if there is, the exposure allowed would surely be lower. A hand-held ionization chamber can be used to measure the radiation from both the microscope and the color monitors. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
Hi Michael, I believe that the article in Microscopy Today that you are looking for is "Preparation And Use Of Needles and Micropipets For Handling Very Small Particles" by Anna Teetsov of McCrone Research Institute. It is a 4-pager that I am faxing to you. I would be pleased to fax copies to anyone else on the list that would like a copy. Best to all, Don Grimes, Microscopy Today
-----Original Message----- } From: Roberson, Michael (M) [mailto:MRoberson-at-dow.com] Sent: Friday, June 15, 2001 8:07 AM To: 'Microscopy-at-MSA.Microscopy.Com'
Does anyone happen to know in what issue of microscopy Today the following information on sharpening W needles was found? (or how to make them) If so please send me a note at, MRoberson-at-dow,com
I don't know about the current crop of FE cathodes, but in the past an electrolytic cell was set up with the W needle as part of the circuit. The needle was slowly dipped and removed several times from the NaOH electrolyte. Essentially, it was the same technique as was (and is) used to produce micomanipulator needles. I think the set-up was shown and described well in a Microscopy Today article within the past 12 months.
Michael Roberson (517)636-8656 SMX Analytical Sciences Midland, MI 48667
I have some problems with our EM chillers. The water in EM cooling circulation is bad contaminated by algae, bacteria, maybe more microorganisms. Does somebody here know some water cleaner for chillers of EM? Appreciate for the help.
Dear Haixin, We use dichlorophene to prevent the growth of micro-organisms, but you may also need to remove those which are already in your system. The appropriate cleaner will depend on what the components of the system are made of. If you can disassemble and clean out the hoses, lenses, and chiller separately, you would probably be able to get things cleaner, but it could be a big effort. Cu++ is toxic to algae, and will not usually damage metals, so you might want to circulate a solution of CuSO4, pH adjusted to ~8 through the lenses if the lines are constricted within the lenses. Good luck. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
Whew! I am the only one in our EM lab. About 90 - 100 Kidney/surgical/autopsy specimens a year. About 30 or so research specimens a year. Not overworked here, but sure wish I had some backup, as vacations or sick days are a nightmare. I come in all hours, too.
Marilyn Howton EM lab Pathology Department WVUniv. Hospitals
Does anyone have a current email address for Sycon Instruments (makers of film thickness monitors)? Their web site is still up, but with a 1998 date, and the group listed as maintaining the site is apparently defunct. No email addresses are listed on the Sycon web site.
Thanks.
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)
I recently got rid of 2 Zeiss EM 9s2 microscopes and am interested in finding if there is anyone out there who is interested in buying 4489 EM film for a Zeiss 9s2? I have aluminum cassettes, steel cassettes and ~10 boxes of film I wish to sell.
Please respond via my email address!
Cheers! Ken ------------ Ken Tiekotter, Adjunct Professor The University of Portland Dept. of Biology 5000 N. Willamette Blvd. Portland, OR 97203
Fortunately, in my lab, the administration was upstairs so not held in high regard & the original question was for an SEM.
In addition, the rubber stopper solution was meant to be a temporary situation. Given that the maximum 25 - 30 KV X-rays need to penetrate several layers of aluminum foil as well as concrete floor I doubt that it poses a health risk. We had this situation at the last lab I worked & the Safety Department measured the X-ray output and gave it a clean bill of health. Not a good permanent situation but OK for the month or two.
Best Regards,
Earl
----- Original Message ----- } From: {"wft03-at-health.state.ny.us"-at-sparc5.microscopy.com} To: {microscopy-at-sparc5.microscopy.com} Sent: Friday, June 15, 2001 11:33 AM
Hello Microscopists,
We all work very hard at producing beautiful and relevant microscope images. Now is your chance to submit your finest images to the "Small World 2001" contest. This is the 26th year for the famous contest that results in a beautiful calendar showcasing the winners.
The closing date for submitting your images is June 29th, 2001. This can now be accomplished easily and quickly via the http://www.microscopyu.com/smallworld/ website.
There's still time to enter this famous optical photomicrography competition which is open to both professional and amateur microscopists; 35mm slide and/or digital entries of images taken with any kind of light microscope are accepted. Follow this link -
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to learn more about the contest, see the fabulous prizes available, obtain your "Small World Screen Saver" of last years winners and view some of the stunning images that have won in the past.
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Good Luck and Best Regards,
Stan Schwartz Manager, BioScience Dept. Nikon Instruments Inc. 1300 Walt Whitman Rd. Melville, NY 11747 Phone: 631-547-8529 Fax: 631-547-4033 email: sschwartz-at-nikon.net www.nikonusa.com Check out Nikon's Educational Website www.microscopyU.com
A nice plug for Tina's web pages from the NYTimes.
NYTimes 06-14-01
What You Can't See Microscopically
By SHELLY FREIERMAN
If you like to ponder questions on the scale of how many Web sites can fit on the head of a pin, there is a fine collection of electron microscope images to examine at MicroAngela, an online collection (www.pbrc.hawaii.edu/bemf/microangela). The Web site is maintained by Tina M. Carvalho, the supervisor at the Biological Electron Microscope Facility at the University of Hawaii in Manoa.
She has taken the images of tiny bugs, bacteria, mold, parasites and grains of sand, and colored them.
There is a description for each of the objects, which were collected from captured insects; from samples of ocean life, like tubeworms and coral; and even from mold growing on some Romano cheese in Ms. Carvalho's refrigerator.
As indicated earlier (5-13-2001), I am soliciting suggestions for symposia for the year 2002. So far we had very good response and I am confident that the program for 2002 will be excellent. However, before the program will be finalized, I would like to once more ask for your idea for a great topic in any of the areas of
"Physical Sciences" "Biological Sciences" "Advances in Instrumentation and Techniques"
The deadline for suggestions is June 30th 2001. Please contact me with your suggestions before June 30th under the following address:
mm2002-at-ornl.gov
Please forward this request also to your colleagues. Thank you for your support.
With best regards,
Edgar Voelkl Program Chair M&M 2002 --
___________________________
Dr. Edgar Voelkl Program Chair M&M 2002
Oak Ridge National Laboratory P.O. Box 2008 Bldg 4515 Oak Ridge, TN 37830-6064
I've had Epson and Lexmark printers in the past, but can't say enough about the HP Photosmart 1215 I've recently purchased. I've always had paper feed problems with the Epson and Lexmark printers I've used, and noticed that the HP I used on my home computer didn't have those problems. I decided to try an HP printer for my business computer and chose this one. For both color and grayscale images, I find it to be the closest to photographic quality that I've tried (I used to do professional photography in the arts field and did all of my own developing and printing).
The only caveat is that grayscale images should be printed in color mode - black only prints in 600 x 600 resolution while color prints in 2400 x 1200.
This printer includes a separate 4 x 6 inch paper feed tray that is excellent for small prints.
On Friday, June 15, 2001 12:00 AM, treese [SMTP:treese-at-marinebio.mbl.edu] wrote: } ------------------------------------------------------------------------ } } } We are going to replace our old Epson InkJet printer which produces } nice B+W prints of micrographs, when it works. I haven't kept up } with Epson vs HP for electron microscopists and wondered if a clear } preference for one of these, or maybe another mfg or technology has } emerged. We want B+W quality and reliability, and are willing to pay } a bit more,and sacrifice speed and color quality...Thanks...Tom Reese }
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174 voice 630.513.7093 fax 630.513.7092
} } I've had Epson and Lexmark printers in the past, but can't say } enough about the HP Photosmart 1215 I've recently purchased. I've } always had paper feed problems with the Epson and Lexmark printers } I've used, and noticed that the HP I used on my home computer didn't } have those problems. I decided to try an HP printer for my business } computer and chose this one.
Yeah, maybe, but boy, don't extend this to all HP printers.
I used to have at home an HP 400 inkjet, the cheapie, which kept on giving paper-feed problems until I donated it to the repair shop rather than fix it a second time, now, at work, I have a Laserjet 6L that often feeds thru up to 10 sheets at a time!
I can't load up the input stack, have to insert each sheet to be printed, one at a time.
It seems to me that the HP problem is that, in order to keep the footprint small, the paper has to turn almost 180 degrees in a tight circle.
The 6L has at the moment ANOTHER paper jam that I feel too angry to fix, it's been jammed now for two days but I'd rather print to the network printer and walk down the hall to retreive the printout than pull it apart AGAIN.
If I had an outside window I'd be tempted to throw the *!-at-#$% printer through it....................................
cheers
rtch
Dr 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
Thank you all very much for the reply of my question. I think I figoured out how to deal the problem.
Haixn Xu
Biological Sciences/UMBC Baltimore Maryland
On Fri, 15 Jun 2001, Beauregard wrote:
} Hi, } } Chloramine-T can be used to kill or prevent the bacteria but it will cause } corrosion problems with some instruments over time. This is especially } true of } stainless steel fittings where pit corrosion can become a very serious } problem and it was for us. The manufacturer recommended using pure } distilled water in our TEM. We didn't do what the service people said } because we never had the problem with our other instruments in our wing of } the building with C-T. We fixed our problem by using some manufacturer } supplied brass fittings. We also switched to de-ionized water. It worked } fine. } } Anyway, you could kill them with C-T, flush the system completely and then } switch to pure distilled or de-ionized water. Check with your serviceman } for the recommended material to use. } } After flushing, you can monitor the chloride level and the available } chlorine to be sure you have removed the chloramine-T. You can use a } swimming poll test kit for available chlorine and 0.1N silver nitrate } solution for testing for chloride ions. } } After flushing and switching to pure distilled water our corrosion problem } has not surfaced in over 8 years. We have about 10 instruments on our } recirculator system. } } One further note. You can get a blue-green to green color in the water and } inside the water lines. This can be copper ions or a green carbonate, } like copper carbonate. Check any transparent PE or PP lines to see if they } have a } coating inside them of green copper carbonate. I did this on our Edwards } 306 evaporator because it had transparent water lines on the DP. This } color was thought to be algae but it was not algae alone. You can scrape } off some of the coating and under a microscope apply hydrochloric acid. } Look for any bubble generation to confirm the possible presence of copper } carbonate. Of course, the bubbles could be another material like calcium } carbonate but it would not ordinarily be greenish. Remove one of the lines } if you can, add dilute HCl, wait for the line to 'clear' and run the acidic } liquid into a test tube. If it's blue, you probably have copper ions and } most likely copper carbonate deposits. We did. The copper can be } confirmed by atomic absorption if necessary. } } Hope this helps. } } Paul Beauregard } Senior Research Associate } PPG Industries } Monroeville, PA 15601 } } } } At 05:22 PM 6/14/01 -0400, Haixin Xu wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 lister, } } } } I have some problems with our EM chillers. The water in EM cooling } } circulation is bad contaminated by algae, bacteria, maybe more } } microorganisms. Does somebody here know some water cleaner for chillers of } } EM? Appreciate for the help. } } } } Haixin Xu } } } } Biological Sciences } } University of Maryland, Baltimore County } } Baltimore, Maryland } } } } } } } } } }
Hi Don, I would appreciate a copy, if you wouldn't mind. I have a set-up of my own, but it is quite the "homemade" variety.
Using this set-up, I have also produce fine blades by using flattened W wire rather than the rods. Carefully dipping them in a particular way produces a blade on one side and leaves the other side as a support for strength. Works well when slicing nematodes. John Shields EM Lab Univ. of GA, Athens
On 15 Jun 2001, at 15:23, Don Grimes wrote:
} ---------------------------------------------------------------------- } -- The Microscopy ListServer -- Sponsor: The Microscopy Society of
} Hi Michael, } I believe that the article in Microscopy Today that you are looking } for is "Preparation And Use Of Needles and Micropipets For Handling } Very Small Particles" by Anna Teetsov of McCrone Research Institute. } It is a 4-pager that I am faxing to you. I would be pleased to fax } copies to anyone else on the list that would like a copy. Best to all, } Don Grimes, Microscopy Today
Many thanks. I now have the contact information for Sycon Instruments.
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)
} -----Original Message----- } From: "wft03-at-health.state.ny.us"-at-sparc5.microscopy.com } [mailto:"wft03-at-health.state.ny.us"-at-sparc5.microscopy.com] } Sent: Friday, June 15, 2001 3:00 PM } To: microscopy-at-sparc5.microscopy.com } Subject: Re: water cleaner for EM chiller } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } 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 problems with our EM chillers. The water in EM cooling } circulation is bad contaminated by algae, bacteria, maybe more } microorganisms. Does somebody here know some water cleaner } for chillers of } EM? Appreciate for the help. } } Dear Haixin, } We use dichlorophene to prevent the growth of } micro-organisms, but you } may also need to remove those which are already in your system. The } appropriate cleaner will depend on what the components of the } system are } made of. If you can disassemble and clean out the hoses, lenses, and } chiller separately, you would probably be able to get things } cleaner, but } it could be a big effort. Cu++ is toxic to algae, and will } not usually } damage metals, so you might want to circulate a solution of CuSO4, pH } adjusted to ~8 through the lenses if the lines are } constricted within the } lenses. Good luck. } Yours, } } Bill Tivol } Wadsworth Center } Albany NY } (518) 473-7399 WFT02-at-health.state.ny.us } }
We recently acquired an older Link EDS spectrometer and would like to use it on an old ESEM. But we will need the pulse processor and whatever else to run the detector. I have heard of people mixing brands and we have some other EDS systems, but our old Link is presently installed on another scope. Does anyone have an old Link giveaway or have any thoughts on compatibility among the EDS systems? Thanks for any thoughts or assistance. Mike ******************************************************************** Michael L. Boucher Sr. mboucher-at-tc.umn.edu Lab Manager Rm 55 Office Ph 612-624-6590 I.T. Characterization Facility Rm 12 Main Off Ph 612-626-7594 University of MN Fax 612-625-5368 12 Shepherd Labs 100 Union Street S.E. Minneapolis, MN 55455 http://resolution.umn.edu ********************************************************************
This is an update on a possible way of quantifying the orientation of actin filaments within endothelial cells in culture.
I would like any comments on whether this seems reasonable or if there are potential problems.
The fluorenscent labelled cells are captured and saved as grayscale images. A highpass filter is used to enhance the filaments. The filaments are thresholded and skeletonized. then the branch points are selected and deleted. This creates many short linear segments whos "moment angle" or orientation can be measured. There are about a 500 - 1000 measurements per cell. The histogram of the angles have peaks cooresponding to the dominant directions of filaments. I am hoping that control and experimental groups of cells will have appropriatly different histograms.
I have a DURST S-45 enlarger that has not been used for a long time. The mirror is heavily dusted. I want to clean it or want it cleaned. But I am afraid to damage it if clean it not properly and I do not know where and who can do the job for me. I am located in Baltimore. I would very much appreciate your help.
Haixin Xu
Biological Sciences University of Maryland, Baltimore County Baltimore, MD 21250
We are looking for a used Ultracut T that is in good/excellent condition. Please contact Kathleen Greer or Lauren Simmerman if you know of one, -at- Univ. of Nebraska Med. Cen. 402-559-7729 or KathleenGreer-at-nhsnet.org
Paper jams mostly related to the paper quality (if printer itself is OK). You may ask manufacturer which particular type of paper is better for printer or just try different brands. For laser printers it should be mentioned on the paper that this is for laser. Paper for laser and ink -jets are opposite: glossy and "full-body" for laser and more porous (to adsorb inks) for ink-jets. It also important to set correct paper-thickness on the printer. They usually has thick-medium-thin settings. It's mechanical switch, which presents on most models.
We do have some LaserJet B&W printer running for 5 years without any serious problem with paper (heavy loaded, it serves as a network printer for whole Department). We just bought a new Tektronix Color-laser last year and it had jam problems near every day. We did change the paper and it works very good now. Same - for ink-jets.
Sergey
At 05:49 PM 6/18/01 +0000, 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 have a DURST S-45 enlarger that has not been used for a long time. The mirror is heavily dusted. I want to clean it or want it cleaned. But I am afraid to damage it if clean it not properly and I do not know where and who can do the job for me. I am located in Baltimore. I would very much appreciate your help.
Dear Haixin, I would first remove the mirror (if possible), then gently blow as much of the dust off it as possible. The concern, of course, is that some of the dust may consist of mineral particles capable of scratching the mirror. These should be somewhat denser and more easily removed in an air stream than some of the other dust components. I would then rinse the mirror with a stream of water and/or dunk it into a warm detergent solution, then rinse carefully in distilled water followed by alcohol and/or acetone. In none of these steps would I wipe the mirror even with a very soft cloth. After this, I'd check to see how much dust is left. If none, I'd clean the mirror with optic-wipe cloth, if little, I'd re-rinse and do so until as much of the dust was removed as possible. I'd then clean a small area with optic wipe cloth and inspect for scratches, if none, then I'd clean the rest. Good luck. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
Reply to Haixin Xu: Cleaning is done the same as it would be for a front surfaced telescope mirror as follows: 1) Gently let pure water (distilled or deionized) flow over the mirror. 2) dip several "Q-tip" swabs in a 1:100 diluted solution of Triton X 100. 3) allowing only the weight of the wet"Q-tips" to be the downward force gently pass them over the mirror and rinse with more pure water and allow to dry dust-free. If only traces of dust settle on the mirror you can blow them off gently with canned gas or a rubber bulb duster.
Below is the result of your feedback form. It was submitted by (johnsond-at-ns.neiu.k12.pa.us) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, June 18, 2001 at 13:44:52 ---------------------------------------------------------------------------
Email: johnsond-at-ns.neiu.k12.pa.us Name: Debbie Johnson
Organization: Valley View School District
Education: 6-8th Grade Middle School
Location: Archbald. PA USA
Question: What power of magnification would we need to be able to see