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:
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} 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 bacteria cells? They are not very clear using the standard classroom scopes.(400x) Are there microscopes available for classroom use that have more magnification?
I was wondering if anyone out there had any information on a morphometry or image analysis user group that I could get in contact with. Also does anyone have an interest in a confocal microscope MRC 600 that we are going to decommission. I would appreciate any information anyone could offer on any of the above subjects.
Hi Listers, Is there anyone out there who can help with this request? Please reply to plant-tc-at-tc.umn.edu. Thanks, Priscilla *****************************************************************************
----- Original Message ----- } From: Abhay Shendye To: plant-tc-at-tc.umn.edu Sent: Sunday, June 17, 2001 12:05 PM
Hi Mike,
We have had this done the other way (Link have run another maker's detector). They had to change the preamp onwards and ensure that they got the pinout and bias voltage to the crystal correct (obviously). We have also had upgrades and they have always changed the preamp with the pulse processor. What about contacting your local Oxford Instruments rep. to see what they have from recent part exchanges or upgrades?
Good luck, Ron
} 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 } ******************************************************************** } } } } } } }
---------------------- 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
For EM guys it's not big deal to restore even scratched mirror. Only limitation is the size of your vacuum evaporator's vacuum chamber.
1. remove old coating (very likely, it was aluminum, if so, use 1N NaON); 2 rinse in d-H2O and dry in clean place; 3. evaporate a new layer of Al (or Cr) using your vacuum evaporator.
The trick: never touch working surface before and after.
Sergey
P.S. Organic solvents may leave the traces even if they are very pure (alcohol may have the trace of CuSO4 used for dehydratation or something else). Last step in cleaning should be dd-H2O and drying in very clean place. Who wear glasses half of life knows, that any organic solvents leave the traces except detergent and then d-H2O. At least, it's my experience.
At 05:24 PM 6/18/01 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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Dear Listers, I want to take the microstructure of 0.75mm wire, which is used by dentist. Can anyone can guide for the sample preparation of wire for tem.
Arti Harle Regional Sophisticated Instrumentation Center Indian Institute of Technolgy-Bombay Powai, Mumbai- 400076. India. Phone No. 91-22-5767691 Extn. 7694 email: aartih-at-usa.net
____________________________________________________________________ Get free email and a permanent address at http://www.netaddress.com/?N=1
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id IAA26642 for dist-Microscopy; Tue, 19 Jun 2001 08:29:17 -0500 (CDT) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id IAA26639 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Tue, 19 Jun 2001 08:28:47 -0500 (CDT) Received: from mercury.uwe.ac.uk (mercury.uwe.ac.uk [164.11.132.23]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id IAA26632 for {microscopy-at-sparc5.microscopy.com} ; Tue, 19 Jun 2001 08:28:35 -0500 (CDT) Received: from fas655.uwe.ac.uk ([164.11.205.145]) by mercury.uwe.ac.uk (2.0.4/SMS 2.0.4-devel) with SMTP id OAA12760; Tue, 19 Jun 2001 14:25:47 +0100 (BST)
If we assume your bacteria are 0.000001m long then if you magnify them x400 they will appear 0.0004m long ie 0.4mm.
So if you would like them to appear, say 400mm long, you would magnify by x40,000.
There are special microscopes called electron microscopes which can give you this magnification. We look at bacteria at x1,000 - x94,000. Your local university will have some - why not check their web site?
Dave
On Tue, 19 Jun 2001 01:55:56 -0500 johnsond-at-ns.neiu.k12.pa.us wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Below is the result of your feedback form. 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 } bacteria cells? They are not very clear using the standard classroom } scopes.(400x) Are there microscopes available for classroom use that } have more magnification? } } --------------------------------------------------------------------------- }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
To answer the question of compatability. The "older Link EDX" is probably an optical reset FET detector if it is older than 1988. If it is a transistor reset FET ( PentaFet) then it would have an attribute number on the Dewar label. Therefore when you are looking for a compatable analyzer you will need to know which FET is in the detector. The bigger problem is that Link used a software based Restore signal to reset the FET therefore only a Link analyzer system will operate the detector. The only present EDX company using a similar software based Restore signal is Oxford ( Which bought Link Analytical back in the early 1990's ) The pre-amp on the Link detector could be changed so that the Restore signal is generated by the pre-amp. This would allow almost any analyzer system to operate the detector but some compatability issues would still remain.( Inhibit and Gain ) I hope this answers your question. Kind Regards, Jim Nicolino X-Ray Optics / AAT 1816 St. Johns Bluff Road Suite 305 Jacksonville, FL 32246 PH 904 646-3069 FAX 904 646-3131 E-mail JNicolino-at-xraydetectors.com
"Michael L. Boucher" wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 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 } ********************************************************************
Did you try Hexane to clean-up EM parts? Hexane, HPLC grade is very nice for that. As a matter of fact, many organic solvents even very pure has some additives, which is not listed. It includes the traces of compounds, which were used during purification process. Very usual contamination - sort of mineral oil, which comes when you store your stuff in the plastic bottle (or it was stored somewhere before it comes to you in glass bottle) - manufacturers use it in press-forms when they make plastic bottles. Any plastic (even teflon) has some "plastifiers" to make plastic more flexible. Those compounds does not chemically bonded to the plastic matrix, so they erode slowly into stored solution. I had a deal with very precise mirrors from analytical UV centrifuge. Those mirrors has precision comparable with best telescope mirrors. I find, that many organic solvents leave traces even if it never happen before on other surface. The mirror surface, looks like so perfect and reflect easy very tiny imperfection. Of coarse, I am talking about mirrors with reflecting coating up. As for fingerprints - any organic solvent should work, I agree with you. Speaking honestly, in the practical life my precautions may be not necessary. I was talking in general, that scratches on the mirror is not a problem for serious electron microscopist.
Thanks for your reply and have a great day.
Sergey.
At 10:20 AM 6/19/01 -0400, you wrote:
} P.S. Organic solvents may leave the traces even if they are very pure } (alcohol may have the trace of CuSO4 used for dehydratation or something } else). Last step in cleaning should be dd-H2O and drying in very clean } place. Who wear glasses half of life knows, that any organic solvents } leave the traces except detergent and then d-H2O. At least, it's my } experience. } } Dear Sergey, } d-H2O should leave no residue, but I've seen problems using it as the } final rinse--probably because it takes a long time to dry, and our air may } not be that clean. We use acetone as the final rinse on all the metal } parts of the microscope that go in the vacuum, because it's easier to pump } down if there is no residual H2O. We also use 95% EtOH to clean the glass } plates in the Perkin Elmer microdensitometer--it is the best solvent for } fingerprints--and we do not rinse with H2O afterward. I haven't seen any } residue problems from either of these solvents. } Yours, } Bill
Hello fellow microscopists, What are the latest advances in confocal microscopy systems? Any with Dapi or Hoechst detection? What are the pros and cons of PMT vs Camera acquisitions? Aperture, Fixed slits or spinning discs? We would appreciate any input.
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Debbie -
Yes, there are microscopes that will provide an excellent 1000x image. BUT - they are quite a bit more sophisticated (and expensive) than the scope that you are using, and they require careful setup to produce a good image. It's possible that there is a microscopist who reads our ListServer who is near Archibald who would be willing to visit your class to do a demonstration. If you are interested, please write again.
You can find a lot of information about bacteria at the American Society for Microbiology's educational website, http://www.asmusa.org/edusrc/
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 am trying (rather unsuccessfully) to print grayscale images on a large format color printer. The printer does a superb job with color images but generates truly unpredictable, bizarre renderings with grayscale (SEM, TEM) images (greenish/brownish/pinkish tones).
The printer has the option to print using black ink only but, obviously, no color can be used elsewhere. I have made just about every color management adjustment imaginable but have come to the conclusion that it is not possible to print accurate colors and grayscale simultaneously. Is that so?
I need professional help................. (in many ways).
Thank you.
JB
-- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ##############################################################
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I am working on a bacterial protein which are very unstable polymerisation and depolymerization is within very short time. I want to do the Atomic Force microscopy for these proteins What is sample preparation for AFM And how can I stabilise the protein.
Regards Arti Harle
Ms. Arti Harle Regional Sophisticated Instrumentation Center Indian Institute of Technology-Bombay (IIT-B) Powai, Mumbai - 400076. India Phone : 91-22-5767691 Extn 7694 91-22-5720109/ 5721039 (Resi) E-mail : aartih-at-usa.net
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Hi, all,
This question has come up for us at a good time, with this ongoing discussion of printers.
We recently purchased an Epson 880 ink jet printer for our EM/LM lab, and are very happy with it. The black and white prints look excellent. We have tried a number of papers, and find that the Epson "Photo Quality Glossy Paper" and "Photo Paper" give the best results for our purposes.
We are wondering how to go about making our prints more permanent from these printers. If the prints have just been printed and aren't dry yet, or if they are dry and are in contact with any moisture, they smudge quite easily. Is there any coating - lamination or spray, or something else which we can use to protect the surface?
In connection with this, we have also found, especially when we have printed b/w prints, that if we leave them in a room with full spectrum fluorescent lights for a few days, that they begin to change colour, and start to take on a reddish tinge. The instructions that came with the paper say to not leave prints exposed to sunlight - but we're talking about room lights, here! Has anyone else found this happens to them, and is there any way of preventing/minimizing this effect without storing prints in a light tight bag????
Any help or suggestions would be much appreciated. As always, thanks for your help in advance.
Regards,
Paula.
Paula Allan-Wojtas Research Scientist - Food Microstructure Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
Impact Analytical has an opening for a microscopist. Interested candidates are invited to reply.
Job Description for Microscopist IMPACT Analytical seeks an exceptional technical professional to become a member of our Microscopy group. The candidate will be expected to provide outstanding service to our customers on projects in the areas of microtomy, SEM, TEM, and AFM. The candidate will be responsible for the development of approaches to solve customer problems, as well as for projects from the time that samples are received until the final report is written. This will include communication with the customer with regards to clarifying project goals and methods, conducting the analysis, updating the status, generating the report, and following-up on customer requests. Experience in the microscopy of plastics is a plus. The ideal candidate will have at a minimum, a B.S. degree and 2 to 5 years of relevant experience. The candidate should have a sense of urgency, strong organizational skills, be detail oriented, and have very good laboratory technique. The candidate should also have excellent oral, verbal, and interpersonal skills, as well as familiarity with Microsoft based software. Please respond by e-mail to Ms. Cherie Hutter {mailto:hutter-at-mmi.org} or by mail to Ms. Cherie Hutter, Impact Analytical, Job Number 06-01, 1910 West Saint Andrews Road, Midland, MI 48640 to arrive no later than 8 July, 2001.
Kevin Battjes Impact Analytical Michigan Molecular Institute 1910 W. St Andrews Road Midland MI 48640
We've had the same problem on our large format printer. I don't know what the answer is either. I haven't gotten nearly as far as you have in terms of troubleshooting, but my next step would be to call the manufacturer (Epson, in our case). Did your manufacturer have any insight?
Angela V. Klaus
Director, Core Imaging Facility American Museum of Natural History Central Park West at 79th Street New York, NY 10024 USA
Email: avklaus-at-amnh.org Tel: 212-769-5977
On Tue, 19 Jun 2001, John J. Bozzola wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I am trying (rather unsuccessfully) to print grayscale images on a } large format color printer. The printer does a superb job with color } images but generates truly unpredictable, bizarre renderings with } grayscale (SEM, TEM) images (greenish/brownish/pinkish tones). } } The printer has the option to print using black ink only but, } obviously, no color can be used elsewhere. I have made just about } every color management adjustment imaginable but have come to the } conclusion that it is not possible to print accurate colors and } grayscale simultaneously. Is that so? } } I need professional help................. (in many ways). } } Thank you. } } JB } } -- } ############################################################## } John J. Bozzola, Ph.D., Director } I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) } 750 Communications Drive - MC 4402 } Southern Illinois University } Carbondale, IL 62901 U.S.A. } Phone: 618-453-3730 } Fax: 618-453-2665 } Email: bozzola-at-siu.edu } Web: http://www.siu.edu/departments/shops/cem.html } ############################################################## } }
I too have noticed that the cyan ink that Epson uses is not very stable even under fluorescent lights. However, enclosing a print in a polyethylene sheet protector seems to improve the stability quite a bit.
Cheers, Henk
At 07:47 AM 6/20/01 -0400, Paula Allan-Wojtas wrote:
} Hi, all, } } This question has come up for us at a good time, with this ongoing } discussion of printers. } } We recently purchased an Epson 880 ink jet printer for our EM/LM lab, and } are very happy with it. The black and white prints look excellent. We have } tried a number of papers, and find that the Epson "Photo Quality Glossy } Paper" and "Photo Paper" give the best results for our purposes. } } We are wondering how to go about making our prints more permanent from } these printers. If the prints have just been printed and aren't dry yet, } or if they are dry and are in contact with any moisture, they smudge quite } easily. Is there any coating - lamination or spray, or something else } which we can use to protect the surface? } } In connection with this, we have also found, especially when we have } printed b/w prints, that if we leave them in a room with full spectrum } fluorescent lights for a few days, that they begin to change colour, and } start to take on a reddish tinge. The instructions that came with the } paper say to not leave prints exposed to sunlight - but we're talking } about room lights, here! Has anyone else found this happens to them, and } is there any way of preventing/minimizing this effect without storing } prints in a light tight bag???? } } Any help or suggestions would be much appreciated. As always, thanks for } your help in advance. } } Regards, } } Paula. } } Paula Allan-Wojtas } Research Scientist - Food Microstructure } Agriculture and Agri-Food Canada } Atlantic Food and Horticulture Research Centre } Kentville, Nova Scotia Canada B4N 1J5 } } Tel: (902) 679-5566 } FAX: (902) 679-2311 } } email: allanwojtasp-at-em.agr.ca
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.
Title: Optical Microscopy and Imaging in the Biomedical Sciences
When: October 10 - October 19, 2001
Where: Marine Biology Laboratory, Woods Hole, MA, USA
Tuition: $2150 (Includes room and board, text, handouts, supplies)
Application Deadline: July 25, 2001
Admission application and information: Carol Hamel, Admissions Coordinator Marine Biological Laboratory 7 MBL Street Woods Hole, MA 02543-1015 (508) 289-7401 Internet: admissions-at-mbl.edu WWW: http://www.mbl.edu (Application forms available via Adobe Acrobat)
Course Director: Colin S. Izzard, State University of New York -at- Albany Phone: [518] 442 - 4367 EMail: csizzard-at-csc.albany.edu
Course Description:
For Whom: Designed primarily for research scientists, physicians, postdoctoral trainees and advanced graduate students in animal, plant, medical and material sciences. Non-biologists seeking a comprehensive introduction to microscopy and video-imaging will benefit greatly from this course as well. There are no specific prerequisites, but an understanding of the basic principles of optics is desirable. Limited to 24 students.
The eight day course consists of lectures, laboratory demonstrations, exercises and discussions that will enable the participant to obtain and interpret microscope images of high quality, to perform quantitative optical
measurements, and to produce photographic and video records for documentation and analysis.
Topics to be covered include: principles of microscope design and image formation bright field, dark field, phase contrast, polarized light, differential interference contrast, interference reflection, and fluorescence microscopy confocal scanning microscopy, multiphoton excitation fluorescence microscopy and image deconvolution digital image restoration and 3-D reconstruction video imaging, recording, enhancement, and intensification analog and digital image processing and analysis fluorescent probes and ratiometric-imaging laser tweezers and laser scissors
Applications to live cells will be emphasized; other specimens will be covered as well.
Students will have direct hands-on experience with state-of-the-art microscopes, video cameras, recorders and image processing equipment provided by major optical and electronics companies. Instruction will be provided by experienced staff from universities and industry.
Students are encouraged to bring their own biological (primary cultures, cell lines, etc.) and material specimens and to discuss individual
I am trying to repair our Sorvall critical point dryer. I would like to replace the needle valve assemblies as they are starting to leak. Does anyone know if parts are available for this system? If not, we will have to get it replaced.
At 07:37 PM 6/19/01 -0500, John J. Bozzola wrote: } I am trying (rather unsuccessfully) to print grayscale images on a large format color printer. The printer does a superb job with color images but generates truly unpredictable, bizarre renderings with grayscale (SEM, TEM) images (greenish/brownish/pinkish tones). } The printer has the option to print using black ink only but, obviously, no color can be used elsewhere. I have made just about every color management adjustment imaginable but have come to the conclusion that it is not possible to print accurate colors and grayscale simultaneously. Is that so?
Which printer, which operating system, and which program is doing the printing? When you say "color management" what exactly do you mean that you tried?
Offhand, this sounds more like a function of the printer driver and the program driving it. The program you're using to print may only be capable of sending down grey or color at a time. When it sends color, the grey gets treated (and color-dithered) as color. In pure grey mode, no color dithering is introduced.
A more sophisticated printing program, such as a desktop publishing program like Quark, may be capable of driving the printer in a smarter fashion.
Hey, I hear you, been there and done that! (HP - oi!). Finally, in desperation I took a sheet of fine emery or sanding cloth to the rubber rollers that move the paper through the printer - it worked! Try it, be patient and do every roller you can get to. I've had to do this more than once, but not often.I think the rollers pick up a "shine" from paper coatings. Let me know what happens.
Chin up, man!
Gib
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/
} } } } 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 } }
} I am trying (rather unsuccessfully) to print grayscale images on a } large format color printer. The printer does a superb job with color } images but generates truly unpredictable, bizarre renderings with } grayscale (SEM, TEM) images (greenish/brownish/pinkish tones). I think this is why color printing is CMYK and not just CMY. We typically try not to allow the printer to print greyscale images with the color setting on because of the off-tones. I think there really is no way to achieve perfect grey with the color cartridges. I wonder if it would be possible to trick the computer into printing only the greyscale images with the black cartridge and everything else in color. The other alternative would be to do a "two-pass" print with one print through on the grey images, and then reprint again with color. This would be VERY difficult on a poster printer, but a little easier with one of those smaller feedthrough inkjet printers. } } The printer has the option to print using black ink only but, } obviously, no color can be used elsewhere. I have made just about } every color management adjustment imaginable but have come to the } conclusion that it is not possible to print accurate colors and } grayscale simultaneously. Is that so? My only other suggestion is to continue with those color management solutions--do you have all the color profiles configured from monitor to printer? You've adjusted the file to CMYK gamut from RGB before printing? The only other suggestion I can make(short of having the full colorimeter and spectrophotometer calibration) is to make sure that the CMYK color settings on the individual greyscale images are all manually adjusted to absolute neutral.
I am a long time Tem person starting a new job doing both TEM and LSCM. Its very exciting learning LSCM but I need some suggestions for ordering probes. Our lab has a Kr/Ar laser and will be doing neuropathology research, mostly with fixed human brain samples, but also tissue culture cells both live and fixed. A vibratome is in our future. I was thinking of ordering alexa488nm-phalloiden, alexa568- streptavidin, and syto60 from molecular probes to start getting used to the scope and the lasersharp bio-rad software. The immunohistotech in the lab uses an ABC dtection system with Dab. The first probe should contrast the cytoskeleton well(stain F-actin), the second probe will label any primary Ab, and the third will bind to DNA and contrast the nucleus. Will using these three probes together be a good starting point to becoming adept at labeling and producing high quality images on the LSCM including optical sectioning and 3D (and 4D sometime in the future)? I welcome suggestions Thank you
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id NAA00134 for dist-Microscopy; Wed, 20 Jun 2001 13:24:18 -0500 (CDT) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id NAA00126 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Wed, 20 Jun 2001 13:23:48 -0500 (CDT) Received: from gate.pnl.gov (gate.pnl.gov [130.20.64.137]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id NAA00118 for {Microscopy-at-sparc5.microscopy.com} ; Wed, 20 Jun 2001 13:23:35 -0500 (CDT) Received: from vscan2.pnl.gov ([130.20.64.141]) by pnl.gov (PMDF V5.2-33 #42505) with SMTP id {01K4ZKUULF5E8WWCQB-at-pnl.gov} for Microscopy-at-sparc5.microscopy.com; Wed, 20 Jun 2001 11:20:30 PDT Received: from 130.20.128.21 by vscan2.pnl.gov (InterScan E-Mail VirusWall NT) ; Wed, 20 Jun 2001 11:20:28 -0700 (Pacific Daylight Time) Received: by PNLMSE1.pnl.gov with Internet Mail Service (5.5.2650.21) id {NHWFW2AV} ; Wed, 20 Jun 2001 11:20:27 -0700
John,
we used to have the same problem - the B&W images would come out pinkish. We contacted HP Software Support about a year ago, and they pointed out that there isn't any way to isolate a specific image and make it grayscale, leaving the others in color. They suggested making the image a gray scale before importing into the document. So far, when staff have copied the image and pasted it right into the poster (a PowerPoint slide - enlarged to the size of the poster or half the size of the poster) - it seems to work. However, there were some special images that a few people had that didn't work. Good luck,
Alice.
Alice Dohnalkova Dep. of Environmental Microbiology Battelle, Pacific Northwest National Laboratory MS P7-50 Richland, WA 99352 tel. (509) 372-0692 fax (509) 376-1321
-----Original Message----- } From: John J. Bozzola [SMTP:bozzola-at-siu.edu] Sent: Tuesday, June 19, 2001 5:37 PM To: Microscopy-at-sparc5.microscopy.com
I am trying (rather unsuccessfully) to print grayscale images on a large format color printer. The printer does a superb job with color images but generates truly unpredictable, bizarre renderings with grayscale (SEM, TEM) images (greenish/brownish/pinkish tones).
The printer has the option to print using black ink only but, obviously, no color can be used elsewhere. I have made just about every color management adjustment imaginable but have come to the conclusion that it is not possible to print accurate colors and grayscale simultaneously. Is that so?
I need professional help................. (in many ways).
Thank you.
JB
-- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ##############################################################
-----Original Message----- } From: John J. Bozzola [SMTP:bozzola-at-siu.edu] Sent: Tuesday, June 19, 2001 5:37 PM To: Microscopy-at-sparc5.microscopy.com
I am trying (rather unsuccessfully) to print grayscale images on a large format color printer. The printer does a superb job with color images but generates truly unpredictable, bizarre renderings with grayscale (SEM, TEM) images (greenish/brownish/pinkish tones).
The printer has the option to print using black ink only but, obviously, no color can be used elsewhere. I have made just about every color management adjustment imaginable but have come to the conclusion that it is not possible to print accurate colors and grayscale simultaneously. Is that so?
I need professional help................. (in many ways).
Thank you.
JB
-- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ##############################################################
Luminos http://www.lumijet.com has a spray called "Imagesheild" to protect prints. $18/ can. Quoting their website: "LUMIJET IMAGESHIELD will become your favorite print protector. It’s a convenient ozone-friendly low-odor aerosol spray that significantly improves wet-fastness. In addition to protecting delicate inkjet images from moisture, it also shields images from fingerprints and harmful UV rays. This unique product will not only help extend useful print life, it is totally transparent; preserving the original print surface characteristics. From dead matte to glossy, or from canvas to silver foil, you’ll never know it’s there."
} We are wondering how to go about making our prints more permanent } from these printers. If the prints have just been printed and } aren't dry yet, or if they are dry and are in contact with any } moisture, they smudge quite easily. Is there any coating - } lamination or spray, or something else which we can use to } protect the surface?
There were recently (last 6-8 months) some issues with color shift on some Epson papers. Epson determined the shift was from high concentrations of ozone, and not due to exposure to light. Epson recommends Heavyweight Matte or Epson Photo paper for maximum lightfastness.
George
George Laing National Graphic Supply v:(800) 223-7130 X3109 f:(800) 832-2205 email: scisales-at-ngscorp.com } } In connection with this, we have also found, especially when we } have printed b/w prints, that if we leave them in a room with } full spectrum fluorescent lights for a few days, that they begin } to change colour, and start to take on a reddish tinge. The } instructions that came with the paper say to not leave prints } exposed to sunlight - but we're talking about room lights, here! } Has anyone else found this happens to them, and is there any way } of preventing/minimizing this effect without storing prints in a } light tight bag???? }
Epson had a magenta ink fade problem. I have "heard" that it has been improved. Anecdotal reading suggests the paper chemistry vs. ink can have a bearing on colorfastness. Prints I have made in the last year or so on Eposn "photo quality inkjet paper" and Kodak "Picture paper" have held up reasonably well. I have also used Krylon clear acrylic spray to "fix" some prints. ...Not bad for moisture, insufficient data about help with fade. Use many light coats. If you really wet the surface, the paper can become translucent.
WoodyWhite McDermott Technology, Inc
{SNIP} } } In connection with this, we have also found, especially when we } } have printed b/w prints, that if we leave them in a room with } } full spectrum fluorescent lights for a few days, that they begin } } to change colour, and start to take on a reddish tinge. The } } instructions that came with the paper say to not leave prints } } exposed to sunlight - but we're talking about room lights, here! } } Has anyone else found this happens to them, and is there any way } } of preventing/minimizing this effect without storing prints in a } } light tight bag???? } } } }
Below is the result of your feedback form. It was submitted by (aiace_99-at-usa.net) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, June 20, 2001 at 10:45:27 ---------------------------------------------------------------------------
Email: aiace_99-at-usa.net Name: Massimo
Organization: None
Education: Graduate College
Location: Italy
Question: I am writing to enquire about a technical problem. I'd like to see with my microscope some cells coming from a dissected tissue, for instance a vegetable material. So, if I have such small tips into a drop of water and I want to fix, stain and mount in Balsam, how can I perform this operation without to loose that material from the slide? Could you give me some information or a reference where I could find the correct protocol? Thank you Massimo.
I am a long time Tem person starting a new job doing both TEM and LSCM. Its very exciting learning LSCM but I need some suggestions for ordering probes. Our lab has a Kr/Ar laser and will be doing neuropathology research, mostly with fixed human brain samples, but also tissue culture cells both live and fixed. A vibratome is in our future. I was thinking of ordering alexa488nm-phalloiden, alexa568- streptavidin, and syto60 from molecular probes to start getting used to the scope and the lasersharp bio-rad software. The immunohistotech in the lab uses an ABC dtection system with Dab. The first probe should contrast the cytoskeleton well(stain F-actin), the second probe will label any primary Ab, and the third will bind to DNA and contrast the nucleus. Will using these three probes together be a good starting point to becoming adept at labeling and producing high quality images on the LSCM including optical sectioning and 3D (and 4D sometime in the future)? I welcome suggestions Thank you
http://www.resolve3d.com/in_the_news_resview.html Computer Aided Cell and Molecular Biology (CACMB), not medicine, will find the cure for cancer and other diseases. There will always be a need for the trained clinician (MD/RN) but, advanced diagnostic and treatment option selection has become gene based, has moved from the physician's practice to the computerized cell and molecular biology laboratory, and appropriate treatment options should now be based on the personal biology of the patient.
Hi Listers, I have a BioRad Confocal MRC1000 on a Nikon Optiphot (upright scope) and would like to use it on an inverted microscope but the only thing I have is an older Diaphot with the 80/20 prism. Of course this is unacceptable as there is no shutter to prevent someone from inadvertently looking into the eyepieces while scanning. Does anyone know if and how the sliders on the Diaphot can be modified so as to serve as a shutter, i.e. in one position the scanner input is blocked to allow viewing and in another it is directed only to the objective/specimen? I assume it would mean a modification to the internal prisms. Has anyone done this? Source for parts? Procedure? Does anyone do this as a business? I called my local Nikon dealer and was told that the 80/20 can not be converted. Would I be better off to by a new inverted scope? Issues: cost, new infinity corrected optics. What are the issues with this changeover if you have done this. If someone who is on the confocal list could forward this, I would appreciate it. I have not been able to sign on since I changed my email address. Thanks so much for advice and help. Damian Neuberger, PhD Senior Research Scientist Baxter Healthcare Corp. email: damian_neuberger-at-baxter.com Fax: 847/270-5897Phone: 847/270-5888
Along this thread, when we print B&W images on a HP deskjet, we set printer properties to Grayscale, yet when we examine the image with a hand lens, we can see color dots. Why? (perhaps I missed the answer in a previous recent message, sorry). Damian
I got a question from one of the Professors in Pathology here in Tromso, and she wants to know if there are anyone selling seminar boxes with specimens (Light microsopy)from human pathology/anatomy?
Best regards Randi Olsen Department of Electron Microscopy Faculty of Medicine University of Tromso N-9037 TROMSO Norway
I'm not certain but your Sorvall unit may be similar in design to the Polaron CPD units that we service and sell in the U.S. If you could send us a digital photograph and/or copies of any information on the unit that may still be around, we will be happy to do some research to see if we can help you.
Please feel free to contact me off line at mnesta-at-ebsciences.com or by phone at (800) 992-9037.
Mike
Michael R. Nesta General Manager Energy Beam Sciences, Inc. Agawam, MA USA Tel: (413) 786-9322 www.ebsciences.com "Adding Brilliance to Your Vision"
-----Original Message----- } From: swiding [mailto:swiding-at-astro.temple.edu] Sent: Wednesday, June 20, 2001 9:38 AM To: microscopy-at-sparc5.microscopy.com
Hello List,
I am trying to repair our Sorvall critical point dryer. I would like to replace the needle valve assemblies as they are starting to leak. Does anyone know if parts are available for this system? If not, we will have to get it replaced.
We just had a demo of their system from the local sales person, and I must say I was greatly impressed. The company is very young and full of some great ideas. The sales crew seems eager to work with potential customers. The demo was good enough for the powers that be here to approve some "proof of concept" work with some of our own samples. The software is simply amazing in its potential. It seems to have a very user friendly, and intuitive interface that is quite powerful. Obviously the biggest limitation is resolution. Despite the claims of "high resolution", they are limited to typical light microscopy resolutions of about 0.2um/voxel. Another limitation is your material. If you are talking about biological samples, there is no problem, but currently certain materials do present a problem for them.
If you have more specific questions about what I saw at the demo, please feel free to contact me.
Sincerely,
David Bell Scientist Electron Microscopy Lab Millipore Corporation 80 Ashby Road Bedford, MA 01730 (781) 533-2108
sterling stoudenmire To: MICROSCOPY-at-sparc5.microscopy.com {sstouden-at-the cc: links.com} Subject: does anyone have a resolve workstation
06/21/2001 06:42 AM
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if so, how well is it liked?
http://www.resolve3d.com/in_the_news_resview.html Computer Aided Cell and Molecular Biology (CACMB), not medicine, will find the cure for cancer and other diseases. There will always be a need for the trained clinician (MD/RN) but, advanced diagnostic and treatment option selection has become gene based, has moved from the physician's practice to the computerized cell and molecular biology laboratory, and appropriate treatment options should now be based on the personal biology of the patient.
I'm not certain but your Sorvall unit may be similar in design to the Polaron CPD units that we service and sell in the U.S. If you could send us a digital photograph and/or copies of any information on the unit that may still be around, we will be happy to do some research to see if we can help you.
Please feel free to contact me off line at mnesta-at-ebsciences.com or by phone at (800) 992-9037.
Mike
Michael R. Nesta General Manager Energy Beam Sciences, Inc. Agawam, MA USA Tel: (413) 786-9322 www.ebsciences.com "Adding Brilliance to Your Vision"
-----Original Message----- } From: swiding [mailto:swiding-at-astro.temple.edu] Sent: Wednesday, June 20, 2001 9:38 AM To: microscopy-at-sparc5.microscopy.com
Hello List,
I am trying to repair our Sorvall critical point dryer. I would like to replace the needle valve assemblies as they are starting to leak. Does anyone know if parts are available for this system? If not, we will have to get it replaced.
Boeckler/Ventana/RMC/Dupont-Sorvall no longer provide on-site service for old MT2-B microtome, and no support for our MT-5000 microtome. Does anyone know of a good, independent service provider for these instruments in the SE United States?
Julie Piraino Laboratory Manager Smithsonian Maine Station 701 Seaway Drive Fort Pierce, FL 34949 561-465-6630 x143 fax 561-461-8154
I am using the threshold feature of NIH Image to analyze images of cell cultures of individual neurons with processes. We are attempting to quantitate the number of processes in each culture. However, when the threshold function is applied, if a polygon is formed by the processes, NIH Image fills in the space between the "lines" (coverts all the pixels to black = "above threshold"). Since we are trying to measure the number of pixels above threshold, the filled space is counted as part of the number, thus preventing an accurate measurement.
Is there a way to stop the "fill" from occurring?
Thanks.
Paul Madtes Jr. pmadtes-at-mvnc.edu 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
Dear Listers, Does anyone have experience in isolating biofilms from sediment prior to fixation. I thinking of doing a pretreatment with Tween-20 but would appreciate suggestions from anyone who has prepared films for imaging in the LM and SEM. Rosemary -- Rosemary Walsh, Manager The Electron Microscope Facility for the Life Sciences, A Shared Technology Facility, The Life Sciences Consortium 1 South FrearLab Penn State University University Park, PA 16802 (814) 865-0212 rw9-at-psu.edu http://www.lsc.psu.edu/stf/em/home.html
Would anyone who uses a MT-6000, Ultracut E, or NOVA ultramicrotome please comment on reliability, ease-of -use, lighting, etc. for those machines. Our lab is going to purchase a used machine in July.
I found that the learning curve was greatly improved by starting with another Molecular Probe product. I acquired the 6um FocalCheck beads (I got F-14807)(Search MP for document MP 07234) that fit our system - they should be fairly generic. I have made up my own slides. You will probably want one with three colors. The idea is to determine through experience the relationship between Mag Power and Optical Section Thickness. The problem in CSLM is not the x-y plane, it is learning how to get the z-axis right, and how to balance the channels. Choosing the other probes is a question that can only be answered by knowing as much as possible about the spectral characteristics of the excitation and emission filters in your system so that you can minimize crosstalk. NOTE: There is a paucity of information about filters that forces us to be empiric in setting up experiments, at least that's been my experience so far. I keep on asking for the information from the system vendor.
Hope this helps,
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: "woxberry-at-downstate.edu"-at-sparc5.microscopy.com } Sent: Wednesday, June 20, 2001 8:04 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: New Confocal user } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 a long time Tem person starting a new job doing both TEM and } LSCM. Its very exciting learning LSCM but I need some suggestions for } ordering probes. Our lab has a Kr/Ar laser and will be doing } neuropathology research, mostly with fixed human brain samples, but } also tissue culture cells both live and fixed. A vibratome is in our } future. I was thinking of ordering alexa488nm-phalloiden, alexa568- } streptavidin, and syto60 from molecular probes to start getting used to } the scope and the lasersharp bio-rad software. The immunohistotech in } the lab uses an ABC dtection system with Dab. The first probe should } contrast the cytoskeleton well(stain F-actin), the second probe will } label any primary Ab, and the third will bind to DNA and contrast the } nucleus. Will using these three probes together be a good starting } point to becoming adept at labeling and producing high quality images } on the LSCM including optical sectioning and 3D (and 4D sometime in the } future)? } I welcome suggestions } Thank you } } Bill } }
Hello All, I have a Jeol 5900 SEM, and I would like to purchase a BSE detector for it. Does anyone in the group have any suggestions/recommendations/warnings? Is it pretty straight forward to purchase a third-party detector and get it to work with our microscope, or is it best to go through the vendor? Thank you for any input that you can provide.
Brad Johnson Pacific Northwest National Lab P.O. Box 99, K6-24 Richland, WA 99352 voice: 509-372-1635 fax: 509-376-3108
I found that the learning curve was greatly improved by starting with another Molecular Probe product. I acquired the 6um FocalCheck beads (I got F-14807)(Search MP for document MP 07234) that fit our system - they should be fairly generic. I have made up my own slides. You will probably want one with three colors. The idea is to determine through experience the relationship between Mag Power and Optical Section Thickness. The problem in CSLM is not the x-y plane, it is learning how to get the z-axis right, and how to balance the channels. Choosing the other probes is a question that can only be answered by knowing as much as possible about the spectral characteristics of the excitation and emission filters in your system so that you can minimize crosstalk. NOTE: There is a paucity of information about filters that forces us to be empiric in setting up experiments, at least that's been my experience so far. I keep on asking for the information from the system vendor.
Hope this helps,
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: "woxberry-at-downstate.edu"-at-sparc5.microscopy.com } Sent: Wednesday, June 20, 2001 8:04 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: New Confocal user } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 a long time Tem person starting a new job doing both TEM and } LSCM. Its very exciting learning LSCM but I need some suggestions for } ordering probes. Our lab has a Kr/Ar laser and will be doing } neuropathology research, mostly with fixed human brain samples, but } also tissue culture cells both live and fixed. A vibratome is in our } future. I was thinking of ordering alexa488nm-phalloiden, alexa568- } streptavidin, and syto60 from molecular probes to start getting used to } the scope and the lasersharp bio-rad software. The immunohistotech in } the lab uses an ABC dtection system with Dab. The first probe should } contrast the cytoskeleton well(stain F-actin), the second probe will } label any primary Ab, and the third will bind to DNA and contrast the } nucleus. Will using these three probes together be a good starting } point to becoming adept at labeling and producing high quality images } on the LSCM including optical sectioning and 3D (and 4D sometime in the } future)? } I welcome suggestions } Thank you } } Bill } }
I work with ultrasmall nanosized particles, as small as 2 nm, with much lower contrast than metal particles such as gold and silver. These ultrasmall particles don't show up very well on TEM, even with optimal TEM settings. I thought what would help to increase the contrast dramatically would be to use a TEM grid that has very low background signal. I have tried grids with ultrathin carbon film (about 5 nm thick) but the background signal is still too high for these particles to show up clearly. Holey carbon grids is an option except they don't give me enough particles for particle sizing. Any suggestions would be very much appreciated.
In a recent issue of "PEI", May, 2001 (http://www.peimag.com) the following sampling of Inkset Resources was offered in an article entitled: "Inksets for Desktop Inkjet Printers", by Theano Nikitas.
The point of the article was the difference between pigment and dye inks, and those that are specially formulated for archival prints.
I am only using those that are not Printer Manufacturers
1. Cone Editions Press: http://www.inkjetmall.com
2. Luminos Photo Corp: http://www.luminos.com
3. Lyson USA Inc.: http://www.lysonusa.com
4. MIS Associates Inc: http://inksupply.com
5. MediaStreet.com: http://mediastreet.com
6. Xtreme gamut: http://xtremegamut.com
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: Paula Allan-Wojtas } Sent: Wednesday, June 20, 2001 7:47 AM } To: microscopy-at-sparc5.microscopy.com } Subject: Printers and paper } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi, all, } } This question has come up for us at a good time, with this ongoing } discussion of printers. } } We recently purchased an Epson 880 ink jet printer for our EM/LM lab, and } are very happy with it. The black and white prints look excellent. We have } tried a number of papers, and find that the Epson "Photo Quality Glossy } Paper" and "Photo Paper" give the best results for our purposes. } } We are wondering how to go about making our prints more permanent from } these printers. If the prints have just been printed and aren't dry yet, } or if they are dry and are in contact with any moisture, they smudge quite } easily. Is there any coating - lamination or spray, or something else } which we can use to protect the surface? } } In connection with this, we have also found, especially when we have } printed b/w prints, that if we leave them in a room with full spectrum } fluorescent lights for a few days, that they begin to change colour, and } start to take on a reddish tinge. The instructions that came with the } paper say to not leave prints exposed to sunlight - but we're talking } about room lights, here! Has anyone else found this happens to them, and } is there any way of preventing/minimizing this effect without storing } prints in a light tight bag???? } } Any help or suggestions would be much appreciated. As always, thanks for } your help in advance. } } Regards, } } Paula. } } Paula Allan-Wojtas } Research Scientist - Food Microstructure } Agriculture and Agri-Food Canada } Atlantic Food and Horticulture Research Centre } Kentville, Nova Scotia Canada B4N 1J5 } } Tel: (902) 679-5566 } FAX: (902) 679-2311 } } email: allanwojtasp-at-em.agr.ca } } }
I checked out there system from a general interest, "this looks cool" viewpoint. I recall the representative telling me that the technique works by serial sections. I also seem to recall that the images are taken of the remaining block rather than the removed sections.
It sounds like an interesting approach, and it will certainly have some applications. I did not see any immediate applications in my research. I was also unsure what kind of ultimate resolution could be achieved. It seems that the technique would be as limited by the thickness of the sections in the z-direction as it would by microscopy in the x- and y-directions.
At 09:26 AM 6/21/2001 -0400, you wrote: } Hi Sterling, } } We just had a demo of their system from the local sales person, and I must } say I was greatly impressed. The company is very young and full of some } great ideas. The sales crew seems eager to work with potential customers. } The demo was good enough for the powers that be here to approve some "proof } of concept" work with some of our own samples. The software is simply } amazing in its potential. It seems to have a very user friendly, and } intuitive interface that is quite powerful. Obviously the biggest } limitation is resolution. Despite the claims of "high resolution", they } are limited to typical light microscopy resolutions of about 0.2um/voxel. } Another limitation is your material. If you are talking about biological } samples, there is no problem, but currently certain materials do present a } problem for them. } } If you have more specific questions about what I saw at the demo, please } feel free to contact me. } } Sincerely, } } David Bell } Scientist } Electron Microscopy Lab } Millipore Corporation } 80 Ashby Road } Bedford, MA 01730 } (781) 533-2108 } } } From: sterling stoudenmire
In my point of view, it's almost impossible to have gray tones with RGB (3 color scheme) color cartridges. CYMK (4 color) will do right if printer set for gray-scale (to use black cartridge only). To simplify my life I removed color cartridges and replaced them on black one for B&W prints. For may model it's possible to use big black cartridge instead color assembly. I always replace color ribbon on black one in our Tektronix Dye-sub (actually, reverse: black ribbon installed by default and we replace that on color if necessary).
Sergey
At 07:22 AM 6/21/01 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
At 12:53 PM 6/21/2001 -0500, Warren E Straszheim wrote:
I also looked into this a while back, it's an interesting approach. Essentially the sample is fluorescently stained with something like eosin and embedded in an opaque plastic. The surface is imaged, a micron or two is shaved off, the new surface is imaged, and so on. This gives you a (huge) 3D data set that can be viewed at various magnifications and resolutions. You lease/buy a computer system from them, send them the sample and they send you a DVD with the data. The software is quite fast and easy to use. I haven't used their services yet, am still waiting for an appropriate sample.
Tom
} I checked out there system from a general interest, "this looks cool" } viewpoint. I recall the representative telling me that the technique works } by serial sections. I also seem to recall that the images are taken of the } remaining block rather than the removed sections. } } It sounds like an interesting approach, and it will certainly have some } applications. I did not see any immediate applications in my research. I } was also unsure what kind of ultimate resolution could be achieved. It } seems that the technique would be as limited by the thickness of the } sections in the z-direction as it would by microscopy in the x- and } y-directions.
Thomas Moninger (thomas-moninger-at-uiowa.edu) University of Iowa Central Microscopy Research Facility (www.uiowa.edu/~cemrf) View expressed are my own.
I guess my question would be "what is the investigator's definition of a biofilm?" If you detergent-treat sediment, you are likely to remove a substantial majority of the attached bacterial biomass from suspended solids, but this will also contain non-biofilm (porewater) cells and the majority of the spatial information in the sample (relationships of cells to substrata, and of cells to cells) will be lost. Why do you want to "isolate" the biofilms? Why not examine them directly (you are using microscopy - virtually the ONLY way to examine biofilms directly).
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Robert J. Palmer Jr., Ph.D. Natl Inst Dental Craniofacial Res - Natl Insts Health Oral Infection and Immunity Branch Bldg 30, Room 308 30 Convent Drive Bethesda MD 20892 ph 301-594-0025 fax 301-402-0396
I have used GW Electronics BSE systems for many years. I purchased my last system just a few years back. With both the olde and new systems, I had excellent results -performance is great. The equipment is very well made and has been extremely reliable. I have no connection with GW other than being an experineced and satisfied user.
GW can interface to just about any scope.
Woody White McDermott Technology, Inc. http://woody.white.home.att.net
} -----Original Message----- } From: Johnson, Bradley R [mailto:Bradley.Johnson-at-pnl.gov] } Sent: Thursday, June 21, 2001 11:54 AM } To: 'Microscopy-at-MSA.Microscopy.Com' } Subject: SEM: Recommendations for BSE detectors } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Hello All, } I have a Jeol 5900 SEM, and I would like to purchase a } BSE detector for } it. Does anyone in the group have any } suggestions/recommendations/warnings? Is } it pretty straight forward to purchase a third-party detector } and get it to work } with our microscope, or is it best to go through the vendor? } Thank you for any } input that you can provide. } } } Brad Johnson } Pacific Northwest National Lab } P.O. Box 99, K6-24 } Richland, WA 99352 } voice: 509-372-1635 } fax: 509-376-3108 } } Bradley.Johnson-at-pnl.gov }
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Would anyone who uses a MT-6000, Ultracut E, or NOVA ultramicrotome please } comment on reliability, ease-of -use, lighting, etc. for those } machines. Our lab is going to purchase a used machine in July. }
We have had an Ultracut E for 15 years with only occasional and relativley minor repairs needed. When we have needed service, Tek-Net of Lakewood, NJ has been excellent.
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 **********************************************
} Date: Thu, 21 Jun 2001 14:22:08 -0700 } To: "Thearith H. Ung" {tung-at-qdots.com} } From: Sergey Ryazantsev {sryazant-at-ucla.edu} } Subject: Re: TEM-Ultrathin film grid } } Dear Thearith } } 5 nm carbon film is very thick. I am using 1.5-1.8 nm carbon film } on carbon 2 mkm holey film for 1.4 nm for NanoGold particles. You could } also try dark-field mode to enhance the contrast. It's also possible to } decrease the carbon film thickness because it's so strong. I measure the } actual carbon thickness by TM-450 MAXTEK thickness monitor (0.01 nm } resolution). I am using my own design "electron gun" and oil-free vacuum } apparatus for carbon film preparation. } } Sergey } } At 09:51 AM 6/21/01 -0700, 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
Try Bob Ruscica at ETP-USA. He handles Robinson scintillator detectors. Probably the best type made. Very sensitive, versatile and reliable. Not high cost, either. They can be retrofitted to most any scope.
gary g. (highly satisfied Robinson BSE user)
At 08:54 AM 6/21/2001, you wrote:
} Hello All, } I have a Jeol 5900 SEM, and I would like to purchase a BSE } detector for } it. Does anyone in the group have any } suggestions/recommendations/warnings? Is } it pretty straight forward to purchase a third-party detector and get it } to work } with our microscope, or is it best to go through the vendor? Thank you } for any } input that you can provide. } } } Brad Johnson } Pacific Northwest National Lab } P.O. Box 99, K6-24 } Richland, WA 99352 } voice: 509-372-1635 } fax: 509-376-3108 } } Bradley.Johnson-at-pnl.gov
It should works great, thanks for idea. Basically, for my B&W job I am using Tektronix dye-sub which performs fine. Sergey
At 07:13 PM 6/21/01 -0400, you wrote: } look for Quad-Black - it is a four or six grey shade ink set to replace } the color ones - the prints have more grey levels than film. } } } At 03:01 PM 6/21/01, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I used an RMC MT-6000 XL for five years and I am using an Reichert Ultracut E since a couple of weeks. After getting used to the Reichert I like it a lot and think overall it is the better machine. It has some nice features, for example this integral water pump and the specimen trans-illumination etc. and I think the mechanics are more robust. One thing is better about the RMC MT-6000 XL. You can change the angle of the microscope and by that always get the best reflection of the water surface in the knife boat (independent from how high the water level is). The RMC is really not bad but I think a lot of details make the Reichert better.
Hi Craig, As you probably know the machines you are asking about definitely have some age on them. The lab here has a RMC MT-6000XL and an Ultracut E. I really like the Ultracut - it works like a charm and has lighting in every possible direction. I really dislike the design of the RMC MT-6000 and the lighting is either on above or below but not both unless you get a switch installed (we had the switch installed and it makes a big difference to me). The newer RMC ultramicrotomes are very nice but the Ultracuts are still the best ultramicrotomes. Can't comment on the Nova. best regards, Beth
} Would anyone who uses a MT-6000, Ultracut E, or NOVA ultramicrotome please } comment on reliability, ease-of -use, lighting, etc. for those } machines. Our lab is going to purchase a used machine in July.
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
For the more frugal try Johnson's Paste Wax. It works on some inks and papers, not all. It does offer some protection against UV. It does protect against moisture, finger prints, dust and scratches. It does change the finish some.
If the wax causes the ink to smear a fixative spray will usually stop that.
Test it for several weeks before putting it on an expensive display.
Wax also works on regular photographs. The way I discovered it was on B&W prints that were displayed outdoors in the weather for several months.
Gordon
Gordon Couger Stillwater, OK
.From: "George Laing" {scisales-at-ngscorp.com}
Luminos http://www.lumijet.com has a spray called "Imagesheild" to protect prints. $18/ can. Quoting their website: "LUMIJET IMAGESHIELD will become your favorite print protector. It's a convenient ozone-friendly low-odor aerosol spray that significantly improves wet-fastness. In addition to protecting delicate inkjet images from moisture, it also shields images from fingerprints and harmful UV rays. This unique product will not only help extend useful print life, it is totally transparent; preserving the original print surface characteristics. From dead matte to glossy, or from canvas to silver foil, you'll never know it's there."
} We are wondering how to go about making our prints more permanent } from these printers. If the prints have just been printed and } aren't dry yet, or if they are dry and are in contact with any } moisture, they smudge quite easily. Is there any coating - } lamination or spray, or something else which we can use to } protect the surface?
There were recently (last 6-8 months) some issues with color shift on some Epson papers. Epson determined the shift was from high concentrations of ozone, and not due to exposure to light. Epson recommends Heavyweight Matte or Epson Photo paper for maximum lightfastness.
George
George Laing National Graphic Supply v:(800) 223-7130 X3109 f:(800) 832-2205 email: scisales-at-ngscorp.com } } In connection with this, we have also found, especially when we } have printed b/w prints, that if we leave them in a room with } full spectrum fluorescent lights for a few days, that they begin } to change colour, and start to take on a reddish tinge. The } instructions that came with the paper say to not leave prints } exposed to sunlight - but we're talking about room lights, here! } Has anyone else found this happens to them, and is there any way } of preventing/minimizing this effect without storing prints in a } light tight bag???? }
I have used a 'Super Nova' for 11 or 12 years and the only thing that has needed to be replaced/repaired was the fluorescent light source. We are starting to have a niggling problem when switching on the light - it can take several attempts and it looks like there might be some sort of dedicated controller to be fixed. The only other thing that may need to be replaced is the belt (motor drive band) which is beginning to slip when manual raise and lower is used on the specimen arm. It's never needed a service engineer though. The reason we/I opted for the LKB was that I had used them for years and liked their reliability - there is no thin section mechanical feed to go wrong. The 'Super Nova' was a later model than the Nova and has a completely sealed control panel which keeps the dust and probably water out. It can also maintain specimen position accurately when you turn the thermal feed off and the cut stroke is powered rather than just relying on gravity. I don't know if these are standard on the Nova. The Ultracut E has nicer optics, specimen adjustment is superb and it is much less sensitive to external vibration but I cannot swear to it's long term use because I've only used it in demonstrations. I really think it would be worthwhile if you can get a demonstration (especially if you can take a specimen to cut) because these machines are all very good and it may come down to personal taste. One thing you might to want to examine is the availability of spares, accessories and servicing for each machine - in the UK it is becoming difficult to get spares for the 'Super Nova'.
I hope this helps.
The usual disclaimers apply: my opinions etc. and no affiliations to any company.
Malcolm Haswell e.m. unit School of Sciences University of Sunderland UK
craig klotz wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Would anyone who uses a MT-6000, Ultracut E, or NOVA ultramicrotome please } comment on reliability, ease-of -use, lighting, etc. for those } machines. Our lab is going to purchase a used machine in July. } } TIA, } Craig
} } Position: Scientist--EM / Histology (Reference Code 2232) } Company: Alcon / R&D / In Vivo Pharmacology Unit } Location: Global Headquarters in Fort Worth, TX } Position Type: Full Time } } About Alcon... } } Superb facilities. Terrific benefits. Great, long lasting working } relationships. Employee turnover of less than 5% annually. Fifty-three } (53) years of steady, continuous growth. The $2.55 billion global leader } in the ophthalmic and vision care industry. It's no wonder Fortune has } selected Alcon as "one of the 100 Best Companies to work for" for three } consecutive years. Our 10,000 employees worldwide form a unique community } of multi-national, multi-disciplinary, dedicated and successful men and } women who have combined their talents and vision over our 50-year history } of growth and success. The tenure (average 13 years) and caliber of our } employees is a major competitive advantage. } } For much more information about our people, benefits, products and } culture, please visit us at } www.alconlabs.com. } } Position Responsibilities } } * Prepares, processes, examines and provides preliminary } interpretation of animal tissue specimens using various techniques with } light microscopy and/or EM as end-point. } * May provide method development (EM, histology, etc) directed towards } the discovery of new drug candidates. } } Qualifications } } * Bachelor of Science degree in a related discipline } * At least five years of significant light and/or EM experience } related exclusively to human and animal tissue (including tissue } preparation) } * Collaborative and problem solving skills are essential } * Highly refined interpersonal and technical writing skills } * Certification by or eligibility to be certified by the MSA a plus } * Traditional histology tissue preparation knowledge a benefit } * Digital imaging, immunohistochemistry, and experience with ocular } tissues highly desirable } } Alcon is an Equal Opportunity Employer committed to quality through } diversity. M/F/D/V. Pre-employment drug testing. } } If you are interested in submitting your resume for this position, please } email it to helena.loflin-at-alconlabs.com. Please include Reference Code } 2232. } } }
Hi all, We had a short in the objective lens coil a while ago and it was changed. What was happening was that we lost all control of the beam position and focus. We usually work at 15 kV (suitable for most of our needs), but now need to do something at 25. However, when we increase to this acc. voltage, we are finding a similar problem - beam pops out of focus (goes big and diffuse) and shoots off somewhere. We can get it back by pressing the Obj. Pol. button. This normally works if we get the same thing at 20 kV and less, but not at 25. The effect only occurs once an hour at 22 kV. Anyone have an idea why this is happening? By the way, for those who would undoubtably ask, the I is 100 nA for what we are at the the moment. Comments? Malc.
-- Dr MP Roberts Phone: [+27](0)46 603 8313 Dept of Geology Fax: [+27](0)46 622 9715 Rhodes University Cell: 083 4060 262 (try your luck!) 6140 Grahamstown e-mail: m.roberts-at-ru.ac.za SOUTH AFRICA
Dear Bradley, I have used the JEOL BSE detector, the GW BSE detector and the Robinson BSE detector and they all work well and I have had no reliability problems or interface problems. For the third-party detectors, the external signal just goes in through the "EXTERNAL" signal input on the SEM. At 08:54 AM 06/21/2001 -0700, you wrote: } } Hello All, } I have a Jeol 5900 SEM, and I would like to purchase a BSE detector for } it. Does anyone in the group have any suggestions/recommendations/warnings? Is } it pretty straight forward to purchase a third-party detector and get it to work } with our microscope, or is it best to go through the vendor? Thank you for any } input that you can provide. } } } Brad Johnson Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
Jim is right in what he says about the compatibility issues with Oxford/Link detectors and the restore signal being generated by the pulse processor and not the pre-amp as in most detectors. However we are reps for Thomson Scientific Instruments' WINEDS x-ray analyzers and pulse processing electronics in North America. TSI has the DX3000 bias and PX9000 Pulse processor that will interface to the Oxford/Link detectors. We have used it with both the older type and with the Pentafet type without any changes to the detector preamp. The PX9000 is a universal design and can be used with other manufacturers analyzers.
Douglas Connors General Manager TNAS Inc. 7897 Hwy 19, Dane WI 53529 (608)798-2005 voice (608)798-1675 fax tnas1-at-msn.com ----- Original Message ----- } From: "Jim Nicolino" {JNicolino-at-xraydetectors.com} To: "Michael L. Boucher" {mboucher-at-tc.umn.edu} Cc: {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, June 19, 2001 10:30 AM
If you are only imaging the particles, not in context of a particular environment, you can negative stain e.g. with phosphotungstic acid.
Sounds like you're not considering staining them but if the particles are reactive then that is a possibility too.
Richard
} } } "Thearith H. Ung" {tung-at-qdots.com} 6/21/01 9:51:50 AM } } }
Dear colleagues,
I work with ultrasmall nanosized particles, as small as 2 nm, with much lower contrast than metal particles such as gold and silver. These ultrasmall particles don't show up very well on TEM, even with optimal TEM settings. I thought what would help to increase the contrast dramatically would be to use a TEM grid that has very low background signal. I have tried grids with ultrathin carbon film (about 5 nm thick) but the background signal is still too high for these particles to show up clearly. Holey carbon grids is an option except they don't give me enough particles for particle sizing. Any suggestions would be very much appreciated.
I'm fixing small chunks of wood in 4% phosphate buffered paraformaldehyde, and separately in a Karnofsy-type (paraformaldehyde and glutaraldehyde) buffered fixative. I need to store the samples for a year or more - are either of these fixatives appropriate for long-term storage, and if not, what are the problems (e.g., tissue hardening?). Does anyone have suggestions for appropriate storage solutions, or have an idea of how long is OK in either of these fixatives? The samples in paraformaldehyde are used for fluorescnce microscopy, and the Karnofsky fixed samples for TEM. Thanks in advance for your experience/advice.
Rachel
****************************************** Rachel Spicer Biological Laboratories 3119 Organismic and Evolutionary Biology Harvard University 16 Divinity Avenue Cambridge, MA 02138
Thanks for the suggestion. I thought about staining as well, except that these particles tend to get etched by acids. However, it might still work if only a small amount of phosphotunstic acid is used. So it is definitely worthwhile looking into it.
Thearith
-----Original Message----- } From: Richard Thrift [mailto:Richard_Thrift-at-skyepharma.com] Sent: Friday, June 22, 2001 9:57 AM To: tung-at-qdots.com; Microscopy-at-sparc5.microscopy.com
Thearith,
Have you got a possibility to do STEM ? If there are some high Z atoms in your samples, you should get enough contrast to see the particles.
Andreas
} Dear colleagues, } } I work with ultrasmall nanosized particles, as small as 2 nm, with much } lower contrast than metal particles such as gold and silver. These } ultrasmall particles don't show up very well on TEM, even with optimal TEM } settings. I thought what would help to increase the contrast dramatically } would be to use a TEM grid that has very low background signal. I have tried } grids with ultrathin carbon film (about 5 nm thick) but the background } signal is still too high for these particles to show up clearly. Holey } carbon grids is an option except they don't give me enough particles for } particle sizing. Any suggestions would be very much appreciated. } } } Regards, } } Thearith } } } }
Sounds like a contact problem with the OL polarity switch or relay. At higher KV's the lenses need higher current. 22 KV is probably the limit for the contact to cut-out.
Regards,
Earl
----- Original Message ----- } From: "Dr Malcolm Roberts" {m.roberts-at-ru.ac.za} To: "Microscopy discussion group" {Microscopy-at-sparc5.microscopy.com} Sent: Friday, June 22, 2001 6:09 AM
Hi Rachel...
I would imagine that storage in 70% ethanol might be a better bet... brought up through an alcohol series. This is a standard practice for long term storage of museum specimens after fixation, and for most EM labs.
Check out following for reference: Simmons, J.E. 1995. Storage in Fluid Perservatives. In Storage of Natural History Collections: A Preventative Conservation Approach. Vol. 1. C.L. Rose, C.A. Hawks, and H.H. Genoways, eds. Society for the Preservation of Natural History Collections, Iowa City. 161-181.
Best regards,
Angela
Angela V. Klaus
Director, Interdepartmental Laboratories American Museum of Natural History Central Park West at 79th Street New York, NY 10024 USA
Email: avklaus-at-amnh.org Tel: 212-769-5977
On Fri, 22 Jun 2001, Rachel Spicer wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello, } } I'm fixing small chunks of wood in 4% phosphate buffered paraformaldehyde, } and separately in a Karnofsy-type (paraformaldehyde and glutaraldehyde) } buffered fixative. I need to store the samples for a year or more - are } either of these fixatives appropriate for long-term storage, and if not, } what are the problems (e.g., tissue hardening?). Does anyone have } suggestions for appropriate storage solutions, or have an idea of how long } is OK in either of these fixatives? The samples in paraformaldehyde are } used for fluorescnce microscopy, and the Karnofsky fixed samples for TEM. } Thanks in advance for your experience/advice. } } Rachel } } ****************************************** } Rachel Spicer } Biological Laboratories 3119 } Organismic and Evolutionary Biology } Harvard University } 16 Divinity Avenue } Cambridge, MA 02138 } } (617) 496-3580 (phone) } (617) 496-5854 (fax) } spicer-at-oeb.harvard.edu } ****************************************** } } }
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi Richel
If you want to store for long term then better preserve in a deep freezer at -20 to -30 deg as it is and when you want to proceed, allow it to come down at 4 deg and proceed as per normal procedure.
Second option is fix in a fixative and then store in washing buffer at 4 deg celcius, but keep one thing is in mind that you should keep changing this buffer after a short time span, say 15-20 day. Your tissue well perserve in this condition. Like this i have preserved the animal tissues in best of condition for three months.
If you try to preserve in fixattive then their is a chacnes of hardening the material or if you preserves in alchohol 70%, the chances of shrinkage is more.
for best prservation methods Ref. the book by M.A. Hayat, Techniques in Electron Microscopy.
With Best Regards Arti Harle
Angela Klaus {avklaus-at-amnh.org} wrote: ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi Rachel...
I would imagine that storage in 70% ethanol might be a better bet... brought up through an alcohol series. This is a standard practice for long term storage of museum specimens after fixation, and for most EM labs.
Check out following for reference: Simmons, J.E. 1995. Storage in Fluid Perservatives. In Storage of Natural History Collections: A Preventative Conservation Approach. Vol. 1. C.L. Rose, C.A. Hawks, and H.H. Genoways, eds. Society for the Preservation of Natural History Collections, Iowa City. 161-181.
Best regards,
Angela
Angela V. Klaus
Director, Interdepartmental Laboratories American Museum of Natural History Central Park West at 79th Street New York, NY 10024 USA
Email: avklaus-at-amnh.org Tel: 212-769-5977
On Fri, 22 Jun 2001, Rachel Spicer wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello, } } I'm fixing small chunks of wood in 4% phosphate buffered paraformaldehyde, } and separately in a Karnofsy-type (paraformaldehyde and glutaraldehyde) } buffered fixative. I need to store the samples for a year or more - are } either of these fixatives appropriate for long-term storage, and if not, } what are the problems (e.g., tissue hardening?). Does anyone have } suggestions for appropriate storage solutions, or have an idea of how long } is OK in either of these fixatives? The samples in paraformaldehyde are } used for fluorescnce microscopy, and the Karnofsky fixed samples for TEM. } Thanks in advance for your experience/advice. } } Rachel } } ****************************************** } Rachel Spicer } Biological Laboratories 3119 } Organismic and Evolutionary Biology } Harvard University } 16 Divinity Avenue } Cambridge, MA 02138 } } (617) 496-3580 (phone) } (617) 496-5854 (fax) } spicer-at-oeb.harvard.edu } ****************************************** } } }
Ms. Arti Harle Regional Sophisticated Instrumentation Center Indian Institute of Technology-Bombay (IIT-B) Powai, Mumbai - 400076. India Phone : 91-22-5767691 Extn 7694 91-22-5720109/ 5721039 (Resi) E-mail : aartih-at-usa.net
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-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Thearith H. Ung wrote the following: ========================================= I work with ultrasmall nanosized particles, as small as 2 nm, with much lower contrast than metal particles such as gold and silver. These ultrasmall particles don't show up very well on TEM, even with optimal TEM settings. I thought what would help to increase the contrast dramatically would be to use a TEM grid that has very low background signal. I have tried grids with ultrathin carbon film (about 5 nm thick) but the background signal is still too high for these particles to show up clearly. Holey carbon grids is an option except they don't give me enough particles for particle sizing. Any suggestions would be very much appreciated. =============================================== This has become quite a generic problem for many who are working with nano size particles, and many have found the "solution" to be the use of silicon nitride membrane window grids that have been offered by SPI Supplies for several years. Information about these grids can be found on URL http://www.2spi.com/catalog/instruments/silicon-nitride.html
The silicon nitride is completely structureless and featureless and seems to provide excellent electron transparency. The other advantage is that they are sufficiently robust to survive temperatures in excess of 1000° C. And there is no carbon grain to be confused with the nanoparticles of interest.
Disclaimer: SPI Supplies offers silicon nitride membrane window grids for this type of application.
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 ############################ ==================================================
There are other negative staining compounds available - ammonium molybdate and uranyl acetate for instance. However, my first try would be shadow-casting, by evaporating some Pt/C at about 10 degrees to the specimen. Cheers Jim Darley
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On Saturday, June 23, 2001 6:06 AM, Thearith H. Ung [SMTP:tung-at-qdots.com] wrote: } } } } Hi Richard, } } Thanks for the suggestion. I thought about staining as well, except that } these particles tend to get etched by acids. However, it might still work if } only a small amount of phosphotunstic acid is used. So it is definitely } worthwhile looking into it. } } Thearith } } -----Original Message----- } } From: Richard Thrift [mailto:Richard_Thrift-at-skyepharma.com] } Sent: Friday, June 22, 2001 9:57 AM } To: tung-at-qdots.com; Microscopy-at-sparc5.microscopy.com } Subject: Re: TEM-Ultrathin film grid } } } If you are only imaging the particles, not in context of a particular } environment, you can negative stain e.g. with phosphotungstic acid. } } Sounds like you're not considering staining them but if the particles are } reactive then that is a possibility too. } } Richard
Readers, Kindly allow this short reminder that again at the M&M 2001 Conference we will hold a "Just For Fun Micrograph" Contest. The concept being composite images, each of two or more images, one of which must be microscopical in nature. $600 cash in prizes. Contact me direct and I will provide full detail. Don Grimes, Microscopy Today
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This has been discussed before, but... If anyone has a summary regarding contract services that they could share, I would appreciate it. (They seem to be lower priced than electon microscope vendors.) Any recent experience positive or negative with contract services, particularly 1) Advanced Research Systems (Alan Sampson) in Illinois or 2) Sci. Inst. & Applications (V. Feingold) of Duluth, GA or 3) Other EM service providers covering the Midwest. Thanks for your help.
Remember that paraformaldehyde is a reversible fixative, so it isn't appropriate to fix in pform then store in a buffer. I've stored animal tissue for years in pform with no problems, though I'm sure it's not recommended. As for pform/glut, storage after fixation in cacodylate buffer is good, as nothing seems to grow in cacodylate.
Diana
} } Hello, } } I'm fixing small chunks of wood in 4% phosphate buffered paraformaldehyde, } and separately in a Karnofsy-type (paraformaldehyde and glutaraldehyde) } buffered fixative. I need to store the samples for a year or more - are } either of these fixatives appropriate for long-term storage, and if not, } what are the problems (e.g., tissue hardening?). Does anyone have } suggestions for appropriate storage solutions, or have an idea of how long } is OK in either of these fixatives? The samples in paraformaldehyde are } used for fluorescnce microscopy, and the Karnofsky fixed samples for TEM. } Thanks in advance for your experience/advice. } } Rachel --
Diana van Driel Department of Clinical Ophthalmology Sydney University GPO Box 4337 Sydney NSW AUSTRALIA 2001
How about storing part of the sample in buffer with sodium azide to inhibit microorganisms.
Dave
On Fri, 22 Jun 2001 14:38:15 -0400 Rachel Spicer {spicer-at-oeb.harvard.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello, } } I'm fixing small chunks of wood in 4% phosphate buffered paraformaldehyde, } and separately in a Karnofsy-type (paraformaldehyde and glutaraldehyde) } buffered fixative. I need to store the samples for a year or more - are } either of these fixatives appropriate for long-term storage, and if not, } what are the problems (e.g., tissue hardening?). Does anyone have } suggestions for appropriate storage solutions, or have an idea of how long } is OK in either of these fixatives? The samples in paraformaldehyde are } used for fluorescnce microscopy, and the Karnofsky fixed samples for TEM. } Thanks in advance for your experience/advice. } } Rachel } } ****************************************** } Rachel Spicer } Biological Laboratories 3119 } Organismic and Evolutionary Biology } Harvard University } 16 Divinity Avenue } Cambridge, MA 02138 } } (617) 496-3580 (phone) } (617) 496-5854 (fax) } spicer-at-oeb.harvard.edu } ****************************************** } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
I have found that there are a few cacodylate tolerant organisms. Nonetheless I usually take the risk.
Dave
On Mon, 25 Jun 2001 14:28:00 +1000 Diana van Driel {dianavd-at-eye.usyd.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 } -----------------------------------------------------------------------. } } } } Hi Rachel, } } Remember that paraformaldehyde is a reversible fixative, so it isn't } appropriate to fix in pform then store in a buffer. I've stored } animal tissue for years in pform with no problems, though I'm sure } it's not recommended. As for pform/glut, storage after fixation in } cacodylate buffer is good, as nothing seems to grow in cacodylate. } } Diana } } } } } Hello, } } } } I'm fixing small chunks of wood in 4% phosphate buffered paraformaldehyde, } } and separately in a Karnofsy-type (paraformaldehyde and glutaraldehyde) } } buffered fixative. I need to store the samples for a year or more - are } } either of these fixatives appropriate for long-term storage, and if not, } } what are the problems (e.g., tissue hardening?). Does anyone have } } suggestions for appropriate storage solutions, or have an idea of how long } } is OK in either of these fixatives? The samples in paraformaldehyde are } } used for fluorescnce microscopy, and the Karnofsky fixed samples for TEM. } } Thanks in advance for your experience/advice. } } } } Rachel } -- } } } } Diana van Driel } Department of Clinical Ophthalmology } Sydney University } GPO Box 4337 } Sydney NSW } AUSTRALIA 2001 } } Phone 61 2 938 27278/27395 } Mob 0412 165 075 } Fax 61 2 938 27318 }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
Beverly Giammara wrote asking for comments about EM service providers, other than the manufacturers, that cover the Midwest. I would be very appreciative if anyone can offer similar information about EM Service providers that cover Florida.
Sincerely,
Jill
Jill Verlander Reed, D.V.M. Associate Scientist Director, College of Medicine Electron Microscopy Core Facility University of Florida P.O. Box 100215 Gainesville, FL 32610 verlaj-at-medicine.ufl.edu Phone: (352) 846-0820 Fax: (352) 846-3299
We are currently assessing whether or not to replace our old diffraction simulation software and would in this respect be happy to receive the advice of the microscopy community and vendors.
The software should have the following characteristics: * Electron diffraction simulation, kinematical as well as dynamical calculation. * Handle multiple reciprocal lattices in one pattern - that is - several single crystals with different crystallography, size, shape, and orientation relationships, as well as powder diffraction. * Accurate intensity and structure factor determination. * Evaluate double diffraction and crystal shape effects - streaking. * The software should also be able to simulate X-ray diffraction patterns: powder, Laue, Guinier. * Support of common crystallographic/diffraction database formats. * The software should run under Windows NT 4.5 and have a professionally designed GUI. No compiling and linking, and no programming should be necessary but the possibility of doing this (e.g. scripting, ActiveX, Plug-Ins etc.) is a very positive feature.
Sincerely, Paul Baggethun ========================== Paul Baggethun Alcoa Technical Center Alcoa Center, PA 15069 USA (724) 337 1760 paul.baggethun-at-alcoa.com ==========================
Please forgive. My first rule of subjugation is: "They can only learn and do what they are taught!" Thus, one hopes that you will NOT take umbrage with what follows. However, your teachers, ......
As you are in such a "Divine" location, I am hesitant to suggest anything, especially in light of your use of such holy words of biology as "chunks" and "paraformaldehyde". I will, however, and only with great temerity and the deepest humility, suggest that you try to gain access to a tome of non-Harvardian origin entitled, "Principles and Techniques of Electron Microscopy"(3d ed.), Hayat, M.A.[he once worked in Newark, NJ!!!], CRC Press, Boca Raton, FL(1989), pp. 69-70 (sections entitled: "Simultaneous Fixation for Light and Electron Microscopy", and "Tissue Storage".) ISBN: 0-8493-7111-2. My humility is manifested for the divinity, but my temerity is evoked by the ignorance. However, Hayat begins one paragraph with the following sentence, "The size of the tissue specimen and the objective of the study primarily determine the type of the fixative solution to be used." He further mentions 4% formaldehyde and 1% glutaraldehyde in a buffer at pH 7.4 with an osmolality of ~200mo for "medium" pieces (~3mm) for longer times up to 12 months. Plants get specific treatment on pp. 72-73. There are other books, though I must admit, notwithstanding the efforts of Science Librarians, most of them have been 'lifted' by the new wave of dedicated students and others during the past 30 years. Try the 'older' members of the biology faculty - non-molecular, if possible, - and remember that HCHO has been used in some states to INDUCE fluorescence in tissues!
Again, PLEASE forgive. The last time I spoke to a student, a pre-med by-the-by, I didn't understand him either.
We old-timers are severely challenged, because we were trained to:
choose a vocational path to follow, ask questions after we had gone to the library, judge Timothy Leary as the short form of a donkey, understand women as a great mystery of the universe, leave the gloves off when dissecting, PLEASE understand, and NOT ask about the war, use slide rules (to do logs), planimiters (to do areas), and Leroy Lettering Sets (to label figures), understand the divine nature of (inspiration!) discovery take very deep breaths in the lab during days of dissection,
mix formaldehyde ("Formalin") and phenol to make good embalming fluid, squirt phenol on a sponge and wipe to disinfect lab benches in bacteriology, know that et cetera WAS Latin not Siamese(now Thai), take deep breaths in bacteriology lab, accumulate good books while they were relatively inexpensive, know that phenol was hygroscopic, cut sections from any tissue as long as it came from an unlabelled jar, label the jar, remember Latin and Greek roots (On Divinity Avenue, is that "dis-" as in "to separate" or dice-" as in "to chop up"?), do our income taxes on cards, tell students off, treat your fiancée' as the virgin you hoped she was, know the difference between formaldehyde and paraformaldehyde, keep the lab windows open during dissections to keep your eyes from burning, cultivate your wife as your best friend, ignore HIV and gHSV - no one had ever heard of them, be terrified of pregnancy before marriage (the greatest deterrent of STD's in the 20th century!), and know the difference between discovery and invention (a la, polymerase, la la, and the PCR, la la).
We were taught BY the organic chemists to pay very strict attention TO the organic chemists, because they were said to know most of the important secrets. The physical chemists, at the time, were busily unraveling the secrets of life - only one 'A' every five years among hundreds of chemists! Biochemists were chemists. Biologists cultured cells, developed vaccines, cut sections, referenced papers from the 1860's, didn't teach DNA until the early 1960's when protein genes were defeated at the polls, and, just now, may be getting - in another decade - to the point where the calculus may have a place in their courses. In the 1950's, or so went the legend, Harvard Medical School (Heaven's Gate?) sent representatives to Lehigh, where my teachers worked, to discover the secrets of chemical pedagogy that prepared LU graduates so well for the divine medical training some few received at the hands of Harvard's Professeurs! The secret pedagogical trick they discovered at Lehigh consisted of: "NO MERCY!", "NO SYMPATHY!" and "TOTAL RIGOR!", the only means ever devised to overcome the (utter) shame of being considered near to, but nonetheless, NOT, the equal of those "IVY" places.
If you didn't have fun, I wish you had. AND, before you store in buffer, think about it (extraction of fixative and other things) long and hard!
Regards,
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: Rachel Spicer } Sent: Friday, June 22, 2001 2:38 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Q re: long term storage in fixative } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello, } } I'm fixing small chunks of wood in 4% phosphate buffered paraformaldehyde, } and separately in a Karnofsy-type (paraformaldehyde and glutaraldehyde) } buffered fixative. I need to store the samples for a year or more - are } either of these fixatives appropriate for long-term storage, and if not, } what are the problems (e.g., tissue hardening?). Does anyone have } suggestions for appropriate storage solutions, or have an idea of how long } is OK in either of these fixatives? The samples in paraformaldehyde are } used for fluorescnce microscopy, and the Karnofsky fixed samples for TEM. } Thanks in advance for your experience/advice. } } Rachel } } ****************************************** } Rachel Spicer } Biological Laboratories 3119 } Organismic and Evolutionary Biology } Harvard University } 16 Divinity Avenue } Cambridge, MA 02138 } } (617) 496-3580 (phone) } (617) 496-5854 (fax) } spicer-at-oeb.harvard.edu } ****************************************** } } }
Novascan Technologies has a great deal of experience in the area of biochemical AFM and I'm sure we can help you with your project We have been developing AFM products for several years, but our specialty is fabrication of AFM probes and surfaces with specific surface chemistries. These chemistries, for example, can be used to couple molecules to surfaces for imaging and/or molecular characterization. Please contact us at info-at-novascan.com or visit www.novascan.com
Best Wishes,
Raj
At 04:17 AM 06/20/2001 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
********************************************** Dr. Raj Lartius, CEO NovaScan Technologies, Inc. Iowa State University Research Park 2501 North Loop Drive Ames, Iowa USA 50010
Email: rlartius-at-novascan.com Voice: 515-795-3164 Fax: 515-795-4414 ********************************************** "Innovative Tools to Explore the Microworld"
In our lab in Massachusetts, we have a Balzers CPD-020 that is in need of repair. The pressure guage (WIKA, 0-160 bar) is leaking, and we'd like to have it replaced. This unit was made in Switzerland, but the manufacturer has been out-of-business for many years. We'd appreciate input from Balzers CPD owners regarding sources for parts. Any comments concerning safety issues would be welcomed also. Many thanks in advance.
Does anybody have experience with High Resolution Imaging Plate Scanner for Electron Microscopy (http://www.ditabis.de). Instead of conventional EM-films, flexible photostimulable phosphor screens of the same size, the so-called Imaging Plates, are loaded into the TEM magazine. Are they as good as negatives?
Thanks, Julian
_______________________________ Julián Martínez Fernández Senior Research Associate MS 106-5 Ceramic Branch NASA Glenn Research Center Cleveland, OH 44135 Phone: 1-216-433-5512 Fax: 1-216-433-5544
You may find that only the JEOL one will properly fit in, I investigated a GW Electronics one for my JEOL 840, but they didn't have a size that would fit exactly. Mind you, I have a (JEOL) optical microscope fitted, and the BSE detector would have had to fit into a severely constrained space on the end of the OM.
I would ask JEOL first, I find that their prices are sometimes better than those from third-party suppliers anyway.
} } } Hello All, } I have a Jeol 5900 SEM, and I would like to purchase a BSE detector } for } it. Does anyone in the group have any } suggestions/recommendations/warnings? Is it pretty straight forward } to purchase a third-party detector and get it to work with our } microscope, or is it best to go through the vendor? Thank you for } any input that you can provide. }
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
Hello, We just inherited an older model (10 years old) model E-3 Electroscan Environmental SEM. I have some questions in its operating vacuum modes as it has been quite a bit picky since we've installed it.
On going from a standby vacuum state into either Hivac or Wet vacuum modes, it will often have an error of can't pump column EC2. Also, in Wet mode, it will not open valve V1, so you can never see the beam. I've read over the manuals, but I was wondering if anyone out there has some experience with this model ESEM. We are thinking that one of the diffusion pump heater elements needs to be changed. Other times we operate the unit, it will maintain a Hivac vacuum mode for several hours without difficulty.
Any advice or comments, appreciated, Thanks.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
I did study this subject a few years ago and find that this technique is not well optimized for EM. For many reasons:
- cost of the system is similar to the EM digital camera - you require to load/unload Imaging Plates into the cassettes (you waste the time) - it takes from 2 to 5 min to scan one Image Plate (waste time) - I am not sure exactly, but seems to me, that you have to scan Image Plates soon after tacking picture, otherwise the image will deteriorated, and seems to me they are sensitive to light too. - you don't have a chance to adjust your image before recording (as it happens in case of digital cameras). -number of "shots" is limited and Image Plates are princely - Image Plates may be damaged during loading/unloading which may reduce declared number of use (a few thousand times I believe).
The good things about Image Plates are: - Image plate has more pixels than EM digital cameras, so the resolution is better. - you don't have to make any changes in the microscope. - the linearity and dynamic range is similar to the EM digital cameras - easy to use and less problem related to the equipment.
My conclusion was: technically, it's a great stuff, but it does not help in practice. You still need some help from technician to load/unload plates and scan them and you have to pay a 20-40$K for that beauty. Not so promising technology for TEM at least.
This information based on my "study" performed a few years ago and may not reflect modern improvements in this area.
Sergey.
At 03:11 PM 6/25/01 -0400, 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 was talking about particular DITABIS system Julián was mentioning in her original message (with reference to the manufacturer).
All digital cameras including Gatans are approximately 10 times more sensitive than 4489 film. I know it for sure because just returned back from Gatan's demonstration. Image Plate should have very similar characteristics in terms of sensitivity, dynamic range and linearity as modern digital cameras. The best things about Image Plates it's their pixel size: 5000 x 4500 total pixels per plate. It's greater than in digital cameras: 1000x1000/2000 in most cases. It's still less than you could get from film with $800 scanner: 6400x4800 for 1600 optical dpi. BUT!!! You have LOAD/UNLOAD plates into cassettes manually. In the dark. Stack them and load into the scanner (so, you have to have space for the scanner in dark-room). Scanner will process them automatically (and at that time you could not use dark-room for regular things, light from scanner). Load them back into cassettes, degas magazine and load it into microscope. I don't remember exactly, but it seems to me the cost of one plate is something around $100. So, I have two magazines with 50 cassettes each: 100x100$ plus scanner plus salary for technician plus computer and storage media. Ok, I forgot to mention that you have to ERASE plates before use, also, using special eraser. I don't remember, how many plates you could ERASE in one run. May be it's automatic too. Scanning: 3.5 min per plate x 50= 3 h.
As I mentioned before, the technique is great, but not so helpful for regular day-to-day basis EM and does not save time so much. It works fine for molecular biology applications in "phospho-imagers" but people need to scan a few plates per day (even one a week) and they save a lot of time: 10-15 min exposure vs 5-10 days! I just don't see reason to switch to this technique to myself. If I'll have such money, I would buy digital camera. It's more practical solution from my point of view.
Sergey
At 01:36 AM 6/26/01 -0500, you wrote: } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } Sergey, } } I last saw the unit demonstrated at the EUREM meeting in Brno last summer. } It is not as you described it. I won't say it is better but overall it is } comparable to the Gatan product. } } What impressed me the most about it is that } } a) exposure times are comparable to what is now used for conventional film } (the Gatan unit has long exposure times) and } } b) while the scanning step (of the digital plates) takes some time, it does } it all automatically, so there is no babysitting with the scanner. You are } free to do other things including exposing more digital plates. } } They might have one at the MSA meeting in Long Beach. } } Chuck } SPI Supplies } -------- REPLY, Original message follows -------- } } } Date: Monday, 25-Jun-01 08:20 PM } } } } From: Ryazantsev, Sergey, Ph. D. \ Internet: (sryazant-at-ucla.edu) } } To: Julian Martinez-Fernandez \ Internet: } } (julian.martinez-fernandez-at-grc.na) } } To: MICROSCOPY BB \ Internet: } } (microscopy-at-sparc5.microscopy.com) } } } } Subject: Re: High Resolution Imaging Plate Scanner for Electron } Microscopy } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To } } Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On- } Line } } Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } I did study this subject a few years ago and find that this technique is } not } } well optimized for EM. For many reasons: } } } } - cost of the system is similar to the EM digital camera } } - you require to load/unload Imaging Plates into the cassettes (you waste } } the time) } } - it takes from 2 to 5 min to scan one Image Plate (waste time) } } - I am not sure exactly, but seems to me, that you have to scan Image } } Plates soon after tacking picture, otherwise the image will deteriorated, } } and seems to me they are sensitive to light too. } } - you don't have a chance to adjust your image before recording (as it } } happens in case of digital cameras). } } -number of "shots" is limited and Image Plates are princely } } - Image Plates may be damaged during loading/unloading which may reduce } } declared number of use (a few thousand times I believe). } } } } The good things about Image Plates are: } } - Image plate has more pixels than EM digital cameras, so the resolution } is } } better. } } - you don't have to make any changes in the microscope. } } - the linearity and dynamic range is similar to the EM digital cameras } } - easy to use and less problem related to the equipment. } } } } My conclusion was: technically, it's a great stuff, but it does not help } in } } practice. You still need some help from technician to load/unload plates } and } } scan them and you have to pay a 20-40$K for that beauty. Not so } promising } } technology for TEM at least. } } } } This information based on my "study" performed a few years ago and may not } } } reflect modern improvements in this area. } } } } Sergey. } } } } } } } } At 03:11 PM 6/25/01 -0400, you wrote: } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } Does anybody have experience with High Resolution Imaging Plate Scanner } for } } } Electron Microscopy (http://www.ditabis.de). Instead of conventional } } } EM-films, flexible photostimulable phosphor screens of the same size, the } } } } so-called Imaging Plates, are loaded into the TEM magazine. Are they as } } } good as negatives? } } } } } } Thanks, } } } Julian } } } } } } _______________________________ } } } Julián Martínez Fernández } } } Senior Research Associate } } } MS 106-5 Ceramic Branch } } } NASA Glenn Research Center } } } Cleveland, OH 44135 } } } Phone: 1-216-433-5512 } } } Fax: 1-216-433-5544 } } } } } } } _____________________________________ } } } } Sergey Ryazantsev Ph. D. } } Electron Microscopy } } UCLA School of Medicine } } Department of Biological Chemistry } } Box 951737 } } Los Angeles, CA 90095-1737 } } } } Phone: (310) 825-1144 } } Pager: (310) 845-0248 } } FAX (departmental): (310) 206-5272 } } mailto:sryazant-at-ucla.edu } } } } } } } } } } -------- REPLY, End of original message --------
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
I contacted ditabis several months ago. Beyond a confirmation email from the company, I haven't received any from them.
Steve Widing Temple University
On Mon, 25 Jun 2001, Julian Martinez-Fernandez wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Does anybody have experience with High Resolution Imaging Plate Scanner for } Electron Microscopy (http://www.ditabis.de). Instead of conventional } EM-films, flexible photostimulable phosphor screens of the same size, the } so-called Imaging Plates, are loaded into the TEM magazine. Are they as } good as negatives? } } Thanks, } Julian } } _______________________________ } Julián Martínez Fernández } Senior Research Associate } MS 106-5 Ceramic Branch } NASA Glenn Research Center } Cleveland, OH 44135 } Phone: 1-216-433-5512 } Fax: 1-216-433-5544 } }
For electron microscope service try Vitaly Feingold who is based out of Georgia. His references are excellent!! His phone number is 770-232-7785. E-mail address is: vitalylazar-at-worldnet.att.net
-----Original Message----- } From: Jill Verlander Reed [mailto:verlaj-at-medicine.ufl.edu] Sent: Monday, June 25, 2001 12:20 PM To: Microscopy-at-sparc5.microscopy.com
Fellow microscopists -
Beverly Giammara wrote asking for comments about EM service providers, other than the manufacturers, that cover the Midwest. I would be very appreciative if anyone can offer similar information about EM Service providers that cover Florida.
Sincerely,
Jill
Jill Verlander Reed, D.V.M. Associate Scientist Director, College of Medicine Electron Microscopy Core Facility University of Florida P.O. Box 100215 Gainesville, FL 32610 verlaj-at-medicine.ufl.edu Phone: (352) 846-0820 Fax: (352) 846-3299
Dear Rachel, After an entire evening of self-immolation, I have returned to the table to extend my apology for taking advantage of you yesterday. I really do apologize. I looked across the table at my own daughter last evening, and I realized that she might just have failed to see any humor at all in my response had she requested the same information. Having realized that possibility, I spent the rest of the evening in shock at my probable insensitivity. As an explanation, you received, in response to your entirely innocent query, a message composed of every quip I have wanted to make to my children, their teachers, their teachers' parents, my students, and probably a few I can't remember. I thought to make it humorous, and there are some who have decided that I succeeded, but I would suspect that you may have felt somewhat differently. Thus, this message in self-rebuttal. Not nearly as long, and hopefully more to the point. If you had walked into my office with the question you asked, I would have said, "Miss Spicer, we do not use words like "chunks" to describe specimens from which we wish to extract reasonable scientific information." You would have responded with some other nebulous term for the objects with which you were contesting, and I would have cajoled you, with a couple suggestions on precision in thinking and describing, to go to the library and start doing what you are obviously trying to learn how to do - namely, research. So, please let me begin again. My second rule is: "They only learn to think if they are forced." In that regard, I would have requested that you make a list of the steps in the process you are asked implement. For each step in such a process, there must be a purpose. For each step, there must be a prescribed outcome (I have come to despise that word!), and for each step, there must be BACKGROUND/HISTORY. My third rule is this. "The most difficult, and the most successful research for a bench scientist is done in the brain, the office, and/or the library." The work at the bench only certifies or debunks what was done before, but the same principal holds for even the most minor part of the process. You suggest that you must to store plant/wood tissue for a year. Most of us would want to know why? Why must you wait to at least embed. Why would you want to soak/extract the tissue for that length of time before acting on the NEXT step in the procedure. It is apparently not that you wish to archive the tissue/wood. So, with your list in your hand, the question for that step is, "Why?" "What is the purpose/reason for waiting?" I certainly cannot help you to progress in the trek to an answer until I know. Why? I'll give you an example. There was an undergraduate who accosted me in the hall during my first week as a graduate student. He asked me to point him in the direction of distilled water. I asked him what type he wanted. He assumed I was harassing him and told me to knock off the B.S. and just, "point!" I did. Unfortunately, his purpose was to subculture honey bee cells, and the 'distilled' water to which I pointed - the nearest - was made in a chromium plated metal still. Notwithstanding his rudeness, I should have insisted - through instruction, but I was fresh out of the military and I didn't have any time left for stupidity and self-destructiveness, Fourteen weeks later, he blamed me for the failure of his project - my first real interface with a baby boomer of the 'gimmee' type. I only felt sympathy for the instructor, but his advice (he had been in the military too) was to learn to take responsibility for the errors of students, for, someday I would be their teacher.
Over the past thirty years, what was a two-way street in learning when I was a student, has become one-way. I guess the ultimate point is this. If you are working on someone else's project, show him or her my notes. If you are working on a project of your own, and you are willing to exercise patience and learn, then when you ask for information from us - the MSA ListServer - you must provide us with specific information. I can promise you that you failed to receive many responses, because your question lacked three elements:
1. Precise terminology. 2. Purpose/Goal/Reason for need. 3. Trust.
Everyone on the ListServer has a job or values his/her time. Your request was informal in form, because it lacked 1) precise detail, 2) a reason for wanting to delay further processing (that would be important for any specific response), and 3) trust in us to be honest brokers and not steal your project or question its value. Far too many visitors to the Listserver ask a question as if in class, at the end of a lecture. We all came in late.
I hope you want to try again, if what little you have received has not helped. We would like to help as much as possible. We are here for that. Not for entertainment. I am here, because when I read the messages, I learn or have questions that I can address. I am here to be educated, just like you.
With closing apology for wordiness and philosophy, I am
Yours sincerely,
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: Rachel Spicer } Sent: Friday, June 22, 2001 2:38 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Q re: long term storage in fixative } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello, } } I'm fixing small chunks of wood in 4% phosphate buffered paraformaldehyde, } and separately in a Karnofsy-type (paraformaldehyde and glutaraldehyde) } buffered fixative. I need to store the samples for a year or more - are } either of these fixatives appropriate for long-term storage, and if not, } what are the problems (e.g., tissue hardening?). Does anyone have } suggestions for appropriate storage solutions, or have an idea of how long } is OK in either of these fixatives? The samples in paraformaldehyde are } used for fluorescnce microscopy, and the Karnofsky fixed samples for TEM. } Thanks in advance for your experience/advice. } } Rachel } } ****************************************** } Rachel Spicer } Biological Laboratories 3119 } Organismic and Evolutionary Biology } Harvard University } 16 Divinity Avenue } Cambridge, MA 02138 } } (617) 496-3580 (phone) } (617) 496-5854 (fax) } spicer-at-oeb.harvard.edu } ****************************************** } } }
Thanks to all who responded to my post re: long term storage of plant material in fixatives. The responses were as varied as the recommendations in the literature, but I find there is no substitute for hearing from those with first hand experience. If anyone is interested in the specifics I'd be happy to forward a collection of relevant responses. And finally, apologies for my misspelling of "Karnovsky". Thanks again.
Rachel
****************************************** Rachel Spicer Biological Laboratories 3119 Organismic and Evolutionary Biology Harvard University 16 Divinity Avenue Cambridge, MA 02138
Congratulations on your E3 acquisition - they're generally easy-running instruments once you get the preliminary bugs out. I've been running an older E3 for the past few years, but that doesn't make me an expert on them. Your vacuum problems sound a little funny - have you checked that both diff pumps are hot to the touch? If one is cold (the one on the right hand side of the column, viewed from the front, that could explain part of the problem. If it is, try the reset button on its "front" side, and check cooling water flow to make sure it's adequate. Somebody good with electrical stuff (I'm not) can probably determine whether or not your diff pump element is still good. We once had V1 stick a bit, obstructing the beam, but were able to free it up by reversing the compressed air flow momentarily. You can actually watch it open and close if you remove the sheet-metal shroud on the left (the one with the black cylindrical receptacle for the gun for those times when you've pulled the gun - I know, I thought it was a cup holder at first, too, but believe me, it isn't...) The E3 is still supported by FEI, though, and I've always found their guys pretty good about being able to fix things over the phone, even without a service contract. You can get their phone numbers, etc from the site www.feicompany.com. The guys in the Peabody, Mass. office should be able to help you through this one - that's where your E3 was born. Hope this helps.....
Frank Thomas MicroAnalysis Facility Geological Survey of Canada (Atlantic) Bedford Institute of Oceanography Dartmouth, Nova Scotia Canada
----- Original Message ----- } From: "Gordon Vrololjak" {gvrdolja-at-nature.Berkeley.EDU} To: {microscopy-at-sparc5.microscopy.com} Sent: Monday, June 25, 2001 7:11 PM
Julian asked if imaging plates are “as good as negatives”. There are advantages and disadvantages to all imaging systems. In some instances CCD cameras are preferable to film, in other instances film is superior (especially if wide field of view is a factor). What is best to use largely depends on the details of your experiment. I thought I would provide some information on imaging plate systems, and indicate one strong advantage the technology has over other recording media/systems.
Image plate systems use the "photostimulated luminescence" (PSL) phenomenon. Regions in PSL materials that have been previously exposed to a short wave length irradiation, will luminesce when stimulated by a second longer wavelength irradiation. The stored energy is stable until released by the second irradiation. Imaging plates are made using PSL films of barium fluorobromide (= 5 µm grain size), containing a trace amount of bivalent europium as a luminescence center, which are uniformly coated on a polyester support. Imaging plates replace conventional photoemulsion film in the transmission electron microscope and are exposed the same as normal film. To read the recorded image, the imaging plate is latter scanned by a red diode laser and the luminescence is collected through a light collection guide to the photo-multiplier tube (PMT). The analog current produced by the PMT is converted to a 8 to 16 bit depth digital signal. The pixel density can be read at between 50 to 400 pixels/cm. Imaging plates can be fully erased for reuse, but have a finite lifetime.
One main advantage of PSL films in their ultrahigh sensitivity (e.g. 2 x 10-14 - 2 x 10-9 C-cm-2 at 100 kV) to electron irradiation making them valuable for biological work (these are values quoted by a manufacturer Fuji). For example, film emulsions require 0.1 to 1 C-cm-2 to record an optical density S = 1 at 100 k magnification (see Reimer, 1975). Slow scan CCD cameras offer better sensitivity than film, however are inadequate for imaging certain biological specimens. Furthermore, CCD cameras have limited fields of view as compared to film. As an example, Sugi (1997) and coworkers have used a Fuji imaging plate system to record micrographs of myosin head movement induced by ATP hydrolysis in an environmental cell TEM. I avoided using the term “living myosin” since a biomolecule is not strictly alive, but it was biologically active. Electron doses low enough not to damage the delicate biomolecule could be used to record images on an imaging plate.
Reimer, L. (1975) in Physical Aspects of Electron Microscopy and Microbeam Analysis, eds B. M. Siegel, and D. R. Beaman, John Wiley & Sons Publishers, New York, New York, 231-245.
Sugi, H., Akimoto, T., Sutoh, K., Chaen, S., Oishi, N., and Suzuki, S. (1997) Proc. Natl. Acad. Sci. USA 94, 4378-4382.
} } } Does anybody have experience with High Resolution Imaging Plate Scanner } for } } } Electron Microscopy (http://www.ditabis.de). Instead of conventional } } } EM-films, flexible photostimulable phosphor screens of the same size, the } } } so-called Imaging Plates, are loaded into the TEM magazine. Are they as } } } good as negatives? } } } } } } Thanks, } } } Julian } } } } } } _______________________________ } } } Julián Martínez Fernández } } } Senior Research Associate } } } MS 106-5 Ceramic Branch } } } NASA Glenn Research Center } } } Cleveland, OH 44135 } } } Phone: 1-216-433-5512 } } } Fax: 1-216-433-5544 -- _____________________________________________________ Tyrone L. Daulton Director: Marine Geosciences - Electron Microscopy Center Physicist and Senior Electron Microscopist
Naval Research Laboratory Marine Geosciences Division (Code 7400) Stennis Space Center, MS 39529-5004
Thanks so much Charles. I just wonder why I did not know that such grids are available. I use JEOL 200CX TEM to look at nanoparticles. What is the thinnest window grid do you recommend?
Regards,
Thearith
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
This has become quite a generic problem for many who are working with nano size particles, and many have found the "solution" to be the use of silicon nitride membrane window grids that have been offered by SPI Supplies for several years. Information about these grids can be found on URL http://www.2spi.com/catalog/instruments/silicon-nitride.html
The silicon nitride is completely structureless and featureless and seems to provide excellent electron transparency. The other advantage is that they are sufficiently robust to survive temperatures in excess of 1000° C. And there is no carbon grain to be confused with the nanoparticles of interest.
Disclaimer: SPI Supplies offers silicon nitride membrane window grids for this type of application.
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 ############################ ==================================================
We are in the process of purchasing an EDS system for our JEOL 100 CX II TEM microscope. We would like to have the EDS system installed at a high angle but we need a "HAXI" kit in order for the EDS system to be installed correctly.
Does anyone have a "HAXI" kit for purchase?
Thank you,
Lou Bustillos AMA Analytical Services, Inc. lbustillos-at-amalab.com
The Image plate system is awesome for materials applications as well.... !!
It is very easy to obtain diffraction patterns that include varying intensities (e.g., a spot pattern mixed with Kikuchi lines) with just one exposure on a single plate. The plates also show the slightest deviations in contrast which are not discernable using film, a CCD camera, or the TEM viewing screen. For example, differences in contrast in amorphous silicon oxide due to the multiple processing steps in integrated circuits may be easily observed with the plates. The advantage in the plates is evident in the low to intermediate magnification ranges where large variations in image contrast can be manipulated off-line. Each IP file is quite large (25MB) and therefore, contains plenty of information which can be manipulated before reducing its size that is suitable for publication purposes. The disadvantages to the IPs are the cost (although I believe the cost is dropping) and the extra processing time needed (~ 80 minutes for a batch of 32 plates).
The CCD camera is still very useful in the high resolution imaging regime where the intensity is rather uniform throughout the field of view and one can instantly observe the quality of the image. We will be evaluating high resolution CCD images with high resolution images obtained with the IPs in the near future. We are not sure what effect (if any) vibrations from the film mechanism will have on the high resolution images.
******************************************************************* Lucille A. Giannuzzi, Ph.D.
Associate Professor, Mechanical, Materials, and Aerospace Engineering Director, UCF/Cirent Materials Characterization Facility University of Central Florida 12443 Research Parkway, Suite 305 Orlando, FL 32826 email lag-at-mail.ucf.edu phone (407) 882-1500 fax (407) 275-4321 -------------------------------------------------------------------- "Good judgement comes from experience.
Experience comes from making bad judgement."
Mark Twain ********************************************************************
The messages from Professor Monson in reply to a question about long-term tissue storage makes me wonder what has happened to this listserver?
His message was highly entertaining and even if the poster of the original message may have felt "picked on", there was no reason to expect him to post an apology, no matter how informative it was.
Far too many posters use this listserver as a way of finding a quick fix to simple solutions to technical problems that can be easily researched in text books.
Often these messages contain little background information on the technical problem under investigation. Even so, many of these questions get full attention and detailed replies, even if the replies are not posted on the main list for public view.
What has happened to the free and open discussion of issues and protocols that we used to have? Are people so afraid of being accused of being incorrect that they do not post their replies openly? Or maybe people are afraid of being accused of being impolite.
I look forward to controversy and hope that my comments are read carefully and replied to with vigor. Only in this way will I be able to learn new thing, and this is the main reason I still subscribe to this listserver.
If I am factually wrong about something, tell me. It is better to find this out on the listserver than in a more professional (or cruel) environment.
As for discussion subjects: why was there no comment about the unadvisable suggestion made about storing samples in a frozen state? Would anyone working with EM really choose to store their samples at -20 degrees before processing for TEM?
Where is all the discussion about extraction that can occur when storing samples in alcohols or buffers, or on the contamination that I know grows in everyones old cacodylate buffers?
Do we not comment because we know it has all been published in textbooks?
Dear Profesor Monson, do not feel bad about improving our general knowledge and our professional etiquette. I for one support your views and find your comments useful and entertaining. Please keep up your submissions.
Best regards,
Paul Webster
Paul Webster, Ph.D. Scientist II and Director Ahmanson Center for Advanced EM & Imaging House Ear Institute 2100 West 3rd Street Los Angeles CA 90057
Scotty Cornelius wrote: ============================================ In a far corner of my mind, I vaguely remember a set of rare earth glass standards that had been developed and were being offered for sale a few years ago, possibly out of the UK. Does anyone know of such a set, or was it all a pipe dream? I'm not referring to the Drake and Weill standards from the early '70s. ================================================ Could you have in mind our (e.g. SPI Supplies) "15 Rare Earth 'Five Phosphates' Standards" for Microanalysis? They can be found on URL http://www.2spi.com/catalog/standards/reepo/reepo.html
At one point, we had inadvertently listed these as "glasses" when in fact they are crystals. So we might have been ourselves the cause of some confusion.
Chuck
Disclaimer: SPI Supplies offers a range of standards for microanalysis including this unique rare earth "five phosphate" mount.
===============================================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 ############################ ===============================================
We have just been testing the Ditabis IP scanner and were impressed.
It doesnt quite replace film or I think a good small-pixel below-screen CCD camera for resolution, but is much better than a whole field above screen CCD camera for resolution, and for dynamic range is in a class by itself.
We tested with diffraction patterns showing bright and dim diffuse features, convergent beam patterns, wedge-profile ion beam thinned sections, and normal and unstained biological material.
For low dose work, 1-2 seconds exposure at beam intensities barely visible to the eye was no problem. The resolution problem may be avoidable in practice because images may be taken at higher magnifications and lower beam intensities than normal for film - that is just a question of getting used to matching the imaging conditions to a different medium.
PLate handling is easy. The plates are loaded into the TEM magazine in the light. AFter electron exposure they are kept dark, but loaded into the scanner under light as dim as convenient - we used the light from the monitor screen.
Storing the plates for up to five days made only a very small difference to the intensity histogram - we didnt test beyond that. Plates can also be rescanned with very little loss.
The only real inconvienience we found was the impossibility of recording plate number, mag etc on the IP plate, as we are used to on the negative. If we had our own system we would mark the plates permanently in some way that showed on the readout.
IP plates are complementary to CCD cameras in some respects. You dont get the advantages of immediate viewing, but more dynamic range, more pixels, and of course one scanner can take plates from any number of TEMs. Finally, the price was the sticking point for us at this time. But if a cooperative lab somewhere else on this continent gets one we would likely buy a set of plates and work through the postal system until the price drops or somebody comes up with a "must have" application!
regards Sally Stowe
Dr Sally Stowe Facility Coordinator, ANU Electron Microscopy Unit Research School of Biological Sciences Australian National University Canberra ACT0200 AUSTRALIA stowe-at-rsbs.anu.edu.au fax 61 (0)2 6125 3218 ph 02 6125 2743 http://www.anu.edu.au/EMU
} } } Julian Martinez-Fernandez {Julian.Martinez-Fernandez-at-grc.nasa.gov} 06/26/01 05:11AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Does anybody have experience with High Resolution Imaging Plate Scanner for Electron Microscopy (http://www.ditabis.de). Instead of conventional EM-films, flexible photostimulable phosphor screens of the same size, the so-called Imaging Plates, are loaded into the TEM magazine. Are they as good as negatives?
Thanks, Julian
_______________________________ Julián Martínez Fernández Senior Research Associate MS 106-5 Ceramic Branch NASA Glenn Research Center Cleveland, OH 44135 Phone: 1-216-433-5512 Fax: 1-216-433-5544
I have downloaded from Lumijet4s home page information about the Preservation Monochrome Inks they commercialize for jetink printers, a sort of special ink for good image tonality and permanence. Has any one had any experience with that product? or any other like that?
Regular black ink cartridge would be enough por good quality greyscale printing???
Thanks a lot,
Silvia Montoro Centro Regional de Investigacisn y Desarrollo de Santa Fe (CERIDE) G|emes 3450 3000 Santa Fe
We also have an E3 and have had our share of vacuum problems. Last year the vacuum had gotten to the point that we needed to do something. Another employee and I took all the valves off and removed the diffusion pumps. We cleaned up the pumps and added oil. Then we replaced all the o-rings in the couplings and valves and diff pumps. (basically they were all flat) After that the vacuum was as good as when we first got it and pumping down was much faster. This is something that probably should be done every 10 years whether it needs it or not! We also had some mysterious water loss from the chiller for over a year that finally appeared as a major water leak at the diff pump tubing. We cut off the brittle parts and reconnected. Cooling was better and vacuum too. Of course you should be sure all your vacuum pumps are up to the job, particularly the large pump used for the column. The vacuum hose that goes between the 2 diff pumps should have a metal cover on it to protect it from the heat. It does become brittle and should be checked for leaks. We replaced ours as well. As others have mentioned, the E3 is still supported and although we also do not have a service contract, the engineers at FEI versed in the E3 are VERY helpful, patient and willing to troubleshoot a problem over the phone! Good luck! 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 ********************************************************************
If anyone has a Hitachi S-570 with LaB6 that they are getting rid of, we would be interested in any left-over LaB6 crystals mounted in the Wehnelts. Thanks!
Probably today's humor, but hey, it's worth a try.
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)
Paul, While you are correct in stating that the answers to many of the questions asked on this listserver are easily found in a number of available texts, many of the libraries at smaller colleges and universities do not have the funds to maintain an extensive section dedicated to electron microscopy or even microscopy in general. Therefore, those books could be days away via inter-library loan. I always looked at this server as a gathering of some of the great minds in microscopy and a "mother lode" of information based purely on experience, if nothing else. Being a one man show at a small university, I am constantly bombarded with questions about various techniques from investigators throughout the sciences, and have utilized this source many time for answers to those inquiries. There are no stupid questions, only ignorant people asking those questions; and intelligent, informed and timely answers will make those folks somewhat less ignorant. I am now getting down from the soapbox.
Franklin Bailey Electron Microscopy Center University of Idaho Moscow, ID 83844-2204
Our Philips CM-10 transmission electron microscope is now for sale by Michigan State University. For more details, please check the MSU surplus web site:
If you have any other questions, please contact me at: Ph: 517-353-4525. Fax: 517-4325174, Email: fanx-at-msu.edu, We are available for instrument demonstration and examination at any time.
Yours Sincerely, Xudong Fan, Ph.D. Center for Advanced Microscopy Michigan State University East Lansing, MI, 48824
Dear Rachel, Let me add. 1. On storage in buffer: NOT recommended by all my experience and all who have ever advised me. 2. On storage in fixative: NOT recommended as above, always because of unknown variables. 3. On fluorescence microscopy after HCHO: HCHO (dry) has been used to induce fluorescence in so-called biogenic amines (incl., decarboxylated amino acids). One never knows what else it may do during long residence. Ask yourself: Do you want fluorescence of fixed wood or wood? 4. One cannot fix with paraformaldehyde (not your fault). It is a polymer of HCHO and must be depolymerized to yield full methanalic (arch.) power! This (depolymerization) can occur when paraformaldehyde is suspended in a neutral or slightly basic buffer and permitted to stand for a while or heated briefly to 50-60degreesC.
The real identity of paraformaldehyde was the only thing I remember from my failed first course in freight-train organic chemistry [A carload of this and a carload of that yields half a carload of something else!]. A second foray into mechanistic organic went much better.
Summary: Old-timers stored materials to save information. Young folks can't afford the luxury if they want to collect data.
Moral: If you can have some tea while he vents, you might survive to get some solid advice.
Final Regards,
Fred Monson,
} ---------- } From: Rachel Spicer } Sent: Tuesday, June 26, 2001 11:25 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: thanks re: long term storage } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Thanks to all who responded to my post re: long term storage of plant } material in fixatives. The responses were as varied as the recommendations } in the literature, but I find there is no substitute for hearing from } those } with first hand experience. If anyone is interested in the specifics I'd } be happy to forward a collection of relevant responses. And finally, } apologies for my misspelling of "Karnovsky". Thanks again. } } Rachel } } ****************************************** } Rachel Spicer } Biological Laboratories 3119 } Organismic and Evolutionary Biology } Harvard University } 16 Divinity Avenue } Cambridge, MA 02138 } } (617) 496-3580 (phone) } (617) 496-5854 (fax) } spicer-at-oeb.harvard.edu } ****************************************** } } }
A list of independent EM service providers can be found at "www.jcnabity.com/service.htm".
Note that their "service areas" are not necessarily limited to the "location" that is listed.
If anyone has additional information that can be included on this list, please let me know. Any suggestions for independent EM service providers that are based overseas will also be appreciated.
Joe _________________________________________ Joe Nabity, Ph.D. JC Nabity Lithography Systems E-Beam Lithography using Commercial SEMs & STEMs PO Box 5354, Bozeman, MT 59717 USA Voice: (406) 587-0848 FAX: (406) 586-9514 E-mail: info-at-jcnabity.com Web: www.jcnabity.com
In a message dated 6/25/2001 2:59:20 PM Mountain Daylight Time, verlaj-at-medicine.ufl.edu writes:
} Beverly Giammara wrote asking for comments about EM service providers, } other than the manufacturers, that cover the Midwest. I would be very } appreciative if anyone can offer similar information about EM Service } providers that cover Florida. } } Sincerely, } } Jill } } Jill Verlander Reed, D.V.M. } Associate Scientist } Director, College of Medicine Electron Microscopy Core Facility } University of Florida } P.O. Box 100215 } Gainesville, FL 32610 } verlaj-at-medicine.ufl.edu } Phone: (352) 846-0820 } Fax: (352) 846-3299 }
Hello everyone: I have a student who wants to do some immunolocalization of a protein in plant tissue. I have been combing my files, but haven't managed to find a decent protocol that she could work from... (and can't seem to get myself to the library for a more comprehensive search). Can anyone supply us with a "generic" protocol for immunofluorescence in plants?
A customer of mine sent a part that has started showing a failure only recently, although he has manufactured the part for years. The part is a heating element. It consists of a thick SS rod which is the terminal. It has a diameter of about 1 cm. It is joined to an element that is about 2 mm in diameter. The joint is made by 1st drilling the thick stock and then inserting the wire into the hole. Then a notch is made in the thick stock until the thin element is exposed. Then the notched surface is welded without the introduction of any new material. I believe it is a "TIG" weld. A standardless analysis of the terminal shows Fe (82%), Cr(13%), Al(3%), Si(1.6%), and Mn (.3%). He has tried to match the two materials as closely as possible.
The joint is heated repeatedly, as it is used as an element in a glass furnace. The failure starts with a bulk flow of material out the top of the hole, which is about 1.5-2.0 cm from the welded joint/notch, and over the sides of the terminal. I have analyzed this material and it contains principally Fe and O. The area I analyzed was polished so the internal composition of the flowed material contains a great deal of oxygen. No Cr is detected and very little Al.
I feel that this material flow degrades the joint causing the various electrical failures that have been observed.
My questions are:
Is this a typical failure mode with a Fe/Cr SS weld experiencing many heat cycles?
What is the mechanism that results with the crystalline appearance of the iron oxide, leaving the other elements behind?
This failures appears to have started when a new aqueous lubricant was introduced for the initial hole drilling of the terminal. Could this play a role in the failure?
What measures might be taken to reduce the frequency of this failure mode? Joint design? Material compatibility?
Hi, could anyone please give me some information on where I can get my diamond knives (for semi thin sectioning) sharpened? Any such places in Canada? I believe the knives were made in Germany. Any information is highly appreciated. Thank you in advance.
I would like to gather some information from the micromanipulator community. Specifically, this tool will be used for lamella extraction after the FIB work is done. We will be in the market to purchase one in the upcoming months so I'm starting my homework early. Ideally, this will be a dedicated station with an anti vibration table along with the optical microscope. Some questions are: 1) Motorized vs manual 2) Objective lens working distance 3) Probe resolution 4) Sensitivity (microns per degree)
Any comments or suggestions are welcomed!
Thank you,
Jon Hiller
================================================================== Jon M. Hiller Argonne National Laboratory Materials Science Division Electron Microscopy Center Tel: 630-252-9558 Fax: 630-252-4798 Email: hiller-at-anl.gov ==================================================================
There is a wonderful protocol for immunofluorescence in a manual which I use for most of my Arabidopsis work.
Methods in Plant Molecular Biology A laboratory course manual Maliga, Klessig, Cashmore, Gruissem, Varner Cold Spring Harbor Lab Press
Good luck,
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 Wed, 27 Jun 2001, Shea Miller wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello everyone: } I have a student who wants to do some immunolocalization of a protein in plant tissue. I have been combing my files, but haven't managed to find a decent protocol that she could work from... (and can't seem to get myself to the library for a more comprehensive search). Can anyone supply us with a "generic" protocol for immunofluorescence in plants? } } Thanks so much in advance } shea } } } } }
What is the composition of the heating element? The S.Steel terminal is not a common (to me) stainless, given the composition you obtained. I would have to check references to see what it may match. I work mostly with austenitic stainless and nickel super alloys when it comes to "SS".
I suspect the welling up of oxide from the joint/hole is a symptom rather than a cause of the failure. Contact area and/or conductor size reduction will (all else being the same) cause a hot spot. What is the shape of the FeOxide crystals? Sounds like you may have magnetite, a stochiometry often formed under oxygen starved (but still oxidizing) environment.
I would question the customer carefully about process conditions which result in failure. What is the composition of the lube? If residue remains, it could surely play a role in the corrosion. Are there any differences in glass chemistry or temperatures compared to the non-failed parts? What environment is "seen" by the joint? -} In the Glass, near it? Glass chemistry? If near it, what is the atmosphere? Temperatures? Heating power/time curve the same (i.e. any localized I2R heating of joint)? Getting accurate and complete information from customers is sometimes the most difficult part of failure analysis. ...Especially if it is second hand.
Too much weld heat can "sensitize" some alloys (changes carbon/carbides), reducing resistance to corrosives. The failure mode, however, is most often intergranular cracking and (again) I am not familiar with the characteristics of your composition.
So many questions, so few answers, sorry!
Woody
} -----Original Message----- } From: Smartech [mailto:smartech-at-optonline.net] } Sent: Wednesday, June 27, 2001 1:33 PM } To: To all on the list } Subject: Metallurgy, Weld Failure Analysis, SEM Analysis } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } A customer of mine sent a part that has started showing a failure only } recently, although he has manufactured the part for years. } The part is a } heating element. It consists of a thick SS rod which is the } terminal. It } has a diameter of about 1 cm. It is joined to an element } that is about 2 mm } in diameter. The joint is made by 1st drilling the thick } stock and then } inserting the wire into the hole. Then a notch is made in } the thick stock } until the thin element is exposed. Then the notched surface is welded } without the introduction of any new material. I believe it } is a "TIG" weld. } A standardless analysis of the terminal shows Fe (82%), } Cr(13%), Al(3%), } Si(1.6%), and Mn (.3%). He has tried to match the two } materials as closely } as possible. } } The joint is heated repeatedly, as it is used as an element in a glass } furnace. The failure starts with a bulk flow of material out } the top of the } hole, which is about 1.5-2.0 cm from the welded joint/notch, } and over the } sides of the terminal. I have analyzed this material and it contains } principally Fe and O. The area I analyzed was polished so } the internal } composition of the flowed material contains a great deal of } oxygen. No Cr } is detected and very little Al. } } I feel that this material flow degrades the joint causing the various } electrical failures that have been observed. } } My questions are: } } Is this a typical failure mode with a Fe/Cr SS weld } experiencing many heat } cycles? } } What is the mechanism that results with the crystalline } appearance of the } iron oxide, leaving the other elements behind? } } This failures appears to have started when a new aqueous lubricant was } introduced for the initial hole drilling of the terminal. } Could this play a } role in the failure? } } What measures might be taken to reduce the frequency of this } failure mode? } Joint design? Material compatibility? } } } Thanks in advance! } } Ric } } SMARTech } 860-491-3299 } www.semguy.com } 19 Cornwall Drive } Goshen CT 06756 } }
I'm in search of something that will remove immersion oil from slides of
blood smears (no coverslips) without damaging the specimens. A friend told that such a chemical exists, but could not remember the name. It supposedly smells like oranges, and xylene was once used for the same purpose. If you have any ideas as to the name of this mystery chemical,
Does anyone know the best method for embedding 0.45um nitrocellose filter for TEM thin sectioning? We have been filtering vesicle preparations onto these filters and wish to examine them by TEM analysis for immunolabelling. Our 2 previous attempts with Epon mixtures and minimal intermediate solvent changes have lead to poorly polymerized blocks and dissolving membranes. Currently we are trying LR White as a substitute. Any help would be great. Thank you. Ingrid Niesman SRA III Farquhar/Palade Lab UCSD
Dear all, Is the DTSA (Desk Top Spectrum Analyzer) package available for PC or only for Mac? I looked on the NIST web site but couldn't find information (and their internal search engine was broken).
Thanks for any leads you can give.
Ian MacLaren Max Planck Institut für Metallforschung, Seestraße 92, 70174 Stuttgart, Germany Email: ian.maclaren-at-physics.org / maclariz-at-yahoo.co.uk Home page: http://members.tripod.co.uk/IanMacLaren/
I have no idea if it works for your purpose, but the xylene substitute smelling of oranges is called Histoclear.
Dave
On Wed, 27 Jun 2001 17:42:41 -0500 Christine Edly {cedly-at-ou.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 } -----------------------------------------------------------------------. } } } Greetings! } } I'm in search of something that will remove immersion oil from slides of } } blood smears (no coverslips) without damaging the specimens. A friend } told that such a chemical exists, but could not remember the name. It } supposedly smells like oranges, and xylene was once used for the same } purpose. If you have any ideas as to the name of this mystery chemical, } } I'd sure appreciate it. } } Regards, } Christine Edly } University of Oklahoma }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
I have used the 'orangey' smelling solvent under the brandname of Histoclear. It is supposedly extracted large scale from citrus peel and is the composed of the 'zest' that comes out as a spray when we peel oranges.
I couldn't comment on its use for removing immersion oil as you describe. I have only used it as a xylene substitute for tissue processing tissues and also for clearing stained sections prior to mounting.
It is said to be safer than xylene but some of the people I worked with at the time complained of headaches so we stopped using it despite never proving that it was the cause of the headaches.
Hope this helps!
At 17:42 2001-06-27 -0500, Christine Edly wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Med vänliga hälsningar/With best wishes
Gareth
"Close your eyes and look. What you saw at first is there no more; and what you will see next has not yet come to life" Leonardo da Vinci (1452-1519).
Gareth Morgan MPhil MSc FIBMS, Institutionen för Mikrobiologi, Patologi och Immunologi(IMPI), H5, Karolinska Institutet, Avdelningar för biomedicinsk laboratorievetenskap och biomedicinska ämnen, Lindhagensgatan 92, Box 12773, S 112 96, Stockholm Sverige
The point about this server is that it is used by a very heterogeneous public from which a non-exclusive list is the following:
students, professors, scientists/ researchers technical staffs suppliers producers specialists non-specialists etc. etc. etc.
and all these from so many different backgrounds with so many different problems and even from so many different countries... That if we want to teach all of them to behave as we expect before they post a query, the server could go to early retirement.
I guess the philosophical attitude that one should have is to really try and understand both sides of the discussion.
1- on one hand the trust that professor Monson is demanding is not there because our users are a small sample of our society. How many times have we heard that people have had their ideas picked by others and their publications stolen by others and .... in the very scientific community that most of us hoped to be the purest part of the society. Still the scientists are generally speaking very sincere people. So if someone is not having trust to others, it's the consequence of what they have observed in their environment.
2- I do not know if Rachel is a student (may be not), and if she really was not trusting or just did not think to write the reason and so on, so we do not discuss the case on a specific way. But talking about going to the library, there are labs where the students have really not access to the books and there are places where superiors do not give the support the students and other researchers' need, or simply there are not enough books available.
So there are always reasons for the way people behave and the more our society goes towards a very materialistic and "win whatever the situation" attitude, the more the arguments of professor Manson will be applicable because the more people will behave in the way he is not happy about, and I understand and apreciate his point.
All that apart, it is better to listen to people,as often things that are not to be done are not published and are only "known". Of course provided that answers are given generously, as arrogance for giving is so destructive, even for donator. I therefore agree with the fact that we should just be able to ask the question and someone will react to give us an answer.
I think that professor Monson has evoked a point that is of great importance and that is the human aspect of all we do, as "human" is what we are hoping to be at a first place. That needs sensitivity... and continuous effort.
Regards,
Sousan
Paul Webster wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear Colleagues, } } The messages from Professor Monson in reply to a question about long-term } tissue storage makes me wonder what has happened to this listserver? } } His message was highly entertaining and even if the poster of the original } message may have felt "picked on", there was no reason to expect him to post } an apology, no matter how informative it was. } } Far too many posters use this listserver as a way of finding a quick fix to } simple solutions to technical problems that can be easily researched in text } books. } } Often these messages contain little background information on the technical } problem under investigation. Even so, many of these questions get full } attention and detailed replies, even if the replies are not posted on the } main list for public view. } } What has happened to the free and open discussion of issues and protocols } that we used to have? Are people so afraid of being accused of being } incorrect that they do not post their replies openly? Or maybe people are } afraid of being accused of being impolite. } } I look forward to controversy and hope that my comments are read carefully } and replied to with vigor. Only in this way will I be able to learn new } thing, and this is the main reason I still subscribe to this listserver. } } If I am factually wrong about something, tell me. It is better to find this } out on the listserver than in a more professional (or cruel) environment. } } As for discussion subjects: why was there no comment about the unadvisable } suggestion made about storing samples in a frozen state? Would anyone } working with EM really choose to store their samples at -20 degrees before } processing for TEM? } } Where is all the discussion about extraction that can occur when storing } samples in alcohols or buffers, or on the contamination that I know grows in } everyones old cacodylate buffers? } } Do we not comment because we know it has all been published in textbooks? } } Dear Profesor Monson, do not feel bad about improving our general knowledge } and our professional etiquette. I for one support your views and find your } comments useful and entertaining. Please keep up your submissions. } } Best regards, } } Paul Webster } } Paul Webster, Ph.D. } Scientist II and Director } Ahmanson Center for Advanced EM & Imaging } House Ear Institute } 2100 West 3rd Street } Los Angeles } CA 90057 } } pwebster-at-hei.org } p: 213 273 8026 } f: 213 413 6739 } http://www.hei.org/research/depts/aemi/aemi.htm
The micromanipulator we use is a manual hydraulic system (Narishige MNN-1) which is suitable for the plucking process. I do not know how its performance compares of to other makes.
We use a microscope with *50 and * 5, 18 mm working distance lenses.
A couple of general comments about the lift-out process which might be useful.
We use both pulled glass needles and electro-polished metal needles. The yield of plucking using these two types of needles is comparable. An advantage of the metal needles is that they do not shatter if accidentally knocked against the sides of the FIB cuts. An advantage of the glass needles is that if a cross-section has moved up the needle's shank, away from the tip, (which sometimes occurs) then it is possible to see it and therefore, not waste time searching for it in the area surrounding the FIB cuts. (In addition to riding up the needle's shank sometimes a cross-section can also 'flick ' out away from the cuts while it is being plucked).
For cross-sections that have moved up the shank it is possible to re-position them at the needle's tip using a second micro-manipulator. However, it is also possible to get these cross-sections off the needle and onto the support TEM grid by gently tapping the micro-manipulator system while the needle is positioned over the grid.
Instead of using the Formvar or carbon support grids we use copper grids with 7 um sized holes. We have found that only very rarely does a cross-section fall off these. The advantage of these is that there is no support membrane. What can happen is that while a cross-section is being placed onto the support membrane the support membrane can be torn by the tip of the needle and it can then wrap itself around the cross-section.
Also, because of the diversity of use of FIB systems (e.g. for modification or fabrication of optoelectronic devices and micromachining) a FIB discussion list has been set up. The subscribing web page is located at:
http://users.ox.ac.uk/~rml/
Any information, for example details of relevant FIB pages, can be added to the web page if emailed to me.
Regards
Richard
-------------------------------------------------------------- Richard M Langford
Department of Materials, University of Oxford Parks Road, Oxford, OX1 3PH, UK
I agree wholeheartedly with Paul in his comments regarding the mails from Frederick Monson - no need for apology, interesting and informative replies.
Higher education seems to be locked in an apparently paradoxical situation with students being expected to 'take responsibility for their own learning', the use of Student Centred & Problem Based Learning etc. The paradoxical part is that they do not have the appropriate intellectual tools to be able to carry out such assignments. Frustration with their lot then spills out over tutors who are expected to take responsibility when things go wrong (but mostly no feedback when things work).
Ask me a question and I will give you the most appropriate answer I can. The answer may be wrong of course, possibly because I have an incorrect understanding of the concepts involved, possibly because I was asked the wrong question. If I respond to a question with a question of my own it is because I am trying to seek clarification of what EXACTLY is being asked. That, as I see it, is how we work in science.
Paul says - "If I am factually wrong about something, tell me. It is better to find this out on the listserver than in a more professional (or cruel) environment."
I agree absolutely here too - it is easier to fight tooth and claw here and come to some agreement or compromise than if we are addressing 500 people at a conference and one of them asks a simple question to which we have no answer.
My appeal to those posing questions is to give as much background information as possible and to respond clearly to questions on clarification.
My appeal to those answering is to identify clearly what is hearsay "in my experience" "I got that straight from a guy in the pub last night". Identify also the context of the answer/solution to the question/problem and refer to published data whenever possible.
I look forward to a lively, ongoing debate (even though I am going for my summer break tomorrow!).
Med vänliga hälsningar/With best wishes
Gareth
"Close your eyes and look. What you saw at first is there no more; and what you will see next has not yet come to life" Leonardo da Vinci (1452-1519).
Gareth Morgan MPhil MSc FIBMS, Institutionen för Mikrobiologi, Patologi och Immunologi(IMPI), H5, Karolinska Institutet, Avdelningar för biomedicinsk laboratorievetenskap och biomedicinska ämnen, Lindhagensgatan 92, Box 12773, S 112 96, Stockholm Sverige
} My boss just noticed in the 1985 edition of Cellular Pathology Technique, } Culling, Allison and Barr eds., that food oil derivatives, such as } histoclear, will cause "haematoxylin" staining to rapidly fade. Is this } true? I have been having problems with hematoxylin fading for quite some } time and I use histoclear in my paramount all the time. 'Tho I began (8 } years ago) by using xylene, and I think those faded just as much. } } If histoclear does cause the stain to fade, are there any other } substitutes for xylene that would work?
R. D. Lillie made extensive studies of mounting and fading in the 1940s and 1950s (reported in Ch. 5 of his "Histopathologic Technic" book, with references to papers in Stain Technol).
Fading is typically due to either acidity or chemical reduction. Canada balsam is acidic (abietic acid is a major component) but this doesn't matter - the acid can't ionize in the absence of water. Alum-haematoxylin stains keep for many years in Canada balsam though an eosin counterstain may fade a little over the course of 2 years. (This must be from reducing agents in the medium.) Easily reduced colours fade in Balsam (e.g. Prussian blue, acid fuchsine) and are more stable in synthetic media. Easily oxidized colours (notably cobalt sulphide) are well preserved in Canada balsam but fade in a few months in most synthetic resins. (You can revive CoS by removing the coverslip and exposing to ammonium sulphide again.)
Limonene (I think this is what histoclear is) is a reducing agent - two non-conjugated double bonds in the molecule - so its presence in a mounting medium is undesirable. Some is bound to persist, just as some xylene persists when the mounting medium dries. The boiling point of limonene (176C) is a bit higher than xylene (140C), so it might be a bit more persistent.
If your eosin faded equally after xylene or limonene the mounting medium is the probable culprit. Try a wholly synthetic (polystyrene or methacrylate) medium rather than a natural or semi-synthetic one. Synthetic media are the best bet for pretty well everything except cobalt sulphide (from ATPase histochemistry etc). The blue component (alum-haematein) of an H & E stain is very resilient and should not be affected by clearing agents and mounting media. It keeps for decades (perhaps for ever). If yours is fading, something acidic and probably also some alcohol must be passing through into the mounted preparations. This is surely unlikely to occur on a regular basis!
Hope this helps. I'm sure I haven't thought of everything, and have no personal experience with histoclear/limonene, but you should be getting lots of other suggestions too.
John A. Kiernan, Department of Anatomy & Cell Biology, The University of Western Ontario, LONDON, Canada N6A 5C1
Hi
I have used the 'orangey' smelling solvent under the brandname of Histoclear. It is supposedly extracted large scale from citrus peel and is the composed of the 'zest' that comes out as a spray when we peel oranges.
I couldn't comment on its use for removing immersion oil as you describe. I have only used it as a xylene substitute for tissue processing tissues and also for clearing stained sections prior to mounting.
It is said to be safer than xylene but some of the people I worked with at the time complained of headaches so we stopped using it despite never proving that it was the cause of the headaches.
Hope this helps!
At 17:42 2001-06-27 -0500, Christine Edly wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Med vänliga hälsningar/With best wishes
Gareth
"Close your eyes and look. What you saw at first is there no more; and what you will see next has not yet come to life" Leonardo da Vinci (1452-1519).
Gareth Morgan MPhil MSc FIBMS, Institutionen för Mikrobiologi, Patologi och Immunologi(IMPI), H5, Karolinska Institutet, Avdelningar för biomedicinsk laboratorievetenskap och biomedicinska ämnen, Lindhagensgatan 92, Box 12773, S 112 96, Stockholm Sverige
It would be helpful to know what brand the knives are. Most knife manufacturers will take back their own knives for sharpening at the factory. Just find the local distributor for the brand of knife you have and they will arrange things for you. Energy Beam Sciences is a U.S. Company. We recommend and deal primarily with Diatome but we can also help with DDK.
Good luck, Mike
Michael R. Nesta General Manager Energy Beam Sciences, Inc. Agawam, MA USA Tel: (413) 786-9322 "Adding Brilliance to Your Vision"
-----Original Message----- } From: Ping Li [mailto:pli-at-is.dal.ca] Sent: Wednesday, June 27, 2001 4:10 PM To: Microscopy list
Hi, could anyone please give me some information on where I can get my diamond knives (for semi thin sectioning) sharpened? Any such places in Canada? I believe the knives were made in Germany. Any information is highly appreciated. Thank you in advance.
Just wanted to thank everyone who contributed to the discussion and suggested solutions for the problems we were having. It is very much appreciated - we have so many things to try and have learned a lot in the process.
My colleague, who shares the printer with me, was amazed, both with the range of answers and the rapidity of the responses - the perfect combination when you're having trouble in the lab. That's why I keep coming back to the group for answers and have stayed subscribed for so many years - there's no place like it!
Thanks again.
Paula.
Paula Allan-Wojtas Research Scientist - Food Microstructure Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
When we do this sort of thing, we generally use Nuclepore filters, but I think our trick should work for other filters as well. We keep the material on the filters through fixation and change solutions with gentle vacuum. Then we overlay the specimen with a thin layer of low gelling point agarose, pull a gentle vacuum and then chill the filter to solidify the agarose. The entire filter can then be submerged in the next fluid, and the filter is peeled away from the agarose. If all goes well the specimen should be trapped in the agarose and the filter material is no longer a problem. Warning: do not fix agarose in glutaraldehyde. It does not embed well.
At 09:20 PM 6/27/2001 -0700, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Gregory W. Erdos, Ph.D. Assistant Director, Biotechnology Program Scientific Ditrector, ICBR EM CORE University of Florida Ph. 352-392-1295 PO Box 118525 Fax 352-846-0251 Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl
This is a reminder concerning upcoming deadlines for the Microscopy & Microanalysis 2001 meeting from August 5-9 in Long Beach, CA.
1) The deadline for hotel reservations at the meeting rate is this Friday, June 29. The Westin still has some double rooms available for the entire meeting period, but the other meeting hotels are booked. You can reserve a room online by going to www.msa.microscopy.com and following the links to M&M 2001 electronic forms.
2) The deadline for early meeting registration is Friday, July 6. Save some money and register early! Electronic registration forms are also available at the above site. I would also encourage those of you that are interested to register for the Pre-meeting congress (only $10 more with full meeting registration) and workshops ($120-$180) that are available.
3) The deadline for submitting a late breaking poster is Sunday, July 15th. If you would like to submit your work for this session contact Bob Price by email at Price-at-med.sc.edu.
With nearly 700 presentations scheduled, a sold-out exhibit floor and an excellent venue in Long Beach, California, M&M 2001 promises to be an excellent meeting. On behalf of the MSA and MAS Councils, and the Local Arrangements and Program Committees, I hope to see all of you there.
Christine, if it smells like oranges and is used instead of xylene it must be Histoclear. I have mine from Electron Microscopy Sciences, called Xylene substitute, Cat #23410-01. I have never used it for immersion oil removal though (sounds like a good idea), I just use it to clean microtomes of paraffin residue.
Sarka Lhotak Hamilton Regional Cancer Centre McMaster University Hamilton, Ontario, Canada lhotaks-at-mcmaster.ca
} Greetings! } } I'm in search of something that will remove immersion oil from slides of } } blood smears (no coverslips) without damaging the specimens. A friend } told that such a chemical exists, but could not remember the name. It } supposedly smells like oranges, and xylene was once used for the same } purpose. If you have any ideas as to the name of this mystery chemical, } } I'd sure appreciate it. } } Regards, } Christine Edly } University of Oklahoma } }
We are in the market for a freeze-fracture system. Currently there are two models under our consideration: Bal-tec Freeze-Fracture Unit BAF060 and RMC/JEOL 9010. We would appreciate any comments from users of these two systems regarding to their applications, performance, stability, and other features.
Ingrid: It has been a long time since I did this, but my notes show that we have very good embedding using Epon, _but_ I have omitted propylene oxide from my processing schedule (did that more due to contact allergies developed over years of carelessness). Epon (and the various replacements are miscible with EtOH--just make sure that you use freshly opened bottles. (Acetone is another dehydrant that will tend to dissolve the filters, BTW.) Dehydration steps can be shortened (through graded EtOH, including uranyl acetate in 70% if you use a preembedding UA procedure), then go into 2:1, 1:1 and 1:2 EtOH:Epon for infiltration. I also use a minimum of 2 steps in pure before embedding in fresh epoxy. I would guess that you are embedding in the flat molds--best way for filters, IMHO. From personal experience, use of PO and Spurr's are really out for the filters (and my recollection is the same for acetone, as mentioned above). Hope it works out for you.
Roger
On Wed, 27 Jun 2001 21:20:19 -0700, Ingrid R. Niesman wrote:
| ------------------------------------------------------------------------ | The Microscopy ListServer -- Sponsor: The Microscopy Society of America | To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com | On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html | -----------------------------------------------------------------------. | | | Does anyone know the best method for embedding 0.45um nitrocellose filter for TEM | thin sectioning? We have been filtering vesicle preparations onto these filters and | wish to examine them by TEM analysis for immunolabelling. Our 2 previous attempts | with Epon mixtures and minimal intermediate solvent changes have lead to poorly | polymerized blocks and dissolving membranes. Currently we are trying LR White as a | substitute. Any help would be great. | Thank you. | Ingrid Niesman | SRA III Farquhar/Palade Lab | UCSD |
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc. 900 Rigdebury Road Ridgefield, CT 06877 203-798-5448
_______________________________________________________ Send a cool gift with your E-Card http://www.bluemountain.com/giftcenter/
I would appreciate any responses about the following situation:
I have epoxy resin embedded plant leaf (pepper) material infected with a plant pathogenic bacteria (Xanthomonas vessicatoria). We plan to begin with cutting 1-2 micron sections for light microscopy. I know that there are stains using for differential staining of bacteria which are used on fresh tissue or paraffin embedded material. Does any know of stains which would work similarly for epoxy resin embedded material?
Thank you,
Maureen Petersen Dept of Plant Pathology University of Florida Gainesville, FL
Fixation? You shouldn't partially dehydrate or partially clear for EPON, but assuming the membrane system is adequately preserved to survive the alcohol and PropOx, then your problem is poor infiltration, due to poor clearing, due to poor dehydration. This does not mean that you must dehydrate in 30min steps. The times can be reduced empirically until you get to the minimum necessary times for dehydration and clearing. Lots of variables. The filter should not be a problem in any case, though I have no practice with isolated vesicles, only with isolated Eprythrozoon coccoides (a mouse zoonose, something like a mycoplasma, likes RBC's, and 'hibernates') and just morphology. You might want to try GMA for TEM (e.g. SPI) though I have no experience with it, or any other hydrophilic embedment. I assume nano-gold, so you must have been into: Hayat, M.A.(1995), Immunogold-Silver Staining, CRC Press, Boca Raton, FL, ISBN: 0-8493-2449-1(LC Card# 95-1831, LC Loc# QR187.I482H39). Chapter 4 by Krenacs and Krenacs on the subject of: "Which embedding medium?" might be of help.
Good hunting,
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: Ingrid R. Niesman } Sent: Thursday, June 28, 2001 12:20 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: nitrocellulose filters } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Does anyone know the best method for embedding 0.45um nitrocellose filter } for TEM } thin sectioning? We have been filtering vesicle preparations onto these } filters and } wish to examine them by TEM analysis for immunolabelling. Our 2 previous } attempts } with Epon mixtures and minimal intermediate solvent changes have lead to } poorly } polymerized blocks and dissolving membranes. Currently we are trying LR } White as a } substitute. Any help would be great. } Thank you. } Ingrid Niesman } SRA III Farquhar/Palade Lab } UCSD } }
This is very interesting, about the orange or citrus-based stuff that is good for removing immersion oil. There is also a range of products sold for de-greasing bicycle chains and derailleurs, which are based on citrus components. I'm no organic chemist myself, but can one of the chemistry-oriented people out there tell me why something citrus-based should be so good at dispersing oils? Call me curious.....and I hope I'm not too off-topic here..
Frank Thomas Geological Survey of Canada (Atlantic) Bedford Institute of Oceanography Dartmouth, Nova Scotia B3L 4C8
No Protocol, but this was a quick find on Google "Plant immunohistochemistry"
Plant Cell Physiol. 37, 641-649 (1996): Developmental expression of a 23 kDa jasmonate-induced protein (JIP-23) of barley leaves (Hordeum vulgare cv. Salome) was studied by measuring the time-dependent accumulation of transcript and protein during germination. Tissue-specific expression of JIP-23 was analyzed immunocytochemically and by in situ hybridizations, respectively. During seed germination JIP-23 mRNA was found to accumulate transiently with a maximum at 32 h, whereas the protein was steadily detectable after the onset of expression. The occurrence of new isoforms of JIP-23 during germination in comparison to jasmonate-treated leaves suggests, that the JIP-23 gene family of barley is able to express different subsets of isoforms dependent on the developmental stage. JIP-23 and its transcript were found mainly in the scutellum, the scutellar nodule and in lower parts of the primary leaf of 6 days old seedlings. All these tissues exhibited high levels of endogenous jasmonates. In situ hybridization revealed specific accumulation of JIP-23 mRNA in companion cells of the phloem in the nodule plate of the scutellum. In accordance with that, JIP-23 was detected immunocytochemically in phloem cells of the root as well as of the scutellar nodule and in parenchymatic cells of the scutellum. The cell type-specific occurrence of JIP-23 was restricted to cells, which are known to be highly stressed osmotically by active solute transport. This observation suggests, that the expression of this protein might be a response to osmotic stress during development.
} ---------- } From: Shea Miller } Sent: Wednesday, June 27, 2001 12:00 PM } To: microscopy-at-sparc5.microscopy.com } Subject: immunofluorescence on plants } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello everyone: } I have a student who wants to do some immunolocalization of a protein } in plant tissue. I have been combing my files, but haven't managed to } find a decent protocol that she could work from... (and can't seem to get } myself to the library for a more comprehensive search). Can anyone supply } us with a "generic" protocol for immunofluorescence in plants? } } Thanks so much in advance } shea } } } } }
Question from an old exam given to students who demanded MORE MULTIPLE CHOICE!
Which of the following is NOT involved with human reproduction?
a. transcription b. replication c. translation d. osculation e. none of the above
Answer tomorrow.
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: Ian MacLaren } Reply To: Ian MacLaren } Sent: Thursday, June 28, 2001 3:56 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: DTSA } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear all, } Is the DTSA (Desk Top Spectrum Analyzer) package available for PC or only } for Mac? I looked on the NIST web site but couldn't find information (and } their internal search engine was broken). } } Thanks for any leads you can give. } } Ian MacLaren } Max Planck Institut f¸r Metallforschung, Seestrae 92, } 70174 Stuttgart, Germany } Email: ian.maclaren-at-physics.org / maclariz-at-yahoo.co.uk } Home page: http://members.tripod.co.uk/IanMacLaren/ } } }
Please post informational responses to the server! I've been curious about this, too but too lazy to look into it - the dishwashing cleansers sold in "granola" stores (organic, non-toxic to the environment, etc.) are also citrus-oil based (and at least one has almond oil; not sure if that is just for the odor).
Tamara
On Thu, 28 Jun 2001, Frank Thomas wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Listers - } } This is very interesting, about the orange or citrus-based stuff that is } good for removing immersion oil. There is also a range of products sold for } de-greasing bicycle chains and derailleurs, which are based on citrus } components. I'm no organic chemist myself, but can one of the } chemistry-oriented people out there tell me why something citrus-based } should be so good at dispersing oils? } Call me curious.....and I hope I'm not too off-topic here.. } } Frank Thomas } Geological Survey of Canada (Atlantic) } Bedford Institute of Oceanography } Dartmouth, Nova Scotia } B3L 4C8 } } }
|--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward-at-unm.edu |--------------------------------------------------|
Michael Haugh - Director of Operations Resolution - http://www.resolve3d.com 530 Tamal Plaza, Corte Madera, CA 94925 Office:415/945-7360 x13 FAX:415/927-4495 Cell:415/987-9929 Email: mhaugh-at-resolve3d.com
-----Original Message----- } From: Ping Li [mailto:pli-at-is.dal.ca] Sent: Wednesday, June 27, 2001 1:10 PM To: Microscopy list
Hi, could anyone please give me some information on where I can get my diamond knives (for semi thin sectioning) sharpened? Any such places in Canada? I believe the knives were made in Germany. Any information is highly appreciated. Thank you in advance.
Thanks so much for all the info on protocols for plant immunofluorescence... we tried a trial run today on some immature soybeans embedded in paraffin, and got some positive results following the general methods of Tobias Baskin. You have all given great advice, and we have lots to work with to refine our immunoassay.
thanks so much shea
Dr. S. Shea Miller Agriculture & Agri-food Canada Eastern Cereal and Oilseed Research Centre Central Experimental Farm Ottawa, Ontario Canada K1A 0C6 Phone: (613) 759-1760 Fax: (613) 759-1701 Email: millers-at-em.agr.ca
Micronetsters, I am interested in this thread about removing immersion oil, though perhaps diverged a bit from the original post. Over the years, we have often put oil on a slide and then wished to look without. In my hands, this is never easy. I have taken chloroform, acetone, and the like to the coverslip, and wiped and wiped, and it takes truly many wipes to get oil well and truly off. Small beadlets of oil remain tenaciously stuck to the glass. If someone does have a simpler way to get oil off a slide, so well off that there is no residue left behind, I'd love to know about it.
We use Micro Star Technologies for sharpening diamond knives. They do a good job and their turnaround time is very good.
Micro Star Technologies 511 FM 3179 Huntsville, TX 77340-2069 Phone 800/533-2509 FAX 409/294-9861 email mistar-at-msn.com
Mike
Michael Haugh - Director of Operations Resolution - http://www.resolve3d.com 530 Tamal Plaza, Corte Madera, CA 94925 Office:415/945-7360 x13 FAX:415/927-4495 Cell:415/987-9929 Email: mhaugh-at-resolve3d.com
-----Original Message----- } From: Mike Haugh Sent: Thursday, June 28, 2001 1:15 PM To: 'Ping Li'; Microscopy list
Hi, could anyone please give me some information on where I can get my diamond knives (for semi thin sectioning) sharpened? Any such places in Canada? I believe the knives were made in Germany. Any information is highly appreciated. Thank you in advance.
Sparkle! The wonderful purple glass cleaner stuff. Not available in all parts of the US, even - I was given a stash by a student from the midwest. I've heard stories of microscope company service people back east who stockpile the stuff (definitely couldn't find it in New York). I think the guy who gave it to me said it was available in True Value hardware stores; I don't know if they make it.
No financial attachments to Sparkle or True Value, unfortunately.
Tamara
On Thu, 28 Jun 2001, Tobias Baskin wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Micronetsters, } I am interested in this thread about removing immersion } oil, though perhaps diverged a bit from the original post. Over the } years, we have often put oil on a slide and then wished to look } without. In my hands, this is never easy. I have taken chloroform, } acetone, and the like to the coverslip, and wiped and wiped, and it } takes truly many wipes to get oil well and truly off. Small beadlets } of oil remain tenaciously stuck to the glass. If someone does have a } simpler way to get oil off a slide, so well off that there is no } residue left behind, I'd love to know about it. } } Thanks, } Tobias } -- } _ ____ __ ____ Tobias I. Baskin } / \ / / \ / \ \ 109 Tucker Hall } / / / / \ \ \ Biological Sciences } /_ / __ /__ \ \ \__ University of Missouri } / / / \ \ \ Columbia, MO USA } / / / \ \ \ 65211-7400 } / / ___ / \ \__/ \ ____ voice: 573-882-0173 } fax: 573-882-0123 } }
|--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward-at-unm.edu |--------------------------------------------------|
My guess about limonene is that it is non-toxic, biodegradeable, easily detectable by (pleasant) smell, a 'waste' product of the food industry, doesn't need to be mined (environmentally friendly) etc ......
I have seen the bike chain cleaners too and my first reaction was to think Histoclear!
Have a good summer - I'm off to Finland camping on Sunday - Yippeeeeeeee!
At 13:41 2001-06-28 -0300, Frank Thomas wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Med vänliga hälsningar/With best wishes
Gareth
"Close your eyes and look. What you saw at first is there no more; and what you will see next has not yet come to life" Leonardo da Vinci (1452-1519).
Gareth Morgan MPhil MSc FIBMS, Institutionen för Mikrobiologi, Patologi och Immunologi(IMPI), H5, Karolinska Institutet, Avdelningar för biomedicinsk laboratorievetenskap och biomedicinska ämnen, Lindhagensgatan 92, Box 12773, S 112 96, Stockholm Sverige
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Micronetsters, } I am interested in this thread about removing immersion } oil, though perhaps diverged a bit from the original post. Over the } years, we have often put oil on a slide and then wished to look } without. In my hands, this is never easy. I have taken chloroform, } acetone, and the like to the coverslip, and wiped and wiped, and it } takes truly many wipes to get oil well and truly off. Small beadlets } of oil remain tenaciously stuck to the glass. If someone does have a } simpler way to get oil off a slide, so well off that there is no } residue left behind, I'd love to know about it. } } Thanks, } Tobias }
I have try solvents from alcohol to ammonia and thanks to the advice of the local confocal Leica rep. Frank Lie, I am now using Windex with great success for removing immersion oil from oil objectives and COVERSLIPPED microscope slides.
As for using Histoclear, which I have: it is rather strong smelling and I am uncertain whether it will destain uncoverslipped stained histo. preps. Cheers, Ken
-- Ken Tiekotter, Adjunct Professor The University of Portland Department of Biology 5000 N. Willamette Blvd. Portland, OR 97203
At 4:28 PM -0500 6/28/01, Tobias Baskin wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Micronetsters, } I am interested in this thread about removing immersion } oil, though perhaps diverged a bit from the original post. Over the } years, we have often put oil on a slide and then wished to look } without. In my hands, this is never easy. I have taken chloroform, } acetone, and the like to the coverslip, and wiped and wiped, and it } takes truly many wipes to get oil well and truly off. Small beadlets } of oil remain tenaciously stuck to the glass. If someone does have a } simpler way to get oil off a slide, so well off that there is no } residue left behind, I'd love to know about it. } } Thanks, } Tobias } -- *********************
For years, we have used a 1:1 dilution of Windex & water. Use the original formula (blue) Windex. this was recommended to use by our Zeiss representatives and works well on slides and lenses.
lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
} } Hi, could anyone please give me some information on where I can get my } diamond knives (for semi thin sectioning) sharpened? Any such places in } Canada? I believe the knives were made in Germany. Any information is } highly appreciated. Thank you in advance. } } Ping *****************
I believe that almost all of the companies that manufacture & sell diamond knives (Diatome, DDK, Mictostar) will resharpen knives of all makes. All are the US or have US offices.
Lee
-- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
Sparkle glass cleaner is good for cleaning the larger glass surfaces of the microscope. I don't think it would really work well to remove immersion oil, we use acetone for that. Acetone is a low health hazard (fingernail polish remover) and works well for cleaning objectives and eyepieces. Like any cleaner, even Sparkle, you would never soak an optical element. Apply the cleaner to something soft and absorbent and use that to clean the surface. Kimwipes and wooden shafted "Q-tips" work the best. It's true that Sparkle isn't available everywhere. You can order it from their website: http://www.glasscleaner.com/
Dave Burton Optical Specialist University of Washington Scientific Instruments
Just wondering: isn't acetone likely to attack the cements holding the lens elements in place? Or even some plastic components? Apparently you haven't had any problems, but I wonder if anyone else has?
Thanks, Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
-----Original Message----- } From: David Burton [mailto:dburton-at-nwlink.com] Sent: Friday, June 29, 2001 8:40 AM To: Microscopy-at-sparc5.microscopy.com
Sparkle glass cleaner is good for cleaning the larger glass surfaces of the microscope. I don't think it would really work well to remove immersion oil, we use acetone for that. Acetone is a low health hazard (fingernail polish remover) and works well for cleaning objectives and eyepieces. Like any cleaner, even Sparkle, you would never soak an optical element. Apply the cleaner to something soft and absorbent and use that to clean the surface. Kimwipes and wooden shafted "Q-tips" work the best. It's true that Sparkle isn't available everywhere. You can order it from their website: http://www.glasscleaner.com/
Dave Burton Optical Specialist University of Washington Scientific Instruments
Fax support is not built into Outlook 2000 and the newer version of Windows as it was in the older versions. MS does describe the setup on their knowledge base. Basdically, it is still available by going back and installing some add-ons that are not part of the regular Windows or Outlook installation. They are in one of those extra folders that you find on the CDs. I will send you details directly.
At 09:34 AM 6/29/2001 +1000, you wrote:
} Dear colleagues, } } I know that here is some very good experts on computers.. And I cannot } configuration how to do it.. } } I would like to know how to change setting of the fax in Outlook 2000 from } email only to send and receive faxes.. } } Ricardo
---------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
Just a reminder that the hotels are filling up, & the registration rate will be going up next week for TNT 2001 (Segovia, Spain, September 3-7th). There are currently some 250 participants from all over the globe, from the fields of physics, microscopy, FIB, biotech, materials and the semiconductor industry, as well as dedicated Nanotechnologists and a smattering of venture capitalists.
More information is at www.cmp-cientifica.com/tnt2001
Regards
Tim
***************************************************************** Tim E. Harper CEO CMP Cientifica s.l. Space & NanoTechnology Division Phone +34 91 640 71 85 Fax +34 91 640 71 86 http://www.cmp-cientifica.com/
1) Avery 9x12" Self - Adhesive Laminating sheets, 50/ pack. #461253 / 73601 The result is a nice glossy finish, and an image that doesn't fade.
2) archival printer ink. Check the Epson web site for more info about archival inks and printers like the Epson StylusPhoto 2000P. Wilheim research does the most testing for archivalness of photographic media. They may have some pointers on the best ink-paper combos for light stability. http://www.wilheim-research.com
3) enclosing a print in a polyethylene sheet protector (archival grade)
4) printer specific paper -Stay with the papers recommended for use with the new Epson inks
- try Epson's Heavyweight Matte paper It's cheap ($9/50 sheets from a discount sppplier), delivers the Epson long-life guarantee, & it has a robust surface right out of the printer. And beautiful prints.
-There were recently (last 6-8 months) some issues with color shift on some Epson papers. Epson determined the shift was from high concentrations of ozone, and not due to exposure to light. Epson recommends Heavyweight Matte or Epson Photo paper for maximum lightfastness.
-if the "Photo Quality Glossy Paper" isn't new, it may be the paper's fault - it's been reformulated to deal with this problem. See http://www.luminous-landscape.com/1280.htm & http://www.wilhelm-research.com/
5) printer settings - make sure that when you print B&W prints, set the printer to "B&W". Colors can appear when "color" is selected, and the color printing mimics B&W
6) protective sprays - Luminos http://www.lumijet.com has a spray called "Imagesheild" to protect prints. $18/ can. Quoting their website: "LUMIJET IMAGESHIELD will become your favorite print protector. It*s a convenient ozone-friendly low-odor aerosol spray that significantly improves wet-fastness. In addition to protecting delicate inkjet images from moisture, it also shields images from fingerprints and harmful UV rays. This unique product will not only help extend useful print life, it is totally transparent; preserving the original print surface characteristics. From dead matte to glossy, or from canvas to silver foil, you*ll never know it*s there."
-Krylon clear acrylic spray Use many light coats. If you really wet the surface, the paper can become translucent.
-Johnson's Paste Wax. It works on some inks and papers, not all. It does offer some protection against UV. It does protect against moisture, finger prints, dust and scratches. It does change the finish. If the wax causes the ink to smear a fixative spray will usually stop that. Test it for several weeks before putting it on an expensive display.
Comments: Do not use the printer for archival printing. For long lasting prints use a silver halide digital printer (or dye sub) or traditional dark room processing.
Ink-jet prints have never been designed for long term preservation. the inks used in those printers are made of dyes, which are very sensitive to U.V electromagnetic waves present in fluorescent lamps and sun rays. Dyes are not chemically fixed to the fibers, so can cause the smudging problems. The use of a fixative to stabilize the colour is only a temporary and partial solution: the polymers spayed on the surface of the print might partly protect the dyes by filtering some U.V rays but doing so, will tend to yellow with time and give a yellowish cast to your images. Digitizing images is not a complete longterm solution either. For best preservation of images, save many copies on CDs, and check them often for deteroiration and update the formats as needed to be sure they are always accessible (tiff uncompressed files) and keep it in a cool place, with low humidity (30% RH), or rent a place in a specialised data keeping system firm.
Paula Allan-Wojtas Research Scientist - Food Microstructure Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
You are absolutely right, and most of the time we behave just as you describe us. Patient, etc. One purpose of my tirade was to point out that there are differences between today and yesterday that give ALL of us choices to make. I know a high school student who failed the year, because he wanted to punish his parents. When he told me what he had done, I responded that I knew a young man just like him when I was in high school. His solution was to get straight A's so he could become more quickly independent. Science is our own, very human, endeavor. Those of us who do science must constantly work to remove the temptations to embellish our results, to twist the statistics and to 'jump' to conclusions. But, and here's where I differ perhaps slightly. Sometimes, there is a good reason to yell! My wife hates my streak of sarcasm! But I was raised on the stuff. When I did something 'dumb,' everyone who loved me, joined in the derisive repair. I learned to think at my dinner table before I was 10. I did not like to be the but of everyone's attention. My Mom taught, then I taught. I only stopped, because no one [too few] wanted to learn any more [bad generalization]! Go to college today, and what do you find? Young people unprepared for the rigor. Young people unprepared for the intellectual dance - as we used to call it. Young people who don't know what they want to do and are unwilling to work to find out. "They only do what they are taught!" This is my mantra when I look for explanations in the mirror. My older son had to learn - for the test - to name NH3 as "ammonium", because his chemistry teacher would not believe the Handbook of Chemistry and Physics. My chemistry teacher in public high school took chemistry courses at Rutgers every third summer. He was the expert in chemistry. My physics teacher had a PhD in the subject. My biology teacher went on to be the chair of Physiology at a large Southern medical school. If I wanted to complain about a professor at Lehigh, the Dean would show me a list of nearby schools that had openings. The attitude was clearly communicated. The greatest lessons in life are those that teach one to manage the worst parts of it. If I failed a course, it was MY failure - and it was at least MY problem! I wanted to be what I am, once I figured everything out. The only advice I ever received, when I was having problems with learning, was, "Study harder." My Mom just kept on telling me to keep on trying as hard as I could. If I were young now, I'd be in group therapy and zapped on some pharmaceutical, if the system had its way. Given my experience, I hope it is not difficult to understand why I might not appear to understand the problems of others. BUT! If I see a young person in science who is poorly informed about how it is done, then I will act in that person's behalf even if I am thought, and reported, to be insensitive. When I was a graduate student, we were all ordered to write faculty/course evaluations. I did. I wrote analyses of each course, and every one of them was critically positive. Not one word of my analyses were printed. If I had demonized one of my professors, I would have been writing novels today. The people who ordered the writings were ex-faculty administrators who were pandering to the new consumer mentality in education. If I had gone to my undergraduate department chair to complain about a grade, he would have told me that I was wasting his time. Five years later, there were lines of boomers outside of department offices. The generation of manipulators had arrived! While they were standing in line, I was learning. While I was learning, they were just getting warmed up! I have had ideas snitched! Sometimes I even gave them away. On the other hand, I have also had the other side of the experience. For six months of learning followed by 30 minutes of consultation and work, I had the good fortune to become a molecular biologist. For that effort, the individual to whom I gave my best remembered my part - even after he had spent years doing the work - and included me in the author list for a gene(TERE1) that mapped to Chromosome 1 and appeared in print in a Nature publication. He did NOT have to do that. There was NO obligation nor agreement. I helped because he asked. People are basically good, especially scientists, and I always start with TRUST, and I always will. You are still absolutely correct.
Regards for YOUR sensitivity,
Fred Monson
} ---------- } From: Sousan Abolhassani } Sent: Thursday, June 28, 2001 8:07 AM } To: Paul Webster } Cc: MSA listserver submission } Subject: Re: long-term storage OR what has happened? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Paul, } } The point about this server is that it is used by } a very heterogeneous public from which a non-exclusive } list is the following: } } students, } professors, } scientists/ } researchers } technical staffs } suppliers } producers } specialists } non-specialists } etc. } etc. } etc. } } and all these from so many different backgrounds with so } many different problems and even from so many different countries... } That if we want to teach all of them to behave as we expect } before they post a query, the server could go to early retirement. } } I guess the philosophical attitude that one should have is to } really try and understand both sides of the discussion. } } 1- on one hand the trust that professor Monson is demanding } is not there because our users are a small sample of our } society. How many times have we heard that people have had } their ideas picked by others and their publications stolen } by others and .... in the very scientific community that most } of us hoped to be the purest part of the society. Still } the scientists are generally speaking very sincere people. } So if someone is not having trust to others, it's the } consequence of what they have observed in their environment. } } 2- I do not know if Rachel is a student (may be not), and if she } really was not trusting or just did not think to write the reason and } so on, so we do not discuss the case on a specific way. } But talking about going to the library, there are labs } where the students have really not access to the books and there } are places where superiors do not give the support the students } and other researchers' need, or simply there are not } enough books available. } } So there are always reasons for the way people behave } and the more our society goes towards a very materialistic and } "win whatever the situation" attitude, the more the arguments } of professor Manson will be applicable because the more people } will behave in the way he is not happy about, and I understand } and apreciate his point. } } All that apart, it is better to listen to people,as often things } that are not to be done are not published and are only "known". } Of course provided that answers are given generously, as arrogance } for giving is so destructive, even for donator. } I therefore agree with the fact that we should just be able to } ask the question and someone will react to give us an answer. } } I think that professor Monson has evoked a point that is of } great importance and that is the human aspect of all we do, as } "human" is what we are hoping to be at a first place. } That needs sensitivity... and continuous effort. } } Regards, } } Sousan } } } Paul Webster wrote: } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } Dear Colleagues, } } } } The messages from Professor Monson in reply to a question about } long-term } } tissue storage makes me wonder what has happened to this listserver? } } } } His message was highly entertaining and even if the poster of the } original } } message may have felt "picked on", there was no reason to expect him to } post } } an apology, no matter how informative it was. } } } } Far too many posters use this listserver as a way of finding a quick fix } to } } simple solutions to technical problems that can be easily researched in } text } } books. } } } } Often these messages contain little background information on the } technical } } problem under investigation. Even so, many of these questions get full } } attention and detailed replies, even if the replies are not posted on } the } } main list for public view. } } } } What has happened to the free and open discussion of issues and } protocols } } that we used to have? Are people so afraid of being accused of being } } incorrect that they do not post their replies openly? Or maybe people } are } } afraid of being accused of being impolite. } } } } I look forward to controversy and hope that my comments are read } carefully } } and replied to with vigor. Only in this way will I be able to learn new } } thing, and this is the main reason I still subscribe to this listserver. } } } } If I am factually wrong about something, tell me. It is better to find } this } } out on the listserver than in a more professional (or cruel) } environment. } } } } As for discussion subjects: why was there no comment about the } unadvisable } } suggestion made about storing samples in a frozen state? Would anyone } } working with EM really choose to store their samples at -20 degrees } before } } processing for TEM? } } } } Where is all the discussion about extraction that can occur when storing } } samples in alcohols or buffers, or on the contamination that I know } grows in } } everyones old cacodylate buffers? } } } } Do we not comment because we know it has all been published in } textbooks? } } } } Dear Profesor Monson, do not feel bad about improving our general } knowledge } } and our professional etiquette. I for one support your views and find } your } } comments useful and entertaining. Please keep up your submissions. } } } } Best regards, } } } } Paul Webster } } } } Paul Webster, Ph.D. } } Scientist II and Director } } Ahmanson Center for Advanced EM & Imaging } } House Ear Institute } } 2100 West 3rd Street } } Los Angeles } } CA 90057 } } } } pwebster-at-hei.org } } p: 213 273 8026 } } f: 213 413 6739 } } http://www.hei.org/research/depts/aemi/aemi.htm } }
} ---------- } From: Gareth Morgan } Sent: Thursday, June 28, 2001 8:18 AM } To: Paul Webster; MSA listserver submission } Subject: Re: long-term storage OR what has happened? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Colleagues } } I agree wholeheartedly with Paul in his comments regarding the mails from } Frederick Monson - no need for apology, interesting and informative } replies. } } Higher education seems to be locked in an apparently paradoxical situation } with students being expected to 'take responsibility for their own } learning', the use of Student Centred & Problem Based Learning etc. The } paradoxical part is that they do not have the appropriate intellectual } tools to be able to carry out such assignments. Frustration with their lot } then spills out over tutors who are expected to take responsibility when } things go wrong (but mostly no feedback when things work). } } Ask me a question and I will give you the most appropriate answer I can. } The answer may be wrong of course, possibly because I have an incorrect } understanding of the concepts involved, possibly because I was asked the } wrong question. If I respond to a question with a question of my own it is } because I am trying to seek clarification of what EXACTLY is being asked. } That, as I see it, is how we work in science. } } Paul says - "If I am factually wrong about something, tell me. It is } better } to find this out on the listserver than in a more professional (or cruel) } environment." } } I agree absolutely here too - it is easier to fight tooth and claw here } and } come to some agreement or compromise than if we are addressing 500 people } at a conference and one of them asks a simple question to which we have no } answer. } } My appeal to those posing questions is to give as much background } information as possible and to respond clearly to questions on } clarification. } } My appeal to those answering is to identify clearly what is hearsay "in my } experience" "I got that straight from a guy in the pub last night". } Identify also the context of the answer/solution to the question/problem } and refer to published data whenever possible. } } I look forward to a lively, ongoing debate (even though I am going for my } summer break tomorrow!). } } } } } Med v?nliga h?lsningar/With best wishes } } Gareth } } "Close your eyes and look. What you saw at first is there no more; and } what } you will see next has not yet come to life" Leonardo da Vinci (1452-1519). } } http://www.ki.se/biomedlab e-mail Gareth.Morgan-at-impi.ki.se } } Tel +46 8 728 3734 } Fax +46 8 728 3688 } } Gareth Morgan MPhil MSc FIBMS, } Institutionen f^r Mikrobiologi, Patologi och Immunologi(IMPI), H5, } Karolinska Institutet, } Avdelningar f^r biomedicinsk } laboratorievetenskap och } biomedicinska ?mnen, } Lindhagensgatan 92, Box 12773, } S 112 96, Stockholm } Sverige } } OBS! Bes^ksadress: Lindhagensgatan 92 } NB! Visiting address:Lindhagensgatan 92 } }
} ---------- } From: Gareth Morgan } Sent: Thursday, June 28, 2001 7:45 AM } To: Christine Edly; Microscopy-at-sparc5.microscopy.com } Subject: Re: immersion oil remover? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi } } I have used the 'orangey' smelling solvent under the brandname of } Histoclear. It is supposedly extracted large scale from citrus peel and is } the composed of the 'zest' that comes out as a spray when we peel oranges. } } I couldn't comment on its use for removing immersion oil as you describe. } I } have only used it as a xylene substitute for tissue processing tissues and } also for clearing stained sections prior to mounting. } } It is said to be safer than xylene but some of the people I worked with at } the time complained of headaches so we stopped using it despite never } proving that it was the cause of the headaches. } } Hope this helps! } } } } At 17:42 2001-06-27 -0500, Christine Edly wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Greetings! } } } } I'm in search of something that will remove immersion oil from slides of } } } } blood smears (no coverslips) without damaging the specimens. A friend } } told that such a chemical exists, but could not remember the name. It } } supposedly smells like oranges, and xylene was once used for the same } } purpose. If you have any ideas as to the name of this mystery chemical, } } } } I'd sure appreciate it. } } } } Regards, } } Christine Edly } } University of Oklahoma } } } } } Med v?nliga h?lsningar/With best wishes } } Gareth } } "Close your eyes and look. What you saw at first is there no more; and } what } you will see next has not yet come to life" Leonardo da Vinci (1452-1519). } } http://www.ki.se/biomedlab e-mail Gareth.Morgan-at-impi.ki.se } } Tel +46 8 728 3734 } Fax +46 8 728 3688 } } Gareth Morgan MPhil MSc FIBMS, } Institutionen f^r Mikrobiologi, Patologi och Immunologi(IMPI), H5, } Karolinska Institutet, } Avdelningar f^r biomedicinsk } laboratorievetenskap och } biomedicinska ?mnen, } Lindhagensgatan 92, Box 12773, } S 112 96, Stockholm } Sverige } } OBS! Bes^ksadress: Lindhagensgatan 92 } NB! Visiting address:Lindhagensgatan 92 } }
I bet we would all like to see some results when you get them.
Regards,
Fred Monson
} ---------- } From: Shea Miller } Sent: Thursday, June 28, 2001 4:58 PM } To: microscopy-at-sparc5.microscopy.com } Subject: plant immunofluorescence-thanks } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Thanks so much for all the info on protocols for plant } immunofluorescence... we tried a trial run today on some immature soybeans } embedded in paraffin, and got some positive results following the general } methods of Tobias Baskin. You have all given great advice, and we have } lots to work with to refine our immunoassay. } } thanks so much } shea } } } } Dr. S. Shea Miller } Agriculture & Agri-food Canada } Eastern Cereal and Oilseed Research Centre } Central Experimental Farm } Ottawa, Ontario } Canada K1A 0C6 } Phone: (613) 759-1760 } Fax: (613) 759-1701 } Email: millers-at-em.agr.ca } } }
The classic blood stains are the dried salts of cationic(methylene blue) and anionic(eosin) dyes dissolved in methanol. So, acid will remove one and base will remove the other, and the aromatic organic is what we no longer want to use. How about a non-ionic detergent? Is that what's in the "Orange" mystery?
Well here's my offering for something 'orangy'. I acquired it a Home Depot. "Commercial ZEP Citrus All Purpose Cleaner". The Directions say that you can use it full strength or diluted. Probably doesn't have any of THOSE aromatics. Nothing on the label. Looks like a typical proprietary scientific mixture. Just like "Histoclear" and "Paraplast." [Why does that period look so out of place?] "Removes grease, oil and soils [Sic!] from concrete, metal and other hard surfaces."
Glass is hard unless one has a very long perspective. I wonder. If I had some old slides, I could try it. Full strength or a dilution series? Ten-fold or two-fold? Water or alcohol as a vehicle? Wonder whether "triphosphate" would work. Did a fine job on the driveway. Time for lunch! I'll stop by Home Depot on the way back.
Respectfully submitted in partial fulfillment of requirements for.......,
Fred Monson [Is he this way all the time? Only when awake.] } ---------- } From: Christine Edly } Sent: Wednesday, June 27, 2001 6:42 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: immersion oil remover? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Greetings! } } I'm in search of something that will remove immersion oil from slides of } } blood smears (no coverslips) without damaging the specimens. A friend } told that such a chemical exists, but could not remember the name. It } supposedly smells like oranges, and xylene was once used for the same } purpose. If you have any ideas as to the name of this mystery chemical, } } I'd sure appreciate it. } } Regards, } Christine Edly } University of Oklahoma } }
I am very grateful to everybody for your help with regards to finding suitable TEM grid film for small particles. The number of responses were really overwhelming. Special thanks to those who were willing to send me grids to try out. Thank you all once again.
Hi, everyone. I am posting this for someone else. However replies can be either posted on the server or sent to my email address directly. I will forward all to her. Thank you in advance.
Hong
Hi Hong,
Okay... here is a paragraph or so about what we are trying to see that you can paste onto the list serv.
We are attempting to visualize the polysaccharide capsule of Neisseria meningitidis serogroup B by electron microscopy. The capsule is (alpha2-8)-linked polysialic acid with about 200-300 residues per polymer. We have tried staining the negatively charged capsule with Ruthenium red, followed by embedding, sectioning, and counterstaining. However, it appears that only the inner and outer membranes were stained and not the capsule (we did capsule + and capsule - controls). In a previous paper that effectively stained capsule of E. coli K5, the cells were washed with capsule-specific antibody before fixation to stabilize the capsule. We have not attempted the procedure with the antibody stabilization yet. Please let me know if this would be recommended or if there are any other suggestions. Thank you.
Hi again; we are having a wee bit of trouble distinguishing our signal from the autofluorescence in our immunoassay on immature soybean seeds/seed coats. So far, we have used a light stain of toluidine blue, but I'm not completely happy, and am considering Evans Blue instead. What do others use to quench the yellow autofluorescence (our Ab is FITC labelled) in plant tissues, and when do you actually apply the counterstain? Before the Ab incubations, or after??
thanks so much again in advance shea
Dr. S. Shea Miller Agriculture & Agri-food Canada Eastern Cereal and Oilseed Research Centre Central Experimental Farm Ottawa, Ontario Canada K1A 0C6 Phone: (613) 759-1760 Fax: (613) 759-1701 Email: millers-at-em.agr.ca
I never heard that before. After 36 years of marriage, and I find this out NOW?
Fred Monson } ---------- } From: Nicol Aitken } Sent: Friday, June 29, 2001 10:43 AM } To: 'Monson, Frederick C.' } Subject: RE: Today's Real Humor } } I'll answer e, even though strictly speaking osculation need not occur! } Nick } } -----Original Message----- } From: Monson, Frederick C. [mailto:fmonson-at-wcupa.edu] } Sent: Thursday, June 28, 2001 1:30 PM } To: 'Microscopy Listserver' } Subject: Today's Real Humor } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Question from an old exam given to students who demanded MORE MULTIPLE } CHOICE! } } Which of the following is NOT involved with human reproduction? } } a. transcription } b. replication } c. translation } d. osculation } e. none of the above } } Answer tomorrow. } } } } 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. } }
Perhaps I'm missing something from the biological side with which I am not familiar. However, for my purposes, it has always been very easy to remove immersion oil using a non-polar organic such as one of the following: hexane, iso-octane, VM&P Naphtha (Varnish Maker's and Printer's Naphtha from the hardware store), "petroleum ether" (not really a C-O-C ether but a mixed alkane distillate), ligroine, etc.
These have the following attributes - they are extremely low in viscosity so they penetrate any porosity quickly and dissolve the oil; they evaporate quickly so that any residual oil quickly becomes apparent, they do not swell or dissolve most mounting materials (limonene - the environmentally friendly base of many "green" solvents, is a potent stripper of polymers); and their high vapor pressure makes them easy to eliminate prior to SEM examination. I have routinely used immersion oil to temporarily coverslip geological thin sections for optical microscopy prior to microprobe work and then removed the oil with these solvents. Several drops in succession, less than a milliliter total, is sufficient to clean one slide.
John Twilley
Tamara Howard wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Please post informational responses to the server! I've been curious about } this, too but too lazy to look into it - the dishwashing cleansers sold in } "granola" stores (organic, non-toxic to the environment, etc.) are also } citrus-oil based (and at least one has almond oil; not sure if that is } just for the odor). } } Tamara
As we have been instructed for the past twenty years, nothing is NOT involved with that stuff.
So, 'e' is the correct answer. My students learned to hate multiple choice as much as I did.
Regards to all, with advice to my son. Always brush your teeth first, so the kisses are sweet. And he asked, "Mint or wintergreen?"
Have a nice weekend and remember, we can all get away with this because someone once worked on the 4th of July!
Fred Monson
} ---------- } From: Monson, Frederick C. } Sent: Thursday, June 28, 2001 1:30 PM } To: 'Microscopy Listserver' } Subject: Today's Real Humor } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Question from an old exam given to students who demanded MORE MULTIPLE } CHOICE! } } Which of the following is NOT involved with human reproduction? } } a. transcription } b. replication } c. translation } d. osculation } e. none of the above } } Answer tomorrow. } } } } 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. } }
Here's an old list, checked and amplified, that came to me from "Microscopy Today" I have checked all of these URL's
www.home.earthlink.net/~veilcs/ (EM+ special in Philips) (west Coast - USA) www.southernmicro.com (Georgia) (Research LM) www.dscoptical.com/services.htm (Boston?) (LM) www.caleywhitmore.com (Boston?) (LM) www.opticalexpertise.com (NY) (connection failure?) www.mwrn.com/product/microscope/repair.htm (LM and EM) www.sciscope.com/field-service.htm (30+ states) (up to Advanced LM) www.svms.com/services/index.html (CA) (LM) www.meyerinst.com (TX) (up to advanced LM) www.microresource.com/services2.htm (IL) (LM) www.allometrics.com/microscope_serv.htm (LA)(LM) www.labworksinc.com (San Francisco) (Limited to specialized LM scope accessories) www.mikronet.com (San Diego) (general LM services) www.uams.edu/oas/OES/oesilab.htm (AR) (University of Arkansaw LM support facility) www.uni.edu/pur/mms/equipment.html(U. N. Iowa) (Wide range of equipment including EM's) www.dominionmicroscope.com/services.htm (Richmond, VA) (LM and associated equipment) www.eosltd.co.uk/ (UK.COM) www.emsys.co.uk/ (UK.COM) www.hii.hitachi.com/svcem.htm (Hitachi) www.leo-usa.com/ (LEO) www.feic.com/products/index.htm (FEI - Philips) www.jeol.com/locations/service/service.html (JEOL Service, USA)
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.
Some faculty here are entertaining the idea of developing a confocal core facility. There is currently a single confocal microscope on campus in an individual PI's lab and it is not as available to others as they might like due to that lab's workload, location, etc.
I have been invited to join the group investigating the possibility of a second, more general access microscope, probably supported by several departments. I am currently the person responsible for the EM lab on campus, we have a TEM and an SEM, plus most of the standard prep. equipment. We also do some digital imaging. The EM business is not booming, save for a few hardcore users.
Since I am one of the 'Mr. Microscopes' here, I would like to join this group with a running start. I ask for your sage advice regarding the potential good and bad of this proposal and any comments you might offer on the state of confocal and such a lab.
I know a lot about microscopes, less specifically about confocals, but I could learn. If the microscope was put into the existing EM lab, I have a separate room, bench space, hoods etc. Just about anything needed to do microscopy. The only drawback would be that we are in a different building than most of the users. Its not a bad walk, but how big a deal is it to prepare specimens away from one's own lab? It would be pretty fair for the multiple users, since none of them are all in the same building either, they would all have to do some traveling to get here.
I am pretty sure there will be the usual squabbling that goes along with central facilities, like who gets to use it if there is competition, who will pay for it, etc. Is that likely to be any worse then with any shared equipment? Am I asking for trouble to have it added to the EM Lab?
Do you have an experience with the kind of day to day issues that a lab like this might generate. Are there lots of supplies that need to be ordered and kept on hand? Is the equipment reliable and are there any special features of one particular type of instrument that would make it more appropriate for such a general use facility vs. something used by an individual lab?
Your comments will be carefully considered and as always, greatly appreciated.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
The department I worked for took a different approach to the students not "ready for the dance". Every Thursday night the lab was open a faculty member from assistant professor to department head was there from 7:00 pm until at least 11:00 pm to help them with what ever problems they had not just Ag Engineering courses. I worked a research computer programmer and we didn't teach any programming courses and I had not teaching duties at all but any student could come to me any time with a computer problem and the same was true for any member of the staff or faculty for any problem the students had.
No grad students taught courses or labs.
The policy paid off real well. I don't remember when one of our students didn't get a job after graduation and many have gone on to very good things. This years senior design class is applying for patent on part of their class project.
I realize that farm kids are a good deal better prepared for life than most students but their education is usually a good deal poorer in science and math than the kids from the city and they have a good deal of catching up to do.
You are short changing your students if you just present the knowledge and walk away.
I realize that the approach we used is difficult to use in departments with large number of students. Turning education into a mass market commodity is IMO much of what is wrong with it.
Gordon
Gordon Couger
Stillwater, OK
----- Original Message ----- } From: "Monson, Frederick C." {fmonson-at-wcupa.edu} } } } You are absolutely right, and most of the time we behave just as you } describe us. Patient, etc. One purpose of my tirade was to point out that } there are differences between today and yesterday that give ALL of us } choices to make. I know a high school student who failed the year, because } he wanted to punish his parents. When he told me what he had done, I } responded that I knew a young man just like him when I was in high school. } His solution was to get straight A's so he could become more quickly } independent. } Science is our own, very human, endeavor. Those of us who do } science must constantly work to remove the temptations to embellish our } results, to twist the statistics and to 'jump' to conclusions. But, and } here's where I differ perhaps slightly. Sometimes, there is a good reason } to yell! My wife hates my streak of sarcasm! But I was raised on the } stuff. When I did something 'dumb,' everyone who loved me, joined in the } derisive repair. I learned to think at my dinner table before I was 10. I } did not like to be the but of everyone's attention. My Mom taught, then I } taught. I only stopped, because no one [too few] wanted to learn any more } [bad generalization]! Go to college today, and what do you find? Young } people unprepared for the rigor. Young people unprepared for the } intellectual dance - as we used to call it. Young people who don't know } what they want to do and are unwilling to work to find out. "They only do } what they are taught!" This is my mantra when I look for explanations in } the mirror. My older son had to learn - for the test - to name NH3 as } "ammonium", because his chemistry teacher would not believe the Handbook of } Chemistry and Physics. My chemistry teacher in public high school took } chemistry courses at Rutgers every third summer. He was the expert in } chemistry. My physics teacher had a PhD in the subject. My biology teacher } went on to be the chair of Physiology at a large Southern medical school. } If I wanted to complain about a professor at Lehigh, the Dean would show me } a list of nearby schools that had openings. The attitude was clearly } communicated. The greatest lessons in life are those that teach one to } manage the worst parts of it. If I failed a course, it was MY failure - and } it was at least MY problem! I wanted to be what I am, once I figured } everything out. The only advice I ever received, when I was having problems } with learning, was, "Study harder." My Mom just kept on telling me to keep } on trying as hard as I could. If I were young now, I'd be in group therapy } and zapped on some pharmaceutical, if the system had its way. } Given my experience, I hope it is not difficult to understand why I } might not appear to understand the problems of others. BUT! If I see a } young person in science who is poorly informed about how it is done, then I } will act in that person's behalf even if I am thought, and reported, to be } insensitive. } When I was a graduate student, we were all ordered to write } faculty/course evaluations. I did. I wrote analyses of each course, and } every one of them was critically positive. Not one word of my analyses were } printed. If I had demonized one of my professors, I would have been writing } novels today. The people who ordered the writings were ex-faculty } administrators who were pandering to the new consumer mentality in } education. If I had gone to my undergraduate department chair to complain } about a grade, he would have told me that I was wasting his time. Five } years later, there were lines of boomers outside of department offices. The } generation of manipulators had arrived! While they were standing in line, I } was learning. While I was learning, they were just getting warmed up! } I have had ideas snitched! Sometimes I even gave them away. On the } other hand, I have also had the other side of the experience. For six } months of learning followed by 30 minutes of consultation and work, I had } the good fortune to become a molecular biologist. For that effort, the } individual to whom I gave my best remembered my part - even after he had } spent years doing the work - and included me in the author list for a } gene(TERE1) that mapped to Chromosome 1 and appeared in print in a Nature } publication. He did NOT have to do that. There was NO obligation nor } agreement. I helped because he asked. People are basically good, } especially scientists, and I always start with TRUST, and I always will. } You are still absolutely correct. } } Regards for YOUR sensitivity, } } Fred Monson
with Nikon AFX-II camera control assembly using a Polaroid 4x5 film holder with a 4X marked on it.
Focus for images was done through the monocular eyepiece on the camera assembly.
I assume that my magnification though the microscope eyepiece would be 30X I assume that the magnification to the camera back would be 24X
Sorry if this seems like a silly question but I do not do much optical microscopy and got to thinking that it may be more than just the sum of the optics magnification.
Thanks
Roy Beavers Southern Methodist University Dept. of Geological Sciences Electron Microprobe Lab P.O. Box 750395 Dallas, Tx 75275 voice: 214-768-2756 fax: 214-768-2701 E-mail: rbeavers-at-mail.smu.edu
The House Ear Institute, a Los Angeles-based, non-profit research and education center dedicated to the hearing-impaired, has experienced tremendous growth over the past 50 years. We currently have an exceptional opportunity for a motivated and innovative self-starter.
You will support research projects in the cell and molecular biology of hearing. You will also perform advanced research procedures, such as immunoelectron microscopy and microwave processing, and maintain records. Requires a detail-oriented, flexible and enthusiastic team player with a BS in the biomedical field, as well as knowledge of TEM and SEM, specimen preparation for TEM and SEM, darkroom developing/printing, and digital image capture/manipulation. Must have efficient communication skills. Familiarity with electron microscopes, mechanical/electronic equipment, vacuum systems, and Unix, PC or Mac is ideal. Experience in cell culture, microbiology or protein purification is useful but not essential.
The Ahmanson Advanced Electron Microscopy and Imaging Center was recently established to perform independent research and provide support to a cell and molecular biology program in hearing research. The Center has a CM 120 BioTwin TEM and XL30 SFEG SEM. Specimen preparation equipment include ultramicrotomes for resin and cryosectioning, sputter coaters, vacuum evaporator, freeze-substitution, freeze-fracture and rapid freezing devices, a microwave processor, digital and stereologucal imaging capabilities and an SGI octane with 3-D reconstruction software.
The House Ear Institute, located in the downtown Los Angeles area, is a non-profit research and education organization dedicated to improving the lives of people who have hearing and hearing related disorders. The House Ear Institute is an Equal Opportunity Employer.
For consideration, please submit resume and cover letter to: House Ear Institute, Human Resources, 2100 W. 3rd St., Los Angeles, CA 90057-1922. FAX: (213) 483-8789. E-mail: jobs-at-hei.org.
When applying, please state that you saw this position advertised on the MSA listserver.
Regards,
Paul Webster
Paul Webster, Ph.D. Scientist II and Director Ahmanson Center for Advanced EM & Imaging House Ear Institute 2100 West 3rd Street Los Angeles CA 90057
this is some kind of follow-up question to Thearith's grid problem. We are looking at about 2 nm features in a polymer matrix. The features have very low contrast with respect to the all-carbon matrix due to the fact that they mostly contain O and a little Zn. For sample prep reasons we need to support the samples. Now: Are SiO and Si nitride coatings thin enough to still see such things ? The available thicknesses of these films seem very large compared to conventional C films.
TIA,
Andreas
************************************************* Dr. Andreas Taubert Dept. of Materials Science and Engineering 3231 Walnut Street University of Pennsylvania Philadelphia PA 19104-6272 tel: +1 215 898 2700 fax: +1 215 573 2128
Physical Chemistry is everything for which 1/T is linear ... *************************************************
Not thinking today, in my previous message I made assumptions about total magnification being the sum of the objectives but should have been the product.
Thanks
Roy Beavers Southern Methodist University Dept. of Geological Sciences Electron Microprobe Lab P.O. Box 750395 Dallas, Tx 75275 voice: 214-768-2756 fax: 214-768-2701 E-mail: rbeavers-at-mail.smu.edu
Yes, Fred, we published this list in Microscopy Today. We got it from a lady from Australia, whose name currently escapes me. Due to the interest in this topic, allow me to request that all readers contact companies that they know that does microscopy (all techniques)independent service. Ask them to contact me by email with the following: 1) Name of organization. 2) Contact info (tel. number and web site) 3) Products serviced 4) Area served (states, region, whatever) I will organize and publish the full listing, with Nestor's blessing, on this server. Best to all, Don Grimes, Microscopy Today
-----Original Message----- } From: Monson, Frederick C. [mailto:fmonson-at-wcupa.edu] Sent: Friday, June 29, 2001 12:23 PM To: 'service-at-jcnabity.com' Cc: 'Microscopy Listserver'
Here's an old list, checked and amplified, that came to me from "Microscopy Today" I have checked all of these URL's
www.home.earthlink.net/~veilcs/ (EM+ special in Philips) (west Coast - USA) www.southernmicro.com (Georgia) (Research LM) www.dscoptical.com/services.htm (Boston?) (LM) www.caleywhitmore.com (Boston?) (LM) www.opticalexpertise.com (NY) (connection failure?) www.mwrn.com/product/microscope/repair.htm (LM and EM) www.sciscope.com/field-service.htm (30+ states) (up to Advanced LM) www.svms.com/services/index.html (CA) (LM) www.meyerinst.com (TX) (up to advanced LM) www.microresource.com/services2.htm (IL) (LM) www.allometrics.com/microscope_serv.htm (LA)(LM) www.labworksinc.com (San Francisco) (Limited to specialized LM scope accessories) www.mikronet.com (San Diego) (general LM services) www.uams.edu/oas/OES/oesilab.htm (AR) (University of Arkansaw LM support facility) www.uni.edu/pur/mms/equipment.html(U. N. Iowa) (Wide range of equipment including EM's) www.dominionmicroscope.com/services.htm (Richmond, VA) (LM and associated equipment) www.eosltd.co.uk/ (UK.COM) www.emsys.co.uk/ (UK.COM) www.hii.hitachi.com/svcem.htm (Hitachi) www.leo-usa.com/ (LEO) www.feic.com/products/index.htm (FEI - Philips) www.jeol.com/locations/service/service.html (JEOL Service, USA)
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.
Ok, I can't resist... I haven't seen anyone mention the way I learned to do it in 1962...
Cut lens paper into strips a bit wider and longer than the slide. Place an end of the paper over the end of the slide and add a drop or so of your favorite liquid to the paper on the slide...just enough to spread over the width of the slide. Grasp the edge of the paper and pull slowly down the length of the slide so that the excess liquid is wicked into the remaining paper as it is pulled across the slide. Try to run out of paper as you reach the other end of the slide.
As Ever, Orin Keplinger ----- Original Message ----- } From: "Tobias Baskin" {BaskinT-at-missouri.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, June 28, 2001 4:28 PM
Hi there fellow microscopists,
Here in the Pathology EM lab we do not do rocket science, but standard thin-section TEM of renal biopsies.
In the last two weeks though I have been getting some ka ka (technical term) on my grids from my UA stain. I have already made up two new batches of UA stain and use .45 micron filters just before use. Question is that all of the sudden I am getting UA stain artifact on my grids. I have not have any trouble since I have been here. I have changed everything, new staining plate, cleaned and scrubbed everything that I use for staining. I still get some UA artifact. Is this normal and should I just forget about it or is there something else wrong I am missing now. I mix up a 2% UA stain of 50% methanol and 50% water. I stain for usually 10-12 minutes. the grids before staining or clean, but after UA stain they are a bit dirty...
Any suggestions?
Thanks,
Eric A. Rosen UCLA Medical Center Dept. Pathology EM Lab
Fellow colleagues,I am in a dilema. I just found out that RMC(whom our lab had Maintenance Agreement on two MT2 and one MT2B) has been taken over by first Ventana and at a later date Biobeck? Later company is not offereing maintenance service contracts for "old" MT2 or MT2B. Is there any one out there that could refer me to someone who works on these great workhorses in EM? Thanking you in advance, Teresa
Listers: I've been following the discussion of lens cleaing and would like insert my two cents worth. I'm from far outside of generally biological tenor of this group; an industrial metallographer ( microscopy of metal and alloys). After using an oil immersion lens, I clean it with Kodak lens cleaner fluid after blotting up the excess oil with a lens tissue, also Kodak. After 30 years of use, my lenses are clear and clean with no apparent deterioration. I did use xylene at first but was dissuaded by knowledgeable microscope salesman.
Sam Purdy Technical Center National Steel Corp. Trenton MI
Hello, Many thanks to everyone for their helpful comments on the Electrscan E3 SEM. We are still having vacuum problems in wet mode only. It works fine in High vacuum mode though. We checked the diffusion pumps, tried moving a rotary pump closer to the column, but no improvement. We are slowly changing o-rings to see if that helps.
Does anyone have any detailed repair manuals or diagrams with o-rings for this instrument? We have the customer maintenance manual, but it is rather limited. It mainly covers how to change apertures and doesn't go too deep into troubleshooting the vacuum system.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
I don't use this scope but I could venture the following guestimates.
If the photo system and light path does not include a separate photo objective (photo eyepiece) then the film plane magnification is the objective x 4. This would then be 20 x 4 = 80X.
Usually, with a 35mm camera body, the system is set up to be 1:1 with the oculars. This would mean 20 x 10 = 200X. If this field of view is different with the 4x5 cone, then there is some intermediary lens element. Usually, it is a 2.5X objective. For photo work, the oculars won't make any difference.
Perhaps some Optiphot experts can help you better.
gary g.
At 02:16 PM 6/29/2001, you wrote:
} Need some help trying to figure the magnification of some photos taken with } the following: } } Nikon Optiphot-Pol Microscope } CF eyepiece 10X } CF objective 20X } } with Nikon AFX-II camera control assembly using a Polaroid 4x5 film holder } with a 4X marked on it. } } Focus for images was done through the monocular eyepiece on the camera } assembly. } } I assume that my magnification though the microscope eyepiece would be 30X } I assume that the magnification to the camera back would be 24X } } Sorry if this seems like a silly question but I do not do much optical } microscopy and got to thinking that it may be more than just the sum of the } optics magnification. } } Thanks } } Roy Beavers } Southern Methodist University } Dept. of Geological Sciences } Electron Microprobe Lab } P.O. Box 750395 } Dallas, Tx 75275 } voice: 214-768-2756 } fax: 214-768-2701 } E-mail: rbeavers-at-mail.smu.edu
We have found staining artifacts to be one of the most frustrating, intermittent mysteries of all. A while back we began having trouble with UA and Pb contamination where no trouble had occured before---same stains, same processing, same everything, but suddenly there was goop everywhere. We mixed new stains, changed all of our 0.2 um filters, spun everything down, and filtered again, used NaOH religiously, held our breath while staining, etc., etc., etc. Finally, the only thing we could think of was water, so we called in our water system maintainer (let's just call them "Hey Mystery Man!!!!) who had been servicing our reverse osmosis, deionization system forever.
Well, they said, your system's shot. The diaphragm's bad, the cartridges are way expired, and the filter is one big bucket of rust. You need to maintain this better. We informed them that we paid them a princely sum to maintain it for us and what the h... were they doing every month when they came in to fuss with it?
Turns out that they thought we owned the system and had only been swapping out some tank or other, and they assumed we were doing the rest. When we produced documentation of our service contract on their leased system that we had been paying on since about 1735 A.D., there was an awkward pause. Then we fired them.
We purchased a Millipore tabletop water purification system that solved our problems overnight. It's a small, low-volume gizmo that produces a few litres of ultra-high purity water a day. Costs about $1200, as I recall, and requires annual filter changes to the tune of $200-300.
Moral of the story: Check the water when all else fails. Worked for us, anyway. Couldn't hurt to check.
We have no stake in Millipore whatsoever, except as relieved users of one of their products.
Randy Tindall Electron Microscopy Core University of Missouri
-----Original Message----- } From: Eric To: 'Microscopy Listserver' Sent: 6/29/01 5:24 PM
Hi there fellow microscopists,
Here in the Pathology EM lab we do not do rocket science, but standard thin-section TEM of renal biopsies.
In the last two weeks though I have been getting some ka ka (technical term) on my grids from my UA stain. I have already made up two new batches of UA stain and use .45 micron filters just before use. Question is that all of the sudden I am getting UA stain artifact on my grids. I have not have any trouble since I have been here. I have changed everything, new staining plate, cleaned and scrubbed everything that I use for staining. I still get some UA artifact. Is this normal and should I just forget about it or is there something else wrong I am missing now. I mix up a 2% UA stain of 50% methanol and 50% water. I stain for usually 10-12 minutes. the grids before staining or clean, but after UA stain they are a bit dirty...
Any suggestions?
Thanks,
Eric A. Rosen UCLA Medical Center Dept. Pathology EM Lab
I met some problems during I prepared the samples for GaN/sapphire plan-view using tripod wedge technique. After polishing, the sample was thinned by pips in one side. However, The other side usually was covered by copper, as a result the sample became dark. It seems the unthinned side should be covered something to prevent sputtered materials from growing on it. Do you have some knowledge on it?
Thanks in advance.
Best regards.
Feng ********************************************** Dr. Feng Wu Dept. of Materials Engineering Ben-Gurion University of the Negev Beer-Sheva 84105, Israel
Mica! 1stly core drill a 3mm disc. Then use sharp tweezers to peel a thin piece from dozens or even hundreds of layers in the mica disc depending how good you are to separate them. Anyway you just need one thin piece to cover the other side, and you may be able to reuse that thin piece.
********************************* Chaoying Ni, PhD Electron Microscopy Laboratory Materials Science and Engineering College of Engineering University of Delaware Newark, DE 19716 *********************************
On Sat, 30 Jun 2001, Feng Wu wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear All, } } I met some problems during I prepared the samples for GaN/sapphire plan-view } using tripod wedge technique. After polishing, the sample was thinned by pips } in one side. However, The other side usually was covered by copper, as a result } the sample became dark. It seems the unthinned side should be covered something } to prevent sputtered materials from growing on it. Do you have some knowledge } on it? } } Thanks in advance. } } Best regards. } } Feng } ********************************************** } Dr. Feng Wu } Dept. of Materials Engineering } Ben-Gurion University of the Negev } Beer-Sheva 84105, Israel } } voice 972-8-6461473 } fax 972-8-6472944 } fwu-at-bgumail.bgu.ac.il } ********************************************** } } }
Dr. Feng Wu wrote: =============================================== I met some problems during I prepared the samples for GaN/sapphire plan-view using tripod wedge technique. After polishing, the sample was thinned by pips in one side. However, The other side usually was covered by copper, as a result the sample became dark. It seems the unthinned side should be covered something to prevent sputtered materials from growing on it. Do you have some knowledge on it? ================================================= There are two materials that to the degree we have been able to determine, are essentially equivalent for this purpose: One is called Microshield™ and the other Lacomit™.
SPI Supplies has offered Microshield for this purpose and more information about its use and application can be found on the SPI Supplies website at URL http://www.2spi.com/catalog/chem/microshield.html
Disclaimer: SPI Supplies has not done any careful analysis to prove that Microshield and Lacomit are identical, only that for this particular TEM application, they do seem to be equivalent. If there is information to the contrary, we would appreciate hearing about it. Suggestion: Use in conjunction with the Microshield remover.
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 ############################ ===============================================
} Need some help trying to figure the magnification of some photos taken with } the following: } } Nikon Optiphot-Pol Microscope } CF eyepiece 10X } CF objective 20X } Hi Roy,
If you still have access to the microscope, put a specimen on the stage which has a large feature, one you can accurately measure. Use the lowest power lens available. Remove the film cassette from the camera and stretch a piece of Scotch Magic tape across the film plane. Focus on the specimen, lower the room lights and measure the size of the image on the scotch tape, which will now be clearly visible. You could also measure some feature on the original slide and compare it to the photograph.
I would guess the mag factor is 80x, but you can confirm that using this process.
Dave Burton Optical Specialist University of Washington
Dr. Andreas Taubert asked: ============================================= this is some kind of follow-up question to Thearith's grid problem. We are looking at about 2 nm features in a polymer matrix. The features have very low contrast with respect to the all-carbon matrix due to the fact that they mostly contain O and a little Zn. For sample prep reasons we need to support the samples. Now: Are SiO and Si nitride coatings thin enough to still see such things ? The available thicknesses of these films seem very large compared to conventional C films. ============================================= If the thinnest thickness of the nitride should still be too much, which is presently 30 nm, but soon to be reduced to the 10-20 nm range, the present silicon nitride membrane grids can be furnished "holey", that is, with a matrix of holes that are about as small as you want them, in just about any arrangement that you might want them. In that kind of situation, there is no nitride underneath the sample being analyzed, so long as you are looking "through" the holes. The only carbon (or oxygen) present is that which is coming from the sample. The typical compliment of holes approximately doubles the cost of the grid, because it is a separate operation done by micromachining techniques. This is explained in greater detail at URL http://www.2spi.com/catalog/instruments/silicon-nitride.html
Disclaimer: SPI Supplies offers silicon nitride membrane window grids, both with and without the holes.
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 ############################ ==============================================
The ion beam forms a cloud of debris, which settles on the reverse side of the specimen and this needs to be covered. A double beam ion gun system avoid that problem because the specimen is thinned on both sides simultaneously. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Saturday, June 30, 2001 5:06 PM, Feng Wu [SMTP:fwu-at-bgumail.bgu.ac.il] wrote: } } Dear All, } } I met some problems during I prepared the samples for GaN/sapphire plan-view } using tripod wedge technique. After polishing, the sample was thinned by pips } } in one side. However, The other side usually was covered by copper, as a } result } the sample became dark. It seems the unthinned side should be covered } something } to prevent sputtered materials from growing on it. Do you have some knowledge } } on it? } } Thanks in advance. } } Best regards. } } Feng } ********************************************** } Dr. Feng Wu } Dept. of Materials Engineering } Ben-Gurion University of the Negev } Beer-Sheva 84105, Israel } } voice 972-8-6461473 } fax 972-8-6472944 } fwu-at-bgumail.bgu.ac.il } **********************************************
i am trying to find a way into the literature on experimental techniques appropriate to study 511 photon detection and capture vs properties of scintillating materials. Is there a standard way to do this?
Can someone point me at a good place to begin a literature survey?' sorry, my question is not better phrased, but I am just beginning to look at this?
sterling
Computer Aided Cell and Molecular Biology (CACMB), not medicine, will find the cure for cancer and other diseases. There will always be a need for the trained clinician (MD/RN) but, advanced diagnostic and treatment option selection has become gene based, has moved from the physician's practice to the computerized cell and molecular biology laboratory, and appropriate treatment options should now be based on the personal biology of the patient.
The nearest program in the PC world is JEMS. I recently examined what was available from PC's particularly a program that could simulate electron diffraction.
I purchased Pierre Stadelman's JEMS - runs in JAVA - and it is beautiful for both images and diffraction. Runs very nicely on a Compaq 1.6 GHz machine with 512 MB and will also run on a Mac or Sun. I have already set up all the amphibole minerals mainly for electron diffraction work. Very useful so far. JEMS also plays well with triclinic phases something many other programs have difficulty with. Something to do with right angles I guess.
Check it out at {http://cimewww.epfl.ch/people/stadelmann}
MacTempas and CrystalKit together were(?) unique however in how they were able to treat interfaces such as twins. I don't think JEMS does this very easily.
I found however that if you are interested in looking at crystal structures neither CrystalKit nor JEMS can compete with David Palmer's CrystalMaker program which currently runs only on a Mac. He is however working on a C++ version that will run on PC's.
Check it out at {http://www.crystalmaker.co.uk} and email him that you are anxiously awaiting the PC version.
My advice is purchase an $800 iMac and run the Mac programs and give JEMS a good look for your PC. Can't have too many platforms.
Cheers, Gordon Nord US Geological Survey - Emeritus
Maria Lada wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear ALL, } } Is there a version of MAcTempas/CrystalKit for a Windows/PC platform? } } Is there nay other windows platform Package doing a similat job? } } with thanks } } Maria Lada } EEE department } University of Sheffield
HMDS *MUST* be used in a hood. I'm at home as well, so don't have the MSDS, but there is no question about this.
Phil
} Hi Liststers! } I would appreciate any information on the safety of } hexamethyldisilazane (HMDS). I didn't get much out of the data } sheet. I would like to know what I am working with... Is it } hazardous to one's health? I don't have a hood in my room, but I } could move to another room which does have a hood. Thanks!
-- }}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{ 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)
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html -----------------------------------------------------------------------.
Maria, Thierry Epicier has written SIMPLY-S code for HREM image simulation (and more) for the PC. The simulation part is based on the original SHRLI (simulated high-resolution lattice image) code. Since MacTempas is a port to the Mac of the TEMPaS code (developed from SHRLI to run on a Vax system with a Tektronics GUI), the results of SIMPLY-S and MacTempas should be the same. You can reach Thierry at Thierry.Epicier-at-insa-lyon.fr Mike O'Keefe
----- Original Message ----- } From: "Maria Lada" {m.lada-at-sheffield.ac.uk}
Hi Barbara,
I received my HMDS from Agar Scientific and it came with a 6 page material safety data sheet.
In short:
it is highly flammable, toxic, harmful by inhalation, in contact with skin and if swallowed, irritating to eyes, causes severe skin irritation, is readily absorbed through the skin - and the target organ is nerves.
In case of contact with eye, rinse immediately with plenty of water and seek medical advice. Wear suitable protective clothing.
You find MSDS in the internet by doing a keyword search and the major EM suppliers have MSDS of their products on their web sites.
I always use it in the fume cupboard, with gloves - and make sure that their is no hot plate near by. (in fact I handle it as I would handle Propylen oxide).
Regards
Claudia
Date sent: Fri, 06 Jul 2001 16:18:12 -0700 } From: Barbara Plowman {Bplowman-at-sfmail.dental.uop.edu} To: Microscopy {Microscopy-at-sparc5.microscopy.com}
Hi Liststers! I would appreciate any information on the safety of hexamethyldisilazane (HMDS). I didn't get much out of the data sheet. I would like to know what I am working with... Is it hazardous to one's health? I don't have a hood in my room, but I could move to another room which does have a hood. Thanks!
Dr. C. Hayward-Costa School of Life Sciences Kingston University Penrhyn Road, Kingston upon Thames Surrey KT1 2EE, UK 44(0)208 547 2000 x 2240 Email: c.hayward-at-kingston.ac.uk Fax: 44(0)208 547 7562
Baresh, there is a very easy, quick and effective way if you have Adobe PhotoShop:
1) Create a new layer above your image. 2) Select the region you want to colour. 3) Fill it with the colour you want. 4) Change the layer properties from 'Normal' to Overlay. Done!
You may do step (2) by drawing round the region with the 'freehand select' or 'polygon select' tool, or on the lower layer using the magic wand.
e-mail richard.beanland-at-marconi.com Tel. +44 1327 356363 Fax. +44 1327 356398 ============================================================== "The information contained in this message is legally privileged and confidential information intended for the eyes of the individual or entity named above. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution or copying of this message is strictly prohibited. If you have received this message in error, please notify us immediately by telephone." Marconi Optical Components Limited Registered in England No. 4113798 Registered Office: One Bruton Street London W1J 6AQ
A BSE standard can be made from almost any materials (suitable for SEM). The std should be ground and polished to remove topography contrast. What materials you choose will depend on the examination goals.
This brings me to a point. I would not stress BSE "calibration" as a generally useful technique. Under VERY limited circumstances, such calibration/standardization can be beneficial. Routine use of this technique will guarantee that sooner or later an important phase will be missed when examining an unknown. When examining with BSE, I find it imperative to be "all over the map" with the BSE B&C to be sure I am not missing something. It is not unheard of to need two or more images to define what is happening. Sometimes there is simply not enough dynamic range presented in one image to perceive all variations in composition.
Regards, Woody White, McDermott Technology, Inc.
} -----Original Message----- } From: Schumacher, Elaine [mailto:efschuma-at-uop.com] } Sent: Friday, July 06, 2001 12:27 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Brightness/Contrast Calibration Standards } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Fellow Microscopists, } } We're looking for suggestions for SEM standards to calibrate } brightness/contrast settings (for example, setting an auto } B/C feature in } the SEM software, so that the desired repeatable B/C level is } obtained by } multiple users). Our customers' goal is to do } correction/matching of gray } scale histograms between images of different samples, and } they would like to } make the process of adjusting B/C levels at the start of } image acquisition } to be as standardized as possible. The imaging might be done } in secondary } or backscattered mode, so suggestions for both would be appreciated. } } Thanks vey much for you input, } } Elaine } } Elaine F. Schumacher } Analytical Specialist } UOP LLC } 25 East Algonquin Road } Des Plaines, IL 60017-5017 } Phone: 847-391-3403 } Fax: 847-391-3719 } Email: efschuma-at-uop.com } }
Dear Smartech, If someone published such an article comparing EDS detectors as to their ability to detect light elements it should be the SEM manufacturers. The light element sensitivity is determined by the geometry of the detector not the entrance window. Basically the higher the take-off angle of the emerging photon, the better the sensitivity due to the absorption of the matrix. The high voltage of the electron beam also changes the sensitivity due to the depth of penetration and therefore the depth of the x-ray source. Operating a EDX repair facility, I work on all of the EDX manufacturer detectors. With one possible exception, all the manufacturers use the same entrance window material, using various trade mark names. There also seems to be very little difference is the Si(Li) crystals or FET's which are the two internal components of the detectors. Since all of the manufacturers use a resolution specification for Fe55 photons and a peak to background ratio for the 1.0 Kev channel, there would be no comparison specifications for light element sensitivity. I hope this answers your question. Jim Nicolino
I concur with Woody's comments, and offer a couple of more.
It is often possible for the same sample type to set BSE brightness to be consistent between images, and there are many time you would want to do that. We do image analysis of concrete and use calcite and graphite to repeatably set our signal. Some images look good that way, but even then, variation in polishing can greatly affect what the final image looks like. Sometimes it is not so good.
Last week I was looking at samples of a Cr-Al alloy. (It required much different contrast than did the concrete.) Some areas of some samples had three phases present (Al, Al-Cr, and Cr); others had only two (Al and Al-Cr). My inclination was naturally to first turn up the contrast to spread the signal throughout the grayscale to show the character of what was present. I later went back to take some images at consistent signal levels to show that the Cr-rich phase was missing.
Now "calibration" is a reasonable process for BSE. It is much less tractable for SE. SE signals are just not that consistent. Since they are so sensitive to topography, I find myself changing contrast all the time. I expect to need to reduce contrast as I increase magnification as more details come into view. That is the nature of it.
I wonder, do you have a waveform monitor on your scope to help in setting contrast? We use ours religiously and I have been surprised to hear that some scopes don't have them. (I guess the one we had on our JEOL U3 was an add-on, but since they have been part of our spec.) We have determined where white and black are on our waveform display, and before each photo we check the waveform to be sure that the signal is within our recordable range.
There is still some experience involved in setting the level so the picture looks good afterwards. A picture may be completely within the recordable range and still "look" too dark or light. I tend to prefer my background away from full black or white even if there is nothing to see in it. That just takes practice since there is such a wide variety of samples out there.
Digital imaging with gamma control after the fact has done a lot to make sure the first picture will be a keeper. If you don't have that yet, please consider it, or figure that your operators will just have to practice more.
Cheers, Warren
At 07:18 AM 7/9/2001 -0500, you wrote: } Elaine, } } A BSE standard can be made from almost any materials (suitable for SEM). } The std should be ground and polished to remove topography contrast. What } materials you choose will depend on the examination goals. } } This brings me to a point. I would not stress BSE "calibration" as a } generally useful technique. Under VERY limited circumstances, such } calibration/standardization can be beneficial. Routine use of this } technique will guarantee that sooner or later an important phase will be } missed when examining an unknown. When examining with BSE, I find it } imperative to be "all over the map" with the BSE B&C to be sure I am not } missing something. It is not unheard of to need two or more images to } define what is happening. Sometimes there is simply not enough dynamic } range presented in one image to perceive all variations in composition. } } Regards, } Woody White, McDermott Technology, Inc. } } } -----Original Message----- } } From: Schumacher, Elaine [mailto:efschuma-at-uop.com] } } Sent: Friday, July 06, 2001 12:27 PM } } To: Microscopy-at-sparc5.microscopy.com } } Subject: Brightness/Contrast Calibration Standards } } } } Fellow Microscopists, } } } } We're looking for suggestions for SEM standards to calibrate } } brightness/contrast settings (for example, setting an auto } } B/C feature in } } the SEM software, so that the desired repeatable B/C level is } } obtained by } } multiple users). Our customers' goal is to do } } correction/matching of gray } } scale histograms between images of different samples, and } } they would like to } } make the process of adjusting B/C levels at the start of } } image acquisition } } to be as standardized as possible. The imaging might be done } } in secondary } } or backscattered mode, so suggestions for both would be appreciated. } } } } Thanks vey much for you input, } } } } Elaine } } } } Elaine F. Schumacher } } Analytical Specialist } } UOP LLC ---------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
The main tricks are to low-angle mill both sides of the specimen and to not use Cu grids. We switched to Mo grids years ago and we rarely see back sputtered material on samples milled on both sides (from the substrate direction, of course). If you want to use up your stock of Cu grids, cut the back side of the grid away so that there is no grid material beyond the specimen to sputter from. Grid becomes the "C" shaped grid used for FIB prepped specimens. Sometimes some sputtered material persists after very low angle milling. In that case, pop the sample into a conventional high-angle mill and mill at 25 degrees for 30 seconds at 2 or 3 KeV to clean off the sputter artifact.
Ron Anderson, IBM, Hopewell Jct., New York, USA. anderron-at-us.ibm.com
IBM Analytical Services; http://www.chips.ibm.com/services/asg
Richard Beanland +44 1327 356363 {richard.beanland-at-marconi.com} on 07/02/2001 07:02:41 AM
To: Feng Wu {fwu-at-bgumail.bgu.ac.il} cc: Microscopy Listserver {microscopy-at-sparc5.microscopy.com}
Feng, the trick is not to have a hole in your specimen where the ions can go through and back-sputter the Cu on to your sample. So make your specimen big enough to cover up the hole in the support grid and stop ion milling immediately when a hole appears. The problem will go away to some extent if you use the two-sided specimen holder. Alternatively, use an inert grid and dissolve the Cu with an acid after specimen prep. (Most acids are unlikely to affect GaN or sapphire.)
Richard
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear All, } } I met some problems during I prepared the samples for GaN/sapphire plan-view } using tripod wedge technique. After polishing, the sample was thinned by pips } in one side. However, The other side usually was covered by copper, as a result } the sample became dark. It seems the unthinned side should be covered something } to prevent sputtered materials from growing on it. Do you have some knowledge } on it? } } Thanks in advance. } } Best regards. } } Feng } ********************************************** } Dr. Feng Wu } Dept. of Materials Engineering } Ben-Gurion University of the Negev } Beer-Sheva 84105, Israel } } voice 972-8-6461473 } fax 972-8-6472944 } fwu-at-bgumail.bgu.ac.il } ********************************************** } }
e-mail richard.beanland-at-marconi.com Tel. +44 1327 356363 Fax. +44 1327 356398 ============================================================== "The information contained in this message is legally privileged and confidential information intended for the eyes of the individual or entity named above. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution or copying of this message is strictly prohibited. If you have received this message in error, please notify us immediately by telephone." Marconi Optical Components Limited Registered in England No. 4113798 Registered Office: One Bruton Street London W1J 6AQ
Another potential cause of variation is dye concentration. To be certified by the Biological Stain Commission, toluidine blue dye lots must have at least 50% dye content. Unless corrected each time, toluidine blue solutions will vary in dye content.
Gary Gill
-----Original Message----- } From: Rosemary White [mailto:Rosemary.White-at-pi.csiro.au] Sent: Friday, July 06, 2001 5:04 AM To: microscopy-at-sparc5.microscopy.com
Continuing on this thread, I had always thought Richardson's stain was the methylene blue/azure II combination, but not so, it seems, since the 1960 paper refers to both this stain and to tol blue. Someone emailed that methylene blue must be used with azure II, but I don't think this is necessary, though it may give better staining.
Re metachromicity, as Fred Monson and Jan Coetzee note, it depends quite a bit on pH of the tol blue (for plant material at least), and used to depend on the source of stain but I guess this is not such a problem these days(?). We generally use a benzoate buffered solution of tol blue at pH 4.5 for fresh tissue and tissue embedded in hydrophilic resins like LR White and other methacrylates, and a pH 10 solution (buffered, if necessary, in sodium carbonate) for sections of hydrophobic epoxy resins - e.g. Spurr's. Tol blue metachromicity isn't so good at high pH (at least in my hands). The pH of tap water is usually slightly acidic, so aqueous tol blue is generally OK for fresh tissue and methacrylate resins. Since borate buffers are alkaline, perhaps this was the original reason they were used for the hydrophobic resins(?) - and they do produce a much more permanent stain.
Re. "old" references and books, most people here do molecular/genetic type work and literature older than about 1995 is considered venerable..... I am amazed (and appalled) at the great old histochemistry and cytochemistry books thrown out by libraries - the source of most of my "old" books.
Friday night, burning a few CDs..... Looked up another reference on histochemistry - A.G. Everson Pearse (1968) Histochemistry Vol 1 3rd edn (SBN 7000-1326-1 for Fred Monson - perhaps ISBN came after this time....) - a method from 1947 is given that uses a solution of 0.25% tol blue in 0.25% borax followed by 0.5% potassium dichromate in sat'd mercuric chloride, for permanent staining of paraffin sections prior to dehydration and mounting in Canada Balsam. Hess M, Hollander F (1947) J Lab Clin Med 32:905
Mallory, whose name used to be associated with various staining protocols, put it all together in a book: Mallory FB (1938) Pathological Technique. WB Saunders, Philadelphia. This is where Mallory's borax methylene blue comes from - 1% methylene blue plus 1% azure II in 1% borax. Lillie refers to it in his book - Histopathologic Technic - we have the 2nd edition, published in 1954, LC number 53-5622 - again, pre-ISBN.
Will finish now, in case this is getting too bor-ing!
cheers, Rosemary
Rosemary White Microscopy Centre CSIRO Plant Industry GPO Box 1600 Canberra, ACT 2601 Australia
Yes geometry, accelerating voltage, sample composition and thickness have a huge effect, but you have to compare things correctly. Start with the detector and its window, if it isn't configured properly to start with then the sample and operating conditions can be irrelevant.
There are different windows out there, and not all windows are identical even if they are made from the same nominal material. Variation in thickness of the different components can make a significant difference in the performance of a window. This was shown in a round robin test run by Larry Thomas a long while back
Comparison of UTW/WL Detectors.... L.E. Thomas etal Analytical Electron Microscopy - 1984 San Francisco Press. pge 333-336.
It is true that the windows are more uniform today than they were back then, but there is still variation. In addition any contamination deposited on the window by the instrument environment (or ice formation inside the cryostat), and of course electronic noise will significantly deteriorate a detectors low energy performance.
If you wish to see a plot of detection efficiency vs detector window type (which DOES make a difference if the windows are different) lookup the following reference
N.J. Zaluzec , Evolutionary Developments in X-ray and Electron Energy Loss Microanalysis Instrumentation .... MicroBeam Analysis, 1990 San Francisco Press. pge 330-333.
Some of the figures from these two papers can also be found in this PDF documnet
Hello, My protein sample does not adsorb to the grid even after glow discharg, it is a membrane protein complex with the membrane lipids froming vescicles. Nothing show up after negative staining. What should I do to improve the adsorption? I have tried quick dring with filter paper after put the sample on the grid, but it does not work.
perfect microscope for home or garage or for any small lab needind an addition al imaging microscope. This scope is in great shape and needs a home with someone who can appreciate the fantastic imaging capabilities and the user serviceable nature of the scope!
Please check the Central Michigan University Purchasing Department Web site for details: http://www.purchasing.cmich.edu/frametemp/sales_main.html (go to the sealed bid items under "Pop Stops" or check the link below)
Nestor Zaluzec has a PPT presentation about EDS on the Argonne National Lab website that he advertised a few weeks back.
Part of that presentation deals with sensitivities for the various window materials and detector types. There are significant differences. No one detector is best for all situations and all elements. You may have to determine which detector is best for your combination of elements.
There should be similar critiques available through any number of sources, but they will probably have to be a general discussion. Manufacturers have a way of changing things and there may be something new, or at least slightly different on the market this week.
Warren
At 06:46 PM 7/6/2001 -0400, you wrote:
} Does anybody know of a good article that compares EDS detectors, from a } variety of manufacturers, w/ respect to their light element sensitivity? } } SMARTech } 860-491-3299 } www.semguy.com } 19 Cornwall Drive } Goshen CT 06756
---------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
Hello, My protein sample does not adsorb to the grid even after glow discharg, it is a membrane protein complex with the membrane lipids froming vescicles. Nothing show up after negative staining. What should I do to improve the adsorption? I have tried quick dring with filter paper after put the sample on the grid, but it does not work.
Dear Mike, Unfortunately, the appropriate conditions for optimal coverage vary from sample to sample. You can change the hydrophobicity/hydrophyllicity of a thin carbon layer by "aging" over a few days for more hydrophobic carbon, glow discharging for more hydrophyllic carbon, and glow discharging in the presence of a volatile amine (we have amyl amine) for even more hydrophyllicity. You can also use either the side of the thin carbon--if, indeed, that is your substrate--which was in contact with mica--if the carbon was vacuum-evaporated onto freshly-cleaved mica--or the side away from the mica. Each of these has somewhat different properties. If you are using carbon-coated formvar as a substrate, you may still be able to alter its properties, but not in as many ways as with a thin carbon film. You do not say whether you are testing a preparation for cryo-EM by looking at a negatively-stained grid or whether you just want to look at a neg-st specimen. If the former, try blotting from one side only. Blotting from the back should leave more of the vesicles on the grid (although the thickness of the specimen when frozen can be hard to control). Also, you need to assure yourself that you can identify your specimen after mock cryo preparation followed by warming up and staining; it may look very little like a typical neg-st prep. If the latter, try letting the specimen with the stain sit on the grid for several minutes before blotting. This kind of preparation can be very tricky, and you just need to try different combinations of conditions until you get good coverage. Good luck. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
I guess that my original post was too curt. We have gone away from Tol Blue, to methylene blue due to safety reasons. There is no toxicology data on the MSDS for Tol Blue, but there is for Meth Blue. As I have a pregnant technician, I feel better knowing the LD 50 information, which is on the MSDS for Meth Blue. But, methylene blue is not as good for a general stain as Tol Blue, so I was just curious if there was an overriding reason to use it. I am presently experimenting with food based stains, which eliminates the issue completely.
The issue of pregnant (or possibly pregnant) workers in the lab has turned out to be fairly complicated. If anyone else out there has beem through this situation, I would like to know how you dealt with it.
Bill
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: Monson, Frederick C. [mailto:fmonson-at-wcupa.edu] Sent: Thursday, July 05, 2001 10:20 AM To: 'Sally Stowe' Cc: 'Microscopy Listserver'
Dear Sally, To address the REAL attribution issue, one of us is going to have to go to a REALLY good library that has these OLD journals back to the late '50's. We may have to divide the labor, but the question is sufficiently interesting and meritorious that we ALL should get it right, when asked. Someone was responsible for the use of borax (Sodium tetraborate) with Toluidine blue O (1% in 1%). Here are all the citations that Pease provides and his [attributions]:
1. Trump, Schmuckler and Benditt (1961), J. Ultrastruc Res., 5: 343 [ 0.1% Toluidine blue O in 2.5% carbonate at pH 11 (confirmed in Lewis and Knight below)] 2. Charles and Meek , G.R. separately in a discussion of paper by Mercer(1963), J Microscp Sci., 81:179. [mentions 1% Toluidine blue O in 1% borax used at 85 degrees C] 3. Pease, D.C.(1964), Histological Techniques for Electron Microscopy (2d), Academic Press, NY, NY, pp 256-262. [mentions on p. 260 in 2st paragraph the exact method that I use for staining as 'his' method - "...flood the slide, cook until [edge] of drop [froths/]steams [or dries]....". [This is a long discussion to which Pease devotes an entire section, but one can just FEEL his impatience with the unimportance to HIM of this issue. The book, after all, is NOT about light microscopy. The semi-thin section is for orientation only, and hardly deserves any more notice - especially if the light microscopists cannot SEE what is to be seen in the best material ever handed them.] 4. In Kay, D.H.(ed)(1965), Techniques for electron Microscopy(2d), F.A. Davis, Philadelphia, ISBN: [NOT given - is this book pre-historic?!] The following is cited as THE reference for using Toluidine blue O: Benscome, Stone, Latta, and Madden (1959), J. biophys. biochem. cytol, 5: 508.
I have just looked in Geoffrey Meek's book (1970), Practical Electron Microscopy for Biologists, Wiley_Interscience, NY, NY, p. 456-457. ISBN: 0-471-59030-4. 1% in 1% with no citation, though in this book he has purposely NOT used citations at all.
I have also consulted the Sorvall manual on "Thin Sectioning"(3d Ed)(1973) as well as the SerVall manual "Thin Sectioning and Associated Technics for Electron Microscopy"(1959) which apparently was distributed with the MT-1. In the latter, only staining of Methacrylate sections was given and no mention of 1% *bor* of any kind. In the former, 1% borate at pH 4.5 is mentioned, but that is not relevant to this discussion.
I would bet that you might have some of these references. If you do not, let me know, and I will dig them up, in old Philadelphia, in Ben Franklin's old stacks (at the U. of P(ennsylvania) as we call it). I will do it anyway, because I want to know, but I will do it quicker if you tell me I'm keeping you waiting.
Regards,
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: Sally Stowe } Sent: Tuesday, July 3, 2001 3:37 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Tol blue/borax } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Somebody just asked me the reference for toluidine blue/sodium borate (1% } each) which is our routine LM stain - I cant find an attribution in any of } about 10 books. Can anybody help? This person should not be forgotten! } One suggestion was Alsop? } } thanks } } Sally } } } } Dr Sally Stowe } Facility Coordinator, } ANU Electron Microscopy Unit } Research School of Biological Sciences } Australian National University } Canberra ACT0200 } AUSTRALIA } stowe-at-rsbs.anu.edu.au fax 61 (0)2 6125 3218 } ph 02 6125 2743 } http://www.anu.edu.au/EMU } } } } }
Woody, I certainly agree with you, but it seems Elaine was talking about qualitative or even quantitative comparison of the specimens histograms (I hope the specimens are flat). I think, the most straightforward thing is to use one of the measured specimens with wide brightness and contrast range. It makes no sense to calibrate BSE image for range from Si to Pb just to look at specimens containing, say, Cu and Au. The same for SE image. Polished specimen has much narrower signal range for brightness and contrast than fractured specimen. And, of course, for a microscope with a tungsten gun it is better to work well in the saturation range to insure brightness stability.
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} -----Original Message----- } From: White, Woody N. [mailto:nwwhite-at-mcdermott.com] } Sent: Monday, July 09, 2001 7:19 AM } To: 'Schumacher, Elaine'; Microscopy-at-sparc5.microscopy.com } Subject: RE: Brightness/Contrast Calibration Standards } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Elaine, } } A BSE standard can be made from almost any materials } (suitable for SEM). } The std should be ground and polished to remove topography } contrast. What } materials you choose will depend on the examination goals. } } This brings me to a point. I would not stress BSE "calibration" as a } generally useful technique. Under VERY limited circumstances, such } calibration/standardization can be beneficial. Routine use of this } technique will guarantee that sooner or later an important } phase will be } missed when examining an unknown. When examining with BSE, I find it } imperative to be "all over the map" with the BSE B&C to be } sure I am not } missing something. It is not unheard of to need two or more images to } define what is happening. Sometimes there is simply not } enough dynamic } range presented in one image to perceive all variations in } composition. } } Regards, } Woody White, McDermott Technology, Inc. } } } -----Original Message----- } } From: Schumacher, Elaine [mailto:efschuma-at-uop.com] } } Sent: Friday, July 06, 2001 12:27 PM } } To: Microscopy-at-sparc5.microscopy.com } } Subject: Brightness/Contrast Calibration Standards } } } } } } -------------------------------------------------------------- } } ---------- } } The Microscopy ListServer -- Sponsor: The Microscopy Society } } of America } } To Subscribe/Unsubscribe -- Send Email to } } ListServer-at-MSA.Microscopy.Com } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -------------------------------------------------------------- } } ---------. } } } } } } Fellow Microscopists, } } } } We're looking for suggestions for SEM standards to calibrate } } brightness/contrast settings (for example, setting an auto } } B/C feature in } } the SEM software, so that the desired repeatable B/C level is } } obtained by } } multiple users). Our customers' goal is to do } } correction/matching of gray } } scale histograms between images of different samples, and } } they would like to } } make the process of adjusting B/C levels at the start of } } image acquisition } } to be as standardized as possible. The imaging might be done } } in secondary } } or backscattered mode, so suggestions for both would be appreciated. } } } } Thanks vey much for you input, } } } } Elaine } } } } Elaine F. Schumacher } } Analytical Specialist } } UOP LLC } } 25 East Algonquin Road } } Des Plaines, IL 60017-5017 } } Phone: 847-391-3403 } } Fax: 847-391-3719 } } Email: efschuma-at-uop.com } } } } } }
Sputter deposition of Cu on the back side of holed samples was a common problem in one-sided thinning with the PIPS. The Cu came from the sample support posts that the sample was waxed onto. As Richard Beanland suggested, a simple acid cleaning (e.g., dilute nitric acid) removes the Cu effectively if the sample can take this treatment. Alternatively, you could vapor deposit a thin coating of rocksalt on the sample and later clean it off with water. Two-sided milling will avoid gross sputter contamination, but--
As a general rule in ion milling, it's highly recommended to avoid using any material in the sample fabrication or holder that you can't tolerate having in your sample. Whether you use a Cu or Mo grid, or fabricate samples by sandwiching the sample between Si blocks, for example, the nature of the ion milling process just about guarantees some of this material will collect in the sample cracks and crevices.
Larry Thomas Pacific Northwest National Laboratory, Richland, WA, USA larry.thomas-at-pnl.gov
--------- From: Ronald Anderson Sent: Monday, July 9, 2001 6:45 AM To: richard.beanland-at-marconi.com Cc: Feng Wu; Microscopy Listserver Subject: Re: GaN/Sapphire thinned by PIPS
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The main tricks are to low-angle mill both sides of the specimen and to not use Cu grids. We switched to Mo grids years ago and we rarely see back sputtered material on samples milled on both sides (from the substrate direction, of course). If you want to use up your stock of Cu grids, cut the back side of the grid away so that there is no grid material beyond the specimen to sputter from. Grid becomes the "C" shaped grid used for FIB prepped specimens. Sometimes some sputtered material persists after very low angle milling. In that case, pop the sample into a conventional high-angle mill and mill at 25 degrees for 30 seconds at 2 or 3 KeV to clean off the sputter artifact.
Ron Anderson, IBM, Hopewell Jct., New York, USA. anderron-at-us.ibm.com
IBM Analytical Services; http://www.chips.ibm.com/services/asg
Richard Beanland +44 1327 356363 {richard.beanland-at-marconi.com} on 07/02/2001 07:02:41 AM
To: Feng Wu {fwu-at-bgumail.bgu.ac.il} cc: Microscopy Listserver {microscopy-at-sparc5.microscopy.com} Subject: Re: GaN/Sapphire thinned by PIPS
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Feng, the trick is not to have a hole in your specimen where the ions can go through and back-sputter the Cu on to your sample. So make your specimen big enough to cover up the hole in the support grid and stop ion milling immediately when a hole appears. The problem will go away to some extent if you use the two-sided specimen holder. Alternatively, use an inert grid and dissolve the Cu with an acid after specimen prep. (Most acids are unlikely to affect GaN or sapphire.)
Richard
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear All, } } I met some problems during I prepared the samples for GaN/sapphire plan-view } using tripod wedge technique. After polishing, the sample was thinned by pips } in one side. However, The other side usually was covered by copper, as a result } the sample became dark. It seems the unthinned side should be covered something } to prevent sputtered materials from growing on it. Do you have some knowledge } on it? } } Thanks in advance. } } Best regards. } } Feng } ********************************************** } Dr. Feng Wu } Dept. of Materials Engineering } Ben-Gurion University of the Negev } Beer-Sheva 84105, Israel } } voice 972-8-6461473 } fax 972-8-6472944 } fwu-at-bgumail.bgu.ac.il } ********************************************** } }
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Hello, My protein sample does not adsorb to the grid even after glow discharg, it is a membrane protein complex with the membrane lipids froming vescicles. Nothing show up after negative staining. What should I do to improve the adsorption? I have tried quick dring with filter paper after put the sample on the grid, but it does not work.
Thank you ......was written by a student with good intent and fair grammar/spelling. I apologize, lesson learned. Thanks for the help.
We as microscopists often receive requests for information about specific chemicals or chemical derivatives which we are requested to investigate or required to use in studies that are requested. The amount of information available is staggering, but relevant information is not always immediately available, and, for many of us, the means of access is not always apparent. The one datum that will provide you with the quickest access to information is the Chemical Abstracts Registry Number(CAS Reg. or just CAS). It is usually located in the catalog from which a chemical was ordered and can be obtained from the supplier and used to search any chemical database with specificity and speed. What follows is what I believe to be a reasonable strategy for free, rapid, and reasonable access to information about any chemical that raises a question. I have limited the list to four, because I have found what I need using no more. There are other sources, but my object was speed and utility.
A. NIST Chemistry WebBook(ver. 2000): http://webbook.nist.gov/
B. ToxLine Hazardous Substance Database: http://toxnet.nlm.nih.gov/cgi-bin/sis/htmlgen?HSDB
C. INTERNATIONAL CHEMICAL SAFETY CARDS (ICSCs) (NIOSH/WHO): http://www.cdc.gov/niosh/ipcs/nicstart.html (US National Version)
D. Material Safety Data Sheets: http://siri.org/msds/index.html (This is the only site that is NOT supported by a government agency!)
For masters and students, I believe that the three URL's listed above provide access to the best information about chemicals that are considered hazardous.
DISCLAIMER: To assume that any chemical is harmless is foolish and probably wrong. Many years ago there was a news report of a pre-adolescent who woke up on 1November with diabetic acidosis after eating a grocery bag full of candy collected the night before. Even NaCl or KCl can be toxic!!! The Moral Is: If you get no return from any of the four above URL's for a chemical name with which you are NOT familiar, check the name and require a copy of the MSDS (Material Safety Data Sheet) BEFORE you permit it in your lab.
To date, I have used Google for over a year to search for info on everything including many organic chemicals. I have always received returns on chemical searches.
Respectfully submitted,
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.
Below is the result of your feedback form. It was submitted by (drcline-at-flash.net) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, July 9, 2001 at 17:07:43 ---------------------------------------------------------------------------
Email: drcline-at-flash.net Name: Diane Cline
Organization: George Washington University
Education: Graduate College
Location: Washington DC
Question: My grandmother, Mrs. Ethel Schwartz, is trying to find an appropriate museum or library to donate about 500 slides of my grandfather's, Dr. J.B. Robbins, who developed the technique of photography through a triocular microscope of chemical crystals on slides in the late 196o's through early 1970's. Might you be able to suggest a collection (museum, university archive, or Library) that might be especially interested? Thank you, Sincerely Diane Harris Cline
I am interested in counting the total number of glomeruli in the kidneys in } rats with obstructive uropathy. To perform stereology, I have to cut thick } sections at 20 micrometers. I found that the kidney itself breaks down and } get very damaged at this thickness (kidneys fixed in 10% formalin and } embedded in parrafin), probably because the kidney is much harder than the } paraffin. In the litterature, some people embed kidneys in glycol } methacrylate. It might be better (harder), but then I do not know how to cut } my sections which are about 2.5 to 3 cm long, if I need a glass/diamond knife } that long. Any ideas about cutting thick sections in paraffin?, or is it } worth re-embedding in glycolmethacrylate, and any ideas about cutting my } serial sections at 20 micrometers in that resin? } } Thank you a lot for your help! } } F. Cachat } University of Virginia } fc6b-at-virginia.edu
Bill - here is the animal toxicity data according to the MSDS sheets available from our site. Methylene Blue: Oral rat LD50: 1180mg/kg Mutation references cited. Animal reproductive effects cited Toluidine Blue O: IPR-Rat LD50: 215mg/kg; IVN-Rat LD50: 28930?g/kg; IVN-Mus LD50: 45mg/kg; UNR-Mus LD50: 28mg/kg; IVN-Rbt LD50: 13440?g/kg. In an ideal world that data would be comparable, but this is what we have and I assume that its based on some realities. I would not overly worry about the LD50 figures, unless they indicate great toxicity. More concern is warranted if a toxin is cumulative and if its carcinogenic or mutagenic. Mutagenicity is indicated for methylene blue and not for toluidine blue O. Since mutagenicity is one of the great fears during pregnancy, I would suggest to revert to toluidine blue O. I think that most people in this forum would understand that carcinogenic and mutagenic molecules are ingested daily and in huge numbers. Clearly the body wards off all of these and it is a very rare event that a cancer actually forms. It is well known that a person with a healthy life-style and on a good diet (particularly one rich in the anti-oxidant vitamins) is more likely to avoid cancer. That healthy life-style includes minimizing, but never excluding contact with "adverse" substances. I suggest that one should be circumspect, but avoid paranoia. On balance, I regard a well run EM lab as one of the safer workplaces about; it would be nice to have comparative data of sick days or better still accident data, but I doubt that they exist. While on the topic of safety and carcinogens I like to tell a little story. It happened at an unmentionable university about 15 years ago. The ceiling in one large public room was found to contain fibres, which were confirmed to be asbestos. After some agitation the university's response was to invite an "expert", who was used by various organisations to "resolve" asbestos problems. The expert soon had his first meeting with union officials during which he insisted that asbestos was not very dangerous. To make his point, he opened a jar allegedly containing powdered asbestos and threw a handful into the air. Even 15 years ago, such action identified that man as an idiot. Educated perhaps, but an idiot nonetheless. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Tuesday, July 10, 2001 5:05 AM, William McManus [SMTP:billemac-at-biology.usu.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 } -----------------------------------------------------------------------. } } } To Sally and all: } } I guess that my original post was too curt. We have gone away from Tol } Blue, to methylene blue due to safety reasons. There is no toxicology } data on the MSDS for Tol Blue, but there is for Meth Blue. As I have a } pregnant technician, I feel better knowing the LD 50 information, which } is on the MSDS for Meth Blue. But, methylene blue is not as good for a } general stain as Tol Blue, so I was just curious if there was an } overriding reason to use it. I am presently experimenting with food } based stains, which eliminates the issue completely. } } The issue of pregnant (or possibly pregnant) workers in the lab has } turned out to be fairly complicated. If anyone else out there has beem } through this situation, I would like to know how you dealt with it. } } Bill } } 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: Monson, Frederick C. [mailto:fmonson-at-wcupa.edu] } Sent: Thursday, July 05, 2001 10:20 AM } To: 'Sally Stowe' } Cc: 'Microscopy Listserver' } Subject: RE: Tol blue/borax } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 Sally, } To address the REAL attribution issue, one of us is going to } have to } go to a REALLY good library that has these OLD journals back to the } late } '50's. We may have to divide the labor, but the question is } sufficiently } interesting and meritorious that we ALL should get it right, when asked. } Someone was responsible for the use of borax (Sodium tetraborate) with } Toluidine blue O (1% in 1%). Here are all the citations that Pease } provides } and his [attributions]: } } 1. Trump, Schmuckler and Benditt (1961), J. Ultrastruc } Res., 5: } 343 [ 0.1% Toluidine blue O in 2.5% carbonate at pH 11 (confirmed in } Lewis } and Knight below)] } 2. Charles and Meek , G.R. separately in a discussion of } paper } by Mercer(1963), J Microscp Sci., 81:179. [mentions 1% Toluidine blue O } in } 1% borax used at 85 degrees C] } 3. Pease, D.C.(1964), Histological Techniques for Electron } Microscopy (2d), Academic Press, NY, NY, pp 256-262. [mentions on p. 260 } in } 2st paragraph the exact method that I use for staining as 'his' method - } "...flood the slide, cook until [edge] of drop [froths/]steams [or } dries]....". [This is a long discussion to which Pease devotes an } entire } section, but one can just FEEL his impatience with the unimportance to } HIM } of this issue. The book, after all, is NOT about light microscopy. The } semi-thin section is for orientation only, and hardly deserves any more } notice - especially if the light microscopists cannot SEE what is to be } seen } in the best material ever handed them.] } 4. In Kay, D.H.(ed)(1965), Techniques for electron } Microscopy(2d), F.A. Davis, Philadelphia, ISBN: [NOT given - is this } book } pre-historic?!] The following is cited as THE reference for using } Toluidine } blue O: Benscome, Stone, Latta, and Madden (1959), J. biophys. biochem. } cytol, 5: 508. } } I have just looked in Geoffrey Meek's book (1970), Practical } Electron Microscopy for Biologists, Wiley_Interscience, NY, NY, p. } 456-457. } ISBN: 0-471-59030-4. 1% in 1% with no citation, though in this book he } has } purposely NOT used citations at all. } } I have also consulted the Sorvall manual on "Thin Sectioning"(3d } Ed)(1973) as well as the SerVall manual "Thin Sectioning and Associated } Technics for Electron Microscopy"(1959) which apparently was distributed } with the MT-1. In the latter, only staining of Methacrylate sections } was } given and no mention of 1% *bor* of any kind. In the former, 1% borate } at } pH 4.5 is mentioned, but that is not relevant to this discussion. } } I would bet that you might have some of these references. If } you do } not, let me know, and I will dig them up, in old Philadelphia, in Ben } Franklin's old stacks (at the U. of P(ennsylvania) as we call it). I } will } do it anyway, because I want to know, but I will do it quicker if you } tell } me I'm keeping you waiting. } } Regards, } } 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: Sally Stowe } } Sent: Tuesday, July 3, 2001 3:37 AM } } To: Microscopy-at-sparc5.microscopy.com } } Subject: Tol blue/borax } } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } Somebody just asked me the reference for toluidine blue/sodium borate } (1% } } each) which is our routine LM stain - I cant find an attribution in } any of } } about 10 books. Can anybody help? This person should not be } forgotten! } } One suggestion was Alsop? } } } } thanks } } } } Sally } } } } } } } } Dr Sally Stowe } } Facility Coordinator, } } ANU Electron Microscopy Unit } } Research School of Biological Sciences } } Australian National University } } Canberra ACT0200 } } AUSTRALIA } } stowe-at-rsbs.anu.edu.au fax 61 (0)2 6125 3218 } } ph 02 6125 2743 } } http://www.anu.edu.au/EMU } } } } } } } } } }
Hi Jan, Just to keep the historical stuff together on this thread, the well-known Giemsa blood stain was first published, as a footnote, in "...an obscure journal...." in (see Gray, P.(1954), The Microtomist's Formulary and Guide, Blakiston, NY,NY. (reprinted by Krieger(1975), ISBN 0-88275-247-2). The dry Giemsa stain is an eosinate of methylene blue [acid + base = insoluble salt (in HOH but not in MeOH! Later, on the slide, soluble (mass action) in large excess of MeOH-water or buffer mix yields differential dyeing].
I searched the Krieger site (http://www.web4u.com/krieger-publishing/) for ISBN and came up with the following:
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THE MICROTOMIST'S FORMULARY AND GUIDE Peter Gray, , 0-88275-247-2
Pages: 808, Binding: Cloth, Price: $81.50
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This is a known and recognized source reference work. The book includes a treatise on the art of making microscopic slides from biological specimens, as well as a classified list of the formulas and techniques used in this art.
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NOTE: I am not connected except by long-familiarity (30+ years). Ouch!!!
Fred Monson
} ---------- } From: Jan Coetzee } Reply To: janc-at-ccnet.up.ac.za } Sent: Friday, July 6, 2001 3:55 AM } To: microscopy-at-sparc5.microscopy.com } Subject: Re: Tol blue/borax } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 } } To contribute to the search for the roots of the Toluidine blue } procedure: } } There is an interesting paper by Kramer & Windrum (J. Histochem } Cytochem, 1955, Vol 3, 227-237, The Metachromic Staining Reaction) } that probably was instrumental in initiating the wider use of Tol } Blue. In this paper the effect of stain concentration and residual } water is shown (amongst many other factors) to be important for the } development of the multiplicity of colours. } } The use of Toluidine Blue as a stain for monitor sections of epoxy- } and acrylic-embedded tissue probably stems from these metachromic } effects that this stain produces, with the result that some deductions } regarding the chemical composition of the sample can be made after a } simple Tol Blue staining. } } Even though we, in this lab, have since forever used Tol Blue for the } staining of monitor sections of resin-embedded tissues, every batch of } Tol Blue that we make up seems to have slightly different staining } characteristics. This was such a source of irritation that Chris van } der Merwe recently decided to systematically investigate some of the } variables. } } He found that the most important parameters are: } 1. Tol blue concentration (0.1 or 0.5%) } 2. Buffer or vehicle type, pH and concentration (0.1 - 1% } Na-carbonate, Na-benzoate buffer at pH4, Na-tetraborate (borax) at pH } 8.8) } 3. Embedding resin formulation (Spurr, Quetol, Polarbed 812, LR-White) } 4. Section thickness. } } All the above have an effect on the intensity, as well as the degree } of metachromicity, of the stain. } Resin formulation is probably the major influencer of the results. } } This may be a reason for the existence of the wide variety of Tol Blue } recipes that abound in the literature. } } More information on Tol Blue staining can be had directly from Chris } at cvdmerwe-at-scienti.up.ac.za } } } Regards } } Jan Coetzee } Lab for Microscopy and Microanalysis } University of Pretoria, South Africa. } Tel: 012-420-2075, Fax 012-362-5150 } www.up.ac.za/academic/electron/emunit1.htm } } } } "Monson, Frederick C." wrote: } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } Dear Sally, } } To address the REAL attribution issue, one of us is going to } have to } } go to a REALLY good library that has these OLD journals back to the } late } } '50's. We may have to divide the labor, but the question is } sufficiently } } interesting and meritorious that we ALL should get it right, when asked. } } Someone was responsible for the use of borax (Sodium tetraborate) with } } Toluidine blue O (1% in 1%). Here are all the citations that Pease } provides } } and his [attributions]: } } } } 1. Trump, Schmuckler and Benditt (1961), J. Ultrastruc } Res., 5: } } 343 [ 0.1% Toluidine blue O in 2.5% carbonate at pH 11 (confirmed in } Lewis } } and Knight below)] } } 2. Charles and Meek , G.R. separately in a discussion of } paper } } by Mercer(1963), J Microscp Sci., 81:179. [mentions 1% Toluidine blue O } in } } 1% borax used at 85 degrees C] } } 3. Pease, D.C.(1964), Histological Techniques for Electron } } Microscopy (2d), Academic Press, NY, NY, pp 256-262. [mentions on p. 260 } in } } 2st paragraph the exact method that I use for staining as 'his' method - } } "...flood the slide, cook until [edge] of drop [froths/]steams [or } } dries]....". [This is a long discussion to which Pease devotes an } entire } } section, but one can just FEEL his impatience with the unimportance to } HIM } } of this issue. The book, after all, is NOT about light microscopy. The } } semi-thin section is for orientation only, and hardly deserves any more } } notice - especially if the light microscopists cannot SEE what is to be } seen } } in the best material ever handed them.] } } 4. In Kay, D.H.(ed)(1965), Techniques for electron } } Microscopy(2d), F.A. Davis, Philadelphia, ISBN: [NOT given - is this } book } } pre-historic?!] The following is cited as THE reference for using } Toluidine } } blue O: Benscome, Stone, Latta, and Madden (1959), J. biophys. biochem. } } cytol, 5: 508. } } } } I have just looked in Geoffrey Meek's book (1970), Practical } } Electron Microscopy for Biologists, Wiley_Interscience, NY, NY, p. } 456-457. } } ISBN: 0-471-59030-4. 1% in 1% with no citation, though in this book he } has } } purposely NOT used citations at all. } } } } I have also consulted the Sorvall manual on "Thin Sectioning"(3d } } Ed)(1973) as well as the SerVall manual "Thin Sectioning and Associated } } Technics for Electron Microscopy"(1959) which apparently was distributed } } with the MT-1. In the latter, only staining of Methacrylate sections } was } } given and no mention of 1% *bor* of any kind. In the former, 1% borate } at } } pH 4.5 is mentioned, but that is not relevant to this discussion. } } } } I would bet that you might have some of these references. If } you do } } not, let me know, and I will dig them up, in old Philadelphia, in Ben } } Franklin's old stacks (at the U. of P(ennsylvania) as we call it). I } will } } do it anyway, because I want to know, but I will do it quicker if you } tell } } me I'm keeping you waiting. } } } } Regards, } } } } 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: Sally Stowe } } } Sent: Tuesday, July 3, 2001 3:37 AM } } } To: Microscopy-at-sparc5.microscopy.com } } } Subject: Tol blue/borax } } } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } -----------------------------------------------------------------------. } } } } } } } } } } } } Somebody just asked me the reference for toluidine blue/sodium borate } (1% } } } each) which is our routine LM stain - I cant find an attribution in } any of } } } about 10 books. Can anybody help? This person should not be } forgotten! } } } One suggestion was Alsop? } } } } } } thanks } } } } } } Sally } } } } } } } } } } } } Dr Sally Stowe } } } Facility Coordinator, } } } ANU Electron Microscopy Unit } } } Research School of Biological Sciences } } } Australian National University } } } Canberra ACT0200 } } } AUSTRALIA } } } stowe-at-rsbs.anu.edu.au fax 61 (0)2 6125 3218 } } } ph 02 6125 2743 } } } http://www.anu.edu.au/EMU } } } } } } } } } } } } } } } } }
You may have better luck using a vibratome. The kidney, being sharply demarcated and well encapsulated, lends itself to processing in this manner. I would fix before sectioning, (dependent on species donating the kidney-probably not bigger than rat) but I have seen it work either way. The fixative is dictated by stains/probes of interest. Using a cryotome and a water soluble embedding media has also worked well for me in the past. good luck
Ramin
-----Original Message----- } From: "CachatF-at-aol.com"-at-sparc5.microscopy.com [mailto:"CachatF-at-aol.com"-at-sparc5.microscopy.com] Sent: Monday, July 09, 2001 11:19 PM To: Microscopy-at-sparc5.microscopy.com
I am interested in counting the total number of glomeruli in the kidneys in } rats with obstructive uropathy. To perform stereology, I have to cut thick
} sections at 20 micrometers. I found that the kidney itself breaks down and
} get very damaged at this thickness (kidneys fixed in 10% formalin and } embedded in parrafin), probably because the kidney is much harder than the
} paraffin. In the litterature, some people embed kidneys in glycol } methacrylate. It might be better (harder), but then I do not know how to cut } my sections which are about 2.5 to 3 cm long, if I need a glass/diamond knife } that long. Any ideas about cutting thick sections in paraffin?, or is it } worth re-embedding in glycolmethacrylate, and any ideas about cutting my } serial sections at 20 micrometers in that resin? } } Thank you a lot for your help! } } F. Cachat } University of Virginia } fc6b-at-virginia.edu
At 1:04 PM -0600 7/9/01, William McManus wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
When I was pregnant, 9 years ago, I was the entire staff of the EM facility here (still was until a month ago)and so could not avoid things. The way I handled safety concerns was to be extra cautious: I worked in the hood, wore double gloves when working with fixatives & resins and gloves for other times when i would not usually wear them( like using Tol Blue), safety glasses if splashing was a possibility, washed my hands almost compulsively. In short, I used common sense. I was, and am, lucky that (UrAc aside) I don't have to deal with radioactivity. colleagues of mine who were also pregnant at the time (we had a rash of pregnancies that year) who did need to work with isotopes minimized their use, got new shields, and at times convinced labmates to do those steps for them!
We all, thankfully had healthy babies who are now all approaching adolescence and driving us nuts, but that's another issue for another forum!
Lee
-- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
} I am interested in counting the total number of glomeruli in the kidneys in } } rats with obstructive uropathy. To perform stereology, I have to cut thick } } sections at 20 micrometers. I found that the kidney itself breaks down and } } get very damaged at this thickness (kidneys fixed in 10% formalin and } } embedded in parrafin), probably because the kidney is much harder than the } } paraffin. In the litterature, some people embed kidneys in glycol } } methacrylate. It might be better (harder), but then I do not know how to } cut } } my sections which are about 2.5 to 3 cm long, if I need a glass/diamond } knife } } that long. Any ideas about cutting thick sections in paraffin?, or is it } } worth re-embedding in glycolmethacrylate, and any ideas about cutting my } } serial sections at 20 micrometers in that resin? } } } } Thank you a lot for your help! } } } } F. Cachat } } University of Virginia } } fc6b-at-virginia.edu
How about frozen sections? Fix in formalin, cryoprotect with ascending concentrations of sucrose (10%, 20%, 30%) then snap freeze and section at 20 microns on a sliding microtome. Collect sections in ice-cube trays to keep them in order. In my experience, thick paraffin sections are much more difficult than thinner ones. Embedding tissue in methacrylates (and other plastics) is usually intended for thinner sections (1-3 microns).
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 **********************************************
We read your message with great interest.i routinely fix kidney of approximately the same size but usually cut for stereological work -at-10u. you do not state any info regarding processing times etc but if your tissue is brittle this could be due to numerous factors over fixation leaving in alcohol/clearing agent too long (especially chloroform as clearing agent) ( i come from a laboratory that hand processes, which gives more control over times processed compared to that from machines.) i also would question the fixative i always use Bouins fixative with kidney and have never had any problems in sectioning. i would suggest that you do not embed in glycolmethacrylate as this is for cutting thin's and also there is no way you would be able to get diamond knife to cut the size if tissue you want to. you could make ralph knives but again this will probably not work on the thickness you are after.
my last suggestion to you is regarding you 'already brittle blocks' try bakers soaking fluid (mixture of alcohol/glycerine) this should cure your problem but it is slow going as you will only get a few sections before they will need resoaking
i hope this works and is helpful to you yours
Phil
Mr P R Mutch Senior Imaging/Research Technician Nottingham University School of Biomedical Sciences E Floor, Medical School Queens Medical Centre Clifton Boulevard Nottingham NG7 2UH UK Tel 0115 9709415 E-mail philip.mutch-at-nottingham.ac.uk
Mr P R Mutch Senior Imaging/Research Technician Nottingham University School of Biomedical Sciences E Floor, Medical School Queens Medical Centre Clifton Boulevard Nottingham NG7 2UH UK Tel 0115 9709415 E-mail philip.mutch-at-nottingham.ac.uk
Hello, my name is Chen Chen, in Johns Hopkins University School of Medicine. Mike DeLannoy introduced me to subscribe to this group. Thank you Chen Chen
This is what I have done so far: 1. Removed all stored chemicals from the lab. 2. Moved all chemical mixing into fume hood. 3. Met for 3 hours with environmental safety officer. 4. Met for 2 hours with radiation safety officer. 5. Eliminated all protocols which involve teratogens from pregnant workers routine. 6. Established a personal hygiene plan for pregnant worker, which is being signed of by worker, myself, director of the lab, safety officer, and her physician. 7. Reiterated safety protocols for worker, coat , gloves, safety glasses, etc. 8. Had lab monitored for radiation, passed. 9. Had lab monitored for formaldehyde gas, passed. 10. Planning to monitor xylene exposure to worker when thin sectioning (cannot do this in the hood, obviously). Heat pen( modified soldering iron) has not worked well so far, any ideas how to eliminate xylene? 11. Still looking for a non toxic general 1 micron stain for Spurrs and LR White. Any ideas? I am experimenting with food related stains (grape juice, etc.). 12. Employee has completed, lab safety training course, formaldehyde training course and radiation training course. 13. Will continue to monitor lab for chemical and radation exposure.
To some this may seem like over kill, but I believe that it is very important that every stone is turned in regards to safety. Another issue that I face is that I have numerous grad students coming through the lab, and there is always a possibility that they are pregnant and do not realize it or keep it from me. I also have a nursing mother who was visiting the lab and never bothered to tell me, until the subject of why the lab was reorganized came up. I now demand that all female student, techs, visitors, ect. tell me of their "motherhood" status.
Bill
-----Original Message----- } From: Leona Cohen-Gould [mailto:lcgould-at-med.cornell.edu] Sent: Tuesday, July 10, 2001 7:28 AM To: William McManus; Monson, Frederick C.; Sally Stowe Cc: Microscopy Listserver
Every summer at our institute we sponsor a summer camp program called Adventures in Science. We invite inner-city fifth graders to come and hopefully see how much fun science can be. It has been rather successful and we all seem to enjoy the week.
One of the scientists here wants to give a demonstration involving vortices and fluids. He has a few short demonstrations to show the kids, but I told him I think he needs more to keep their interest and to keep them busy. He has five groups of 6 children for about an hour.
I told him about the MSA newsgroup and how helpful you all are and said I would post this question. I know this is not a microscopy question but I thought some of you might have some suggestions.
Thanks in advance for any help.
Peggy Bisher --
Margaret E. Bisher
NEC Research Institute 4 Independence Way Princeton, NJ 08540. Tel.: (609) 951-2629 Fax: (609) 951-2496 e-mail: peggy-at-research.nj.nec.com
I commend you for your concern about safety. I went through two pregnancies as a biological electron microscopist. Since I was the solitary person doing TEM, I didn't have the choice of delegating anything. I actually chose to delay an R&D project for my first trimester because I didn't want to use uranyl acetate, osmium tetroxide/ruthenium tetroxide and especially lead citrate and tannic acid stains all involved in TEM. Other than that, I just used extra caution, especially when weighing/handling powdered chemicals. I never paid much attention to the Tol. blue - you have obviously looked into this in great detail.
I missed your original post. Are you trying to stain epoxy-embedded sections for light microscopy? I've tried a number of food dyes in the past and had very limited success. Also just because they are FD&C or D&C approved doesn't guarantee safety. I've worked with Erioglaucine (Brilliant Blue FCF, FD&C Blue #1) and the MSDS states it is a possible carcinogen.
I've tried some non-traditional dyes on epoxy sections of mammalian tissue. The standard Tol. Blue, "Epoxy Tissue Stain" (mixture with Basic Fuchsin) and/or Azur II always worked the best. BTW, my MSDS's listed Toluidine Blue as an irritant, but Methylene Blue as an irritant and a mutagen. Therefore I would think Tol. Blue was a little safer.
Excerpts from an experiment I did in '97 trying different stains on ~1micron sections of skin embedded in Spurr's resin (also not a good safety choice): Dyes tested: 0.1% aqueous Phloxine B. [related to eosin, a useful D&C approved dye for fresh tissue and fluorescence. ] 0.15% Sudan Black B in 70% ethanol. 0.5% aqueous Neutral Red. [a fairly nontoxic dye (although MSDS states possible mutagen) that we use a lot in fresh tissue.] Zymed Methyl Green. All were done on a hotplate for 30-60 seconds
Staining Results: Phloxine B seemed to stain only the dermal collagen with a fairly bright pink stain. Sudan Black B produced an interesting pattern of staining small elongated strips throughout the dermis, as well as intracellular staining of basal epidermal cells. The fat cell staining was non-uniform. Neutral Red stained in a very similar pattern to Sudan Black B in the dermis, only much more faint. Methyl Green produced a strong general stain similar but not exactly the same as Toluidine Blue.
Hope this is useful, Karen -------------------------------------------------------------------------------- Karen S. Zaruba BioMaterials Technology Center 3M Center Bldg. 270-1S-01 Tel: (651) 737-2971 Fax: (651) 737-5645
To: "Monson, Frederick C." {fmonson-at-wcupa.edu} "Sally Stowe" {STOWE-at-rsbs.anu.edu.au} cc: "Microscopy Listserver" {microscopy-at-sparc5.microscopy.com} (bcc: Karen S. Zaruba/US-Corporate/3M/US) Subject: RE: Tol blue/borax
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
To Sally and all:
I guess that my original post was too curt. We have gone away from Tol Blue, to methylene blue due to safety reasons. There is no toxicology data on the MSDS for Tol Blue, but there is for Meth Blue. As I have a pregnant technician, I feel better knowing the LD 50 information, which is on the MSDS for Meth Blue. But, methylene blue is not as good for a general stain as Tol Blue, so I was just curious if there was an overriding reason to use it. I am presently experimenting with food based stains, which eliminates the issue completely.
The issue of pregnant (or possibly pregnant) workers in the lab has turned out to be fairly complicated. If anyone else out there has beem through this situation, I would like to know how you dealt with it.
Bill
William McManus Supervisor Electron Microscopy Facility Department of Biology Utah State University Logan UT 84322-5305
What I do is after they are hired, or at the beginning of the class, in front of witnesses, I explain that for their own safety that I must know if they are pregnant, so that all the appropriate precautions are taken. I also tell them that this information will be kept in strict confidence. It is pretty easy for people to figure out what is going on, however, once you start treating one person differently than the rest.
Bill
-----Original Message----- } From: Chuck Butterick [mailto:cbutte-at-ameripol.com] Sent: Tuesday, July 10, 2001 1:31 PM To: William McManus
This is what I have done so far: 1. Removed all stored chemicals from the lab. 2. Moved all chemical mixing into fume hood. 3. Met for 3 hours with environmental safety officer. 4. Met for 2 hours with radiation safety officer. 5. Eliminated all protocols which involve teratogens from pregnant workers routine. 6. Established a personal hygiene plan for pregnant worker, which is being signed of by worker, myself, director of the lab, safety
officer, and her physician. 7. Reiterated safety protocols for worker, coat , gloves, safety glasses, etc. 8. Had lab monitored for radiation, passed. 9. Had lab monitored for formaldehyde gas, passed. 10. Planning to monitor xylene exposure to worker when thin sectioning (cannot do this in the hood, obviously). Heat pen( modified soldering iron) has not worked well so far, any ideas how to eliminate xylene? 11. Still looking for a non toxic general 1 micron stain for Spurrs and LR White. Any ideas? I am experimenting with food related stains (grape juice, etc.). 12. Employee has completed, lab safety training course, formaldehyde training course and radiation training course. 13. Will continue to monitor lab for chemical and radation exposure.
To some this may seem like over kill, but I believe that it is very important that every stone is turned in regards to safety. Another issue that I face is that I have numerous grad students coming through the lab, and there is always a possibility that they are pregnant and do not realize it or keep it from me. I also have a nursing mother who was visiting the lab and never bothered to tell me, until the subject of why
the lab was reorganized came up. I now demand that all female student, techs, visitors, ect. tell me of their "motherhood" status.
Bill
-----Original Message----- } From: Leona Cohen-Gould [mailto:lcgould-at-med.cornell.edu] Sent: Tuesday, July 10, 2001 7:28 AM To: William McManus; Monson, Frederick C.; Sally Stowe Cc: Microscopy Listserver
At 1:04 PM -0600 7/9/01, William McManus wrote: } -----------------------------------------------------------------------
- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id QAA08039 for dist-Microscopy; Tue, 10 Jul 2001 16:51:38 -0500 (CDT) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id QAA08029 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Tue, 10 Jul 2001 16:51:07 -0500 (CDT) Received: from www.psrc.usm.edu (www.psrc.usm.edu [131.95.98.20]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id QAA08021 for {microscopy-at-sparc5.microscopy.com} ; Tue, 10 Jul 2001 16:50:55 -0500 (CDT) Received: from localhost (lssomlai-at-localhost) by www.psrc.usm.edu (8.8.7/8.8.7) with ESMTP id RAA27733 for {microscopy-at-sparc5.microscopy.com} ; Tue, 10 Jul 2001 17:07:41 -0500
Hi,
I'm looking for anyone who may have images of red blood cells, white blood cells, and platelets. I'm also looking for an illustration of haemoglobin binding. My mother-in-law is working on an educational web page about blood for grades 4-6. All images will be referenced and properly credited. If you can help, I would greatly appreciate it.
Postdoctoral position(s): Dynamics of biological machines
Postdoctoral position(s) are available in the McGough lab at Purdue University to study biological machines by a variety of microscopic, computational, and biochemical approaches. Current projects in the lab include structure-function studies of the cytoskeleton and bacteriophage assembly.
Successful candidate(s) will work in close collaboration other laboratories at Purdue in cross-disciplinary studies using a variety of approaches including protein biochemistry, video light microscopy, time-resolved electron cryomicroscopy, atomic force microscopy, magnetic tweezers, and molecular modeling. Highly qualified candidates will be encouraged to develop new collaborations involving members of the cytoskeleton, structural biology, and single molecule groups at Purdue.
Applicants should have a Ph.D. in engineering or in one of the physical or life sciences. The ideal candidate will be a hard-working, enthusiastic individual with strong communication skills, experience in protein biophysics, and a commitment to applying an integrated structure-function approach to study biological machines.
Salary is negotiable.
Interested individuals should contact:
Dr. Amy McGough Department of Biological Sciences Purdue University West Lafayette, IN 47907-1392 USA phone: +1 (765) 496-7501 fax: +1 (765) 496-7501 amcgough-at-bilbo.bio.purdue.edu http://bilbo.bio.purdue.edu/~amcgough/
Purdue is an equal access/equal opportunity university.
} On the other hand, if newer technologies could improve your performance } significantly, then you might want to consider upgrading. The } downside? When the PC that is running the latest system becomes obsolete } (3 years? 5 years?) and the rest of your system (vacuum, High Voltage, } etc.) is still fine, what are you going to do? Try to find a new 486 with } Win 3.1 to replace a crashed but fairly recent Amray, for example. Their } boards and software won't run on anything newer. I'm not picking on them, } I'm just familiar with them.
I'm running an Amray 1910FE with PC control over the Nibblenet. The system came with a Dell 486 and 250MB hard drive. I am actually using a Pentium III/450 with Win95 or Win98 (either works fine). The 8741 MPU on the Nibblenet board in the PC needs the latest firmware version (2.3 or 2.4) to run with a Pentium. The board is hardwired to IRQ9. With a simple PC system, there are no IRQ conflicts.
I do not use the Image Control software/hardware (Matrox IP8) but rather use Soft Imaging ADDA II for active beam control and image capture and the GrabBit frame grabber to capture TV video. Console electronics are un-changed. PC-10 video goes to one of the GrabBit RS-190 inputs. The other input is from the chamber view camera.
So far, KLA/Amray is still supporting the FE systems.
} One of the scientists here wants to give a demonstration involving } vortices and fluids. He has a few short demonstrations to show the } kids, but I told him I think he needs more to keep their interest and } to keep them busy. He has five groups of 6 children for about an } hour.
Hi, Peggy --
I'll give you a great idea that takes about ten bucks worth of parts and a source of compressed air. How about a gizmo made of two 1/2-inch (OK, Ok, 12mm) pipes, a washer, and a ball valve which appears to contain Maxwell's demon. Blow compressed air in the middle and presto, hot air comes out one pipe and cold air comes out the other! It's called a Hilsch tube, and here's a link:
http://www.visi.com/~darus/hilsch/
If you do a search on "hilsch tube" you can find some commercial suppliers of ready-made units.
I remember seeing it in an old issue of Scientific American, the Amateur Scientist column. An outfit called Tinker's Guild, which I believe to be the creation of Shawn Carlson, current author of the Amateur Scientist, is offering a CD containing *every Amateur Scientist column written since 1928* for a snappy $70. Quite the bargain:
http://www.tinkersguild.com
Any of you who are science educators should get a copy of this! No commercial interest, just a fascinated customer who remembers actually trying quite a few of these as a kid.
We have a student who has been looking at biofilms on artificial membranes and in the process found a strange-looking inclusion. If there's anyone out there who has the time to have a look at this (it is displayed on http://www.ru.ac.za/emu/lechbug.htm) and give any suggestions this will be much appreciated.
Regards
Rob
======================================================= Robin H Cross (Mr) Director : Electron Microscopy Unit Rhodes University, PO Box 94, Grahamstown, South Africa tel: +27 46 603 8168/9, fax: +27 46 622 4377 email: r.cross-at-ru.ac.za http://www.ru.ac.za/emu/index.htm
** remember ICEM-15 in Durban in September 2002 **
} From: "Rick Mott" {rickmott-at-alumni.princeton.edu} } } Peggy Bisher wrote: } } } One of the scientists here wants to give a demonstration involving } } vortices and fluids. He has a few short demonstrations to show the } } kids, but I told him I think he needs more to keep their interest and } } to keep them busy. He has five groups of 6 children for about an } } hour. } } Hi, Peggy -- } } I'll give you a great idea that takes about ten bucks worth of parts } and a source of compressed air. How about a gizmo made of two } 1/2-inch (OK, Ok, 12mm) pipes, a washer, and a ball valve which } appears to contain Maxwell's demon. Blow compressed air in } the middle and presto, hot air comes out one pipe and cold air } comes out the other! It's called a Hilsch tube, and here's a link: } } http://www.visi.com/~darus/hilsch/ } } If you do a search on "hilsch tube" you can find some commercial } suppliers of ready-made units. } } I remember seeing it in an old issue of Scientific American, the } Amateur Scientist column. An outfit called Tinker's Guild, which } I believe to be the creation of Shawn Carlson, current author of } the Amateur Scientist, is offering a CD containing *every Amateur } Scientist column written since 1928* for a snappy $70. Quite } the bargain: } } http://www.tinkersguild.com } } Any of you who are science educators should get a copy of } this! No commercial interest, just a fascinated customer who } remembers actually trying quite a few of these as a kid. } The Amateur Scientist is no more. It got cut along with most of the other columns. It looks like a very large short fall in advertising revenues are a problem. The adds are almost all consumer advertising. Not much that match the readership. With some of the politically correct stuff they have been publishing the last few years I expect that the scientific readership is being run off and it is over the lay readerships head. I was particularly put off by their piece on terrorism in the USA with out a any mention of environmental terrorism.
The Amateur Scientist got a great number of us started on the road to science it is a shame that it is being let go.
I have heard Fry's has the CD-ROM on sale for $25 with a $25 mail in rebate. Now that's a deal.
Cold Spring Harbor Laboratory is seeking an experienced and responsible microscopy technician for the laboratory's core biological microscopy facility. The individual should have practical expertise in transmission electron microscopy, confocal and widefield fluorescence microscopy, and digital imaging. The successful candidate will be involved in designing and carrying out experimental protocols for users, training individuals in the use of various microscopes, and aligning microscopes and keeping the facility operating at an efficient and high level of productivity. Interested individuals should send their resume, including a description of their expertise and the names and addresses of 3 references to: Dr. David L. Spector, Cold Spring Harbor Laboratory, One Bungtown Road, Cold Spring Harbor, New York 11724, email: spector-at-cshl.org -- David L. Spector Cold Spring Harbor Laboratory One Bungtown Road Cold Spring Harbor, New York 11724 Tel. (516) 367-8456 Fax (516) 367-8876 email: spector-at-cshl.org
} Dear Sir, } } I am a media artist based in Canada. I am presently working on a low } budget, art funded (Canada Council for the Arts) DVD project destined for } non-profit art gallery use. } } The work is a philosophic and poetic look at evolution and bio-diversity. } } I am very interested in getting and using some of the video your members may } have made on primitive life forms such as viruses, bacteria, etc. } I would be also interested in } acquiring from you a high quality beta-sp or preferable dv tape you might } have of these studies as well as other studies you may have of microscopic } life forms/elements dna, diatoms, etc. Full credit to your organization } (and the videographer if desired) would be given in the final piece. I } would pay for all shipping and handling costs. } } } } The video and images would be a small component of a much longer work. } } My last work recently screened at the Canadian Embassy in Washington, D.C. } and I have had shows at the Museum of Modern Art in New York City and at } many other prestigious venues around the world. I include my bio. below. } } Please get back to me regarding my request. } } } Best Wishes, } Oliver Hockenhull } } BIO - 2001 Oliver Hockenhull } } Home Site: http://www.crosswinds.net/~ideoplastic/ } (From this site link to various works in VRML, hypertext, etc.) } } Oliver Hockenhull is an award-winning media artist, director, communication } theorist, and lecturer. } } He has completed five feature length works and numerous dramatic and } experimental shorts. The films have shown internationally at such film } festivals and venues as -(selected) The Kerala-India International Film } Festival, The Montréal World Film Festival, The Leipzig International } Documentary Film Festival, The Vancouver International Film Festival, The } European Media Arts Festival, The Nouveau Cinema Festival , The } International Documentary Film Festival of Amsterdam, The Museum of Modern } Art in New York City, the National Gallery of Art in Washington, D.C., and } LACE (Los Angeles Contemporary Exhibitions).
Gregory W. Erdos, Ph.D. Assistant Director, Biotechnology Program Scientific Ditrector, ICBR EM CORE University of Florida Ph. 352-392-1295 PO Box 118525 Fax 352-846-0251 Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl
A neat topic is eddy currents. There are a number of ways to experiment with them.
My son did a project that I helped him with. I first showed him the effect by getting a magnet and tying a string to it and used it as a pendulum. We counted the number of periods for it to stop. Then we got a an aluminum tray and put it under the swinging magnet and counted again. We changed the distance of the tray to see the effect.
Then we got fancy. We built a swinging pendulum in which we could put sheets of metals that we got from the hobby store. We used aluminum, brass, and titanium. The sheets could pass through magnets that we set up in epoxy on a board that fit onto the base of the pendulum with pegs so that they put in the same place every time. We made two sets of magnets with different gaps. The magnets were bought at Radio Shack. We experimented with different thickness of sheets, and stacks of sheets to make the same thickness. We experimented with sheets of paper that isolated the sheets of metal. It was really a nice little project that illustrated a number of effects.
To get even more fancy, you could do the same things with disks of metal rotating on an inexpensive dc motor. You could then monitor the current with a meter to measure the power change with different magnetic fields.
-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: Peggy Bisher [mailto:peggy-at-research.nj.nec.com] Sent: Tuesday, July 10, 2001 12:07 PM To: Microscopy-at-sparc5.microscopy.com
Dear Microscopists,
Every summer at our institute we sponsor a summer camp program called Adventures in Science. We invite inner-city fifth graders to come and hopefully see how much fun science can be. It has been rather successful and we all seem to enjoy the week.
One of the scientists here wants to give a demonstration involving vortices and fluids. He has a few short demonstrations to show the kids, but I told him I think he needs more to keep their interest and to keep them busy. He has five groups of 6 children for about an hour.
I told him about the MSA newsgroup and how helpful you all are and said I would post this question. I know this is not a microscopy question but I thought some of you might have some suggestions.
Thanks in advance for any help.
Peggy Bisher --
Margaret E. Bisher
NEC Research Institute 4 Independence Way Princeton, NJ 08540. Tel.: (609) 951-2629 Fax: (609) 951-2496 e-mail: peggy-at-research.nj.nec.com
} Sorry, forgot to include his email. } } } } Can anyone help this guy?? } } } } } Dear Sir, } } } } I am a media artist based in Canada. I am presently working on a low } } budget, art funded (Canada Council for the Arts) DVD project destined for } } non-profit art gallery use. } } } } The work is a philosophic and poetic look at evolution and bio-diversity. } } } } I am very interested in getting and using some of the video your members may } } have made on primitive life forms such as viruses, bacteria, etc. } } I would be also interested in } } acquiring from you a high quality beta-sp or preferable dv tape you might } } have of these studies as well as other studies you may have of microscopic } } life forms/elements dna, diatoms, etc. Full credit to your organization } } (and the videographer if desired) would be given in the final piece. I } } would pay for all shipping and handling costs. } } } } } } } } The video and images would be a small component of a much longer work. } } } } My last work recently screened at the Canadian Embassy in Washington, D.C. } } and I have had shows at the Museum of Modern Art in New York City and at } } many other prestigious venues around the world. I include my bio. below. } } } } Please get back to me regarding my request. } } } } } } Best Wishes, } } Oliver Hockenhull hereticfilms-at-home.com
} } BIO - 2001 Oliver Hockenhull } } } } Home Site: http://www.crosswinds.net/~ideoplastic/ } } (From this site link to various works in VRML, hypertext, etc.) } } } } Oliver Hockenhull is an award-winning media artist, director, communication } } theorist, and lecturer. } } } } He has completed five feature length works and numerous dramatic and } } experimental shorts. The films have shown internationally at such film } } festivals and venues as -(selected) The Kerala-India International Film } } Festival, The Montréal World Film Festival, The Leipzig International } } Documentary Film Festival, The Vancouver International Film Festival, The } } European Media Arts Festival, The Nouveau Cinema Festival , The } } International Documentary Film Festival of Amsterdam, The Museum of Modern } } Art in New York City, the National Gallery of Art in Washington, D.C., and } } LACE (Los Angeles Contemporary Exhibitions). } } Gregory W. Erdos, Ph.D. } Assistant Director, Biotechnology Program } Scientific Ditrector, ICBR EM CORE } University of Florida Ph. 352-392-1295 } PO Box 118525 Fax 352-846-0251 } Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl }
Gregory W. Erdos, Ph.D. Assistant Director, Biotechnology Program Scientific Ditrector, ICBR EM CORE University of Florida Ph. 352-392-1295 PO Box 118525 Fax 352-846-0251 Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Colleagues } } We have a student who has been looking at biofilms on artificial } membranes and in the process found a strange-looking inclusion. If } there's anyone out there who has the time to have a look at this (it } is displayed on http://www.ru.ac.za/emu/lechbug.htm) and give any } suggestions this will be much appreciated. } } Regards } } Rob } } } ======================================================= } Robin H Cross (Mr) } Director : Electron Microscopy Unit } Rhodes University, PO Box 94, Grahamstown, South Africa } tel: +27 46 603 8168/9, fax: +27 46 622 4377 } email: r.cross-at-ru.ac.za } http://www.ru.ac.za/emu/index.htm } } ** remember ICEM-15 in Durban in September 2002 ** }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
} Hello listers, } I recall seeing that thread late last week about the business opportunity. } This just came in from a Canadian news source, CBC. } Nick } RCMP make arrests in international phone scam } WebPosted Tue Jul 10 17:27:29 2001 } TORONTO - The RCMP, along with the FBI and the U.S. Secret Service, have } cracked a multi-million dollar scam originating in Nigeria and arrested } three people in the Toronto area. } More than 300 people around the world - most of them Americans - were } duped into giving money to a fake organization. The RCMP say some people } lost more than $50,000. Others lost millions. } Police say the fraud was sophisticated, high tech and organized. } Typically, victims received a letter either by mail or fax allegedly from } officials of a large Nigerian institution, such as the Nigerian National } Petroleum Corporation or the Central Bank of Nigeria. } The letters would contain an urgent request for help in transferring } millions of dollars out of Nigeria to financial institutions in the } victim's country. If the victims would help, they'd be in line for a } handsome profit, in some cases as much as 30 per cent of the monies being } transferred. } The letter says foreign accounts are needed and victims were asked to wire } personal banking information and $10,000 US to cover administration fees. } 'Boiler room' in Toronto } Then, victims were contacted by a person posing as a merchant banker of } the Central Bank of Nigeria. They were told the funds were now in a } clearing house or mercantile bank in North America. } Phone numbers given for the fake banks actually rang in a "boiler room" in } the Toronto area. } The scam continued with the victim asked to pay taxes, duties and other } fees, all with the promise of a windfall, until the victim was bankrupt. } The RCMP began a co-ordinated effort with U.S. police forces in 1998. } According to an RCMP news release, police detachments in the Toronto area } have received at least one complaint a week for the past eight years from } victims who lost money in the scam. } Staff Sergeant Darryl Ross says victims lost more than money. Some } suffered health problems, mental breakdowns, divorce and the break-up of } their families. } }
Can anyone recommend a good/cost-effective type of wall shielding to eliminate 60 and 180 cycle interferences coming into an SEM room from recently-installed large motors on the opposite side of the wall (that can't be moved now)?
Much obliged, Dee
*************************************************************** Dee Breger Mgr. SEM/EDX Facility Lamont-Doherty Earth Observatory 61 Route 9W Palisades, NY 10964 USA T: 845/365-8640 F: 845/365-8155
http://www.ldeo.columbia.edu/micro http://www.discovery.com/area/science/micro/micro1.html http://www.lsc.org/antarctica/front.html Journeys in Microspace (Columbia University Press, 1995)
As chairperson of the Traveling Poster Exhibit I am looking for judges for the Poster Award Program. Anyone who will be attending the MSA meeting at Long Beach and would be interested in helping please contact me. I need five judges for each discipline, Biological and Material Sciences. Thank you, Caroline Miller
Dear Goeff, Paraffin embedding works best on tissues that have some internal support, such as a collagen matrix (e.g., skin, etc.). If you think about the matter, the treatment to which tissues are subjected is harsh. Kidney tends to dehydrate, clear and infiltrate more slowly. Further, any imperfection in those steps can lead to a hard, fractious tissue. I understand what you want to do, and, while I should not overextend, I feel if I'm going to help with one, I may take the liberty of helping with all.
First. Whole kidney is very hard to infiltrate with paraffin without either segmenting it or removing the capsule (which is not always easy). I always, at least, opened the capsule. This is probably necessary, because as dehydration, clearing and infiltration proceed the capsule shrinks with the organ. If the capsule shrinks more, which is my conclusion on the matter, the internal pressure might be sufficient to retard the processes. Second. If 'First' is true, then the heat will essentially dry the parenchyma of the organ and leave you with the impression that it is 'hard'. It is interesting, that such blocks, when faced into the tissue always respond positively to immersion to glycerinated water (10-20%). Of course, all this manipulation does not leave one in the mood to try to finish with something quantitative, so some means to avoid the problem must be devised.
Solutions. 1. I agree with the frozen section idea, if you can take serial sections, then you can dye/stain and count every section if you wish or some statistical sample designed for glomeruli. 2. I would try a milder dehydration routine and vacuum embedding/infiltrating, at the lowest temperature, for 48 hours. My experience has been with whole organs, and these can cause problems. A milder dehydration does not take more than 1.5 the time but softens the steps (e.g., instead of 30,70,95,100 EtOH, benzene you could use 20% steps in EtOH followed by 1:2,1:1,2:1 (Benzene:EtOH) and 1:1(Benzene:Paraffin) sub-steps (no longer than 1.25hr) and all but the benzene:paraffin steps at 4oC.
NOTE: Tt has been said that the most arduous transition is to 70%EtOH, thus, if you accept that, then one ought to be able to leave tissue for long periods at higher concentrations. I once experimented with the final dehydration step in 100EtOH (and I mean 100%(200 Proof)) by placing a piece of tissue in absolute EtOH in my -10oC freezer with a pint bottle of 200 Proof. I changed the fluid each day for a week and then finished the processing. I saw no difference between the tissue processed at RT and that dehydrated at -10oC, but if I had to try a whole kidney, I would remember that brief experiment in the execution of the art of tissue processing if I had infiltration problems.
Solution (My own approach). I would take control kidneys, since rat kidney is unipolar as I recall, and take transverse segments of known thickness (single-edge razor blade) through the fixed kidney (laying on its side or held 'up' with the 'ends of the bean' down) being careful to isolate the segments in order. I would then process each segment between leaves of fine mesh, using vacuum infiltration for the shortest time and at the lowest temperature. Following installation of each segment in a block, I would carefully orient each block to permit me the largest return of serial sections {10um. The ribbons of serial sections would be placed on poster board in order. Knowing the mean diameter of glomeruli, I would choose a sub-serial sample of sections from each segment and mount them on slides, in order. Meanwhile, my notebook would be filling with the numbers of the sections from the series of each segment (A4-20-1)=Slide Label = Segment A, Start w/section 4, every 20th section, slide 1), etc. Each slide with a mounted series would be marked with a diamond scribe as 1,2,... and then as above - record in notebook. I would then stain each slide with the PAS (periodic acid Schiff) reaction for carbohydrates (BUT in kidney, really does a great job on glomeruli, and one can use a green filter to count and photograph). A nuclear counterstain is optional depending on your purpose, BUT then, there are hundreds of other, adjacent sections waiting for such elaboration if you need them. If the PA(to generate aldehydes from vicinyl glycols) and the S(especially!) are fresh and tested (20ml of S with a drop of formaldehyde to prove the color change from none to intense magenta), the PAS is almost entirely predictable and effortless(?). I would then count the glomeruli in each section or in a statistically predetermined sub-sample, multiply, and get ready for the pathologic specimens. NOTE: Tissue dryness can occur even with the best laid plans and execution, so I have one more suggestion - which will, of course, eliminate the PAS as an appropriate stain. Consider using Polyethylene Glycol(PEG) as an embedment. For those who have learned the method, it is easy and productive, though I know of no reconstructive or statistical methods that have been based on sections of PEG-infiltrated material. NOTE: In any morphometric analysis, it is the sampling method that makes the entire process meaningful or not. Take for example the alternate experiment on a sheep. Would you have the same plan as you proposed to us? What if you wanted to compare mouse and elephant with respect to the structure of a single cell type. What are the statistical implications of the problem? [RHETORICAL QUESTION!!!!!!!!] What you might really like is to be able to take 10 random sections from each serially sectioned segment of a kidney and demonstrate not only the differences between normal and pathologic organs but also the presence or absence of any variation in the distribution of pathology in any organ. Would you expect the same result in a mouse or rabbit which also have unipolar kidneys? What about your expectation in a multipolar kidney like yours? How about a horseshoe multipolar like mine?
I have a protocol for PAS which you may have for the asking, but it is to be found in any version of Pearse, A.G.E. and many others for the looking. And by elaborating as I have done, I remain open to further questions that will be welcome.
Regards,
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: Geoff McAuliffe } Sent: Tuesday, July 10, 2001 10:47 AM } To: CachatF-at-aol.com } Cc: Microscopy-at-sparc5.microscopy.com } Subject: Re: stereology on thick sections } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } "CachatF-at-aol.com"-at-sparc5.microscopy.com wrote: } } } I am interested in counting the total number of glomeruli in the kidneys } in } } } rats with obstructive uropathy. To perform stereology, I have to cut } thick } } } sections at 20 micrometers. I found that the kidney itself breaks down } and } } } get very damaged at this thickness (kidneys fixed in 10% formalin and } } } embedded in parrafin), probably because the kidney is much harder than } the } } } paraffin. In the litterature, some people embed kidneys in glycol } } } methacrylate. It might be better (harder), but then I do not know how } to } } cut } } } my sections which are about 2.5 to 3 cm long, if I need a } glass/diamond } } knife } } } that long. Any ideas about cutting thick sections in paraffin?, or is } it } } } worth re-embedding in glycolmethacrylate, and any ideas about cutting } my } } } serial sections at 20 micrometers in that resin? } } } } } } Thank you a lot for your help! } } } } } } F. Cachat } } } University of Virginia } } } fc6b-at-virginia.edu } } How about frozen sections? Fix in formalin, cryoprotect with ascending } concentrations of sucrose (10%, 20%, 30%) then snap freeze and section at } 20 } microns on a sliding microtome. Collect sections in ice-cube trays to keep } them } in order. } In my experience, thick paraffin sections are much more difficult than } thinner } ones. Embedding tissue in methacrylates (and other plastics) is usually } intended } for thinner sections (1-3 microns). } } 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 } ********************************************** } } } }
Research Associate position(s) available immediately to study roles of calcium dynamics during somite formation and myofibril assembly. Primary training in real-time digital fluorescence microscopy (both wide-field and confocal) using calcium tools, FP-tagged constructs, and FRET approaches. Experience in protein biochemistry or molecular biology highly desirable. Salaries commensurate with qualifications and experience, start date is negotiable. Research and position descriptions can be found at http://biology.uark.edu/ferrari. Research and resources at the University of Missouri - Kansas City School of Biological Sciences can be reviewed at http://sgi.bls.umkc.edu/research.html. Please send cover letter, CV, and names of three references to either address below or, preferably, attach via e-mail: ferrari-at-uark.edu. The University of Missouri-Kansas City is an affirmative action/equal opportunity employer.
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Michael B. Ferrari Department of Biological Sciences University of Arkansas Fayetteville, Arkansas 72701 Ph: 501-575-6372 Fax: 501-575-5349 http://biology.uark.edu/ferrari
*****Note address change effective Jan. 2002***** School of Biological Sciences University of Missouri - Kansas City Kansas City, Missouri 64110-2499 Ph: 816-235-2247 Fax: 816-235-5595
} Peggy Bisher wrote: } } } One of the scientists here wants to give a demonstration involving } } vortices and fluids. He has a few short demonstrations to show the } } kids, but I told him I think he needs more to keep their interest and } } to keep them busy. He has five groups of 6 children for about an } } hour. } Hi, Peggy -- } The Exploratorium, an excellent hands-on science museum in San Francisco, sells manuals on building interactive demo equipment; do a web search. For example, here's a description of their optics manual (from the MICRO bibliography - URL below): ---------------------- Doherty, P. and Rathjen, D. 1995 The Magic Wand and other Bright Experiments on Light and Color 125pp, 7.5x9", $10.95. ISBN 0-471-11515-0 John Wiley & Sons, NY The Exploratorium in San Francisco is famous for its interactive science exhibits. They have published a series of "science snackbooks" that give adults detailed instructions on how to duplicate those exhibits for classrooms or science museums. This one collects 25 easy-to-construct optics demonstrations that investigate reflection, refraction, polarization, and magnification. Many could be good science fair projects. All grade levels. RECOMMENDED ----------------------
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
Is anyone familiar with the Golgi-Cox Method for LM of brain? Any help would be appreciated! The method I was referred to uses Amfix(?) to decolorise the background. Has anyone heard of this and where can I order it? Or if anyone has a better method that has worked for them, please let me know.
Thanks! Peggy Sherwood -- Peggy Sherwood Lab Associate, Photopathology Wellman Laboratories of Photomedicine (W224) Massachusetts General Hospital 50 Blossom Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) sherwood-at-helix.mgh.harvard.edu
We've recent acquired a Bomar SPC-900/EX Critical Point Dryer, but of course it didn't come with an operator's manual. Does anyone out there have a manual for this instrument, and if so, would you be willing to send or fax me a copy? Is this company still in business?
Thanks in advance,
Steve
Steven Verhey, Ph.D Assistant Professor Department of Biology Central Washington University Ellensburg, WA 98926 509.963.3431 office 509.963.2730 fax verheys-at-cwu.edu
Unfortunately, good and cost effective may not be go together when speaking of large area magnetic shielding. The best material for magnetic shielding is known as Mu-Metal. This material is typically supplied fully annealed which provides the most effective shielding. FWIW, If strained by forming, it is common to re-anneal. This probably won't affect you if you simply "wallpaper" with sheet stock using contact cement. I would bond joints well (pop rivets, solder?). The web site for a supplier of Mu-Metal is: http://www.advancemag.com/
A poor second choice might be fully annealed silicon steel used in electrical transformers.
Regards, Woody White McDermott Technology, Inc. ---------------------------------------------------------------------
} -----Original Message----- } From: Dee Breger [mailto:micro-at-ldeo.columbia.edu] } Sent: Wednesday, July 11, 2001 11:07 AM } To: microscopy-at-sparc5.microscopy.com } Subject: magnetic field shielding } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } 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 colleagues, } } Can anyone recommend a good/cost-effective type of wall shielding to } eliminate 60 and 180 cycle interferences coming into an SEM room from } recently-installed large motors on the opposite side of the wall (that } can't be moved now)? } } Much obliged, } Dee } } } } } *************************************************************** } Dee Breger } Mgr. SEM/EDX Facility } Lamont-Doherty Earth Observatory } 61 Route 9W } Palisades, NY 10964 USA } T: 845/365-8640 } F: 845/365-8155 } } http://www.ldeo.columbia.edu/micro } http://www.discovery.com/area/science/micro/micro1.html } http://www.lsc.org/antarctica/front.html } Journeys in Microspace (Columbia University Press, 1995) } } }
I remember receiving the same scam from the mail. I am surprised that people can and will succumb to these things. Oh well, stupid is as stupid does.
Earl ----- Original Message ----- } From: "Nicol Aitken" {nicol-at-semiconductor.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, July 11, 2001 7:05 AM
} } Peggy Bisher wrote: } } } } } One of the scientists here wants to give a demonstration involving } } } vortices and fluids. He has a few short demonstrations to show the } } } kids, but I told him I think he needs more to keep their interest and } } } to keep them busy. He has five groups of 6 children for about an } } } hour. } } } Hi, Peggy -- } } } The Exploratorium, an excellent hands-on science museum in San Francisco, } sells manuals on building interactive demo equipment; do a web search. For } example, here's a description of their optics manual (from the MICRO } bibliography - URL below):
I would like to second Caroline's recommendation for the exploratium. They have put a lot of the interactive demos on their website,check out: http://www.exploratorium.edu/snacks/snacksbysubject.html There are other great informal science education tools on their website, try using their search engine to find what you need.
Bomar is a name that will live in infamy.....at least in this facility. More than 15 yrs ago, the former director returned a CPD unit to Bomar for repair. After not hearing from them for many months, and after her phone calls were not returned, she demanded the return of the unit. No response. The company had gone bankrupt, its assets were seized and our unit was gone. We did file a lawsuit against the company but were told to forget pursuing the matter since it would cost more than the unit was worth. If you do find that they are in business again, let me know. I will look around to see if any information on the CPD.
Good luck......
JB
} We've recent acquired a Bomar SPC-900/EX Critical Point Dryer, but } of course it didn't come with an operator's manual. Does anyone out } there have a manual for this instrument, and if so, would you be } willing to send or fax me a copy? Is this company still in business? } } Thanks in advance, } } Steve } } Steven Verhey, Ph.D } Assistant Professor } Department of Biology } Central Washington University } Ellensburg, WA 98926 } 509.963.3431 office } 509.963.2730 fax } verheys-at-cwu.edu
-- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ##############################################################
We routinely cut 20-50 micron sections from methacrylate blocks for stereology and as far as we are concerned it is the only way to go for stereology studies. We use technovit 7100 kits and deal with samples as large as whole ovaries from sheep.If the whole ovary is to large we simply cut it in half and process and section seperately, for stereology studies we simply count the 2 halves seperately and the sum of the 2 halves = the whole organ.The cutting is a bit of an art but with practice becomes fairly easy, fortuneately we are blessed here with an artist and the following is a description of how she does the cutting.We have also used a number of different stains on the thick sections, if you decide to go down this path and need any further info don't hesitate to e-mail us, alternatively look for any references by HJG Gundersen, in my opinion the guru of stereology, a good review article is in APMIS 96:1988 p 379-394 and p857-881.
The following is how we cut thick sections (30 Microns), using Technovit Resin. Before cutting thick sections, blocks have to be completely hard, either been made for months or hardened in a 37 degree oven for 3-5 days. Unlike paraffin sections which have a right and wrong side, Resin/Plastic sections can be placed in the waterbath on either side, so for this reason I cut the right hand corner off the block, so I can regulate the mounting of sections onto slides. I use a Microm Rotary Microtome HM 350 with a Ralph Glass Knife with a concave knife profile, knives are made on a LKB Histo Knifemaker. The microtome is programmed with a cutting window (selected according to the size of the block) and a cutting speed (selected speed is 2.2). As I mainly do serial sections I have a continuous stroke programmed on the microtome (this means that I pick up a section every 5-10seconds), the microtome is controlled with the foot/hand switch. My waterbath is a large glass staining dish with distilled water at room temperature which is placed in a black box (so that you can see the sections on the bottom of the waterbath). I also have a small glass beaker with distilled water and a pipette, this is so that I can put water onto the glass knife as the section is being cut. Cut sections are taken to forceps and placed into the waterbath were they rest on the bottom of the waterbath until they are picked up onto cleaned glass slides and placed into a 60 degree oven for at least 30 minutes (sections are very wrinkled at first but straighten out with the heat from the oven).
Hope this is of some help
Peter Smith Reproduction Group AgResearch Wallaceville Research Centre P.O. Box 40063, Upper Hutt, New Zealand
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I am not familiar with the Golgi-Cox method, but if it involves silver in any shape or form, then the "Amfix" most probably refers to a photographic rapid fixer (ammonium thiosulphate). As far as I know this is still available, but Agfa Agefix and Ilford Hypam would be equivalent. Kodak Unifix would probably work as well, but it does contain a photographic emulsion hardener.
Jan Coetzee
Peggy Sherwood wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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! } } Is anyone familiar with the Golgi-Cox Method for LM of brain? Any } help would be appreciated! The method I was referred to uses } Amfix(?) to decolorise the background. Has anyone heard of this and } where can I order it? Or if anyone has a better method that has } worked for them, please let me know. } } Thanks! } Peggy Sherwood } -- } Peggy Sherwood } Lab Associate, Photopathology } Wellman Laboratories of Photomedicine (W224) } Massachusetts General Hospital } 50 Blossom Street } Boston, MA 02114 } 617-724-4839 (voice mail) } 617-726-6983 (lab) } 617-726-3192 (fax) } sherwood-at-helix.mgh.harvard.edu
-- Prof Jan Coetzee Lab for Microscopy and Microanalysis University of Pretoria, South Africa. Tel: 012-420-2075, Fax 012-362-5150 www.up.ac.za/academic/electron/emunit1.htm
Dee Breger wrote: Can anyone recommend a good/cost-effective type of wall shielding to eliminate 60 and 180 cycle interferences coming into an SEM room from recently-installed large motors on the opposite side of the wall (that can't be moved now)?
Hi, Have you as an alternative considered to use the magnetic field cancellation systems that are available nowadays? The first I found by searching the web for "magnetic field cancellation" was:
http://www.spicerconsulting.com/sc12.htm
but there are quite a few other producers.
Best regards, Jorgen.
{:::} ¤ {:::} ¤ {:::} ¤ {:::} ¤ {:::} ¤ {:::}
Joergen B. Bilde-Soerensen Senior Research Scientist, Ph. D. Materials Research Department Risoe National Laboratory DK-4000 Roskilde Denmark
I am too confront with this problem. But we can move the SEM (witch we have just moved... in a electromagnetic "dirty" room!).
As Woody said it, mu-metal is the best way, but with terrible difficulties and cost. One of our collegues, magnetism specialist, proposed to use thick aluminium or cooper plates (10 mm or more thick). Aluminium (dural) has the big advantage to be light, copper is better for electrical continuity, but heavier and expensive (with 10 to 15 mm thick...), soft iron or silicon iron is not too expensive, but heavy. You must build a box around the SEM, with a door on entry lock side and a good electrical continuity. It looks nice ! It's not a simple problem, but as consolation, remember that you can recycle the plates when the interferences are eliminated. But of course the best way is to eliminate the source of interferences.
By the way, did you mesure the AC field ? In case, how much do you have ? I am interested on the solution you will find. Our building is VERY noisy.
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess 67037 Strasbourg CEDEX France
Unfortunately, good and cost effective may not be go together when speaking of large area magnetic shielding. The best material for magnetic shielding is known as Mu-Metal. This material is typically supplied fully annealed which provides the most effective shielding. FWIW, If strained by forming, it is common to re-anneal. This probably won't affect you if you simply "wallpaper" with sheet stock using contact cement. I would bond joints well (pop rivets, solder?). The web site for a supplier of Mu-Metal is: http://www.advancemag.com/
A poor second choice might be fully annealed silicon steel used in electrical transformers.
Regards, Woody White McDermott Technology, Inc. ---------------------------------------------------------------------
} -----Original Message----- } From: Dee Breger [mailto:micro-at-ldeo.columbia.edu] } Sent: Wednesday, July 11, 2001 11:07 AM } To: microscopy-at-sparc5.microscopy.com } Subject: magnetic field shielding } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } 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 colleagues, } } Can anyone recommend a good/cost-effective type of wall shielding to } eliminate 60 and 180 cycle interferences coming into an SEM room from } recently-installed large motors on the opposite side of the wall (that } can't be moved now)? } } Much obliged, } Dee } } } } } *************************************************************** } Dee Breger } Mgr. SEM/EDX Facility } Lamont-Doherty Earth Observatory } 61 Route 9W } Palisades, NY 10964 USA } T: 845/365-8640 } F: 845/365-8155 } } http://www.ldeo.columbia.edu/micro } http://www.discovery.com/area/science/micro/micro1.html } http://www.lsc.org/antarctica/front.html } Journeys in Microspace (Columbia University Press, 1995) } } }
Title: Optical Microscopy and Imaging in the Biomedical Sciences
When: October 10 - October 19, 2001
Where: Marine Biology Laboratory, Woods Hole, MA, USA
Tuition: $2150 (Includes room and board, text, handouts, supplies)
Application Deadline: July 25, 2001
Admission application and information: Carol Hamel, Admissions Coordinator Marine Biological Laboratory 7 MBL Street Woods Hole, MA 02543-1015 (508) 289-7401 Internet: admissions-at-mbl.edu WWW: http://www.mbl.edu (Application forms available via Adobe Acrobat)
Course Director: Colin S. Izzard, State University of New York -at- Albany Phone: [518] 442 - 4367 EMail: csizzard-at-csc.albany.edu
Course Description:
For Whom: Designed primarily for research scientists, physicians, postdoctoral trainees and advanced graduate students in animal, plant, medical and material sciences. Non-biologists seeking a comprehensive introduction to microscopy and video-imaging will benefit greatly from this course as well. There are no specific prerequisites, but an understanding of the basic principles of optics is desirable. Limited to 24 students.
The eight day course consists of lectures, laboratory demonstrations, exercises and discussions that will enable the participant to obtain and interpret microscope images of high quality, to perform quantitative optical
measurements, and to produce photographic and video records for documentation and analysis.
Topics to be covered include: principles of microscope design and image formation bright field, dark field, phase contrast, polarized light, differential interference contrast, interference reflection, and fluorescence microscopy confocal scanning microscopy, multiphoton excitation fluorescence microscopy and image deconvolution digital image restoration and 3-D reconstruction video imaging, recording, enhancement, and intensification analog and digital image processing and analysis fluorescent probes and ratiometric-imaging laser tweezers and laser scissors
Applications to live cells will be emphasized; other specimens will be covered as well.
Students will have direct hands-on experience with state-of-the-art microscopes, video cameras, recorders and image processing equipment provided by major optical and electronics companies. Instruction will be provided by experienced staff from universities and industry.
Students are encouraged to bring their own biological (primary cultures, cell lines, etc.) and material specimens and to discuss individual
At 3:53 PM -0400 7/11/01, Peggy Sherwood wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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! } } Is anyone familiar with the Golgi-Cox Method for LM of brain? Any } help would be appreciated! The method I was referred to uses } Amfix(?) to decolorise the background. Has anyone heard of this and } where can I order it? Or if anyone has a better method that has } worked for them, please let me know. } } Thanks! } Peggy Sherwood } -- *****************
Hi Peggy, Here is the Golgi-Cox protocol, with formulae. Its out of:"Histological & Histochemical Methods: Theory & Practice" by JA Kiernan (1981)Pergamon Press.
A. Golgi-Cox Fixative-prepared from 3 stable stock solutions: (i) 5% aqueous mercuric chloride (HgCl2) (ii) 5% aqueous potassium dichromate (K2Cr2O7) (iii)5% aqueous potassium chromate (K2CrO4)
Working s olution} Mix, IN ORDER (i) 20ml (ii) 20ML Water: 40ml (iii) 16ml Mix before useing.
B. Alkaline developer Strong ammonia solution: (17% NH3) 5 ml Water 95 ML Mix just before using.
Procedure:
1. fix pieces of nervous tissue (not more than 10mm thick) in the working sol'n of the golgi-Cox fix for 6-8 weeks in a tightly capped container at 37 deg. C. Pour off fix and replace with fresh after first day. 2. Wash in water, many changes, for 6-8 hours, then dehydrate and embed in nitrocellulos. Cut sections at 100 microns thick. 3. treat sections with the alkaline developer (sol'n B) for 2-3 minutes. 4. Wash in water, dehydrate, clear & mount in a resinous medium. (see NOTE)
Result: some neurons & neuroglial cells, including cytoplasmic processes--black. Background, pale yellow.
NOTE: A mixture of 33ml chloroform, 33ml xylene and 33ml abs,. alcohol is preferred as the lat stage of dehydration. This can be followed by clearing for 10-15 min in creosote or in cedarwood oil, followed by a rinse in xylene.
thae black color fades with time if a coverslip is applied, so it is usual to mount sections in THICK canada Balsam, and allow to set at 40-45 deg C without a coverslip. Such preps. are stable for many years. If a synthetic mounting medium is used (such as DPX), coverslips may be applied, but fading occurs after about 6 months.
Its a lot of work, using reagents most of us are not familiar with, and which will make your Health & Safety officers cringe! good luck, Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
We are looking for a Philips 400 series hot stage. The intent is to make some modifications to it so that even one that is broken (depending upon what is broken) might be acceptable. If you have one that you might be willing to part with, please communicate to me its state and under what conditions you might let it go.
Richard
Richard Vanfleet Assistant Professor Advanced Materials Processing and Analysis Center (AMPAC) and Department of Physics
Mail: AMPAC University of Central Florida 12443 Research Parkway, Suite 404 Orlando, FL 32826
I thought this might be worth drawing to your attention if you scan e.m. negatives in with a flat bed scanner.
We currently use an Epson Perfection 1200 with transparency adapter. I know the spec is considered to be pretty low in terms of O.D., greyscale and DPI but it produces very good pictures about 98% of the time. One problem that I have encountered, however, from time to time is one or rarely more than one extremely fine dark lines appears on the scanned image. This can be very annoying because they are only visible if you look very carefully at an enlarged image. I had assumed that there was a problem with one or two points on the CCD scanner detector array because the lines were straight and at right angles to the scanner head.
The lines are actually caused because this particular scanner has a small reference slot to calibrate the CCDs for fluctuations in the light. If a small particle of dust falls in this area of the glass top it causes the the scanner to overcompensate for the CCD on the corresponding point of the scanner head. This usually results in a very fine white line on the negative image or black on the positive image. I confirmed this using a series of varying density dots on film and got a perfect series of lines.
I'm sorry if people knew this already but thought it just might save someone tearing their hair out or throwing a perfectly functional scanner away. I don't know if some of the other flatbed glass topped scanners with transmitted light sources use a similar calibration system but it's worth taking extra care if they do.
Malcolm Haswell e.m. unit School of Sciences University of Sunderland SUNDERLAND UK
One other very interesting aspect of the scam. The letters sent via snailmail used counterfeit (forged) stamps. So many of these letters were sent that copies of these fakes still show up in packets of stamps sold to collectors. And the envelopes with stamps on them (called covers in the hobby) are now considered to be relatively valuable collectors' items. (Oh, you mean it's obvious that I'm one of those stamp collectors? Too true.)
Roger On Wed, 11 Jul 2001 15:38:24 -0700, Earl Weltmer wrote:
| ------------------------------------------------------------------------ | The Microscopy ListServer -- Sponsor: The Microscopy Society of America | To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com | On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html | -----------------------------------------------------------------------. | | | I remember receiving the same scam from the mail. | I am surprised that people can and will succumb to these things. | Oh well, stupid is as stupid does. | | Earl | ----- Original Message ----- | } From: "Nicol Aitken" {nicol-at-semiconductor.com} | To: {Microscopy-at-sparc5.microscopy.com} | Sent: Wednesday, July 11, 2001 7:05 AM | Subject: RE: more on that scam | | | } ------------------------------------------------------------------------ | } The Microscopy ListServer -- Sponsor: The Microscopy Society of America | } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com | } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html | } -----------------------------------------------------------------------. | } | } | } | } | } | } } Hello listers, | } } I recall seeing that thread late last week about the business | opportunity. | } } This just came in from a Canadian news source, CBC. | } } Nick | } } RCMP make arrests in international phone scam | } } WebPosted Tue Jul 10 17:27:29 2001 | } } TORONTO - The RCMP, along with the FBI and the U.S. Secret Service, have | } } cracked a multi-million dollar scam originating in Nigeria and arrested | } } three people in the Toronto area. | } } More than 300 people around the world - most of them Americans - were | } } duped into giving money to a fake organization. The RCMP say some people | } } lost more than $50,000. Others lost millions. | } } Police say the fraud was sophisticated, high tech and organized. | } } Typically, victims received a letter either by mail or fax allegedly | from | } } officials of a large Nigerian institution, such as the Nigerian National | } } Petroleum Corporation or the Central Bank of Nigeria. | } } The letters would contain an urgent request for help in transferring | } } millions of dollars out of Nigeria to financial institutions in the | } } victim's country. If the victims would help, they'd be in line for a | } } handsome profit, in some cases as much as 30 per cent of the monies | being | } } transferred. | } } The letter says foreign accounts are needed and victims were asked to | wire | } } personal banking information and $10,000 US to cover administration | fees. | } } 'Boiler room' in Toronto | } } Then, victims were contacted by a person posing as a merchant banker of | } } the Central Bank of Nigeria. They were told the funds were now in a | } } clearing house or mercantile bank in North America. | } } Phone numbers given for the fake banks actually rang in a "boiler room" | in | } } the Toronto area. | } } The scam continued with the victim asked to pay taxes, duties and other | } } fees, all with the promise of a windfall, until the victim was bankrupt. | } } The RCMP began a co-ordinated effort with U.S. police forces in 1998. | } } According to an RCMP news release, police detachments in the Toronto | area | } } have received at least one complaint a week for the past eight years | from | } } victims who lost money in the scam. | } } Staff Sergeant Darryl Ross says victims lost more than money. Some | } } suffered health problems, mental breakdowns, divorce and the break-up of | } } their families. | } } | } } | } | |
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc. 900 Rigdebury Road Ridgefield, CT 06877 203-798-5448
_______________________________________________________ Send a cool gift with your E-Card http://www.bluemountain.com/giftcenter/
} } Is anyone familiar with the Golgi-Cox Method for LM of brain? Any } } help would be appreciated! The method I was referred to uses } } Amfix(?) to decolorise the background. Has anyone heard of this and } } where can I order it? Or if anyone has a better method that has } } worked for them, please let me know.
I am not familiar with any need to decolorize the background in most of the Golgi impregnation methods, the Cox variant included. OTOH, my experience has not been with wax embedding but only with the harder (celloidin, araldite, etc.) media do not require any clearing.
There is, however, a need to add an oxident, like NaOH, to the alkalinization step after impregnation and before embedding to prevent background blackening.
I have a Sorvall JB4 microtome that is not advancing at all. If anyone has any suggestions, please contact me. Thanks.
Phoebe
Phoebe J. Doss Manager/Adjunct Instructor Electron Microscope Lab Department of Physiological Sciences 264 McElroy Hall Oklahoma State University Stillwater, OK 74078 405-744-6765
My two cent's worth: as a former Spicer Consulting SC12 user, I was highly satisfied with the unit. We too had EMI from large power cables for water pumps and some other unknown source affecting our SEM images and the SC12 cleared it up. I don't remember how much the unit cost, but remember that is was less than a room full of Mu-Metal. With the added advantages of easy installation and if the SEM ever changes locations, the EMI shielding (the unit) can go with it.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dee Breger wrote: } Can anyone recommend a good/cost-effective type of wall shielding to } eliminate 60 and 180 cycle interferences coming into an SEM room from } recently-installed large motors on the opposite side of the wall (that } can't be moved now)? } } Hi, } Have you as an alternative considered to use the magnetic field cancellation } systems that are available nowadays? The first I found by searching the web } for "magnetic field cancellation" was: } } http://www.spicerconsulting.com/sc12.htm } } but there are quite a few other producers. } } Best regards, } Jorgen. } } {:::} ¤ {:::} ¤ {:::} ¤ {:::} ¤ {:::} ¤ {:::} } } Joergen B. Bilde-Soerensen } Senior Research Scientist, Ph. D. } Materials Research Department } Risoe National Laboratory } DK-4000 Roskilde } Denmark } } e-mail: j.bilde-at-risoe.dk } phone: +45 4677 5802 (direct) } phone: +45 4677 4677 (switchboard) } fax: +45 4677 5758 } website: http://www.risoe.dk/afm/Personal/jqbi/jqbi.htm
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-598-1291 (pager) DSPS Packaging Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
I am intrigued by the recommendation of the referenced magnetism specialist to use aluminum and/or copper plate. The principle in using high conductivity non ferromagnetic shielding (such as Al and Cu) is that of eddy current shielding. This is very effective for high frequency magnetic fields, but becomes worthless as you approach DC (except for the special case of super conducting material, which does shield DC fields). Line-frequency magnetic fields (50-60Hz) are typically dealt with via ferromagnetic shielding because of the relatively low frequency.
Presumably your specialist has done the calculations and feels that eddy current shielding will do the job. This suggests one of three things to me: (1) he knows the problems are due to relatively high frequencies; (2) your fields are very severe and he is only trying to knock them down to a more tractable level; or (3) he knows something I don't. I would personally be interested in learning more about how he arrived at this solution. Specifically, what level of attenuation does he anticipate achieving for 50 Hz magnetic fields with this kind of shielding?
Fred Schamber
Faerber Jacques wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 too confront with this problem. But we can move the SEM (witch we } have just moved... in a electromagnetic "dirty" room!). } } As Woody said it, mu-metal is the best way, but with terrible difficulties } and cost. One of our collegues, magnetism specialist, proposed to use } thick aluminium or cooper plates (10 mm or more thick). Aluminium (dural) } has the big advantage to be light, copper is better for electrical } continuity, but heavier and expensive (with 10 to 15 mm thick...), soft } iron or silicon iron is not too expensive, but heavy. You must build a box } around the SEM, with a door on entry lock side and a good electrical } continuity. It looks nice ! It's not a simple problem, but as } consolation, remember that you can recycle the plates when the } interferences are eliminated. But of course the best way is to eliminate } the source of interferences. } } By the way, did you mesure the AC field ? In case, how much do you have ? } I am interested on the solution you will find. Our building is VERY noisy. } } J. Faerber } IPCMS-GSI } (Institut de Physique et Chimie des Matériaux de Strasbourg } Groupe Surface et Interfaces) } 23, rue de Loess } 67037 Strasbourg CEDEX } France } } Tel 00 33(0)3 88 10 71 01 } Fax 00 33(0)0 88 10 72 48 } E-mail Jacques.Faerber-at-ipcms.u-strasbg.fr } } ---------- Forwarded message ---------- } Date: Wed, 11 Jul 2001 16:49:16 -0500 } } From: "White, Woody N." {nwwhite-at-mcdermott.com} } To: 'Dee Breger' {micro-at-ldeo.columbia.edu} , microscopy-at-sparc5.microscopy.com } Subject: RE: magnetic field shielding } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 Dee, } } Unfortunately, good and cost effective may not be go together when speaking } of large area magnetic shielding. The best material for magnetic shielding } is known as Mu-Metal. This material is typically supplied fully annealed } which provides the most effective shielding. FWIW, If strained by forming, } it is common to re-anneal. This probably won't affect you if you simply } "wallpaper" with sheet stock using contact cement. I would bond joints well } (pop rivets, solder?). } The web site for a supplier of Mu-Metal is: http://www.advancemag.com/ } } A poor second choice might be fully annealed silicon steel used in } electrical transformers. } } Regards, } Woody White } McDermott Technology, Inc. } --------------------------------------------------------------------- } } } -----Original Message----- } } From: Dee Breger [mailto:micro-at-ldeo.columbia.edu] } } Sent: Wednesday, July 11, 2001 11:07 AM } } To: microscopy-at-sparc5.microscopy.com } } Subject: magnetic field shielding } } } } } } -------------------------------------------------------------- } } ---------- } } The Microscopy ListServer -- Sponsor: The Microscopy Society } } 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 colleagues, } } } } Can anyone recommend a good/cost-effective type of wall shielding to } } eliminate 60 and 180 cycle interferences coming into an SEM room from } } recently-installed large motors on the opposite side of the wall (that } } can't be moved now)? } } } } Much obliged, } } Dee } } } } } } } } } } *************************************************************** } } Dee Breger } } Mgr. SEM/EDX Facility } } Lamont-Doherty Earth Observatory } } 61 Route 9W } } Palisades, NY 10964 USA } } T: 845/365-8640 } } F: 845/365-8155 } } } } http://www.ldeo.columbia.edu/micro } } http://www.discovery.com/area/science/micro/micro1.html } } http://www.lsc.org/antarctica/front.html } } Journeys in Microspace (Columbia University Press, 1995) } } } } } }
Our supplier of negative films (Agfa) for the Philips CM-200 EM in the Microscopy Service of the University of Seville does not fabricate these negatives any more. According to Philips Spain, Kodak does not sell those negatives in Spain (Kodak Electron Microscopy 4489, Dimensions: 6.5 x 9 cm) and they do not give us an immediate solution (?).
Could somebody help us with this silly problem and let us know of a supplier close to Spain?
May be I am wrong, but aluminum is "transparent" to the magnetic fields (as well as copper). Since the physics course in the middle school, I had strong feelings that soft iron is a good material for shielding. In Russia we had special requirements for EM rooms (like "floating" basement and proper magnetic/electric shielding). As I remember, our EM room has iron mesh immersed into the walls/ceilings/floors. That mesh was properly grounded (copper sheet 1x1 m buried 5 meters deep into the ground, 20 meters away of the building with very thick copper "conductor"; this "conductor" was mounted across the perimeter of the room and was connected to the mesh at many points). It also seems to me that the thickness of shielding material do not so important. I think, described "shielding" schedule was designed mostly against electric fields, but it should works against magnetic fields as well. I would be happy to listen the comments of the specialist on this matter.
Sergey.
At 10:29 AM 7/12/01 +0200, 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
Does anyone know who makes them and/or sells them on a regular basis?
rtch
} } } } Any opinions of this $195 device? } http://cgi.ebay.com/aw-cgi/eBayISAPI.dll?ViewItem&item=1164152801 } } - John } P.S. I have no association with this auction. } }
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
Its the Kodak dealer that has chosen not to stock that size. We no longer stock boxes of 100 sheets because there was too little demand and its very expensive when film goes out of date. ProSciTech and half a dozen other EM suppliers would be delighted to supply you. Just ask for a quote including freight. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Friday, July 13, 2001 6:30 AM, Julian Martinez-Fernandez [SMTP:Julian.Martinez-Fernandez-at-grc.nasa.gov] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear colleges: } } Our supplier of negative films (Agfa) for the Philips CM-200 EM in } the Microscopy Service of the University of Seville does not fabricate } these negatives any more. According to Philips Spain, Kodak does not sell } those negatives in Spain (Kodak Electron Microscopy 4489, Dimensions: 6.5 x } 9 cm) and they do not give us an immediate solution (?). } } Could somebody help us with this silly problem and let us } know of a supplier close to Spain? } } Many thanks, } Julian
Thanks to all who replied to my question about the DTSA software package. I have downloaded it onto a Mac here. As yet, I don't have it fully working as it doesn't want to import the EMSA files exported by our Noran Voyager system. I'm still trying to working out what the problem is, but in principle it seems to be working (except with my files).
As helpfully pointed out by Scott Wight at NIST, the latest version resides on their server at http://www.cstl.nist.gov/div837/Division/outputs/DTSA/DTSA.htm
Thanks again
Ian MacLaren Max Planck Institut für Metallforschung, Seestraße 92, 70174 Stuttgart, Germany Email: ian.maclaren-at-physics.org / maclariz-at-yahoo.co.uk Home page: http://members.tripod.co.uk/IanMacLaren/
I need information about the deep focus of objetives:
Plan Neofluar, 2.5/0.075, inf/0.17
Ph1 Plan Neofluar, 10x/0.30,inf/0.17
Ph2 Plan neofluar, 20x/0.50,inf/0.17
Ph2 plan neofluar, 40x/0.75, inf/0.17
thank you for your help
___________________________________
Dr. José Luis Pech Pacheco PhD. Marine Ecology Image & Vision D. Instituto de Óptica, CSIC. Madrid, Spain Fax 0034915645557 _____________________________________
} Thanks to all who replied to my question about the DTSA software package. I } have downloaded it onto a Mac here. As yet, I don't have it fully working } as it doesn't want to import the EMSA files exported by our Noran Voyager } system. I'm still trying to working out what the problem is, but in } principle it seems to be working (except with my files).
FYI, there are two DTSA plug-ins for EMSA import. A version 1.0 and version 1.1. Check to make sure your are using the correct one. If you still have problems, You can also e-mail me some of your Voyager EMSA files and I can tell you what is the problem. I'm well versed with the internals of DTSA and that of EMSA file formats.
Scott
----------------------------------------------------------------------- Scott D. Davilla Phone: 919 489-1757 ext 13 (tel) 4pi Analysis, Inc. Fax: 919 489-1487 (fax) 3500 Westgate Drive, Suite 403 email: davilla-at-4pi.com Durham, North Carolina 27707-2534 web: http://www.4pi.com
Group, I just received a product release on a new product, one that I had not heard of before, that may be of interest to a number of you. The product, called ScanScope T50, from Aperio Technologies converts all information from a microscope slide into a common digital format. They say, for example, that a typical tissue section can be digitized at 54,000 pixels per inch. Their web site is www.aperiotech.com Obvioulsy I (unfortunately)have no financial or other interests in this company or its products. Don Grimes, Microscopy Today
I had seen this item a while back and may have said something to the listserver. At the time, I thought that it would have worked nicely on the eyepiece on a microprobe and perhaps on the binoculars of a TEM. I had gotten some information on it and here is the contact information:
International Micro-Vision Inc. 667 El Camino Real Redwood City, CA 94063-1317
I went to the web site. Not very impressive. Their big product digitizes all the info on a slide without a conventional microscope. OK, nice idea but don't you think they could have posted at least one image! No data of any type at the site.
} ------------------------------------------------------------------------ } 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 have not solved our problems completely but will share some things I have "heard". If you decide to go with the mu-metal, it should be easier and cheaper to build a small enclosure around the actual source (motors) than to shield the EM or entire room. This assumes you are confident that the new motors are the source; i.e., the SEM was fine before their installation. Anyway, this is how I was advised to proceed; unfortunately we could not pinpoint a single source, so I haven't actually tried it. Often, the advice you may recieve from "magnetism experts" and shielded-room experts is better suited to the Earth's steady field than it is to A/C fields.
Matt
Matthew J. Lynn Center for Advanced Microscopy University of Miami (305)284-4736 mlynn-at-miami.edu
On Wednesday, July 11, 2001 11:07 AM, Dee Breger [SMTP:micro-at-ldeo.columbia.edu] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi colleagues, } } Can anyone recommend a good/cost-effective type of wall shielding to } eliminate 60 and 180 cycle interferences coming into an SEM room from } recently-installed large motors on the opposite side of the wall (that } can't be moved now)? } } Much obliged, } Dee } } } } } *************************************************************** } Dee Breger } Mgr. SEM/EDX Facility } Lamont-Doherty Earth Observatory } 61 Route 9W } Palisades, NY 10964 USA } T: 845/365-8640 } F: 845/365-8155 } } http://www.ldeo.columbia.edu/micro } http://www.discovery.com/area/science/micro/micro1.html } http://www.lsc.org/antarctica/front.html } Journeys in Microspace (Columbia University Press, 1995) }
In a message dated 7/11/2001 3:14:47 PM Mountain Daylight Time, micro-at-ldeo.columbia.edu writes:
} Can anyone recommend a good/cost-effective type of wall shielding to } eliminate 60 and 180 cycle interferences coming into an SEM room from } recently-installed large motors on the opposite side of the wall (that } can't be moved now)?
Can you be more specific on what you mean by cost-effective by providing an actual dollar amount that you could afford? Otherwise, many of the suggestions being made may be completely impractical based on funding concerns.
The reason I suggest this is that the last I heard, the commercial magnetic field canceling systems cost from ~$10k to ~$25k. Also, one person has said that covering a room in mu-metal may be of a similar cost.
A less expensive mu-metal solution may be to build a box around the column, or even just around the chamber. Years ago, JEOL sold such a mu-metal box that surrounded just the chamber of a 840A SEM (on 4 full sides plus the top and bottom where it stopped at the perimeter of the column and chamber, respectively). At the time, it cost about $6k from JEOL. Using the JEOL mu-metal box (which was about 2mm thick, if I recall correctly), reduced the interference on the beam by about a factor of 3. (We were doing lithography at the time, so it was easy to compare the amplitude of the interference in the patterns that were written.)
I know one person who designed a similar box at a substantially lower cost using an in-house machine shop. The key in that case is to properly anneal the mu-metal after it has been fabricated.
Regarding the idea of using Cu or Al, I agree with the previous postings that question its usefulness for line frequency magnetic fields, although I haven't done any calculations to evaluate the suggestion.
I have also heard of soft iron being used, which should be relatively inexpensive, but will be very heavy because of the increased thickness that would be required to match the shielding of mu-metal.
Could you build a heavy, ugly, and cheap box around the motors???
The lowest cost solution may be to move the SEM, if you have a suitable room available. A professional move may cost thousands, but doing it yourself is a reasonable option, if you know about shipping bolts and scintillators.
Joe _________________________________________ Joe Nabity, Ph.D. JC Nabity Lithography Systems E-Beam Lithography using Commercial SEMs & STEMs PO Box 5354, Bozeman, MT 59717 USA Voice: (406) 587-0848 FAX: (406) 586-9514 E-mail: info-at-jcnabity.com Web: www.jcnabity.com
Try mandel Scientific , Canada if you cannot find it closer. e-mail info-at-mandel.ca
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
} } } Julian Martinez-Fernandez {Julian.Martinez-Fernandez-at-grc.nasa.gov} 07/12 4:30 PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear colleges:
Our supplier of negative films (Agfa) for the Philips CM-200 EM in the Microscopy Service of the University of Seville does not fabricate these negatives any more. According to Philips Spain, Kodak does not sell those negatives in Spain (Kodak Electron Microscopy 4489, Dimensions: 6.5 x 9 cm) and they do not give us an immediate solution (?).
Could somebody help us with this silly problem and let us know of a supplier close to Spain?
In a message dated 7/13/01 10:55:26 AM, PhillipsT-at-missouri.edu writes:
} I went to the web site. Not very impressive. Their big product } digitizes all the info on a slide without a conventional microscope. } OK, nice idea but don't you think they could have posted at least one } image! No data of any type at the site.
I agree that there is no info whatever on the site. Also, note that 54000 points per inch is 1/2 micron. Certainly that is not real resolution, and one wonders how with anything approaching that optical resolution they can assure good focus. It takes some serious mechanical engineering and software to scan a whole slide to collect and tile images. I've seen it done, using good microscopes and stages with well designed optics and a digital camera. I'd have to see some real examples before believing anything from this outfit.
I am a newcomer to this field, trying to differentiate between autofluorescent lignin, callose, cutin and suberin in LR-white embedded plant material. My question is: Is there a way of identifying one of these substances by selectively quenching the autofluoescence of the others ? Can the colour of the emission be used as a tool for differentiation ? I tried Berberinsulfat but it stains lignin and suberin. Neutralred stains the hydrophobic domain of suberin but is also used as a fluorochrome for lipids in general and therefore probably stains cutin as well.
Below is the result of your feedback form. It was submitted by (tcote-at-medicacorp.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Friday, July 13, 2001 at 12:35:36 ---------------------------------------------------------------------------
Email: tcote-at-medicacorp.com Name: Tyler Cote
Organization: Medica Corporation
Education: Undergraduate College
Location: Bedford, MA 01730
Question: Is is it possible to get clear images of white and red blood cells using an air objective ?
I am curently working on a hematalogical assistant that uses a digital camera to image white and red cells. The red cells seem to be acting like little lenses. Most red cells appear to have a small dark band around them. Upon further investigation it appears this band is actually a series of colored bands.
When I wet the slide with a liquid that has an index of refraction closer to that of the cells, the bands disapear. I am still using an air objective, just looking at a wetted slide.
Is this common ?
Is there a way to get around having to wet the slides ?
Any help you can give is greatly appreciated. Thank you.
Below is the result of your feedback form. It was submitted by (joelgerardo-at-correoweb.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, July 11, 2001 at 16:22:19 ---------------------------------------------------------------------------
Email: joelgerardo-at-correoweb.com Name: joel gerardo diaz M.D.
Organization: medicine faculty unam
Education: Graduate College
Location: mexico city , fedeeral district City, State, Country
Question: I need a manual of Quality Control in the manufacturation of Colposcopes and Microscopes ,I will be very grateful for your attention, sincerely. Joel gerardo Diaz Sanchez M.D.
First, I must precise again that I am not a specialist of magnetisme ! I am only confronted with the same problem of electromagnetic interference. So I asked this collegue which has answered and calculated only with the hands.
What he said, and others have said it too, is than mu-metal is the best, soft iron could work too. About Al or Cu, it's of course only efficient with AC field. The goal is to disipate the AC electromagnetic filed in a good conductor by Foucauld current. 50 or 150 Htz is not DC, even it is not "high frequency". As a test is sometimes as good as much calculations, we have tested with a 10 mm thick Al plate, 500 x 500 mm large, aproching the SEM, and we have seen the distorsion reducing, but not been supressed. I don't know if an Al box would be sufficient to put down enought the interfernce. As we can move the SEM in an other room, we didn't try further. The advice of my collegue was : "If you can't supress the source, try to move the SEM ; shielding is often risky and expensive". That what other have soon said on the list.
So about your suppositions (mail from F. Schamber):
} } Presumably your specialist has done the calculations and feels that eddy } } current shielding will do the job. This suggests one of three things to } } me: (1) he knows the problems are due to relatively high frequencies; } } (2) your fields are very severe and he is only trying to knock them down } } to a more tractable level; or (3) he knows something I don't. I would } } personally be interested in learning more about how he arrived at this } } solution. Specifically, what level of attenuation does he anticipate } } achieving for 50 Hz magnetic fields with this kind of shielding?
it is the second witch is right ! We had 20-25 mGauss, 50-150 htz at the sample level . That is much more that the 3 mG max recommended.
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess 67037 Strasbourg CEDEX France
In a message dated 7/11/2001 3:14:47 PM Mountain Daylight Time, micro-at-ldeo.columbia.edu writes:
} Can anyone recommend a good/cost-effective type of wall shielding to } eliminate 60 and 180 cycle interferences coming into an SEM room from } recently-installed large motors on the opposite side of the wall (that } can't be moved now)?
In my previous reply, I forgot to mention that a listing of suppliers for vibration and magnetic field canceling systems as well as suppliers for mu-metal enclosures can be found at "www.jcnabity.com/links.htm".
(If anyone knows of other companies that should be listed, please let me know.)
Joe _________________________________________ Joe Nabity, Ph.D. JC Nabity Lithography Systems E-Beam Lithography using Commercial SEMs & STEMs PO Box 5354, Bozeman, MT 59717 USA Voice: (406) 587-0848 FAX: (406) 586-9514 E-mail: info-at-jcnabity.com Web: www.jcnabity.com
Ian; I had the same import problem with Noran files. It seemed that the Noran interpretation of the EMSA file spec is different than the interpretation by the authors of DTSA. --John Mardinly
-----Original Message----- } From: Ian MacLaren [mailto:maclaren-at-hrem.mpi-stuttgart.mpg.de] Sent: Friday, July 13, 2001 4:54 AM To: Microscopy Discussion Group
Thanks to all who replied to my question about the DTSA software package. I have downloaded it onto a Mac here. As yet, I don't have it fully working as it doesn't want to import the EMSA files exported by our Noran Voyager system. I'm still trying to working out what the problem is, but in principle it seems to be working (except with my files).
As helpfully pointed out by Scott Wight at NIST, the latest version resides on their server at http://www.cstl.nist.gov/div837/Division/outputs/DTSA/DTSA.htm
Thanks again
Ian MacLaren Max Planck Institut für Metallforschung, Seestraße 92, 70174 Stuttgart, Germany Email: ian.maclaren-at-physics.org / maclariz-at-yahoo.co.uk Home page: http://members.tripod.co.uk/IanMacLaren/
I am currently evaluating EDS packages for my Philips XL-30 ESEM with LaB6 and wanted to poll the list members for their opinions, be they positive or negative. How do members deal with the beam spread issues? Can we trust the results? What about water vapor and ice build up? As a multi-user facility how easy is it to get the data out to the researcher these days? What about service quality?
I will post a summary so if you prefer your responses excluded, please let me know.
Thanks
Scott Whittaker SEM Lab Manager National Museum of Natural History Smithsonian Institution 10th & Constitution Ave, NW Washington DC 20560-0104 202-357-1651
The NORAN EMSA files follow the specifications fully and support all varieties of EMSA options available. The problem is that DTSA only reads one type of EMSA file format (X,Y pairs of data) which is not the default condition of the VOYAGER files. The default format for VOYAGER stores only the channel data in 5 columns. The default file format for VANTAGE was changed to be X,Y pairs which is consistant with DTSA.
There is a solution for VOYAGER. You can save the EMSA format compatible with DTSA by using the shell command: emsa_io -x=1 -w={path to store to} -s={spectrum name to store} example: emsa_io -x=1 -w=$HOME/spectra/myspec.emsa -s=spectrum1
When the next version of VOYAGER software is released the default condition for the emsa_io program will be converted to the same as VANTAGE which will make VOYAGER's default version of EMSA file format consistant with DTSA.
Greg Fritz
} -----Original Message----- } } From: Mardinly, John [mailto:john.mardinly-at-intel.com] } Sent: Monday, July 16, 2001 11:08 AM } To: 'Ian MacLaren'; Microscopy Discussion Group } Subject: RE: DTSA } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Ian; } I had the same import problem with Noran files. It seemed that the } Noran interpretation of the EMSA file spec is different than the } interpretation by the authors of DTSA. } --John Mardinly } } -----Original Message----- } } From: Ian MacLaren [mailto:maclaren-at-hrem.mpi-stuttgart.mpg.de] } Sent: Friday, July 13, 2001 4:54 AM } To: Microscopy Discussion Group } Subject: DTSA } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Thanks to all who replied to my question about the DTSA software package. I } have downloaded it onto a Mac here. As yet, I don't have it fully working } as it doesn't want to import the EMSA files exported by our Noran Voyager } system. I'm still trying to working out what the problem is, but in } principle it seems to be working (except with my files). } } As helpfully pointed out by Scott Wight at NIST, the latest version resides } on their server at } http://www.cstl.nist.gov/div837/Division/outputs/DTSA/DTSA.htm } } Thanks again } } Ian MacLaren } Max Planck Institut f¸r Metallforschung, Seestraþe 92, } 70174 Stuttgart, Germany } Email: ian.maclaren-at-physics.org / maclariz-at-yahoo.co.uk } Home page: http://members.tripod.co.uk/IanMacLaren/ } } }
I am not sure if anyone mentioned this, but there is a difference in using DTSA and the EMSA file handler plug-ins versions 1 and 2. Two did not work for me at all in any of the EMSA formatted files I tried, but version 1 did. I was using the power version of DTSA.
-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: Mardinly, John [mailto:john.mardinly-at-intel.com] Sent: Monday, July 16, 2001 12:08 PM To: 'Ian MacLaren'; Microscopy Discussion Group
Ian; I had the same import problem with Noran files. It seemed that the Noran interpretation of the EMSA file spec is different than the interpretation by the authors of DTSA. --John Mardinly
-----Original Message----- } From: Ian MacLaren [mailto:maclaren-at-hrem.mpi-stuttgart.mpg.de] Sent: Friday, July 13, 2001 4:54 AM To: Microscopy Discussion Group
Thanks to all who replied to my question about the DTSA software package. I have downloaded it onto a Mac here. As yet, I don't have it fully working as it doesn't want to import the EMSA files exported by our Noran Voyager system. I'm still trying to working out what the problem is, but in principle it seems to be working (except with my files).
As helpfully pointed out by Scott Wight at NIST, the latest version resides on their server at http://www.cstl.nist.gov/div837/Division/outputs/DTSA/DTSA.htm
Thanks again
Ian MacLaren Max Planck Institut für Metallforschung, Seestraße 92, 70174 Stuttgart, Germany Email: ian.maclaren-at-physics.org / maclariz-at-yahoo.co.uk Home page: http://members.tripod.co.uk/IanMacLaren/
I am not sure if anyone mentioned this, but there is a difference in using DTSA and the EMSA file handler plug-ins versions 1 and 2. Two did not work for me at all in any of the EMSA formatted files I tried, but version 1 did. I was using the power version of DTSA.
-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: Mardinly, John [mailto:john.mardinly-at-intel.com] Sent: Monday, July 16, 2001 12:08 PM To: 'Ian MacLaren'; Microscopy Discussion Group
Ian; I had the same import problem with Noran files. It seemed that the Noran interpretation of the EMSA file spec is different than the interpretation by the authors of DTSA. --John Mardinly
-----Original Message----- } From: Ian MacLaren [mailto:maclaren-at-hrem.mpi-stuttgart.mpg.de] Sent: Friday, July 13, 2001 4:54 AM To: Microscopy Discussion Group
Thanks to all who replied to my question about the DTSA software package. I have downloaded it onto a Mac here. As yet, I don't have it fully working as it doesn't want to import the EMSA files exported by our Noran Voyager system. I'm still trying to working out what the problem is, but in principle it seems to be working (except with my files).
As helpfully pointed out by Scott Wight at NIST, the latest version resides on their server at http://www.cstl.nist.gov/div837/Division/outputs/DTSA/DTSA.htm
Thanks again
Ian MacLaren Max Planck Institut für Metallforschung, Seestraße 92, 70174 Stuttgart, Germany Email: ian.maclaren-at-physics.org / maclariz-at-yahoo.co.uk Home page: http://members.tripod.co.uk/IanMacLaren/
} To All following this lead, Is anyone concerned about the probability that the treatment of women described by several of the listers is illegal? Treating a woman like a criminal or a delinquent child who needs to be closely monitored as to her "motherhood" status is not only ridiculous, but descriminatory and hopefully illegal. It is the responsibility of each individual in a laboratory to watch out for his or her own safety and therefore the business and responsibility of each woman in the lab to watch out for the safety of her unborn child. We are all well past the kindergarten stage and should no longer require a hand to hold while crossing the street. Incensed in Iowa, Kristen
} to all out there: } } This is what I have done so far: } 1. Removed all stored chemicals from the lab. } 2. Moved all chemical mixing into fume hood. } 3. Met for 3 hours with environmental safety officer. } 4. Met for 2 hours with radiation safety officer. } 5. Eliminated all protocols which involve teratogens from pregnant } workers routine. } 6. Established a personal hygiene plan for pregnant worker, which is } being signed of by worker, myself, director of the lab, safety } officer, and her physician. } 7. Reiterated safety protocols for worker, coat , gloves, safety } glasses, etc. } 8. Had lab monitored for radiation, passed. } 9. Had lab monitored for formaldehyde gas, passed. } 10. Planning to monitor xylene exposure to worker when thin sectioning } (cannot do this in the hood, obviously). Heat pen( modified soldering } iron) has not worked well so far, any ideas how to eliminate xylene? } 11. Still looking for a non toxic general 1 micron stain for Spurrs and } LR White. Any ideas? } I am experimenting with food related stains (grape juice, etc.). } 12. Employee has completed, lab safety training course, formaldehyde } training course and radiation training course. } 13. Will continue to monitor lab for chemical and radation exposure. } } To some this may seem like over kill, but I believe that it is very } important that every stone is turned in regards to safety. Another } issue that I face is that I have numerous grad students coming through } the lab, and there is always a possibility that they are pregnant and do } not realize it or keep it from me. I also have a nursing mother who was } visiting the lab and never bothered to tell me, until the subject of why } the lab was reorganized came up. I now demand that all female student, } techs, visitors, ect. } tell me of their "motherhood" status. } } Bill } } -----Original Message----- } } From: Leona Cohen-Gould [mailto:lcgould-at-med.cornell.edu] } Sent: Tuesday, July 10, 2001 7:28 AM } To: William McManus; Monson, Frederick C.; Sally Stowe } Cc: Microscopy Listserver } Subject: RE: Tol blue/borax } } } At 1:04 PM -0600 7/9/01, William McManus wrote: } } ----------------------------------------------------------------------- } - } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------
Kristen A. Lennon, Ph.D. Department of Plant Pathology 351 Bessey Hall Iowa State University Ames, IA 50011 515-294-8854 kalen-at-iastate.edu www.baumlab.org
The Associate Director position at the University of Missouri Molecular Cytology Core (LM imaging facility) remains open.
This position involves training clients, developing new methodologies, maintaining instrumentation and managing junior staff members. Although many applicants will have a PhD in life sciences, we would be happy to consider individuals without a PhD but who have extensive experience in the relevant techniques. Opportunities for individuals with PhD's to conduct independent or collaborative research can be negotiated but the position is not tenure-track. The salary range for this position is $30,257 -$54,663.00 and is 100% benefit eligible. Opportunities for professional development will be encouraged.
Our facility includes a BioRad Radiance 2000 Confocal Laser Scanning Microscope system equipped with a Krypton/Argon laser (488 nm & 568 nm excitation lines) and Red laser diode (638 nm) laser mounted for highest sensitivity in the Keller configuration (bottom port) of an IX 70 Olympus inverted microscope. Simultaneous acquisition of three different fluorescent labels, as well as a high quality Nomarski image, is possible. Appropriate objectives, including water immersion and long working distance objectives, are available. An adjacent PC is equipped with the popular image analysis software package MetaMorph (Universal Imaging). For wide-field fluorescence work, we have another Olympus IX-70 inverted microscope equipped with a Photometrics Sensys cooled CCD camera, a Ludl filter wheel on the excitation side and a Ludl z axis stage motor, and a host of Chroma HQ single, double and triple fluorochrome filters. The Windows-based PC that controls this system includes the Vaytek MicroTome deconvolution software package that does Nearest Neighbor, No Neighbor, Constrained Iterative and Measure PSF deconvolution algorithms. It runs under Image Pro Plus (Media Cybernetics) for the image acquisition and 2D image analysis. For 3D work, we use MetaMorph. This system is also equipped with a CRI color filter to allow the acquisition of high quality color bright field images. Objectives and condenser apertures for high resolution phase contrast and differential interference contrast (DIC, Nomarski) are available. We also have a Nikon Optiphot upright photomicroscope with bright field, phase, DIC and fluorescence optics. This system is coupled to a Spot2 chilled CCD digital camera and a second copy of the MetaMorph image analysis and processing software. The main laboratory of the MCC includes a Reichert Frigocut 2800 cryostat for generating 2-100 µm thick frozen sections and a Reichert UltraCut S ultramicrotome for preparation of semi-thin (0.5 -1 µm) LM sections and ultra-thin EM sections. The MCC also contains two Apple workstations with extensive software for image manipulation and presentation graphics. For publication quality output, the Core is equipped with a Polaroid ProPallette 7000 film recorder and a Fuji Pictrography 3000 for photographic quality color prints. In the last year, our facility attracted researchers from the laboratories of over 100 different principal investigators located in 26 different departments or units on our campus.
Although an ideal candidate would have experience in confocal scanning laser microscopy, bright field microscopy, wide field fluorescence microscopy, deconvolution, image processing/analysis, all types of microtomy, immunocytochemistry, in situ hybridization and Adobe Photoshop, candidates with extensive experience in selected areas and who have the desire and capacity to learn the additional areas will be considered. Excellent oral and written communication skills are essential. Experience in a multi-user core facility would be viewed positively. Electron microscopy is not a component of this core facility. Women and minority candidates are especially encouraged to apply. Review of applications will begin immediately and continue until an appropriate candidate is hired. This position has a salary range of $30,257 $54,663.00 and is 100% benefit eligible. Opportunities for professional development will be encouraged. Columbia is a great, inexpensive place to live.
The Core's web site can be viewed at http://www.biotech.missouri.edu/mbp/cores/index.html
Address applications (CV and 3 letters of reference) or inquires to:
Thomas E. Phillips, Ph.D. Division of Biological Sciences 3 Tucker Hall, University of Missouri Columbia, MO 65211-7400. 573-882-4712 PhillipsT-at-missouri.edu
-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
I agree in principal. But, we live in a very litigious, cover-your-ass type of world. It behooves employers and supervisors to document the training and warnings that women (and men) receive with respect to chemical hygiene in the laboratory environment -especially with respect to pregnancy. I would wager to bet that even the most idealist feminist would be looking for litigation relief if they were exposed to hazardous materials during their pregnancy even if it was her own fault with respect to educating herself to the dangers.
I think that people should be warned of the dangers and that warning should be documented. Their impending motherhood (or lack of) should be their own business. If a woman is pregnant and the unborn child can be potentially exposed to hazardous chemicals, the employer can only make adjustments to the woman's work environment if they are told. If they are not, they should not be held accountable. To protect the employer, there should also be documentation that determines when a woman first tells the company. Even when the training and documentation is there and the employee chooses to ignore them, a company can still be held responsible.
And another thing: this thread is about responsible scientists doing things to protect beings that can not protect themselves. They approached this problem in a responsible, caring and concerned way, not to subjugate women. It is nothing to get incensed about.
CYA rules.
These are just my humble opinions.
-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: Kristen Lennon [mailto:kalen-at-iastate.edu] Sent: Monday, July 16, 2001 5:30 PM To: Microscopy-at-sparc5.microscopy.com
} To All following this lead, Is anyone concerned about the probability that the treatment of women described by several of the listers is illegal? Treating a woman like a criminal or a delinquent child who needs to be closely monitored as to her "motherhood" status is not only ridiculous, but descriminatory and hopefully illegal. It is the responsibility of each individual in a laboratory to watch out for his or her own safety and therefore the business and responsibility of each woman in the lab to watch out for the safety of her unborn child. We are all well past the kindergarten stage and should no longer require a hand to hold while crossing the street. Incensed in Iowa, Kristen
} to all out there: } } This is what I have done so far: } 1. Removed all stored chemicals from the lab. } 2. Moved all chemical mixing into fume hood. } 3. Met for 3 hours with environmental safety officer. } 4. Met for 2 hours with radiation safety officer. } 5. Eliminated all protocols which involve teratogens from pregnant } workers routine. } 6. Established a personal hygiene plan for pregnant worker, which is } being signed of by worker, myself, director of the lab, safety } officer, and her physician. } 7. Reiterated safety protocols for worker, coat , gloves, safety } glasses, etc. } 8. Had lab monitored for radiation, passed. } 9. Had lab monitored for formaldehyde gas, passed. } 10. Planning to monitor xylene exposure to worker when thin sectioning } (cannot do this in the hood, obviously). Heat pen( modified soldering } iron) has not worked well so far, any ideas how to eliminate xylene? } 11. Still looking for a non toxic general 1 micron stain for Spurrs and } LR White. Any ideas? } I am experimenting with food related stains (grape juice, etc.). } 12. Employee has completed, lab safety training course, formaldehyde } training course and radiation training course. } 13. Will continue to monitor lab for chemical and radation exposure. } } To some this may seem like over kill, but I believe that it is very } important that every stone is turned in regards to safety. Another } issue that I face is that I have numerous grad students coming through } the lab, and there is always a possibility that they are pregnant and do } not realize it or keep it from me. I also have a nursing mother who was } visiting the lab and never bothered to tell me, until the subject of why } the lab was reorganized came up. I now demand that all female student, } techs, visitors, ect. } tell me of their "motherhood" status. } } Bill } } -----Original Message----- } } From: Leona Cohen-Gould [mailto:lcgould-at-med.cornell.edu] } Sent: Tuesday, July 10, 2001 7:28 AM } To: William McManus; Monson, Frederick C.; Sally Stowe } Cc: Microscopy Listserver } Subject: RE: Tol blue/borax } } } At 1:04 PM -0600 7/9/01, William McManus wrote: } } ----------------------------------------------------------------------- } - } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------
Kristen A. Lennon, Ph.D. Department of Plant Pathology 351 Bessey Hall Iowa State University Ames, IA 50011 515-294-8854 kalen-at-iastate.edu www.baumlab.org
Can anyone recommend a vendor for a confocal microscope for materials science use? We would like to discuss a materials science application with a knowledgeable vendor..
Thanks, Steve Bradley UOP 50 E Algonquin Road Des Plaines, IL 60017 1-847-391-3321
This year we will be hosting the 7th Annual RMC Materials Microtomy Short Course in Tucson, Arizona from October 23 - 26.
For those of you residing in colder climes Tucson has just the weather to chase away the onset of those winter-time blues.
Join us, and our internationally renowned course faculty, in sunny Tucson to participate in this unique event. This short course is designed specifically for researchers in the field of materials science who wish to gain exposure to advances in specimen preparation for electron microscopy.
email stacy-at-boeckeler.com to receive full details and a course brochure
Dave Roberts Director - RMC EM Products Boeckeler Instruments, Inc 4650 South Butterfield Drive Tucson, AZ 85714 Tel: (520) 745-0001 Fax: (520) 745-0004 email: dave-at-boeckelr.com website: www.rmcproducts.com
I've registered for M & M 2001 in Long Beach, but I'm temporarily a single parent during the event, and need to make arrangements for my six-year-old child. One option is to bring her with me...but I need to make sure she is looked after during the short courses and some of the scientific symposia. If some local listers could give me any suggestions for care, or if there's anyone else in the same position who would be willing to pool expenses, I would be very grateful if you could contact me (you can e-mail me directly).
Thanks in advance,
Rick Powell
***************************************************************************************** Richard D. Powell * rpowell-at-nanoprobes.com * www.nanoprobes.com NANOPROBES, Incorporated 95 Horse Block Road, Yaphank, NY 11980-9710, USA
Sign up for our newsletter: http://www.nanoprobes.com/Newsletter.html *****************************************************************************************
"I am currently evaluating EDS packages for my Philips XL-30 ESEM with LaB6 .................What about water vapor and ice build up? ....
I acquired a (Si) detector from Oxford for our ESEM 2020 in 1995. At that moment half a dozen detectors had been sold by Oxford to ESEM users, half a dozen by Noran, and one by EDAX. Oxford offered in its software package the possibility of "conditioning" the detector, a partial warm-up to remove ice etc., which takes two hours only. The other companies did not offer that possibility at that time, probably because it was patented. I do not know whether they offer it by now. It is very easy. I have used it for 7 years now, without problems so far. I believe that regular "conditioning" in combination with other measures has allowed this. Other measures: 1. a protective tube in which the detector is retracted when not operating (or during hot-stage operation, to protect from hot dust); and 2. maintaining a low pressure in the sample chamber (1 Torr instead of the regular 5 T), when the ESEM is not used.
As to : " How do members deal with the beam spread issues? " ,
X-ray mapping is a good method to eliminate the effects of beam spreading, and check particle analyses. Furthermore, beam spreading can be reduced by working at the lowest pressure possible without getting charging (I use 0.9-1 Torr routinely) and by using the detector set-up (the elongated detector/bullit in the ESEM 2020) with the shortest path in the higher-presssure surrounding, i.e. the sample chamber. I know that for quantitative analysis new software programmes exist which allow e.g. to extrapolate the results from a higher-pressure analysis + a lower-pressure analysis to zero pressure, or use some method of background/surroundings subtraction (several methods have been mentioned in the literature), but my (older) software does not offer that possibility. With best regards, Emond de Roever
We are a hospital EM Unit performing surgical pathology examinations and virology exams on clinical specimens. We perform EM on about 130 surgical cases per year (works out to about 150 specimens, nearly half of which are kidneys, the remainder are solid tissue tumors, muscle, nerve). We perform direct EM on about 300 stools/vesicle fluids for virus examination per year.
We have 1.4 FTEs (full-time equivalents). Our .4FTE covers leaves and off-sets workload. We are a manual service.
Our TAT for pathology is 3 working days from processing (day 1), thick sectioning/consult with pathologist/thin sectioning, staining/scoping (by technologist-specialist)/photography (day 2), photographic printing and reporting to pathologist (day 3).
Has anyone ever worked out an estimate of how many "man staff-hours" are involved in processing a biopsy specimen, manually (pardon me again!), for EM? This includes scoping, interpretation and reporting.
Thank you,
C. Powell EM Unit Saint John Regional Hospital NB, Canada powca-at-reg2.health.nb.ca
We have been critical point drying scallop specimens successfully for SEM to examine them as they grow. We are now at a point where the specimens no longer fit in the CPD chamber. With only a couple of specimens to go we thought to try HMDS. The samples are approximately 30mm in diameter. Does someone have a protocol they have used for larger specimens that has worked for them? You can send them directly to me and I can summarize for the list. Thanks in advance.
David O'Neil Institute for Marine Biosciences National Research Council of Canada 1411 Oxford St. Halifax, NS B3H 3Z1 ph. 902-426-8258 fax 902-426-9413 david.o'neil-at-nrc.ca http://www.imb.nrc.ca/micros_e.html
I have lost the e-mail address of Lesley Bechtold of Jackson Labs. Can some one help me out? Thanks, Tim
Timothy Schneider Director of Electron Microscopy Department of Pathology Thomas Jefferson University Room 238 JAH 1020 Locust Street 19107 215-513-4798 Work 610-613-8170 Cell
I can add a few points to this reply. Since you have the XL30, you will generally be leaving it in Hivac overnight and when not in use for ESEM. Ice buildup should be a lesser problem for you; depending of course on the nature of your work. I have the Oxford system on our XL30 ESEM-FEG and have had good results with the package and with the service (rarely needed). I did not purchase the package myself so did not evaluate competitors' offerings. I don't know which detectors you have but the lower-pressure ones are best for EDS: Large Field Detector or CP+ insert with Backscattered ED will operate at much less than 1T. If you need higher pressures; use the extended cone X-ray detectors. I have used the conditioner on occassion; I believe this has been a feature on Link systems since the early 90's at least. Monitor low-end sensitivity with NiLa vs. NiKa.
I find that beam skirt is a minor issue; generally the ESEM samples (customers) are not using EDS. When you need both, spot-check surrounding areas, or run a few maps at low resolution, and be less "quantitative" about your conclusions unless you need to get into the corrections mentioned by Emond.
The new Oxford system is significantly different from mine, so I won't talk about ease-of-use and portability of data.
Matt
Matthew J. Lynn Center for Advanced Microscopy University of Miami (305)284-4736 mlynn-at-miami.edu
On Tuesday, July 17, 2001 3:48 AM, Emond W F de Roever [SMTP:ederoever-at-ONDEO-Nalco.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 } -----------------------------------------------------------------------. } } } } Scott, } } Responding to your question: } } "I am currently evaluating EDS packages for my Philips XL-30 ESEM with } LaB6 .................What about water vapor and ice } build up? .... } } I acquired a (Si) detector from Oxford for our ESEM 2020 in 1995. At that } moment half a dozen detectors had been sold by Oxford to ESEM users, half a } dozen by Noran, and one by EDAX. Oxford offered in its software package the } possibility of "conditioning" the detector, a partial warm-up to remove ice } etc., which takes two hours only. The other companies did not offer that } possibility at that time, probably because it was patented. I do not know } whether they offer it by now. It is very easy. I have used it for 7 years } now, without problems so far. I believe that regular "conditioning" in } combination with other measures has allowed this. Other measures: 1. a } protective tube in which the detector is retracted when not operating (or } during hot-stage operation, to protect from hot dust); and 2. maintaining a } low pressure in the sample chamber (1 Torr instead of the regular 5 T), } when the ESEM is not used. } } As to : " How do members deal with the beam spread issues? " , } } X-ray mapping is a good method to eliminate the effects of beam spreading, } and check particle analyses. Furthermore, beam spreading can be reduced by } working at the lowest pressure possible without getting charging (I use } 0.9-1 Torr routinely) and by using the detector set-up (the elongated } detector/bullit in the ESEM 2020) with the shortest path in the } higher-presssure surrounding, i.e. the sample chamber. I know that for } quantitative analysis new software programmes exist which allow e.g. to } extrapolate the results from a higher-pressure analysis + a lower-pressure } analysis to zero pressure, or use some method of background/surroundings } subtraction (several methods have been mentioned in the literature), but my } (older) software does not offer that possibility. With best regards, Emond } de Roever } } Ondeo-Nalco Europe, Oegstgeest, the Netherlands } }
On my three year old XL30 I usually work at 6 Torr (to keep specimens really wet) and working distance 7.5 mm. It is routine work to get decent maps or line scans of Ca, P, S (4-10% consentration). If you do not need trace elements or quantitative analysis, EDS on ESEM will do pretty good work.
If you samples are not wet, just not conductive, then you can go to higher voltage, lower pressure (1 Torr or less) and use BSE, which is less influenced by charging and then it will be difficalt to find difference with conventional high vacuum EDS analysis.
My detector have never had ice build up, but oil contamination became substantial in a few weeks after cleaning. As far as I know others do not have this particular problem, and I am loosing my battle with EDS and microscope manufacturers.
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} -----Original Message----- } From: Scott Whittaker [mailto:Whittaker.scott-at-nmnh.si.edu] } Sent: Monday, July 16, 2001 1:16 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: xl_30 ESEM and new EDS } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } I am currently evaluating EDS packages for my Philips XL-30 ESEM with } LaB6 and wanted to poll the list members for their opinions, be they } positive or negative. How do members deal with the beam spread } issues? Can we trust the results? What about water vapor and ice } build up? As a multi-user facility how easy is it to get the data out } to the researcher these days? What about service quality? } } I will post a summary so if you prefer your responses excluded, } please let me know. } } Thanks } } Scott Whittaker } SEM Lab Manager } National Museum of Natural History } Smithsonian Institution } 10th & Constitution Ave, NW } Washington DC 20560-0104 } 202-357-1651 } } }
I seen several comments on this problem. It is a legal minefield. It is important for companies to get an expert attorney's advice on this matter. Several years ago a car battery manufacturer tried to limit the exposure of pregnant women to lead by re-assigning them to different jobs. The case went to the U.S. Supreme Court and the company lost. The Court said in an unanimous opinion that the company must make the workplace safe for all employees equally. A better approach is to give all employees, regardless of gender, the same safety education at the time of hiring and to periodically update this as needed. In other words men would also be told of the hazards for pregnant individuals. The best approach is to simply make the workplace safe for pregnant women. Get proper legal advice. This is not for amateurs.
I can ask around with some of our friends who watch our children to see if anyone would be interested in heading up to Long Beach. We are about a 1 hour drive from the convention center so I would guess they would want to do a whole day thing rather than just a few hours. If you think that would help, let me know and I will see what I can arrange.
Best regards-
David
Rick Powell at Nanoprobes wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello Microscopists: } } I've registered for M & M 2001 in Long Beach, but I'm temporarily a single } parent during the event, and need to make arrangements for my six-year-old } child. One option is to bring her with me...but I need to make sure she is } looked after during the short courses and some of the scientific symposia. } If some local listers could give me any suggestions for care, or if there's } anyone else in the same position who would be willing to pool expenses, I } would be very grateful if you could contact me (you can e-mail me directly). } } Thanks in advance, } } Rick Powell } } ***************************************************************************************** } Richard D. Powell * rpowell-at-nanoprobes.com * www.nanoprobes.com } NANOPROBES, Incorporated } 95 Horse Block Road, Yaphank, NY 11980-9710, USA } } US Toll-free: (877) 447-6266 * Tel: (919) 845-6324 * Fax: (631) } 980-3608 } } Sign up for our newsletter: http://www.nanoprobes.com/Newsletter.html } *****************************************************************************************
-- 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 am working with a TEM and I would like to know the best way to disperse Titanium Oxide particles for analysis. I am currently using a methanol solution, and it is not working very well.
Even though I am a female, I have to agree with Scott on this one. People will seize any opportunity to sue, no matter how great their understanding of the risks involved ahead of time - I have seen it myself in a lab situation. I would love to think that people would take responsibility for their own actions, but that just isn't the reality these days - the lure of easy money always ends up being too great. So, as Scott says, you CYA as best you can and keep your fingers crossed.
Jan Markham Motorola
Kristen Lennon wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } To All following this lead, } Is anyone concerned about the probability that the treatment of women } described by several of the listers is illegal? Treating a woman like a } criminal or a delinquent child who needs to be closely monitored as to her } "motherhood" status is not only ridiculous, but descriminatory and } hopefully illegal. It is the responsibility of each individual in a } laboratory to watch out for his or her own safety and therefore the } business and responsibility of each woman in the lab to watch out for the } safety of her unborn child. We are all well past the kindergarten stage and } should no longer require a hand to hold while crossing the street. } Incensed in Iowa, } Kristen } } } to all out there: } } } } This is what I have done so far: } } 1. Removed all stored chemicals from the lab. } } 2. Moved all chemical mixing into fume hood. } } 3. Met for 3 hours with environmental safety officer. } } 4. Met for 2 hours with radiation safety officer. } } 5. Eliminated all protocols which involve teratogens from pregnant } } workers routine. } } 6. Established a personal hygiene plan for pregnant worker, which is } } being signed of by worker, myself, director of the lab, safety } } officer, and her physician. } } 7. Reiterated safety protocols for worker, coat , gloves, safety } } glasses, etc. } } 8. Had lab monitored for radiation, passed. } } 9. Had lab monitored for formaldehyde gas, passed. } } 10. Planning to monitor xylene exposure to worker when thin sectioning } } (cannot do this in the hood, obviously). Heat pen( modified soldering } } iron) has not worked well so far, any ideas how to eliminate xylene? } } 11. Still looking for a non toxic general 1 micron stain for Spurrs and } } LR White. Any ideas? } } I am experimenting with food related stains (grape juice, etc.). } } 12. Employee has completed, lab safety training course, formaldehyde } } training course and radiation training course. } } 13. Will continue to monitor lab for chemical and radation exposure. } } } } To some this may seem like over kill, but I believe that it is very } } important that every stone is turned in regards to safety. Another } } issue that I face is that I have numerous grad students coming through } } the lab, and there is always a possibility that they are pregnant and do } } not realize it or keep it from me. I also have a nursing mother who was } } visiting the lab and never bothered to tell me, until the subject of why } } the lab was reorganized came up. I now demand that all female student, } } techs, visitors, ect. } } tell me of their "motherhood" status. } } } } Bill } } } } -----Original Message----- } } } From: Leona Cohen-Gould [mailto:lcgould-at-med.cornell.edu] } } Sent: Tuesday, July 10, 2001 7:28 AM } } To: William McManus; Monson, Frederick C.; Sally Stowe } } Cc: Microscopy Listserver } } Subject: RE: Tol blue/borax } } } } } } At 1:04 PM -0600 7/9/01, William McManus wrote: } } } ----------------------------------------------------------------------- } } - } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to } } ListServer-at-MSA.Microscopy.Com } } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } ----------------------------------------------------------------------- } } Kristen A. Lennon, Ph.D. } Department of Plant Pathology } 351 Bessey Hall } Iowa State University } Ames, IA 50011 } 515-294-8854 } kalen-at-iastate.edu } www.baumlab.org
Other organizations have child care options. I believe that the AVS has had something available and the ICMCTF meeting has been talking about it. I have a colleague who used it at the AVS and I believe that she said it was on the expensive side, but that it did enable her to go to the meeting. I know of other colleagues who have not attended meetings because of conflicting schedules with their spouses.
Perhaps this is a topic that should be discussed at the MSA and MAS business meetings. Since my kids are grown, it doesn't affect me, but I can understand the problem.
Another option is to use a forum such as the listserver to organize a child care pool or to pool the resources of those members that need such a service.
-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: Rick Powell at Nanoprobes [mailto:rpowell-at-nanoprobes.com] Sent: Tuesday, July 17, 2001 2:21 AM To: Microscopy-at-sparc5.microscopy.com
Hello Microscopists:
I've registered for M & M 2001 in Long Beach, but I'm temporarily a single parent during the event, and need to make arrangements for my six-year-old child. One option is to bring her with me...but I need to make sure she is looked after during the short courses and some of the scientific symposia. If some local listers could give me any suggestions for care, or if there's anyone else in the same position who would be willing to pool expenses, I would be very grateful if you could contact me (you can e-mail me directly).
Thanks in advance,
Rick Powell
***************************************************************************************** Richard D. Powell * rpowell-at-nanoprobes.com * www.nanoprobes.com NANOPROBES, Incorporated 95 Horse Block Road, Yaphank, NY 11980-9710, USA
Sign up for our newsletter: http://www.nanoprobes.com/Newsletter.html *****************************************************************************************
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Along this vane, We became concerned about safety issues quite a number of years ago. We try to train users of this facility as to the dangers involved and appropriate scientific procedures...regardless of sex, age, etc. We have made it a point to provide sufficent hoods, etc so that users can work safely. We also make a point of the fact that it is the researcher's responsibility to inform themselves of hazards and to ask questions if they are not sure of appropriate procedure. Material safety data sheets are also available in an indexed binder so that anyone can easily check on specific reagents.
In addition we have them sign a sheet when they first start using the facility which says that they understand the potential risks (broad categories of possible hazards are listed on the sheet) and assume the responsibility for their own actions. This may not hold up completely in a court of law but it does show an attempt on our part to make the user aware of their own responsibility for their actions.
Safety is everyone's concern regardless of sex or biological conditions. Whatever policies are in place should be there for everyone, not just that occasional special situation. To have a lab manager or supervisor demand private medical information about users is most certainly an invasion of privacy. Debby
Debby Sherman Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-099e Whistler Building West Lafayette, IN 47907
--------------------------------------
I agree in principal. But, we live in a very litigious, cover-your-ass type of world. It behooves employers and supervisors to document the training and warnings that women (and men) receive with respect to chemical hygiene in the laboratory environment -especially with respect to pregnancy. I would wager to bet that even the most idealist feminist would be looking for litigation relief if they were exposed to hazardous materials during their pregnancy even if it was her own fault with respect to educating herself to the dangers.
I think that people should be warned of the dangers and that warning should be documented. Their impending motherhood (or lack of) should be their own business. If a woman is pregnant and the unborn child can be potentially exposed to hazardous chemicals, the employer can only make adjustments to the woman's work environment if they are told. If they are not, they should not be held accountable. To protect the employer, there should also be documentation that determines when a woman first tells the company. Even when the training and documentation is there and the employee chooses to ignore them, a company can still be held responsible.
And another thing: this thread is about responsible scientists doing things to protect beings that can not protect themselves. They approached this problem in a responsible, caring and concerned way, not to subjugate women. It is nothing to get incensed about.
CYA rules.
These are just my humble opinions.
-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: Kristen Lennon [mailto:kalen-at-iastate.edu] Sent: Monday, July 16, 2001 5:30 PM To: Microscopy-at-sparc5.microscopy.com
} To All following this lead, Is anyone concerned about the probability that the treatment of women described by several of the listers is illegal? Treating a woman like a criminal or a delinquent child who needs to be closely monitored as to her "motherhood" status is not only ridiculous, but descriminatory and hopefully illegal. It is the responsibility of each individual in a laboratory to watch out for his or her own safety and therefore the business and responsibility of each woman in the lab to watch out for the safety of her unborn child. We are all well past the kindergarten stage and should no longer require a hand to hold while crossing the street. Incensed in Iowa, Kristen
} to all out there: } } This is what I have done so far: } 1. Removed all stored chemicals from the lab. } 2. Moved all chemical mixing into fume hood. } 3. Met for 3 hours with environmental safety officer. } 4. Met for 2 hours with radiation safety officer. } 5. Eliminated all protocols which involve teratogens from pregnant } workers routine. } 6. Established a personal hygiene plan for pregnant worker, which is } being signed of by worker, myself, director of the lab, safety } officer, and her physician. } 7. Reiterated safety protocols for worker, coat , gloves, safety } glasses, etc. } 8. Had lab monitored for radiation, passed. } 9. Had lab monitored for formaldehyde gas, passed. } 10. Planning to monitor xylene exposure to worker when thin sectioning } (cannot do this in the hood, obviously). Heat pen( modified soldering } iron) has not worked well so far, any ideas how to eliminate xylene? } 11. Still looking for a non toxic general 1 micron stain for Spurrs and } LR White. Any ideas? } I am experimenting with food related stains (grape juice, etc.). } 12. Employee has completed, lab safety training course, formaldehyde } training course and radiation training course. } 13. Will continue to monitor lab for chemical and radation exposure. } } To some this may seem like over kill, but I believe that it is very } important that every stone is turned in regards to safety. Another } issue that I face is that I have numerous grad students coming through } the lab, and there is always a possibility that they are pregnant and do } not realize it or keep it from me. I also have a nursing mother who was } visiting the lab and never bothered to tell me, until the subject of why } the lab was reorganized came up. I now demand that all female student, } techs, visitors, ect. } tell me of their "motherhood" status. } } Bill } } -----Original Message----- } } From: Leona Cohen-Gould [mailto:lcgould-at-med.cornell.edu] } Sent: Tuesday, July 10, 2001 7:28 AM } To: William McManus; Monson, Frederick C.; Sally Stowe } Cc: Microscopy Listserver } Subject: RE: Tol blue/borax } } } At 1:04 PM -0600 7/9/01, William McManus wrote: } } ----------------------------------------------------------------------- } - } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------
Kristen A. Lennon, Ph.D. Department of Plant Pathology 351 Bessey Hall Iowa State University Ames, IA 50011 515-294-8854 kalen-at-iastate.edu www.baumlab.org
I assume that you have tried other solvents as well, including water. You might consider adding a small surfactant.
One method that has worked for me for diamond particles is to ultrasonic a suspension of water and let it set for a few moments and then pour some of the liquid into a nebulizer. I then use the nebulizer over the surface that I want to disperse the diamond particles on while it is on a hot plate at about 120C.
-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: Christopher M May [mailto:cmmay-at-US.ibm.com] Sent: Tuesday, July 17, 2001 12:52 PM To: microscopy-at-sparc5.microscopy.com
Hello,
I am working with a TEM and I would like to know the best way to disperse Titanium Oxide particles for analysis. I am currently using a methanol solution, and it is not working very well.
Sometimes hotels have child care services or nannies who will come to rooms. Perhaps several folks could go in together and pool resources. Try calling your hotel concierge.
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
} Safety is everyone's concern regardless of sex or biological conditions. Whatever policies are in place should be there for everyone, not just that occasional special situation. To have a lab manager or supervisor demand private medical information about users is most certainly an invasion of privacy. } Debby } } Debby Sherman Phone: 765-494-6666 } Life Science Microscopy Facility FAX: 765-494-5896 } Purdue University E-mail: dsherman-at-purdue.edu } S-099e Whistler Building } West Lafayette, IN 47907
Hello- I am a grad student at the University of Idaho and would like some input on a protocol for fixing rat tissue in order to view their spinal ganglia on the TEM. First heparin is injected intracardiac, then perfusion via the heart with saline (37C) followed by paraformaldehyde (4%, 15C). The vertebral column is then removed and immersed in 4% paraformaldehyde for several hours before dissecting the spinal ganglia. This protocol was given to me by my advisor, but I have been unable to find a similar method cited in the literature. Also, I have used this method and found all the mitochondria have already lost their cristae. Any suggestions and comments would be appreciated. Thank you. --Meghan Towner
Of course, your technique will be selecting for the smaller particles as the larger ones as well as the agglomerates will be dropping out while it sets before you decant into the nebulizer. Nebulizing will also have issues about particle selection.
The original requester didn't say what the size range of the particles are but I assume that they must be submicron or nanometer range as he is using the TEM. Was he trying to size them, study their morphology, or what? I usually end up trying a variety of suspending fluids and also use on ocassion water with surfactant. But there are other techniques to use depending on what we're trying to find out about the particles.
Damian Neuberger Senior Research Scientist Particle Science and Technology Baxter Healthcare Corp
} I assume that you have tried other solvents as well, including water. You might consider adding a small surfactant. } } One method that has worked for me for diamond particles is to ultrasonic a suspension of water and let it set for a few moments and then pour some of the liquid into a nebulizer. I then use the nebulizer over the surface that I want to disperse the diamond particles on while it is on a hot plate at about 120C. } } -Scott }
Below is the result of your feedback form. It was submitted by (cokendolpher-at-aol.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, July 17, 2001 at 19:33:06 ---------------------------------------------------------------------------
Email: cokendolpher-at-aol.com Name: James Cokendolpher
Organization: Midwestern State University
Education: Graduate College
Location: Lubbock, Texas USA
Question: I have used an AO150 for my home research for the past 20 or so years. With the advent of good digital cameras I am now thinking of buying an SL3D microscope and puting my camera lucida in a museum. Do you know of any other microscope with a greater depth of field for under $3000? Do you know of any articles that are good or bad about the SL3D; other than those on their website? My work involves the description and illustration of minute arthropod parts. Thanks.
Hi! I have a question regarding the removal of Spurrs resin from sections on slides. I want to use aniline blue as a callose stain for plant tissue. I have sections (embedded in Spurrs) on slides. The protocol I have to remove the resin uses sodium methoxide and benzene (Mayor, Hampton and Rosario, 1961). I was wondering if anyone can suggest a protocol that is less dangerous than using the sodium methoxide. Or, alternatively, has anyone used this technique, or a similar one and had any problems/success with it? Thanks, Rose
{center} {bold} {bigger} {bigger} ANALYTICAL GROUP {/center}
{center} ELECTRON MICROSCOPIST {/center}
{flushboth} {/bold} {smaller} {smaller} Applications are invited from suitably= qualified and experienced candidates for the above post. The successful candidate will be responsible for providing an electron microscopy service in support of the Institute's research programme and on a contract basis to commercial organisations. {/flushboth}
{flushboth} Candidates should possess a degree in an appropriate subject an= d be experienced in the analysis of a wide range of materials by scanning electron microscopy. {/flushboth}
{flushboth} Starting salary, depending upon qualifications and experience, will be normally within the range =A318,500 - =A321,400 per annum (performance related to a maximum of =A3 26,000). Non- contributory Superannuation Scheme. {/flushboth}
MLURI receives funding from the Scottish Executive Environment and Rural Affairs Department.
{flushboth} Further particulars and application forms can be obtained from Personnel, The Macaulay Land Use Research Institute, Craigiebuckler, Aberdeen, AB15 8QH Tel 01224-498200, Fax 01224- 311556, E-mail { HYPERLINK mailto:e.cockburn-at-mluri.sari.ac.uk } {underline} = {color} {param} 0000,0000,FF00 {/param} e.cockburn-at-macaulay.ac.uk {/underline} {= color} {param} 0100,0100,0100 {/param} to whom completed application forms must be returned by 20 July 2001. {/flushboth}
{center} {bold} {FontFamily} {param} Book Antiqua {/param} {bigger} {bigger} MACAU= LAY INSTITUTE {/bold} {color} {param} 0100,0100,0100 {/param} {smaller} {smaller} = {/center}
{center} {bold} {bigger} {bigger} ANALYTICAL GROUP {/center}
{center} ELECTRON MICROSCOPIST {/center}
{flushboth} {/bold} {smaller} {smaller} Applications are invited from suitably= qualified and experienced candidates for the above post. The successful candidate will be responsible for providing an electron microscopy service in support of the Institute's research programme and on a contract basis to commercial organisations. {/flushboth}
{flushboth} Candidates should possess a degree in an appropriate subject an= d be experienced in the analysis of a wide range of materials by scanning electron microscopy. {/flushboth}
{flushboth} Starting salary, depending upon qualifications and experience, will be normally within the range =A318,500 - =A321,400 per annum (performance related to a maximum of =A3 26,000). Non- contributory Superannuation Scheme. {/flushboth}
MLURI receives funding from the Scottish Executive Environment and Rural Affairs Department.
{flushboth} Further particulars and application forms can be obtained from Personnel, The Macaulay Land Use Research Institute, Craigiebuckler, Aberdeen, AB15 8QH Tel 01224-498200, Fax 01224- 311556, E-mail { HYPERLINK mailto:e.cockburn-at-mluri.sari.ac.uk {FontFamily} = {param} Times New Roman {/param} } {underline} {color} {param} 0000,0000,FF00 {/pa= ram} {FontFamily} {param} Book Antiqua {/param} e.cockburn-at-macaulay.ac.uk {/unde= rline} {color} {param} 0100,0100,0100 {/param} to whom completed application forms must be returned by 20 July 2001. {/flushboth}
Quote Ref MA17/01
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Rose wrote: } Hi! } I have a question regarding the removal of Spurrs resin from sections on } slides. I want to use aniline blue as a callose stain for plant tissue. I } have sections (embedded in Spurrs) on slides. The protocol I have to } remove the resin uses sodium methoxide and benzene (Mayor, Hampton and } Rosario, 1961). I was wondering if anyone can suggest a protocol that is } less dangerous than using the sodium methoxide. Or, alternatively, has } anyone used this technique, or a similar one and had any problems/success } with it? } Thanks, } Rose
Rose, I use sodium ethoxy (a saturated NaOH solution in absolute ethanol) for the Spurr removal in plant tissues. Use 5 minutes for a semithin section with 0.35 micrometers. Wash shortly in ethanol and, after, distilled water. The technique gives good results and it does not use methanol (dangerous to your health). I think that the protocol can be used by you without problems. Dr. Rinaldo Pires dos Santos E-mail: rinaldop-at-uol.com.br Plant Anatomy Laboratory - Dept. of Botany UFRGS - Porto Alegre - RS Brazil
Applications are invited from suitably qualified and experienced candidates for the above post. The successful candidate will be responsible for providing an electron microscopy service in support of the Institute's research programme and on a contract basis to commercial organisations.
Candidates should possess a degree in an appropriate subject and be experienced in the analysis of a wide range of materials by scanning electron microscopy.
Starting salary, depending upon qualifications and experience, will be normally within the range £18,500 - £21,400 per annum (performance related to a maximum of £ 26,000). Non- contributory Superannuation Scheme.
MLURI receives funding from the Scottish Executive Environment and Rural Affairs Department.
Further particulars and application forms can be obtained from Personnel, The Macaulay Land Use Research Institute, Craigiebuckler, Aberdeen, AB15 8QH Tel 01224-498200, Fax 01224- 311556, E-mail { HYPERLINK mailto:e.cockburn-at-mluri.sari.ac.uk }e.cockburn-at-macaulay.ac.uk to whom completed application forms must be returned by 20 July 2001. Quote Ref MA17/01 Web Site { HYPERLINK http://www.mluri.sari.ac.uk }http://www.macaulay.ac.uk
I am going to wade in on this, despite the potential for being said to be beating a dead horse:
During a graduate virology course a few years ago it became apparent that one of our students was pregnant. Out of concern for her and her child, we took special interest in providing information to her, and reducing her exposure to chemicals. We run a safe ship, but everyone should acknowledge that during gestation a developing child is vulnerable to things the mother may be exposed to. In this case, the students worked in pairs, and it was a simple matter the have her partner handle chemicals of concern. Since she was a new graduate student, we felt it was responsible to provide her with all the information we could for her safety and her child's. Her learning experience was not hindered, and on a personal level I think she appreciated our concern.
Information and education are critical to learning to make responsible decisions regarding safety, pregnant or not.
-----Original Message----- } From: Chris Jeffree [mailto:c.jeffree-at-ed.ac.uk] Sent: Tuesday, July 17, 2001 7:38 PM To: Debby Sherman Cc: microscopy-at-sparc5.microscopy.com
Hear hear!
} Safety is everyone's concern regardless of sex or biological conditions. Whatever policies are in place should be there for everyone, not just that occasional special situation. To have a lab manager or supervisor demand private medical information about users is most certainly an invasion of privacy. } Debby } } Debby Sherman Phone: 765-494-6666 } Life Science Microscopy Facility FAX: 765-494-5896 } Purdue University E-mail: dsherman-at-purdue.edu } S-099e Whistler Building } West Lafayette, IN 47907
Your protocol is for immunoEM with perfusion-fixation. I think perfusion-fixation is necessary for your experiment. But if your purpose is to see the ultrastructure, you should use 2% glutaraldehyde in stead of 4% paraformaldehyde as the fixative for the primary fixation. Your protocol for the perfusion is okay. Just try gluta.
Greg Ning
Gang Ning, M.D., Ph.D. Director EM Facility Medical College of Wisconsin
Meghan Towner wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello- } I am a grad student at the University of Idaho and would like some input on a protocol for fixing rat tissue in order to view their spinal ganglia on the TEM. First heparin is injected intracardiac, then perfusion via the heart with saline (37C) followed by paraformaldehyde (4%, 15C). The vertebral column is then removed and immersed in 4% paraformaldehyde for several hours before dissecting the spinal ganglia. This protocol was given to me by my advisor, but I have been unable to find a similar method cited in the literature. Also, I have used this method and found all the mitochondria have already lost their cristae. Any suggestions and comments would be appreciated. } Thank you. } --Meghan Towner
Your solution depends on what type of TiO2 you have and the particle size of the TiO2 you want to disperse. It can depend on whether you want to see ultimates, agglomerates or aggregates (primary structural units). You gave no indication of the type of TiO2 you want to disperse or if it was milled.
R900, 901, 100, &101 Dupont equivalents will have to be dispersed differently from something like micronized TiO2 (very agglomerated) or something like Nanotech TiO2.
Dispersion can depend on the TiO2 being rutile (I assume this is your type (coated)), anatase (uncoated), or a mixture of both, like Nanotech's spherical TiO2.
The technique we used for TiO2, and still use, was written up and given at the University of Michigan to the president of JEOL (Dr. Ito) and Prof. Wil Bigelow of the University of Michigan in the 1960s.
The presentation was given in the about 1963? by Robert Fear and was called A Versatile Mulling Technique (or Method) for Dispersing Fine Powders (or Pigments). Bob says it was sent to the EMSA / MSA educational archives. It was complete with colored 35mm slides. We have no idea where this material is currently at the MSA.
Paul Beauregard / Robert Fear Sr. Research Associate PPG Industries Monroeville Technical Center Monroeville, PA 15601
Any opinions are ours and not those of PPG Industries as a whole.
-----Original Message----- } From: Christopher M May [mailto:cmmay-at-US.ibm.com] Sent: Tuesday, July 17, 2001 12:52 PM To: microscopy-at-sparc5.microscopy.com
Hello,
I am working with a TEM and I would like to know the best way to disperse Titanium Oxide particles for analysis. I am currently using a methanol solution, and it is not working very well.
Good morning, I am planning to complete the microscopical characterization of some selected waxes using a hot stage on a light mcroscope. There does not seem to be much information in the literature on this subject and I thought there must be some microscopists well acqainted with this topic. We have completed DSC thermograms and other than looking for melting/crystallization temperatures, degree of crystallinity, is there anything else to be characterized? Can anyone provide further guidance? Thanks.
Paul
Paul J. Gerroir Microscopy Materials Characterization Xerox Research Centre of Canada 2660 Speakman Drive Mississauga, Ontario L5K 2L1
Readers, A question many may be asking, how do I get from the L.A. airport to Long Beach? I am advised that there is a (blue) bus system called SuperShuttle which leaves from outside the airline locations to Long Beach. Cost is $15 a head and it takes around a half hour and, typically, makes 3 stops. The taxi fare is around $45. Don Grimes, Microscopy Today
I don't understand Greg's comment (below). Power DTSA v 2.5.1, with the MSA 1.0 plug-in, will happily read EDAX Phoenix MSA files, which have a single column of spectral data as integers, and Link ISIS MSA files, which have the Y-data in 5 columns of reals. There is a tag, #DATATYPE, which, if set to "Y", tells the system to expect just the y-data, and not data pairs.
The problem with MSA files is that the MSA file specification is not a "format" - i.e., two people writing the same data to an MSA file can end up with different files, each of which is completely according to the specification. The problem for the reader is to be able to handle all the different possibilities.
One way that you can sometimes use to force a file to read in is to use the target system to write an MSA file (which, presumably, it will read back in) using the same channel specifiacations (number of channels, range, etc) as the source spectrum. Then use a text editor to copy the header from the target spectrum to the source spectrum. Tags like the #NCOLUMNS tag can be edited as required. Even this doesn't work if the format of the data itself is different, or if the target system will only accept a limited range of options (e.g. #NPOINTS must be a power of 2 in some systems), and the source doesn't match the required option.
The MSA reader should, of course, at least exit gracefully if it can't accept a spectrum, but not all do!
Tony Garratt-Reed.
} } The NORAN EMSA files follow the specifications fully and support all } varieties of EMSA options available. The problem is that DTSA only reads } one type of EMSA file format (X,Y pairs of data) which is not the default } condition of the VOYAGER files. The default format for VOYAGER stores only } the channel data in 5 columns. The default file format for VANTAGE was } changed to be X,Y pairs which is consistant with DTSA. } } There is a solution for VOYAGER. You can save the EMSA format compatible } with DTSA by using the shell command: } emsa_io -x=1 -w={path to store to} -s={spectrum name to store} } example: emsa_io -x=1 -w=$HOME/spectra/myspec.emsa -s=spectrum1 } } When the next version of VOYAGER software is released the default condition } for the emsa_io program will be converted to the same as VANTAGE which will } make VOYAGER's default version of EMSA file format consistant with DTSA. } } Greg Fritz }
** 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 **
Hi Meghan, Try almost any of the introductory books on EM by Hayat, M.A.. Your library or advisor, or EM book shelf should have a copy. Most problems such as you describe are due to the failure to initiate fixation in a timely manner, the osmolarity of the vehicle, and the postfixation time and component sequence. Most in situ, perfusion fixations include a postfix with glutaraldehyde following the initial stabilization with HCHO (made FROM paraformaldehyde). Also, it is my experience that one can 'freeze' thin muscle bands with 4% HCHO at 4oC in 10 min. But one cannot satisfactorily FIX the material for subsequent light(via paraffin) or EM (via plastic) viewing in that time. Hayat appears to suggest 24hr postfix but also provides a 4hr fix to block technique, so almost anything, properly designed, can work. Hayat, in a small methods book (HAYAT, M.A., 1972. BASIC ELECTRON MICROSCOPY TECHNIQUES, VAN NOSTRAND REINHOLD, NEW YORK AND LONDON.) references a method by: [Schultz, RL and Case, NM (1970), "A modified aldehyde perfusion technique for preventing certain artifacts in electron microscopy of the central nervous system", J Microscopy, 92:69.] If you can't find either the book or the article, ask and I'll send the method (not brief!)
Good luck,
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: Meghan Towner } Sent: Tuesday, July 17, 2001 7:38 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: tissue preparation for TEM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello- } I am a grad student at the University of Idaho and would like some input } on a protocol for fixing rat tissue in order to view their spinal ganglia } on the TEM. First heparin is injected intracardiac, then perfusion via the } heart with saline (37C) followed by paraformaldehyde (4%, 15C). The } vertebral column is then removed and immersed in 4% paraformaldehyde for } several hours before dissecting the spinal ganglia. This protocol was } given to me by my advisor, but I have been unable to find a similar method } cited in the literature. Also, I have used this method and found all the } mitochondria have already lost their cristae. Any suggestions and comments } would be appreciated. } Thank you. } --Meghan Towner } }
After fooling with EELS data stored in EMSA format from a Gatan system for a while that was stored in rows of five of the Y-only format and trying to extract the data into Excel, I wrote my visual basic program to extract the data and then write the data back in x,y pairs so that I could very easily plot it in Excel. Then I went nuts in Visual Basic 6.0 and wrote the program to plot the data and play around with it. I mentioned it before; I will bring to M&M and make public.
Before I wrote the program, I tried to extract the data with the "rows of five" problem in Excel using macro programming, but gave up. The easiest way that I found to convert the data to single column data and then add the x data was to first use a word processor and get rid of the hard returns using search and replace and add the appropriate character to match the data separator in the row. I then removed the data separators and added a hard return. Then you can save as a text file and open the data file in Excel. You then parse the data correctly and move the column of data over by one. Then you use a series fill to generate the x data. You can then plot the data using Excel (or any other plotting program) to plot the x,y pairs.
If you want to, you can easily change the data headers in the EMSA data file to reflect the change in the way the data is now stored in the file. You can go to the EMMPDL library and find the file with the standard and figure out how to do it relatively easily.
I think that the biggest problem with the EMSA format is allowing the Y-only data to exist in rows of n values. However, I understand why that was done. For the older programming languages, it was easy to format the read/write statement to bring the data in/out that way. However, it is not a major stumbling block.
-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: Anthony J. Garratt-Reed [mailto:tonygr-at-mit.edu] Sent: Wednesday, July 18, 2001 11:32 AM To: Fritz, Greg; microscopy-at-sparc5.microscopy.com
I don't understand Greg's comment (below). Power DTSA v 2.5.1, with the MSA 1.0 plug-in, will happily read EDAX Phoenix MSA files, which have a single column of spectral data as integers, and Link ISIS MSA files, which have the Y-data in 5 columns of reals. There is a tag, #DATATYPE, which, if set to "Y", tells the system to expect just the y-data, and not data pairs.
The problem with MSA files is that the MSA file specification is not a "format" - i.e., two people writing the same data to an MSA file can end up with different files, each of which is completely according to the specification. The problem for the reader is to be able to handle all the different possibilities.
One way that you can sometimes use to force a file to read in is to use the target system to write an MSA file (which, presumably, it will read back in) using the same channel specifiacations (number of channels, range, etc) as the source spectrum. Then use a text editor to copy the header from the target spectrum to the source spectrum. Tags like the #NCOLUMNS tag can be edited as required. Even this doesn't work if the format of the data itself is different, or if the target system will only accept a limited range of options (e.g. #NPOINTS must be a power of 2 in some systems), and the source doesn't match the required option.
The MSA reader should, of course, at least exit gracefully if it can't accept a spectrum, but not all do!
Tony Garratt-Reed.
} } The NORAN EMSA files follow the specifications fully and support all } varieties of EMSA options available. The problem is that DTSA only reads } one type of EMSA file format (X,Y pairs of data) which is not the default } condition of the VOYAGER files. The default format for VOYAGER stores only } the channel data in 5 columns. The default file format for VANTAGE was } changed to be X,Y pairs which is consistant with DTSA. } } There is a solution for VOYAGER. You can save the EMSA format compatible } with DTSA by using the shell command: } emsa_io -x=1 -w={path to store to} -s={spectrum name to store} } example: emsa_io -x=1 -w=$HOME/spectra/myspec.emsa -s=spectrum1 } } When the next version of VOYAGER software is released the default condition } for the emsa_io program will be converted to the same as VANTAGE which will } make VOYAGER's default version of EMSA file format consistant with DTSA. } } Greg Fritz }
** 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 **
Just a brief note to inform the microscopy community of the death of Dr. Lonnie Russell on July 11. Dr. Russell drowned while swimming in the ocean off the coast of Brazil. Lonnie made significant contributions in microscopy (both light and electron) and was an internationally recognized expert in the area of reproductive biology. I have more information about Lonnie, so please contact me personally and I will forward it to you.
-- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ##############################################################
} The problem with MSA files is that the MSA file specification is not a } "format" - i.e., two people writing the same data to an MSA file can end up } with different files, each of which is completely according to the } specification. The problem for the reader is to be able to handle all the } different possibilities. }
The problem we have in implementing a robust MSA file reader is lack of example files from the various MSA file writers. Because there are several flavors in implimenting the writer, many programmers take different paths which should not make a difference to a MSA file reader. When programming the MSA file writer, one sometimes does not think of the other possibilities. It's only when users complain that they cannot read a MSA file from some other system that the problems become known. Then resources have to be allocated to fix a problem that should have been found during the testing phase of the software. Does not seem to be hard but this can be costly (from a company viewpoint).
Something that would help this oversight would be a repository for the various MSA file flavors, something one could download to use as a test case for the robustness of their MSA file reader. The spectral content could be anything but should be real (AlCu calibration for example).
We would be happy to sponser such a location on our web site.
Scott
----------------------------------------------------------------------- Scott D. Davilla Phone: 919 489-1757 ext 13 (tel) 4pi Analysis, Inc. Fax: 919 489-1487 (fax) 3500 Westgate Drive, Suite 403 email: davilla-at-4pi.com Durham, North Carolina 27707-2534 web: http://www.4pi.com
I thought I would forward an example of the kind of madness to which one may be driven by the need to avoid liability in the workplace. The gentleman who formulated the list at the base of this forwarded email had earlier requested information about how others handled pregnancy among workers in electron microscopy laboratories. He then reported his solution as you can see below.
I am inserting without comment a statement from our lawyers, Dumb, Dumber and Dumbest, Esquires, LTD., LLD, LLC, LLB, LLA, LZ, LY, which they returned in response to a question about my potential liability on this matter.
"Understanding that most exposures to hazard occur as a consequence of accident or personal negligence, we have come to the conclusion that it is impossible, if not illegal, to protect anyone in particular, much less someone who is specifically pregnant, from exposure to hazardous materials, especially if the individual is going to take a job that involves such hazard and continue to do THAT job during pregnancy. Thus, it behooves the employer to remove such hazards completely or to reduce them for those at greatest risk so that there is no risk to all, especially those who periodically enter periods of existence during which their risk increases. However, we must state that for us this is a matter of humane treatment of subordinates, especially those weak in mind if not in body, not the law, which in some measure is inscrutable, even to us.
The legal intricacies of this matter in particular border on the insane, and force us to adopt unusual and novel measures and reasoning to protect both the employer and its employees. It appears that the Supreme Court in one of its more enlightened decisions, ruled that a company may NOT discriminate among its employees in the matter of hazardous substance risk. Thus, a battery manufacturer was not permitted to protect pregnant employees by transfer away from lead, when it did not similarly protect the remainder of the work force. In fact, this ruling is quite reasonable given the fact that during the first trimester of pregnancy, the Supreme Court had already ruled that pregnancy, especially during the period mentioned, is in NO respect the business of Society (nor, by any extension, management).
Therefore, if during the first trimester, an accident might occur to damage that which is none of Society's business, there is no recourse set aside for management that can protect that which is none of ITS business during that period. We can all, however, assure the employer that should a pregnant employee decide to continue a pregnancy beyond the first trimester, the employer becomes wholly responsible for any damage that occurred to the object of the pregnancy during the time that the object of the pregnancy was none of its business. It is a quirk of this situation that the object of the pregnancy has no legal standing either during this period, when it is none of Society's or management's business, or, for that matter, during the entire duration of pregnancy. Thus, the employer is uniquely at risk of inflicting damage on an object with no legal standing during a period when that object is none of the employer's business and, legally, does not exist. Of course, one might argue, just as logically, that it is not possible to inflict harm on something that does not exist, but that avoids the practical reality. We continue to expand this reasoning for the purposes of expected appeals should our position prove, in the first instance, legally unreasonable (charges, as stipulated, per hour). The only recourse may be a standard agreement to be signed by ALL employees, that ANY who become pregnant (through physical or ethereal means, regardless of anatomy or lack thereof exhibited by themselves or others related or unrelated to the acts involved or uninvolved, philosophy, political party, voting record, religion, non-religion, sexual disposition or preference, or sexual non-preference or non-disposition, or any absence or presence of preference or disposition and notwithstanding the expected or unexpected effects of genetic predisposition or specific incapacity or capacity) will agree to be transferred to non-hazardous, riskless, activities for the course of that pregnancy, nominally nine months from the moment of conception. If this solution is adopted by the employer, then the employer can be assured that such actions will protect those offspring of employees who have or will have legal standing and for whose well-being the employer can ever be held responsible, if someone decides that such well-being was or is the employer's business.
Since discrimination against the parent is not permitted, it is obvious that all individual distinctions must be suppressed in any safety considerations and actions. For those who refuse such PROTECTION during pregnancy, patently hazardous employment should not be available at any time during which pregnancy (through physical or(see above)....), may or might be improbably possible. The other side of the coin is that all employees, regardless of job, must be treated identically. Thus once safety considerations have been installed in equal consideration of all, all employees will be able and free to enter, with equanimity, their job space by first travelling through those non-discriminating, hazardous, but equally safe and non-hazardous work spaces.
The reasoning for the above solution is simple and straight forward. Since there is no absolute evidence to suggest that true hermaphroditism is not possible in humans, it is reasonable for the employer to subject ALL employees to the above non-discriminatory employment process during the pre-employment interview as well as after employment. The employer, thus, has assumed the responsibility to dictate what it considers hazardous and potentially harmful to this broad group of individual employees. This is reasonable since the employer is at risk for damage to the potential offspring during times when the potentially pregnant employee may bring the potential offspring to the place of employment when such pregnancy is none of the employer's business (but when the employer's potential risk is real). It is not that the employer has rights nor can be said to have responsibility during the time of pregnancy when such pregnancy is none of its business, but the employer can claim the right to protect itself, when the potential offspring, during the time that when its existence is none of the employer's business, is exposed to hazards on the employer's property that may cause harm to the potential offspring when the parent decides to continue the pregnancy beyond that period when it is none of Society's or any employer's business, such that the potential offspring is permitted to continue to develop in such manner that it can potentially claim rights of redress for harm done during that time of its non-existence when it was exposed to hazard when such exposure, by simple logic, was none of the employer's business, and concurrently could not have happened."
It should be clear that the solution recommended by our legal team treats all potential employees in the same potential manner, giving all their potential rights according to Supreme Court ruling and protecting all equally without potential discrimination. Thus, reason, not chaos, will prevail!
Please pardon any errors in the above. I have not had time to review it since it was returned, a second time, edited for clarity and brevity, from the lawyer's office.
Since I cannot, in good conscience, ignore the above reality, I am obliged to pass this information on to you, my superiors, who will also be forced to become 'part of the problem' rather than 'part of the solution'. My apologies to all - indiscriminately! To those pregnant employees who insist that hazardous work must continue without hazard and, thus, constrain their employers to liquidate the workplace, I accord you the sensitivity and concern that I normally reserve for the incoherent. I may wonder what you are saying, but I never try to understand what you are thinking.
Fred Monson, who wouldn't suffer his wife, and thus, their unborn children, to work in the same business as he, but whose daughter has freely chosen it.
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: William McManus } Sent: Tuesday, July 10, 2001 11:30 AM } To: Leona Cohen-Gould; Monson, Frederick C.; Sally Stowe } Cc: Microscopy Listserver } Subject: lab pregnancy } } to all out there: } } This is what I have done so far: } 1. Removed all stored chemicals from the lab. } 2. Moved all chemical mixing into fume hood. } 3. Met for 3 hours with environmental safety officer. } 4. Met for 2 hours with radiation safety officer. } 5. Eliminated all protocols which involve teratogens from pregnant } workers routine. } 6. Established a personal hygiene plan for pregnant worker, which is } being signed of by worker, myself, director of the lab, safety } officer, and her physician. } 7. Reiterated safety protocols for worker, coat , gloves, safety } glasses, etc. } 8. Had lab monitored for radiation, passed. } 9. Had lab monitored for formaldehyde gas, passed. } 10. Planning to monitor xylene exposure to worker when thin sectioning } (cannot do this in the hood, obviously). Heat pen( modified } soldering } iron) has not worked well so far, any ideas how to eliminate } xylene? } 11. Still looking for a non toxic general 1 micron stain for Spurrs and } LR White. Any ideas? } I am experimenting with food related stains (grape juice, etc.). } 12. Employee has completed, lab safety training course, formaldehyde } training course and radiation training course. } 13. Will continue to monitor lab for chemical and radation exposure. } } To some this may seem like over kill, but I believe that it is very } important that every stone is turned in regards to safety. Another } issue that I face is that I have numerous grad students coming through } the lab, and there is always a possibility that they are pregnant and do } not realize it or keep it from me. I also have a nursing mother who was } visiting the lab and never bothered to tell me, until the subject of why } the lab was reorganized came up. I now demand that all female student, } techs, visitors, ect. } tell me of their "motherhood" status. } } Bill } } -----Original Message----- } From: Leona Cohen-Gould [mailto:lcgould-at-med.cornell.edu] } Sent: Tuesday, July 10, 2001 7:28 AM } To: William McManus; Monson, Frederick C.; Sally Stowe } Cc: Microscopy Listserver } Subject: RE: Tol blue/borax } } } At 1:04 PM -0600 7/9/01, William McManus wrote: } } ----------------------------------------------------------------------- } - } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ----------------------------------------------------------------------- } . } } } } } } The issue of pregnant (or possibly pregnant) workers in the lab has } } turned out to be fairly complicated. If anyone else out there has beem } } through this situation, I would like to know how you dealt with it. } } } } Bill } } } } William McManus } } Supervisor } } Electron Microscopy Facility } } Department of Biology } } Utah State University } } Logan UT 84322-5305 } } } **************** } to Bill & All, } } When I was pregnant, 9 years ago, I was the entire staff of the EM } facility here (still was until a month ago)and so could not avoid } things. The way I handled safety concerns was to be extra cautious: } I worked in the hood, wore double gloves when working with fixatives } & resins and gloves for other times when i would not usually wear } them( like using Tol Blue), safety glasses if splashing was a } possibility, washed my hands almost compulsively. In short, I used } common sense. I was, and am, lucky that (UrAc aside) I don't have to } deal with radioactivity. colleagues of mine who were also pregnant } at the time (we had a rash of pregnancies that year) who did need to } work with isotopes minimized their use, got new shields, and at times } convinced labmates to do those steps for them! } } We all, thankfully had healthy babies who are now all approaching } adolescence and driving us nuts, but that's another issue for another } forum! } } Lee } } -- } Leona Cohen-Gould, M.S. } Sr. Staff Associate } Director, Electron Microscopy Core Facility } Manager, Optical Microscopy Core Facility } Joan & Sanford I. Weill Medical College } of Cornell University } voice (212)746-6146 } fax (212)746-8175 } }
Scott, I have a couple of them on my disk and that is included in my program distribution files. I am going to put it in the MSA and MAS libraries. I am bringing it to M&M and I will give you a copy. They include Gatan, Noran, Emispec, and one other that I can't think of at the moment (I think that it was Oxford).
I had asked for examples and I got a few to try in the program.
If anyone would like to send me one of their EMSA formatted ASCII output EDS or EELS spectra files and tell me the vendor associated with source, I could include them with the distribution files.
-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: Scott D. Davilla [mailto:davilla-at-4pi.com] Sent: Wednesday, July 18, 2001 2:25 PM To: Microscopy (E-mail)
} The problem with MSA files is that the MSA file specification is not a } "format" - i.e., two people writing the same data to an MSA file can end up } with different files, each of which is completely according to the } specification. The problem for the reader is to be able to handle all the } different possibilities. }
The problem we have in implementing a robust MSA file reader is lack of example files from the various MSA file writers. Because there are several flavors in implimenting the writer, many programmers take different paths which should not make a difference to a MSA file reader. When programming the MSA file writer, one sometimes does not think of the other possibilities. It's only when users complain that they cannot read a MSA file from some other system that the problems become known. Then resources have to be allocated to fix a problem that should have been found during the testing phase of the software. Does not seem to be hard but this can be costly (from a company viewpoint).
Something that would help this oversight would be a repository for the various MSA file flavors, something one could download to use as a test case for the robustness of their MSA file reader. The spectral content could be anything but should be real (AlCu calibration for example).
We would be happy to sponser such a location on our web site.
Scott
----------------------------------------------------------------------- Scott D. Davilla Phone: 919 489-1757 ext 13 (tel) 4pi Analysis, Inc. Fax: 919 489-1487 (fax) 3500 Westgate Drive, Suite 403 email: davilla-at-4pi.com Durham, North Carolina 27707-2534 web: http://www.4pi.com
} Meghan Towner wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } Hello- } } I am a grad student at the University of Idaho and would like some input on a protocol for fixing rat tissue in order to view their spinal ganglia on the TEM. First heparin is injected intracardiac, then perfusion via the heart with saline (37C) followed by paraformaldehyde (4%, 15C). The vertebral column is then removed and immersed in 4% paraformaldehyde for several hours before dissecting the spinal ganglia. This protocol was given to me by my advisor, but I have been unable to find a similar method cited in the literature. Also, I have used this method and found all the mitochondria have already lost their cristae. Any suggestions and comments would be appreciated. } } Thank you. } } --Meghan Towner
If you want good ultrastructure use gluatarladehyde in the fixative, as little as 1% will make a significant improvement but 2-3% is more common. The buffer used is also important, you did not specify which you are using. Paraformaldehyed alone can give good ultrastructure but you will need to fix for several days at a minimum. If you want to do immunostaining, you may have to skip the glut. and suffer with less than optimal ultrastructure or risk killing off the immunoreactivity. It will depend on the antigen in question. Also, the temperature shift from 37 degrees C for the washout to 15 degrees C for the fixative is likely to induce vasospasm, reducing the amount of fixative delivered to the tissue. Do everything at one temperature, room temperature should be fine. A good basic EM text will be invaluable. Brenda Weakley's "Beginners handbook in biological transmission electron microscopy" is excellent. "Biological techniques in electron microscopy" by Clinton Dawes is also very good but it may be out of print.
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 **********************************************
} Safety is everyone's concern regardless of sex or biological conditions. Whatever policies are in place should be there for everyone, not just that occasional special situation. To have a lab manager or supervisor demand private medical information about users is most certainly an invasion of privacy. } Debby
While Debby's basic premise is correct, it is also a fact that a fetus can be adversely affected by chemicals, or levels of chemicals, that would pose absolutely no threat to adults. I have worked in situations where implementation of all the precautions necessary to prevent such fetal exposure would have made the job impossible. We acknowledge these differences when approving new drugs for the market, and restrict their usage accordingly. Is the lab situation so very different?
In laboratories where large numbers of EM micrographs are produced over the years and researchers may come and go over time, what system do you use to keep track of the micrographs?
We were thinking of implementing a computer database where information about each micrograph (such as date, microscopist, sample, conditions, location of the archived micrograph, etc.) can be stored for future reference. Is there any existing database software (preferably Macintosh based) for this purpose or any opinions on such packages?
Thanks for your help.
- Dennis
------------------------------------------------------------------------ Dennis C. Winkler, PhD. Phone: (301) 496-0131 Laboratory of Structural Biology Research Fax: (301) 480-7629 NIAMS, National Institutes of Health Email: Dennis_Winkler-at-nih.gov Bldg. 6, Room B2-26, MSC-2717 Bethesda, MD 20892-2717, U.S.A.
Try Thumbsplus from Cerious software. (www.cerious.com) PC based. I don't know if they ever came out with the Mac based version, but they did have a beta version at one time.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Dennis C. Winkler [mailto:Dennis_Winkler-at-nih.gov] Sent: Wednesday, July 18, 2001 5:03 PM To: Microscopy-at-sparc5.microscopy.com
Good Day All!
In laboratories where large numbers of EM micrographs are produced over the years and researchers may come and go over time, what system do you use to keep track of the micrographs?
We were thinking of implementing a computer database where information about each micrograph (such as date, microscopist, sample, conditions, location of the archived micrograph, etc.) can be stored for future reference. Is there any existing database software (preferably Macintosh based) for this purpose or any opinions on such packages?
Thanks for your help.
- Dennis
------------------------------------------------------------------------ Dennis C. Winkler, PhD. Phone: (301) 496-0131 Laboratory of Structural Biology Research Fax: (301) 480-7629 NIAMS, National Institutes of Health Email: Dennis_Winkler-at-nih.gov Bldg. 6, Room B2-26, MSC-2717 Bethesda, MD 20892-2717, U.S.A.
Thanks for your input, but let me clarify. I am working with Nanotech TiO2, I am looking at sizes 100nm and smaller. The particles are spherical and what I would eventually like are measurements of their diameters which reflects as accurate a distribution as I can get. I do not want aggolmerates, just the particles. Therefore, any technique that has a selection bias I would rather not use.
Thanks again, Christopher May IBM Analytical Services
----- Forwarded by Christopher M May/Poughkeepsie/IBM on 07/18/2001 11:38 AM -----
"Christopher M May" To: microscopy-at-sparc5.microscopy.com {cmmay-at-US.ibm. cc: com} Subject: Titanium Particle dispersion for TEM
07/17/2001 12:51 PM
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello,
I am working with a TEM and I would like to know the best way to disperse Titanium Oxide particles for analysis. I am currently using a methanol solution, and it is not working very well.
My name is Professor Sabel. Whoever you are, your email is being automatically sent to my account. I think you have a big problem somewhere. Please check.
please find below the announcement for a long-term EELS post-doc position at the group of Electron Microscopy for Materials
Science (EMAT) in Antwerp, Belgium http://www.ruca.ua.ac.be/emat/. May I ask you to distribute this to colleagues that may be interested.
Many thanks,
Nick Schryvers
EELS Post-doc position at EMAT, Antwerp, Belgium
A new post-doctoral position will be available as of January 1, 2002, at
the laboratory of Electron Microscopy for Materials Science (EMAT) in Antwerp, Belgium. The emphasis of the work will be on the combination of
EELS with HRTEM, STEM and EFTEM with the aim to develop advanced detection, processing and interpretation procedures yielding improved resolutions. The work will be performed in the framework of a joint program including a research group on image processing (VISIE) and one specialised in spectral analysis techniques (MiTAC), both from the same university. The ideal candidate should have an extensive experience in performing high precision EELS experiments in the TEM. The experiments will be conducted with two EELS systems (GIF 200(0)), one installed on a
CM30 FEG Ultratwin, designed for ultimate resolution of 0.1nm (using Focus Variation) and another one on a 3000F JEOL FEG machine equipped with a STEM unit with a probe size of 0.4 nm, EDX and FasTEM. At first model systems in the field of CMR and superconducting materials, semiconductor precipitates and diamond like carbon films will be investigated, but the aim is for generic principles useful for a large majority of materials. The position is in principle for one year, but can be extended by one year periods up to a total of 4 years. Salary is arranged following European and local standards and includes the full range of social security benefits (health care, retirement, holiday, …). Actual figures depend on proven experience of the candidate. Please send your extensive resumé to
Prof. Dr. Dominique Schryvers Senior Lecturer Electron Microscopy for Materials Research (EMAT) Department of Physics University of Antwerp, RUCA Groenenborgerlaan 171 B-2020 Antwerp, Belgium Tel: 32-3-2180247 Fax: 32-3-2180257 E-mail: schryver-at-ruca.ua.ac.be, nick.schryvers-at-ua.ac.be
ThumbsPlus is good if you have a digital image to keep track of, but I am not sure if Dennis was referring to that or to physical micrographs. If it is the latter then I would suggest using Microsoft Access and setting up their own forms and reports to suit their particular needs. Greg Erdos
At 05:32 PM 7/18/2001 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Gregory W. Erdos, Ph.D. Assistant Director, Biotechnology Program Scientific Ditrector, ICBR EM CORE University of Florida Ph. 352-392-1295 PO Box 118525 Fax 352-846-0251 Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl
Can anyone advise on removing a coverslip mounted using DPX? I am currently soaking it in xylene, but overnight didn't "cut the mustard"! Any cogent comments? Please?!
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I think the key here is (from the post below):
"No one should be allowed to expose themselves or others to dangers and it is THEIR RESPONSIBILITY to provide data to their managers if such a case exists or might exist."
If someone makes a medical condition known, ANY medical condition, to their supervisor, then it seems that the supervisor should take appropiraite additional action OVER AND ABOVE THE ROUTINE SAFETY PRECAUSTIONS to provide additional information and safety as required.
However, the question is not if we should attempt to make an environment as safe as possible (that does depend on specific individual need and may or may not be sufficiently possible in all situations) but whether a supervisor has the right to demand medical information from someone who may not be willing to share it. Do we have the right to not only demand the information but, upon hearing it, require the employee to observe restrictions when they, after being fully informed of hazards, etc, do not wish to do so.
Also, although this thread has dwelt on pregnancy as the issue of concern, it does take two to tango and concern for healthy sperm is as important as healthy eggs. Do we really know effects, limits, etc on all potential hazards in our labs to these tissues as well as to fetuses?
We can discuss this all day but, not being legal professionals, our thoughts are not necessarily sufficient in a court of law. I think it behooves all of us to provide as safe an environment as possible within reason and then respond to special situations in a way that is agreeable to all parties involved. This may require restrictions or reassignment of personnal if the environment cannot be made as safe an an employee/employer feels necessary (which will no doubt raise all kinds of additional questions/concerns). We are our brother's keepers only to the extent that they want to be kept!
Debby
Debby Sherman Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-099e Whistler Building West Lafayette, IN 47907
--------------------------------------
Dennis,
You've probably already seen the 'Access' part of the MS Office programs as a database, but if not, I use it to keep track of the admin things in our microscopy unit and it works well, has lots of features. Another program that could be useful, although not exactly what you're looking for, is IrfanView, which lets you open up fast all the images you have in a folder, like a contact print of photos - later you can do things like copy, delete etc. on the files. It's freeware and you can download at
Both of these I've found really useful in general organization of the data and files that end up accumulating everywhere.
Good luck!
Mark
******************************************** Mark West, Encargado, (Manager) Unidad de Microscopia(Microscopy Unit), Lab, 301-Sur, Instituto de Fisiologia Celular, Universidad Nacional Autonoma de Mexico, Circuito Exterior, Ciudad Universitaria, Coyoacan, C.P. 04510, Mexico D.F., MEXICO
Pagina de Web (Web page) http://ifcsun1.ifisiol.unam.mx/microscopy.html ********************************************
On Wed, 18 Jul 2001, Dennis C. Winkler wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Good Day All! } } In laboratories where large numbers of EM micrographs are produced } over the years and researchers may come and go over time, what system } do you use to keep track of the micrographs? } } We were thinking of implementing a computer database where } information about each micrograph (such as date, microscopist, } sample, conditions, location of the archived micrograph, etc.) can be } stored for future reference. Is there any existing database software } (preferably Macintosh based) for this purpose or any opinions on such } packages? } } Thanks for your help. } } - Dennis } } ------------------------------------------------------------------------ } Dennis C. Winkler, PhD. Phone: (301) 496-0131 } Laboratory of Structural Biology Research Fax: (301) 480-7629 } NIAMS, National Institutes of Health Email: Dennis_Winkler-at-nih.gov } Bldg. 6, Room B2-26, MSC-2717 } Bethesda, MD 20892-2717, U.S.A. }
I have a JEOL 6320 FEGSEM equipped with a Oxford CT1500 cryo unit which I have used for a couple of years. I recently replaced the Cr foil/al back plate target with a Cr sputtered on aluminum target. Now I get a plasma but no deposition of Cr. (And yes, the target is facing the right direction). Yesterday, I replaced a leaky argon supply valve and metering valve. Still no deposition. I have also tried a full range of current/back pressure settings with no luck. Anyone have any ideas as to what the problem is?
The work is piling up so I am also looking for access to a FEGSEM with a CT1500 or CT2500 cryo unit equipped with a Cr target to get some high priority / high resolution work done. (Preferably somewhere here in the mid-west). Please contact me with the appropriate information.
Thanks, Dave Williams The Dow Chemical Company Midland, MI dmwilliams-at-dow.com
Thanks for all the responses to my search for child care options in Long Beach. For those who wanted to know (and anyone else who is also interested), here's what I found:
The Westin can arrange babystitting, estimated $ 15 - $ 20 per hour.
The Growing Years (1 World Trade Ctr # 199) was recommended. It has been re-named, but can look after children up to 5, telephone (562) 495-6300 (unfortunately not good for me - my child is 6)
Affordable Nannies & Care Givers, (562) 988-6588, is an agency which provides babysitting; usual charges $ 8 - $ 10 per hour plus fee of $ 25 per day.
If you are YMCA members, you can sign up for a few days camp at the YMCA, 1720 N Bellflower Blvd, (562) 96-3394 - it's called 'Sunshine Camp" and costs $ 105 for 1 to 3 days, $ 140 for 4 or 5 days, plus $ 35 registration fee; you need to go there first (i.e. Saturday or earlier) with vaccination records etc.
Using switchboard (www.switchboard.com) to search for businesses (select "child care services" from the list) gave me about 65 hits. One or two day care businesses said they could provide a few days' care, but most only go up to age 5.
Thanks again,
Rick Powell
***************************************************************************************** Richard D. Powell * rpowell-at-nanoprobes.com * www.nanoprobes.com NANOPROBES, Incorporated 95 Horse Block Road, Yaphank, NY 11980-9710, USA
Sign up for our newsletter: http://www.nanoprobes.com/Newsletter.html *****************************************************************************************
Project MICRO (Microscopy In Curriculum - Research Outreach) is MSA's middle school educational outreach program. MICRO frequently sponsors programming at the annual M&M meeting - but because it doesn't fit the basic symposium structure, sometimes it "falls thru the cracks" in program preparation. That happened this year; the MICRO workshop isn't listed in the daily program. Although it's primarily intended for local teachers, it may interest you too.
Repairing School Microscopes: A Workshop for Teachers and Volunteers Tuesday, August 7 10 am - 3 pm, room 203C Led by David Bentley, University of Arizona
If you ask teachers what kind of help they need, a common response is "fix our microscopes"! In this context, "repair" means basic maintenance, rather than things requiring expert attention, and Dave has considerable experience in teaching the subject to teachers. If you want to help with educational outreach but you don't want to be a classroom volunteer, come to this workshop and learn how to help teachers in your local schools.
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
We have inherited a Leitz Metallux 3 microscope with epifluorescence, however, without a user's manual (not a complaint, just a fact) and are wondering whether anyone knows a source for old microscope manuals. There are some quirky things about it, for example, the transmitted light is either yellow or red, never white, and, other than dismantling the entire body, and physically removing the filters, I cannot find a way to fix that. I could take it apart but I'm sure there would be parts left over after reassembly.
Thanks for any help.
Judy Trogadis Bio-Imaging Coordinator St. Michael's Hospital, 8Q 30 Bond St. Toronto, ON M5B 1W8
OK, here you go -- SOP of TEM drop grid preparation for 100nm down or up TiO2 particles:
1) Dilute TiO2 solution to about 1% with DIW or other solvents. I have the luck to use straight H2O in most of the cases involving particles including TiO2. Surfactants sometimes cause more problems than resolve particles. Alcohol or acetone could be good solvents too 2) Sonicate the solution 3) Use a pair of negative tweezers, and pick up a 200 mesh Cu C-film grid WITH Cu-SIDE UP. You should catch as less edge of the Cu grid as possible. 4) Lay the tweezers on a table and place a quadrant of filter paper underneath the grid/tweezes. 5) Use a pipette to transfer a drop of the liquid onto the grid. Wick away extra liquid with half of that quadrant of the filter paper underneath the tweezes 6) Let it dry naturally or put under a light 7) Check with TEM 8) Later, experience will tell you how much concentration to use and how much liquid to drop onto the grid
********************************* Chaoying Ni Electron Microscopy Laboratory Materials Science and Engineering College of Engineering University of Delaware Newark, DE 19716 *********************************
On Wed, 18 Jul 2001, Christopher M May wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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: } } Thanks for your input, but let me clarify. I am working with Nanotech } TiO2, I am looking at sizes 100nm and smaller. The particles are spherical } and what I would eventually like are measurements of their diameters which } reflects as accurate a distribution as I can get. I do not want } aggolmerates, just the particles. Therefore, any technique that has a } selection bias I would rather not use. } } Thanks again, } Christopher May } IBM Analytical Services } } } ----- Forwarded by Christopher M May/Poughkeepsie/IBM on 07/18/2001 11:38 } AM ----- } } "Christopher M } May" To: microscopy-at-sparc5.microscopy.com } {cmmay-at-US.ibm. cc: } com} Subject: Titanium Particle dispersion for TEM } } 07/17/2001 } 12:51 PM } } } } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello, } } I am working with a TEM and I would like to know the best way to disperse } Titanium Oxide particles for analysis. } I am currently using a methanol solution, and it is not working very well. } } } Sincerely, } } Christopher May } IBM Analytical Services } } } } } } }
Clarification: Yes, I am talking about physical micrographs (film/negatives). Sorry about the confusion.
Thanks for all of the suggestions thus far. My first thought was to set up a FileMakerPro database to store the information, but I will look at the commercial packages that have been mentioned.
- Dennis
------------------------------------------------------------------------ Dennis C. Winkler, PhD. Phone: (301) 496-0131 Laboratory of Structural Biology Research Fax: (301) 480-7629 NIAMS, National Institutes of Health Email: Dennis_Winkler-at-nih.gov Bldg. 6, Room B2-26, MSC-2717 Bethesda, MD 20892-2717, U.S.A.
At 9:27 AM -0400 7/19/01, Greg Erdos wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } ThumbsPlus is good if you have a digital image to keep track of, but } I am not sure if Dennis was referring to that or to physical } micrographs. If it is the latter then I would suggest using } Microsoft Access and setting up their own forms and reports to suit } their particular needs. } Greg Erdos }
{x-charset windows-1252} } I have a JEOL 6320 FEGSEM equipped with a Oxford CT1500 cryo unit which I } have used for a couple of years. I recently replaced the Cr foil/al back } plate target with a Cr sputtered on aluminum target. Now I get a plasma but } no deposition of Cr. Yesterday, I replaced a leaky argon supply valve and } metering valve. Still no deposition. Anyone have any ideas as to what the } problem is? } } I am also looking for access to a FEGSEM with a CT1500 or CT2500 cryo unit } equipped with a Cr target to get some high priority work done. (Preferably } somewhere here in the mid-west). Please contact me with the appropriate } information. } Thanks, } } Dave Williams } dmwilliams-at-dow.com {/x-charset}
I am a graduate student at the university of Texas Health Science Center. As part of a project I wish to perform a spectral analysis of a certain medication. The medication is a crystalline powder in the form of small (approx. 1mm diameter) spheres. I want to section the spheres to 10 to 30 microns so as to obtain a spectral analysis of the entire bead from its outer coating to its interior. I have tried OCT embedding and cyrostat sectioning. However, the beads pulverized during sectioning. Can anyone recommend a way to section these spheres so as to preserve the material as much as possible? I should also point out that the material is freely soluble in ethanol and methanol, as well as water.
Thank you in advance for your help and suggestions.
Below is the result of your feedback form. It was submitted by (dkray-at-siu.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Thursday, July 19, 2001 at 14:53:44 ---------------------------------------------------------------------------
Email: dkray-at-siu.edu Name: Dave Krayesky
Organization: SIUC
Education: Graduate College
Location: Carbondale, IL 62901
Question: What is a flat field objective and how does it differ from other types of objectives one might use on a microscope?
Can anyone comment on the retention of copper (in dosage experiments) by "staining" with rubeanic acid? What is the complex seen in light microscopy? Would it be accessible to x-ray microanalysis? ......... after removing a coverslip!
Keith Ryan Marine Biological Association Plymouth UK
Following my posting regarding copper/rubeanic acid, does anyone have information about the "pyroantimonate" method. A visitor to my lab has used it (at my suggestion) in an attempt to retain copper in accumulation/stress experiments from a home-base where cryomethods were not possible.
After a week of looking, we see very little in thin/semi-thin sections with XRMA. It is a method where chemical fixation is combined with potassium pyroantimonate to complex and retain the copper (and any other cations?.
Keith Ryan Marine Biological Association Plymouth, UK
Steve Chapman (Protrain Ltd) will be in Australia this October to give courses at the Electron Microscopy Unit of the Australian National University in Canberra. There will be a total of 8 days covering SEM and TEM, divided into 4 modules -you may enrol in one or more modules. Courses will be taught by Steve with assistance from ANU EMU staff.
Brief description below: details and enrolment on www.anu.edu.au/EMU/workshops/01chapman.html
I. SEM MASTERCLASS Tuesday 16th to Thursday 18th October. Three days: AUS$600
II. Maintaining and Monitoring the SEM. Friday 19th October. Pre-requisite: completion of this or a recent SEM Masterclass, or equivalent. One day: AUS $250
III. TEM Maintenance . Monday 22nd October, Tuesday 23rd October. Pre-requisite: familarity with TEM operation. Two days: AUS $500
IV. TEM Operation. Wednesday 24th October, Thursday 25th October. Two days: AUS $400
Meals and accommodation not included.
See you there - your opportunity to learn about EM instrumentation in a very practical way, and have a great time. This will be the third time we have run Module I - so far the recedivist rate is about 30%.
Sally
Dr Sally Stowe Facility Coordinator, ANU Electron Microscopy Unit Research School of Biological Sciences Australian National University Canberra ACT0200 AUSTRALIA stowe-at-rsbs.anu.edu.au fax 61 (0)2 6125 3218 ph 02 6125 2743 http://www.anu.edu.au/EMU
Cogent comment. If the coverglass was mounted with the minimum of DPX, then the time required will be "over the weekend" or more. This from long experience with removing coverglasses cracked by students and a common excuse, "I thought it was parfocal!" Cogent comment? If the lab is cold/cool, put it all in a screw cap Coplin jar and set IT, not immersed, in a tepid water bath.
Question. I've never used DPX. Is it's solvent xylene? If it's plastic and it dries to plastic, then you might have to use acetone to get by the perimeter seal.
Irrational comment. OK, I won't remove all the chemicals from the lab.
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: Keith Ryan } Reply To: kpr-at-mba.ac.uk } Sent: Thursday, July 19, 2001 9:37 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Removing DPX/cover glass } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Greetings from Plymouth, UK } } Can anyone advise on removing a coverslip mounted using DPX? I am } currently soaking it in xylene, but overnight didn't "cut the mustard"! } Any cogent comments? Please?! } } Keith Ryan } Marine Biological Association } Plymouth } } }
We are close to choosing a new SEM and have come to the conclusion that if we want to view salt crystals with liquid inclusions we must do so under positive chamber pressure in order to reduce explosive cleanups. I have not dealt with such specimens before, so I am relying on common sense when expressing this conclusion. The problem is that my conclusion appears to reduce the market to one. I would like to know if I have this right or not.
Thanks,
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.
Hi All, We have a BioRad/Polaron E-7000 cryo unit (that was attached to a Philips SEM) available to a good home. Please contact me offline. Thanks john shields Univ. of Athens, GA USA jshields-at-cb.uga.edu (706)542-4080
I can't say that I have ever looked at such crystals. Are you hoping to image the inclusions? I know we can see some subsurface voids at higher voltages (20-25kV), but I think you would simply not see much.
How much pressure difference does it take to rupture the crystals? How much internal pressure of the fluid inclusions? Remember that even under absolute vacuum you have only raised the pressure differential by 15 psi. I would think most materials could contain that. Even a 10 torr atmosphere would not spare you much stress. I am not sure what you mean by a scope with positive pressure.
I suppose that there may always be a few inclusions that would blow under vacuum. I would think a pre-pump in a less expensive enclosure would cause those to fail and thus spare the ESEM from harm.
Warren
At 08:52 AM 7/20/2001 -0400, you wrote: } Good Morning Listers, } } We are close to choosing a new SEM and have come to the conclusion } that if we want to view salt crystals with liquid inclusions we must do so } under positive chamber pressure in order to reduce explosive cleanups. } I have not dealt with such specimens before, so I am relying on } common sense when expressing this conclusion. The problem is that my } conclusion appears to reduce the market to one. } I would like to know if I have this right or not. } Thanks, } } Fred Monson } } Frederick C. Monson, PhD } West Chester University of Pennsylvania } Center for Advanced Scientific Imaging (CASI)
---------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
I'm assuming (though you don't actually say) that it is thin sections that you are after. If so, I'm not surprised that embedding and sectioning was unsuccessful. You have one of those challenging specimens that profs seem to love handing to grad students, either with the expectation that specimen prep is trivial, or, knowing it is a beast, seeing if the grad student can come up with a new wrinkle!
You don't actually say that it is absolutely necessary for the analysis that the section hold together. Could you do a thin section, say, 100-200nm, do an area analysis of the (dry-sectioned) pulverized bits (lying on a coated grid on the diamond near the knife edge), then trim very coarse for, say 50 microns, then thin section again, and so on? If there is a 30 degree diamond knife around, you might reduce the 'pulverizing' action.
Alternatively, find someone in the neighbourhood with a focused ion beam (FIB) system, usually in Physics or Engineering. We have access to one and have sectioned (though for SEM analysis so we didn't try thin sectioning) some pretty delicate hydroxide powders with no mechanical damage whatsoever. If your particle is 'reinforced' with resin, a FIB-cut section should still be intact after it is cut through and flops on its side, since the incident Ga ion beam is at near zero degrees. The tricky part will come in getting the section onto a grid with something called a micromanipulator (a very fine glass fibre - electrostatic attraction makes the section 'fly up' onto the tip, hopefully intact, then you move the section over the grid and gently press it down). Most FIB types I know would be intrigued by the demanding nature of your specimen.
Good luck!
Tom Malis malis-at-nrcan.gc.ca
} From: Margaret Miller {MILLERMM-at-uthscsa.edu} } Date: Thu, 19 Jul 2001 22:18:21 -0500 } To: Microscopy-at-sparc5.microscopy.com } Subject: Sectioning a crystalline powder sphere } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } -- } To Listers, } } I am a graduate student at the university of Texas Health Science Center. } As part of a project I wish to perform a spectral analysis of a } certain medication. The medication is a crystalline powder in the } form of small (approx. 1mm diameter) spheres. I want to section the } spheres to 10 to 30 microns so as to obtain a spectral analysis of } the entire bead from its outer coating to its interior. I have tried } OCT embedding and cyrostat sectioning. However, the beads pulverized } during sectioning. Can anyone recommend a way to section these } spheres so as to preserve the material as much as possible? I should } also point out that the material is freely soluble in ethanol and } methanol, as well as water. } } Thank you in advance for your help and suggestions.
Dear all, I have found out the source of my particular problem with importing files into DTSA. It was caused by the way I transferred them over the network to the Mac with Fetch. It did not recognise the files as text files and therefore did not treat them as such. When I explicitly told Fetch that they were Text files, however, they could be opened in DTSA.
So, my problem was not caused by different variations in EMSA file formats, but by the fact that the Mac didn't know that they were text files.
Thanks again for all the suggestions, I seem to have set a huge discussion raging by a seemingly innocent comment about difficulties importing my files!
Best wishes to all.
Ian MacLaren Max Planck Institut für Metallforschung, Seestraße 92, 70174 Stuttgart, Germany Email: ian.maclaren-at-physics.org / maclariz-at-yahoo.co.uk Home page: http://members.tripod.co.uk/IanMacLaren/
} We have inherited a Leitz Metallux 3 microscope with epifluorescence, } however, without a user's manual (not a complaint, just a fact) and are } wondering whether anyone knows a source for old microscope manuals.
Judy -
There's a microscopy book dealer in the UK; try them: http://www.savonabooks.free-online.co.uk
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
Being an SEM guy, I wonder about embedding it and dry sanding it to a cross section with SiC paper. Water is normally used for a lubricant, but you could forego that and just use a few more sheets of paper.
Such a sample would not be good for TEM, but I would think an SEM could handle it well enough.
Warren
At 10:18 PM 7/19/2001 -0500, you wrote: } -- } To Listers, } } I am a graduate student at the university of Texas Health Science Center. } As part of a project I wish to perform a spectral analysis of a certain } medication. The medication is a crystalline powder in the form of small } (approx. 1mm diameter) spheres. I want to section the spheres to 10 to 30 } microns so as to obtain a spectral analysis of the entire bead from its } outer coating to its interior. I have tried OCT embedding and cyrostat } sectioning. However, the beads pulverized during sectioning. Can anyone } recommend a way to section these spheres so as to preserve the material as } much as possible? I should also point out that the material is freely } soluble in ethanol and methanol, as well as water. } } Thank you in advance for your help and suggestions.
---------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
Marbaret: I would be tempted to use a metallographic approach (being a metallographer). Go over to the Materials Science guys for guidance in making an epoxy mount. Talk to Dr. Mark Cavaleri of MMM (mecavaleri-at-mmm.com) about polishing methods. He did a complete section of an M&M without losing any of the sugars.
Good luck,
Sam Purdy Teech. Center National Steel Corp. Trenton MI 48183 spurdy-at-nationalsteel.com
Below is the result of your feedback form. It was submitted by (EvanPaassen) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Friday, July 20, 2001 at 14:32:10 ---------------------------------------------------------------------------
Email: EvanPaassen Name: Ed van Paassen
Organization: WAU
Education: Graduate College
Location: City, State, Country
Question: Does anyone know where I can get a used binocular stereo microscope?
Dennis: Altho' I have to use Access from time to time, it is not the program with which I am most familiar. Several years ago (over 10, believe it or not) I used Paradox to develop a database for the lab work going on--both EM and LM. It has been invaluable, and I have been able to maintain things through upgrades (altho' I think I may have missed the next to last one!). There has never been a problem transferring my data from one version to the next. I personally find Paradox easier to work with than Access, but then I started out using WordPerfect, and I still prefer that program. Another (and, _Free_) program is the 602ProSuite of products. I have only used the wordprocessing package so far, but it seems to have most of what I need/use. The program has received great reviews on the ZDNet website. And it's a fantastic price. Back to Paradox--one of the things I like best is the ability to go back and modify your database without losing (or just as bad, having to reenter everything) data. As projects/experiments have expanded/changed or been linked to other studies (via technique, reagents, etc) new categories can be added. Likewise, the search functions permit cross-database searches for seemingly (or unsuspected) relationships.
Roger
No financial interest--just a very satisfied, long-term user. On Thu, 19 Jul 2001 14:37:41 -0400, Dennis C. Winkler wrote:
| ------------------------------------------------------------------------ | The Microscopy ListServer -- Sponsor: The Microscopy Society of America | To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com | On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html | -----------------------------------------------------------------------. | | | Greg and others, | | Clarification: Yes, I am talking about physical micrographs | (film/negatives). Sorry about the confusion. | | Thanks for all of the suggestions thus far. My first thought was to | set up a FileMakerPro database to store the information, but I will | look at the commercial packages that have been mentioned. | | - Dennis | | ------------------------------------------------------------------------ | Dennis C. Winkler, PhD. Phone: (301) 496-0131 | Laboratory of Structural Biology Research Fax: (301) 480-7629 | NIAMS, National Institutes of Health Email: Dennis_Winkler-at-nih.gov | Bldg. 6, Room B2-26, MSC-2717 | Bethesda, MD 20892-2717, U.S.A. | | | At 9:27 AM -0400 7/19/01, Greg Erdos wrote: | } ------------------------------------------------------------------------ | } The Microscopy ListServer -- Sponsor: The Microscopy Society of | } America To Subscribe/Unsubscribe -- Send Email to | } ListServer-at-MSA.Microscopy.Com | } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html | } -----------------------------------------------------------------------. | } | } | } ThumbsPlus is good if you have a digital image to keep track of, but | } I am not sure if Dennis was referring to that or to physical | } micrographs. If it is the latter then I would suggest using | } Microsoft Access and setting up their own forms and reports to suit | } their particular needs. | } Greg Erdos | } |
Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc. 900 Rigdebury Road Ridgefield, CT 06877 203-798-5448
_______________________________________________________ Send a cool gift with your E-Card http://www.bluemountain.com/giftcenter/
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To all, The annual meetings are quickly approaching. We will again hold a session with discussion centering around topics pertaining to Core Facility Management. The topics and session facilitators are listed at the end of this message. Using the same format as last year, the facilitators will give introductions to the topics and then there will be extensive discussion involving all attending.
The session is scheduled for 8:00AM Monday August 5 in Room 201A. I hope to see lots of early risers as, due to later scheduling which partially conflicts with this session, we will have to start on time and move right along to cover the topics.
Please feel free to bring handouts or overhead if you feel they might be useful in the discussions. Course outlines might be very helpful for the first topic. Illustrations to help clarify questions pertaining to scientific ethics in the performance of our duties might also be helpful for the last topic.
Looking forward to seeing old friends and meeting new fellow microscopists at the meeting. Debby
Debby Sherman Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-099e Whistler Building West Lafayette, IN 47907 ============================================================================
This session is intended to highlight topics that are of concern or interest to managers of microscopy facilities in both academic and industrial settings and with emphasis in life sciences and/or materials areas. There will be three topics discussed at M&M 2001. Each topic will be introduced by one or more facilitators. The introduction will be followed by an extended open discussion with the audience.
Proceedings will be recorded and transcribed so they can be made available to interested persons after the meeting.
1) Training Users: Courses? Workshops? … what works for whom?
Facilitator: John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) Southern Illinois University
2) Calibration of Electron Microscopes: how to do this, how often, pit-falls and problems.
Facilitators: SEM: Michael Postek, Group Leader, Nanometer-scale Metrology National Institute of standards and Technology (NIST)
TEM: John P. McCaffrey National Research Council of Canada
3) Scientific Ethics: what are our responsibilities as facility managers?
Facilitator: Michael Kalichman Director, Research Ethics Program Office of Graduate Studies and Research, 0003 University of California, San Diego
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id RAA22835 for dist-Microscopy; Sun, 22 Jul 2001 17:40:58 -0500 (CDT) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id RAA22832 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Sun, 22 Jul 2001 17:40:27 -0500 (CDT) Received: from avocet.mail.pas.earthlink.net (avocet.mail.pas.earthlink.net [207.217.121.50]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id RAA22825 for {Microscopy-at-sparc5.microscopy.com} ; Sun, 22 Jul 2001 17:40:15 -0500 (CDT) Received: from earthlink.net (cpe-24-221-123-240.az.sprintbbd.net [24.221.123.240]) by avocet.mail.pas.earthlink.net (EL-8_9_3_3/8.9.3) with ESMTP id PAA08040 for {Microscopy-at-MSA.Microscopy.Com} ; Sun, 22 Jul 2001 15:37:24 -0700 (PDT) Message-ID: {3B5B5687.3060704-at-earthlink.net}
Listers,
I am looking for an EDS detector mounting flange for a JEOL JSM-IC845 SEM. The detector to be mounted is a Tracor Northern, with a 40 deg take off angle.
If someone has one of these and would like to part with it, please contact me off line. Thanks.
Bob Roberts EM Lab Services, Inc. 2409 S. Rural Rd Suite C Tempe, Arizona 85282 480.967.3946
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Further clarification.....the date of the session is Monday, August 6 not Monday August 5 as indicated below. Secondly all topics will be covered in order listed below in room 201A. Each topic is assigned 45 min for facilitator's introduction followed by open discussion. I know this may not be sufficient for everyone to speak their piece but it is all the time we were allotted. By keeping on track we can still cover a lot of ground. Perhaps those topics that need more time can be dealt with in more detail at future meetings. Debby
To all, The annual meetings are quickly approaching. We will again hold a session with discussion centering around topics pertaining to Core Facility Management. The topics and session facilitators are listed at the end of this message. Using the same format as last year, the facilitators will give introductions to the topics and then there will be extensive discussion involving all attending.
The session is scheduled for 8:00AM Monday August 5 in Room 201A. I hope to see lots of early risers as, due to later scheduling which partially conflicts with this session, we will have to start on time and move right along to cover the topics.
Please feel free to bring handouts or overhead if you feel they might be useful in the discussions. Course outlines might be very helpful for the first topic. Illustrations to help clarify questions pertaining to scientific ethics in the performance of our duties might also be helpful for the last topic.
Looking forward to seeing old friends and meeting new fellow microscopists at the meeting. Debby
Debby Sherman Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-099e Whistler Building West Lafayette, IN 47907 ============================================================================
This session is intended to highlight topics that are of concern or interest to managers of microscopy facilities in both academic and industrial settings and with emphasis in life sciences and/or materials areas. There will be three topics discussed at M&M 2001. Each topic will be introduced by one or more facilitators. The introduction will be followed by an extended open discussion with the audience.
Proceedings will be recorded and transcribed so they can be made available to interested persons after the meeting.
1) Training Users: Courses? Workshops? … what works for whom?
Facilitator: John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) Southern Illinois University
2) Calibration of Electron Microscopes: how to do this, how often, pit-falls and problems.
Facilitators: SEM: Michael Postek, Group Leader, Nanometer-scale Metrology National Institute of standards and Technology (NIST)
TEM: John P. McCaffrey National Research Council of Canada
3) Scientific Ethics: what are our responsibilities as facility managers?
Facilitator: Michael Kalichman Director, Research Ethics Program Office of Graduate Studies and Research, 0003 University of California, San Diego
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To clarify the previous listing,
The BioRad/Polaron cryo unit was originally attached to a Philips 505 and has been in storage for quite some time. I was told it had problems prior to being boxed up. It will be better suited as spare parts for anyone with an existing unit. Sorry for the previous incompleteness. John Shields EM Lab Univ. of Georgia Athens, GA 30602
I'm not looking to start a flame war here. Please bear with me.
I was going to update my Zeiss Axioplan DIC/phase system to a newer model. I got some major reverberations and feedback that something very bad has happened regarding Zeiss the company. Most feedback seems to be along the lines of bad customer service, no parts, and no support.
My Zeiss systems have been working great for several years and I have not needed any factory support. I ordered some fine clock oil for a condenser and a few other odd and ends and had no problems. My Plan Neofluars don't wear out. So what is the big deal with Zeiss Co. these days?
I have no financial interest in Zeiss. I only have a financial interest in not getting burned. To avoid public acrimony, perhaps private responses off-line are appropriate. I would certainly appreciate some info about what I am obviously missing.
Gary Gaugler, Ph.D. Microtechnics, Inc. 7970 Twin Rocks Rd Granite Bay, CA 95746
Would you confirm us the Core Facility Management session is scheduled for Monday August 6 not Sunday August 5. In case of Aug 5, we have to change our schedule for the flight and hotel.
Thanks
Greg Ning EM Facility Medical College of Wisconsin
Debby Sherman wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } To all, } The annual meetings are quickly approaching. We will again hold a session with discussion centering around topics pertaining to Core Facility Management. The topics and session facilitators are listed at the end of this message. Using the same format as last year, the facilitators will give introductions to the topics and then there will be extensive discussion involving all attending. } } The session is scheduled for 8:00AM Monday August 5 in Room 201A. I hope to see lots of early risers as, due to later scheduling which partially conflicts with this session, we will have to start on time and move right along to cover the topics. } } Please feel free to bring handouts or overhead if you feel they might be useful in the discussions. Course outlines might be very helpful for the first topic. Illustrations to help clarify questions pertaining to scientific ethics in the performance of our duties might also be helpful for the last topic. } } Looking forward to seeing old friends and meeting new fellow microscopists at the meeting. } Debby } } Debby Sherman Phone: 765-494-6666 } Life Science Microscopy Facility FAX: 765-494-5896 } Purdue University E-mail: dsherman-at-purdue.edu } S-099e Whistler Building } West Lafayette, IN 47907 } ============================================================================ } } M&M 2001 } Core Facility Management “Experts” session } } This session is intended to highlight topics that are of concern or interest to managers of microscopy facilities in both academic and industrial settings and with emphasis in life sciences and/or materials areas. There will be three topics discussed at M&M 2001. Each topic will be introduced by one or more facilitators. The introduction will be followed by an extended open discussion with the audience. } } Proceedings will be recorded and transcribed so they can be made available to interested persons after the meeting. } } 1) Training Users: Courses? Workshops? … what works for whom? } } Facilitator: John J. Bozzola, Ph.D., Director } I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) } Southern Illinois University } } 2) Calibration of Electron Microscopes: how to do this, how often, pit-falls and problems. } } Facilitators: } SEM: Michael Postek, } Group Leader, Nanometer-scale Metrology } National Institute of standards and Technology (NIST) } } TEM: John P. McCaffrey } National Research Council of Canada } } } 3) Scientific Ethics: what are our responsibilities as facility managers? } } Facilitator: Michael Kalichman } Director, Research Ethics Program } Office of Graduate Studies and Research, 0003 } University of California, San Diego } } }
Dear Fred, Please refer to the American Vacuum Society site: http://www.aip.org/avsguide/refguide/pressure.html for a fast and useful education in the meaning of pressure. Through the kindness, and forbearance, of Bill Miller you have been saved from exhibiting your own version of ignorance any further. You should apologize to all the non-vacationing listers for exposing yourself in this manner. In future, some information on the basics would probably save you from this kind of embarrassment. First of all, I suspect you would have been helped if the phrase "variable pressure" had been correctly(?) (mis?)stated as "variable vacuum".[there's that period again!] As Bill said, even the best SEM's cannot operate with the chamber open, and you should, with all of your 'smarts', have been able to see that. Once you have your brain switched to the proper channel, I am certain you will see this point and stop bothering these people. Finally, and I'm sure I speak for all, you should take your own advice and look in the books first. But also, please remember what I always tell my students. The only dumb question is the one you don't ask. Well, as you can see, that's not quite the truth, but it's pretty good advice. And, finally, finally, about your question. Perhaps what you should have asked was how users of SEM technology look at water in the liquid state?
Regards and apologies,
Fred Monson
} ---------- } From: Monson, Frederick C. } Sent: Friday, July 20, 2001 8:52 AM } To: 'Microscopy Listserver' } Subject: SEM, Liquid Inclusions in Salt Crystals } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Good Morning Listers, } } We are close to choosing a new SEM and have come to the conclusion } that if we want to view salt crystals with liquid inclusions we must do so } under positive chamber pressure in order to reduce explosive cleanups. } I have not dealt with such specimens before, so I am relying on } common sense when expressing this conclusion. The problem is that my } conclusion appears to reduce the market to one. } I would like to know if I have this right or not. } } Thanks, } } 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. } }
Evening Keith, Check Wilson's disease. rubeanic acid chelates the Cu, Co or Ni Check Merck Index Ray, Xavier(1961), J Indian Chem Soc, 38:535(review) Have no experiece with EM for this problem. Also: http://www.ninds.nih.gov/health_and_medical/disorders/wilsons_doc.htm
Regards and let us know how the marine bio studies go.
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 CASI Home Page: email: fmonson-at-wcupa.edu Schedule: http://bio.wcupa.edu/cgi-bin/webreserve.acgi Please call before visiting.
} ---------- } From: Keith Ryan } Reply To: kpr-at-mba.ac.uk } Sent: Friday, July 20, 2001 3:40 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: copper retention-rubeanic acid } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Hello Listers } } Can anyone comment on the retention of copper (in dosage experiments) by } "staining" with rubeanic acid? What is the complex seen in light } microscopy? Would it be accessible to x-ray microanalysis? ......... } after removing a coverslip! } } Keith Ryan } Marine Biological Association } Plymouth UK } }
I did set up a Filemaker Pro database years ago to log TEM micrographs as I was shooting them. I used the database to make labels for the micrographs (big time-saver for me), print a list of negatives shot at each session and do annual reviews of project work. It was also useful as a searching tool, for instance if I forgot the date that I last examined samples from a certain project. It would be easy to set up a similar database with a field for micrograph storage information.
Note: the initial plan was to set up the database for a whole microscopy lab of mixed PC and Mac-users, but we had filesharing problems over our network system. I did think Filemaker Pro was easier to use than Access (which I converted to after everything went PC). Don't get me wrong, I like using Access as well. But for creating a simple text-based database Filemaker was very straightforward, even if it had some limitations regarding relational comparisons (it's been so long I can't remember what the limitations were, just that they existed!).
I should emphasize that what made the system most convenient for me was having it set up on the computer attached to the TEM. That way I could enter each record as soon as the negative was exposed, the same way one would make an entry in a logbook. It took a few seconds longer than handwriting, but overall it did not feel like an extra chore to do the data entry.
Caveat: I set this up about 8 years ago, and haven't worked with Filemaker Pro for over 3 years. Things may have changed... But if you are interested I'd be glad to send a copy of the database template (if I can still find it). Hope it works out for you, Karen -------------------------------------------------------------------------------- Karen S. Zaruba BioMaterials Technology Center 3M Center Bldg. 270-1S-01 St. Paul, MN 55144 kszaruba1-at-mmm.com
"Dennis C. Winkler" {Dennis_Winkler-at-nih.gov} 07/19/01 01:37 PM
To: Microscopy-at-sparc5.microscopy.com cc: (bcc: Karen S. Zaruba/US-Corporate/3M/US) Subject: RE: Database of Archived EM Micrographs
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Greg and others,
Clarification: Yes, I am talking about physical micrographs (film/negatives). Sorry about the confusion.
Thanks for all of the suggestions thus far. My first thought was to set up a FileMakerPro database to store the information, but I will look at the commercial packages that have been mentioned.
- Dennis
------------------------------------------------------------------------ Dennis C. Winkler, PhD. Phone: (301) 496-0131 Laboratory of Structural Biology Research Fax: (301) 480-7629 NIAMS, National Institutes of Health Email: Dennis_Winkler-at-nih.gov Bldg. 6, Room B2-26, MSC-2717 Bethesda, MD 20892-2717, U.S.A.
To: microscopy-at-msa.microscopy.com,CONFOCAL-at-LISTSERV.ACSU.BUFFALO.EDU
Zeiss has always been a company that has supported their products dating back to almost the stone age. It seems that has come to an end. They broke (terminated) their agreements with their whole dealer base, the independent companies who had been supporting Zeiss products for decades. Now they are not supporting microscopes the have just sold a year or two ago. We have a brand new Zeiss microscope in our Emergency Room Lab that has a bad lens. No parts available to repair it!!! I'm supposed to throw away a new $5000 dual head microscope because the company won't support it?
I no longer recommend purchasing Zeiss products.
There have always been other companies who offer much better value then Zeiss. The redeeming value of Zeiss has been in the past that they offered almost unlimited support for their older products. No more.
Dave Burton Optical Specialist
----- Original Message ----- } From: "Gary Gaugler" {gary-at-gaugler.com} To: "MSA listserver" {Microscopy-at-sparc5.microscopy.com} Sent: Monday, July 23, 2001 7:13 AM
Correction - Rather than being included as attachments to the previous message, contents of the Late Breaking Poster Session and the Corporate Members Platform Session have been added to the meeting web site. You can access these by clicking the Symposia icon on the Microscopy and Microanalysis 2001 web site.
------- Forwarded Message Follows ------- Date sent: Mon, 23 Jul 2001 18:22:05 -0500 To: Microscopy-at-sparc5.microscopy.com } From: "Bob Price" {price-at-dcsmserver.med.sc.edu}
Below is the result of your feedback form. It was submitted by (srinivas.mullapudi-at-uth.tmc.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, July 24, 2001 at 10:07:20 ---------------------------------------------------------------------------
We are attempting to visualize pulmonary vasculature. One method would be to perfuse an animal with a fluorescent dye - for example FITC-dextran - but, since we expect to section the lung, the dye would leak out at the severed edges.
Does anyone know of a compound in which to disolve the dye and perfuse the animal with, that would polymerize into a soft solid and remain within the vessels? I remember reading about something like this used to immobilize suspension cells in a culture dish.
Thank you
Judy Trogadis Bio-Imaging Coordinator St. Michael's Hospital, 8Q 30 Bond St. Toronto, ON M5B 1W8
I wish to measure the grain size of 1 micron thick Al0.5%Cu films on Single Crystal Silicon substrate. These are deposited by magnetron sputtering process. Any suggestions or experience out there? Thanks Rick Eller
What kind of results have some of you had with Kodak MDS 100? The price is getting pretty reasonable for the resolution if the software works well enough to be usable. My particular concern it lack of manual control over exposure.
TIA Gordon
Gordon Couger Stillwater, OK www.couger.com/gcouger
Judy Trogadis wrote: ============================================================== We are attempting to visualize pulmonary vasculature. One method would be to perfuse an animal with a fluorescent dye - for example FITC-dextran - but, since we expect to section the lung, the dye would leak out at the severed edges.
Does anyone know of a compound in which to disolve the dye and perfuse the animal with, that would polymerize into a soft solid and remain within the vessels? I remember reading about something like this used to immobilize suspension cells in a culture dish.
============================================================== I believe the "compound" you are referring to is the Mercox™ Corrosion Casting System, it comes in two different colors (red and green) plus clear (for SEM applications). You can see information about it on our website at URL http://www.2spi.com/catalog/chem/mercox-corrosion-casting.html
You can also see reproduced on the website an article that appeared in Microscopy Today, "Vascular Corrosion Casting Can Provide Quantitative as Well as Morphological Information on the Microvasculature of Organs and Tissues" by Fred E. Hossler, J. H. Quillen College of Medicine, Issue 98-7, Sept. 1998, or see URL http://www.2spi.com/catalog/chem/mercox-casting.html
Disclaimer: SPI Supplies distributes the Mercox kits to researchers worldwide.
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 kind of results have some of you had with Kodak MDS 100? The price is getting pretty reasonable for the resolution if the software works well enough to be usable. My particular concern it lack of manual control over exposure.
Thanks Gordon
Gordon Couger Stillwater, OK www.couger.com/gcouger
Has anyone any experience with the Nikon Epi-Fluorescence Attachment for a Stereo Microscope. For example how does it compare with the Leica MZ FLIII Fluorescence Stereo Microscope?? Any personal responce will be treated in confidence.
Thanks
Ian
Ian Hallett HortResearch Mt Albert Research Centre Private Bag 92 169 Auckland, New Zealand Fax 64-9-815 4201 Telephone 64-9-815 4200 EMail ihallett-at-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. ______________________________________________________
Would anyone have a copy of a Leitz manual for a Dailux 20 with the epifluorescence head? We have one that we want to use and would like information on the filter sets etc..
The first method that comes to mind, is to ion mill from the Si side, and then look in the TEM. I've done this in pre PIPS days on dimpled samples.
I also recall rigging up a chemical method where the Si was etched away (a mixture of nitric acid and HF. I can't recall the concentrations, but it was likely something standard which others might still be using) leaving the surface film, again for TEM.
If you are handy with a microtome, surface slices coud be taken from a bit of the wafer glued to a dummy block. This will give sections thin enough for TEM, but there will be compression artifacts.
And the last "top of my head" suggestion... at 1 micron thick, are the grains large enough to see using channeling contrast in the SEM?
"Eller, Rick" {ellerro-at-tsmd To: "'Microscopy-at-MSA.Microscopy.Com'" .com} {Microscopy-at-sparc5.microscopy.com} cc: 07/24/01 Subject: Grain Size Measurement of Thin Metal Films ? 04:02 PM
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I wish to measure the grain size of 1 micron thick Al0.5%Cu films on Single Crystal Silicon substrate. These are deposited by magnetron sputtering process. Any suggestions or experience out there? Thanks Rick Eller
Microscopists: We are looking for an environmental chamber that will allow us to heat materials to approximately 1100 C under different reactive gasses while viewing in-situ by either SEM (ESEM) or light microscopy. ESEM would be preferred due to the size range of the subject. We would be interested in contracting this work to your lab, "renting" the chamber or purchasing outright, depending on what's available. Please contact us directly with responses so we don't clutter the list!! Thanks in advance. Bill William A. Heeschen Microscopy, Digital Imaging The Dow Chemical Company 1897 Bldg. Midland, MI 48667-1897 waheeschen-at-dow.com voice: 989-636-4005 fax: 989-638-6443
Actually, one merely has to inject Evans Blue into the experimental animals vasculature, and the albumen in the blood will be instantly bonded by the dye. The dye is supposed to fluoresce only when so bonded. The other way, of course is to set up the albumen-Evans Blue first (see Kodani(?) in '60's for testicular barrier studies)followed by suitable dialysis of unbound EVB (stoichiometry is apparently 1:1). Fixation followed by sectioning with fluorescence. FITC-?, though bound with greater avidity than plain fluorescein does not elicit the dye fastness of EVB with albuminoids.
Evans Blue Dye Site (Blood volume): http://www.evansblue.co.uk/index.html
This is everything but the trip to the confocal (see Czymmek, K (2000), "Imaging and volumetric quantitation of vascular corrosion casts with laser scanning confocal microscopy, Proceedings: Microscopy and Microanalysis 2000, Philadelphia, PA, August 13-17, 2000, p 562. (kirk.czymmek-at-mvs.udel.edu). Using Mercox Blue corrosion cast, excite 633nm laser, ~660nm barrier.
Fred
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 CASI Home Page: email: fmonson-at-wcupa.edu Schedule: http://bio.wcupa.edu/cgi-bin/webreserve.acgi Please call before visiting.
} ---------- } From: Judy Trogadis } Sent: Tuesday, July 24, 2001 3:49 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: compound for immobilization } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Fellow Microscopists : } } We are attempting to visualize pulmonary vasculature. One method would be } to perfuse an animal with a fluorescent dye - for example FITC-dextran - } but, since we expect to section the lung, the dye would leak out at the } severed edges. } } Does anyone know of a compound in which to disolve the dye and perfuse the } animal with, that would polymerize into a soft solid and remain within the } vessels? I remember reading about something like this used to immobilize } suspension cells in a culture dish. } } Thank you } } } } } } Judy Trogadis } Bio-Imaging Coordinator } St. Michael's Hospital, 8Q } 30 Bond St. } Toronto, ON M5B 1W8 } } ph: 416-864-6060 x6337 } fax: 416-864-6043 } trogadisj-at-smh.toronto.on.ca } } } }
I want so show that a polymer film has not been etched after exposure to an HCL based environment. The problem is that it is extremely smooth to begin with. I would like to add some sub-micro features as a background so that I am not generating a featureless gray photomicrograph. These features would be essentially a topographic reference. I though of a mono layer of particles dusted on the surface. I stated w/ common baking flour. The particles were too large and not stuck to the surface so well (lots of edge effect) so that one can't effectively have a good contrast of the spheres and the background in the same micrograph. A very fine grid would also be nice, but perhaps expensive. Smaller and flatter particles that were put in solvent and allowed to dried might do the trick. Anybody have any interesting suggestions or materials? I would be glad to share the results.
Good Afternoon! This is a follow up question involving tissue sections prepared for the TEM. My original question dealt with perfusion with physiological saline followed by cold 4% paraformaldhyde. I will be incubating the tissue (dorsal root ganglions) with an antibody. I have spoken to my advisor and Franklin Bailey at the UI EM center about preparing the tissue sections and eventual incubation with antibodies. I have received conflicting advice and am putting this out for more advice. Since I am going to use antibodies, does this change how I perfuse the animal? When incubating with the antibodies, should I do it pre- or postembedding? Immunogold labeling may be used and I have been told that preembedded labeling may damage the knife. I am open to other ways of labeling this tissue. Thank you.
Meghan Towner Graduate Student Biological Sciences University of Idaho
Try smoking some Mo with heat to produce a white smoke and wave your sample in the smoke. You will pick up small particle of MoO3. Alternatively, you could burn Mg and wave your sample in the smoke. Experiment with how high in the smoke you wave your sample.
-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: Smartech [mailto:smartech-at-optonline.net] Sent: Wednesday, July 25, 2001 2:51 PM To: To all on the list
I want so show that a polymer film has not been etched after exposure to an HCL based environment. The problem is that it is extremely smooth to begin with. I would like to add some sub-micro features as a background so that I am not generating a featureless gray photomicrograph. These features would be essentially a topographic reference. I though of a mono layer of particles dusted on the surface. I stated w/ common baking flour. The particles were too large and not stuck to the surface so well (lots of edge effect) so that one can't effectively have a good contrast of the spheres and the background in the same micrograph. A very fine grid would also be nice, but perhaps expensive. Smaller and flatter particles that were put in solvent and allowed to dried might do the trick. Anybody have any interesting suggestions or materials? I would be glad to share the results.
Our company needs to get a microscope for inspection and assembly/disassembly tasks. We would like it to have the following characteristics:
1) Stereo viewing 2) Wide field of view 3) Prefer zoom capabilities, but could take anything that would allow magnifications from about 4X to, say, 20X 4) Good working distance
Would anyone care to make any recommendations or do you have any suggestions for us?
Vendor input is welcome. Contact information follows my sig.
Bruce Girrell Microline Technology Corp. 2397 Traversefield Dr. Traverse City, MI 49686 http://www.microlinetc.com
SMARTech asked the following: =========================================================== I want so show that a polymer film has not been etched after exposure to an HCL based environment. The problem is that it is extremely smooth to begin with. I would like to add some sub-micro features as a background so that I am not generating a featureless gray photomicrograph. These features would be essentially a topographic reference. I though of a mono layer of particles dusted on the surface. I stated w/ common baking flour. The particles were too large and not stuck to the surface so well (lots of edge effect) so that one can't effectively have a good contrast of the spheres and the background in the same micrograph. A very fine grid would also be nice, but perhaps expensive. Smaller and flatter particles that were put in solvent and allowed to dried might do the trick. Anybody have any interesting suggestions or materials? I would be glad to share the results. =========================================================== We faced a similar kind of question involving a Mylar film (some years ago), which also was smooth and featureless at the SEM level and there was the need to look for evidence of etching. In this case, the "etch" was exposure to ozone. Our approach was the following:
a) Apply on one side of the film a special silicone resin; this was an earlier version of the silicone used in our SPI Wet Replica Kit (see URL http://www.2spi.com/catalog/spec_prep/wet_rep_kits.html ) The line between the resin covered side and the side without resin is quite distinct. The resin thickness is about 0.5 to 1 mm.
b) Do your exposure of the film, but the silicone layer protects the one side from exposure and it will not be harmed by the acid environment. Of course, at some point it can ( I presume) be effected by acid so it has to be a matter of time, temperature and pH.
c) After exposure, the silicone can be removed, and if indeed there is some etching, if differences are seen one side of the interface vs. the other, then you could draw some conclusions.
In those days, we found we needed ordinary Pt/C surface replication methods (and TEM) to show the differences, one side to the other. But in the end one is faced with having to prove the null hypothesis: If you see no effect , does that mean no etching or does that mean one has not looked close enough. Today, I would expect that AFM would have a better chance at seeing differences that might not be seen with Pt/C surface replication.
Another approach, if we were to be doing this today, would be to try applying the approach that has been used by Japanese researchers at Nagoya University (if I remember correctly) who have been using thin osmium metal layers, plasma coated, that are not more than ~1 nm thick, to replicate macromolecules on flat surfaces. The method does work well for DNA for example and might work well for a surface with a very subtle etch effect. If you would like more information about this method, I could share additional information with you about how this is done. I could also dig out my notes on the people who are doing this work.
Disclaimer: SPI Supplies offers this type of silicone resin and also, distributes the OPC-60 Osmium Plasma Coater made by Nippon Laser and Electronics Labs, Nagoya, Japan.
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 ############################ ==================================================
ie the anti-backstreaming trap between the mechanical pump and the diff pump:-
is there a consensus out there on what's best?
Molecular seive, activated alumina pellets, or Cu gauze?
I've never been that happy about the first two because of mechanical abrasion releasing abrasive powder into the rotary pump, particularly from those Edwards holders that seem designed to mount directly on the pump, where they get nicely and continuously vibrated.
And how do people regenerate each absorbent?
Heat in air to 600 deg C or so?
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
You might consider an Olympus SZ-40 system with a Fostec fiber optic ACE ring light. There are several sources for these 'scopes as used. The Fostec may be only available as new (I have a near new light source I would sell...no ring light bundle). I like the Fostec very much.
On the surplus market, Nikon stereo zooms come up quite often. Maybe one of these would work for you. Try http://dovebid.com for options.
gary g.
Disclaimer: No financial interest in any company referenced above.
At 02:31 PM 7/25/2001, you wrote:
} Our company needs to get a microscope for inspection and } assembly/disassembly tasks. We would like it to have the following } characteristics: } } 1) Stereo viewing } 2) Wide field of view } 3) Prefer zoom capabilities, but could take anything that would allow } magnifications from about 4X to, say, 20X } 4) Good working distance } } Would anyone care to make any recommendations or do you have any suggestions } for us? } } Vendor input is welcome. Contact information follows my sig. } } 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
Anyone out there who can help me. We want to 'refurbish' an old Leitz DURIMET Microhardness Tester for routine applications by students. However, both objectives are badly damaged and beyond repair. (No wonder, the unit is approximately 50 years old and had been used by numerous students in the past). We would like to 'pick up' or purchase therefore the following types of objectives (used or new): Low Magnification 40x and High Magnification A 0.70 CHM 6.3 40x to suit the DURIMET. Thanks for any advise.
Hans Brinkies Senior Lecturer Swinburne University of Technology School of Engineering and Science Industrial Microscopy Laboratory PO Box 218 - Hawthorn - Victoria - 3122 - Australia Phone: +61 3 9214 8657 - FAX: +61 3 9214 8264
Just my two cents concerning pre-embed labeling with gold damaging the knife. We have done it for years and never noticed any damage.
Bob Underwood Morphology Core U of Washington
On Wed, 25 Jul 2001, Meghan Towner wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Good Afternoon! } This is a follow up question involving tissue sections prepared for the TEM. My original question dealt with perfusion with physiological saline followed by cold 4% paraformaldhyde. I will be incubating the tissue (dorsal root ganglions) with an antibody. I have spoken to my advisor and Franklin Bailey at the UI EM center about preparing the tissue sections and eventual incubation with antibodies. I have received conflicting advice and am putting this out for more advice. Since I am going to use antibodies, does this change how I perfuse the animal? When incubating with the antibodies, should I do it pre- or postembedding? Immunogold labeling may be used and I have been told that preembedded labeling may damage the knife. I am open to other ways of labeling this tissue. Thank you. } } Meghan Towner } Graduate Student } Biological Sciences } University of Idaho } } }
Dear Smartech, Just off the top of my head: 1. clay, ultra-soniced in alcohol and dropped on to dry. 2. the fine aluminum powder used in makeup, dusted on with a paintbrush or dropped from alcohol as in 1. 3. small latex sheres used to calibrate TEMs. Already in suspension, just drop on to dry. I have used all three for various purposes. I have also used a very fine grid (~1000 mesh) laid tight over the sample and then evaporated gold through it. The resulting gold squares are quite visible in the SEM but avoid the problems of edge effect. At 02:50 PM 7/25/01 -0400, you wrote:
} I want so show that a polymer film has not been etched after exposure to an } HCL based environment. The problem is that it is extremely smooth to begin } with. I would like to add some sub-micro features as a background so that I } am not generating a featureless gray photomicrograph. These features would } be essentially a topographic reference. I though of a mono layer of } particles dusted on the surface. I stated w/ common baking flour. The } particles were too large and not stuck to the surface so well (lots of edge } effect) so that one can't effectively have a good contrast of the spheres } and the background in the same micrograph. A very fine grid would also be } nice, but perhaps expensive. Smaller and flatter particles that were put in } solvent and allowed to dried might do the trick. Anybody have any } interesting suggestions or materials? I would be glad to share the results. } } SMARTech
Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
The various types of foreline traps are discussed in some detail on pp. 146 - 149 of my book 'Vacuum Methods in Electron Microscopy" (for a description see http://www.2spi.com/catalog/abooks/book48.html/ and http://www.pup.princeton.edu/titles/6484.html/)
At the time I was writing this book it appeared that perhaps the best method of trapping oil vapors from the roughing pump was to use a Micromaze trap produced by the Lesker company. These traps are provided with a heater to be used for regenerating the trap, and it appeared advantageous to run this heater continuously at about 75% full power. Evidence available then indicated that when hot the ceramic Micromaze material caused the oil molecules to break down into small fragments that could pumped away. See above reference for details.
Maybe some more efficient type of trap has been developed since, but I have not heard of it. -- 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
Does anyone remember the sucrose concentrations needed to form a gradient for separating cells and cell fractions from media debris? We once used this technique, but I have lost all of my references on the procedure. Thanks
Franklin Bailey Electron Microscopy Center University of Idaho Moscow, ID 83844-2204
Dear Folks, I'm in the process of setting up a system for doing time-lapse video microscopy and would love to get information from others on what set-ups work best for them. I'm interested in people's experience with time-lapse VCRs and digital capture. I'm recording in color using a three-chip CCD video camera. We have a Scion capture card on a Mac G4. I will be recording over 24-hour periods. In particular, has anyone been using NIH Image or Imagej to do time-lapse in color?
I am new to this listserv, so my apologies if I'm raking over old territory. I did do a search of the listserv archives and didn't find any postings in this subject. Many thanks, Mary =============================================== Mary_Tyler-at-umit.maine.edu or mtyler-at-maine.edu
Mary S. Tyler Professor of Zoology Dept. of Biological Sciences University of Maine Orono, ME 04469-5751 FAX: 207-581-2537
You can smoke Mo in a number of ways. You can use wires or boats across the leads of an evaporator and do it in air, or you can take a wire and heat it with a Bunsen burner, or propane torch. The smoke should not be that hot, just don't keep your sample in the smoke or near the radiant heat for too long. The wire should continue to smoke a little just as you remove the heat. If you collect the smoke from Mg, you should be able to collect it far from the heat source as the smoke rises. I assumed in my original post that you would need high resolution imaging and small particles distributed with no carrier media to put the stuff on the surface. There was a response to my earlier post that said to scratch the surface, but that would produce features too gross.
-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: Smartech [mailto:smartech-at-optonline.net] Sent: Wednesday, July 25, 2001 8:08 PM To: Walck, Scott D.
I want so show that a polymer film has not been etched after exposure to an HCL based environment. The problem is that it is extremely smooth to begin with. I would like to add some sub-micro features as a background so that I am not generating a featureless gray photomicrograph. These features would be essentially a topographic reference. I though of a mono layer of particles dusted on the surface. I stated w/ common baking flour. The particles were too large and not stuck to the surface so well (lots of edge effect) so that one can't effectively have a good contrast of the spheres and the background in the same micrograph. A very fine grid would also be nice, but perhaps expensive. Smaller and flatter particles that were put in solvent and allowed to dried might do the trick. Anybody have any interesting suggestions or materials? I would be glad to share the results.
POST-DOCTORAL POSITION in Electron Microscopy of Electroceramic Grain Boundaries
A postdoctoral position is available in the area of electroceramics beginning September 1, 2001. The research project focuses on understanding solute segregation at electroceramic grain boundaries, in particular TiO2 and BaTiO3, and the ramifications for electrical transport properties. Aspects of the project will be conducted under the auspices of the Center for Dielectric Studies at the Pennsylvania State University. Through a variety of electron imaging and spectroscopy techniques we aim to quantify solute segregation and grain boundary impedance as a function of doping level and microstructure. For special boundaries we will obtain atomic structure and site-specific segregation information that will be used as the basis for theoretical calculations. The ideal candidates for this position will have experience in HREM, STEM, EDS and/or EELS and have an understanding of grain boundary phenomena in electroceramics. The salary will be commensurate with qualifications and experience.
Please forward questions or send applications to Professor Elizabeth Dickey:
Department of Materials Science and Engineering The Pennsylvania State University 195 Materials Research Institute Building University Park, PA 16802-7003
tel: (814) 865-9067 FAX: (814) 863-8561 email: ecd10-at-psu.edu -- Elizabeth Dickey Department of Materials Science and Engineering Pennsylvania State University 195 Materials Research Institute Building University Park, PA 16802-7003
A colleague has asked if I could help explain why the excitation filter in his DAPI filter set on a Zeiss Axiophot deteriorates after a few months of use.
Excitation filters in other sets last longer and don't show this problem. The symptoms are uneven illumination that shows up in the image and an obvious 'burned ' look to the filter.
His questions are like: Is this normal? If not, what is wrong? How can I avoid this problem?
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
I have a couple Micromaze bakeable foreline traps that Lesker sells. They were on a TEM that I used to own. I almost forgot that I had these. I kept them with the intention of trying to sell them someday. These two were modified for safety reasons and so nothing around them would get damaged by the heat from these. These traps when turned on get pretty hot if I remember right. For these reasons longer stainless pipes were welded on and a cage was built around each one. I would recommend something like this if you use these traps.
Not only are the maze traps better, I think, but they are easier to use. Use to simply plug them into an AC outlet overnight once a week to bake them out. Kept the vacuum system running while bakeing.
Joel McClintock EM Specialist U of Kentucky
At 02:01 PM 7/26/01 GMT+1200, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
It's better to use PERCOLL solution. It's isoosmotic and you could separate cells from microsomal fraction or viruses etc. You may also separate different cell types by this technique. PERCOLL was manufactured by Pharmacia, which is a part of Amersham now, I believe. Good luck. Sergey
At 11:06 AM 7/26/01 -0700, 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 question about using Percoll for TEM - someone told me just last week that you shouldn't use Percoll if you'll be doing thin sections, since it is a silica (can damage knives) - has anyone used it and either seen or not seen gunk in their sections? Scratches &/or damaged a knife? Sea creatures or other beach denizens? (Sorry, I'm tired and couldn't resist)
I'm not planning to use it any time soon, but hearing about it again makes me curious about using it to prep for EM. I've seen nasty sucrose crystals a few times with samples from sucrose gradients, but they are mostly an annoyance.
Thanks!
Tamara
On Thu, 26 Jul 2001, Sergey Ryazantsev wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } It's better to use PERCOLL solution. It's isoosmotic and you could } separate cells from microsomal fraction or viruses etc. You may also } separate different cell types by this technique. PERCOLL was manufactured } by Pharmacia, which is a part of Amersham now, I believe. Good luck. Sergey } } At 11:06 AM 7/26/01 -0700, you wrote:
|--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward-at-unm.edu |--------------------------------------------------|
I agree with the ideas of placing simple structures on the surface of the specimen, Mn smoke or polystyrene latex. However general tips for featureless specimens are -
1. Keep the kV down ( {5kV) - to restrict the probe to a true surface investigation
2. Use high tilt (} 45deg) as the small amount of backscatter present will serve to emphasize topography.
Happy scanning
Steve Chapman Senior Consultant Protrain For professional training in electron microscopy world wide www.emcourses.com
Some time ago we tried to section some epoxy-embedded, Percoll-separated cell fractions. This resulted in very evident damage to glass knife cutting edges - to such an extent that we decided not to jeopardize a diamond knife.
Separating the fractions on a sucrose gradient resulted in blocks that sectioned without any problems.
Jan Coetzee
Lab for Microscopy and Microanalysis University of Pretoria, South Africa. Tel: 012-420-2075, Fax 012-362-5150 www.up.ac.za/academic/electron/emunit1.htm
Tamara Howard wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I have a question about using Percoll for TEM - someone told me just last } week that you shouldn't use Percoll if you'll be doing thin sections, } since it is a silica (can damage knives) - has anyone used it and either } seen or not seen gunk in their sections? Scratches &/or damaged a knife? } Sea creatures or other beach denizens? (Sorry, I'm tired and couldn't } resist) } } I'm not planning to use it any time soon, but hearing about it again makes } me curious about using it to prep for EM. I've seen nasty sucrose crystals } a few times with samples from sucrose gradients, but they are mostly an } annoyance. } } Thanks! } } Tamara } } On Thu, 26 Jul 2001, Sergey Ryazantsev wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } It's better to use PERCOLL solution. It's isoosmotic and you could } } separate cells from microsomal fraction or viruses etc. You may also } } separate different cell types by this technique. PERCOLL was manufactured } } by Pharmacia, which is a part of Amersham now, I believe. Good luck. Sergey } } } } At 11:06 AM 7/26/01 -0700, you wrote: } } |--------------------------------------------------| } Tamara Howard } Department of Cell Biology and Physiology } University of New Mexico - Health Sciences Center } Albuquerque, NM 87131 } thoward-at-unm.edu } |--------------------------------------------------|
School of Mechanical, Materials, Manufacturing Engineering & Management Advanced Materials Research Group University of Nottingham, UK
Professor of Materials
Applications are invited for the above post from candidates with an outstanding record of research to reinforce and add to existing research strengths and providing a third professorial post in the Advanced Materials Group. Candidates should have expertise in one of the following areas: electron microscopy applied to the processing of advanced materials; surface engineering of materials; near-net-shape processing of materials or biomaterials
The person appointed will be expected to contribute to the teaching of courses within the School, which was graded 4A in the Research Assessment Exercise and gained excellent scores for its teaching quality.
Salary will be within the professional range, minimum £41,089 per annum.
Informal enquiries may be addressed to Professor D G McCartney or Professor A B Seddon, tel: 0115 951 3740, Email: Angela.Seddon-at-Nottingham.ac.uk Further details of the School are available on the WWW at: http://www.nottingham.ac.uk/school4m/.
Further details and application forms are available on the WWW at: http://www.nottingham.ac.uk/personnel/ or from the Personnel Office, Highfield House, The University of Nottingham, University Park, Nottingham, NG7 2RD. Tel: 0115 951 3266. Fax: 0115 951 5215. Email: June.Pedlar-at-Nottingham.ac.uk. Please quote ref. MCM/165. Closing date: 15 September 2001.
…………………………………………………………………………………………………………
To be published as follows:
Guardian - Tuesday 24 July THES - Friday 27 July Jobs.ac.uk - w/c Monday 23 July
Back in the early to mid 80's, I used Ficoll-Hypaque to layer out different white cells from peripheral blood. Never had a problem with sugar crystals. Only damage to the diamond knife was (cringe) user error.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
For what its worth:
Some time ago we tried to section some epoxy-embedded, Percoll-separated cell fractions. This resulted in very evident damage to glass knife cutting edges - to such an extent that we decided not to jeopardize a diamond knife.
Separating the fractions on a sucrose gradient resulted in blocks that sectioned without any problems.
Jan Coetzee
Lab for Microscopy and Microanalysis University of Pretoria, South Africa. Tel: 012-420-2075, Fax 012-362-5150 www.up.ac.za/academic/electron/emunit1.htm
Tamara Howard wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I have a question about using Percoll for TEM - someone told me just last } week that you shouldn't use Percoll if you'll be doing thin sections, } since it is a silica (can damage knives) - has anyone used it and either } seen or not seen gunk in their sections? Scratches &/or damaged a knife? } Sea creatures or other beach denizens? (Sorry, I'm tired and couldn't } resist) } } I'm not planning to use it any time soon, but hearing about it again makes } me curious about using it to prep for EM. I've seen nasty sucrose crystals } a few times with samples from sucrose gradients, but they are mostly an } annoyance. } } Thanks! } } Tamara } } On Thu, 26 Jul 2001, Sergey Ryazantsev wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } It's better to use PERCOLL solution. It's isoosmotic and you could } } separate cells from microsomal fraction or viruses etc. You may also } } separate different cell types by this technique. PERCOLL was manufactured } } by Pharmacia, which is a part of Amersham now, I believe. Good luck. Serge y } } } } At 11:06 AM 7/26/01 -0700, you wrote: } } |--------------------------------------------------| } Tamara Howard } Department of Cell Biology and Physiology } University of New Mexico - Health Sciences Center } Albuquerque, NM 87131 } thoward-at-unm.edu } |--------------------------------------------------|
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id LAA11998 for dist-Microscopy; Fri, 27 Jul 2001 11:36:53 -0500 (CDT) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id LAA11994 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Fri, 27 Jul 2001 11:36:23 -0500 (CDT) Received: from thor.oit.pdx.edu (thor.oit.pdx.edu [131.252.120.40]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id LAA11987 for {Microscopy-at-sparc5.Microscopy.Com} ; Fri, 27 Jul 2001 11:36:08 -0500 (CDT) Received: from medusa.oit.pdx.edu (medusa.oit.pdx.edu [131.252.120.41]) by thor.oit.pdx.edu (8.11.1/8.11.1) with ESMTP id f6RGXHU14503 for {Microscopy-at-sparc5.Microscopy.Com} ; Fri, 27 Jul 2001 09:33:17 -0700 (PDT) Received: (from sec-at-localhost) by medusa.oit.pdx.edu (8.11.1/8.11.1) id f6RGXGo23355 for Microscopy-at-MSA.Microscopy.Com; Fri, 27 Jul 2001 09:33:16 -0700 (PDT) X-Authentication-Warning: medusa.oit.pdx.edu: sec set sender to lifengd-at-pdx.edu using -f To: Microscopy-at-sparc5.microscopy.com
Roommate wanted:
I have made a reservation at Hotel Queen Mary in Long Beach, California for the
Microscopy and Microanalysis 2001 Conference from August 5 – 9. It turns out the room has two double beds. If any female colleagues (one female) who want to
share the room with me, please contact me by e-mail (jiaoj-at-pdx.edu). The room is reserved for 4 nights ($115/night for two persons) from August 5 – August 9.
Dr. Jun Jiao Assistant Professor Physics Department Portland State University Portland, Oregon, U.S.A.
I have used Percoll for leydig cell seperations and TEM with no knife damage. After the isolation procedure I have used a series of buffer washes before dehydration and embedding procedures.
Al Kruger Reproduction Research Unit USDA-ARS-MARC Clay Center, NE 68901
Tamara Howard wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I have a question about using Percoll for TEM - someone told me just last } week that you shouldn't use Percoll if you'll be doing thin sections, } since it is a silica (can damage knives) - has anyone used it and either } seen or not seen gunk in their sections? Scratches &/or damaged a knife? } Sea creatures or other beach denizens? (Sorry, I'm tired and couldn't } resist) } } I'm not planning to use it any time soon, but hearing about it again makes } me curious about using it to prep for EM. I've seen nasty sucrose crystals } a few times with samples from sucrose gradients, but they are mostly an } annoyance. } } Thanks! } } Tamara } } On Thu, 26 Jul 2001, Sergey Ryazantsev wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } It's better to use PERCOLL solution. It's isoosmotic and you could } } separate cells from microsomal fraction or viruses etc. You may also } } separate different cell types by this technique. PERCOLL was manufactured } } by Pharmacia, which is a part of Amersham now, I believe. Good luck. Sergey } } } } At 11:06 AM 7/26/01 -0700, you wrote: } } |--------------------------------------------------| } Tamara Howard } Department of Cell Biology and Physiology } University of New Mexico - Health Sciences Center } Albuquerque, NM 87131 } thoward-at-unm.edu } |--------------------------------------------------|
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 CASI Home Page: email: fmonson-at-wcupa.edu Schedule: http://bio.wcupa.edu/cgi-bin/webreserve.acgi Please call before visiting.
} ---------- } From: Mary Tyler } Sent: Thursday, July 26, 2001 3:11 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: LM time-lapse video microscopy } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Folks, } I'm in the process of setting up a system for doing time-lapse video } microscopy and would love to get information from others on what set-ups } work best for them. I'm interested in people's experience with time-lapse } VCRs and digital capture. I'm recording in color using a three-chip CCD } video camera. We have a Scion capture card on a Mac G4. I will be } recording over 24-hour periods. } In particular, has anyone been using NIH Image or Imagej to do time-lapse } in color? } } I am new to this listserv, so my apologies if I'm raking over old } territory. I did do a search of the listserv archives and didn't find any } postings in this subject. Many thanks, Mary } =============================================== } Mary_Tyler-at-umit.maine.edu or mtyler-at-maine.edu } } Mary S. Tyler } Professor of Zoology } Dept. of Biological Sciences } University of Maine } Orono, ME 04469-5751 } FAX: 207-581-2537 } } }
In the old days, filters could be 'burned' out by the high voltage Hg lamp if there wasn't a proper buffer. Is the bulb correct for the illuminator? Sometimes these days, a neutral density filter is placed between the illuminator and the filter set.
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 CASI Home Page: email: fmonson-at-wcupa.edu Schedule: http://bio.wcupa.edu/cgi-bin/webreserve.acgi Please call before visiting.
} ---------- } From: jmkrupp-at-cats.ucsc.edu } Sent: Thursday, July 26, 2001 5:59 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: LM: DAPI excitation filter damage? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi: } } A colleague has asked if I could help explain why the excitation filter in } his DAPI filter set on a Zeiss Axiophot deteriorates after a few months } of } use. } } Excitation filters in other sets last longer and don't show this problem. } The symptoms are uneven illumination that shows up in the image and an } obvious 'burned ' look to the filter. } } His questions are like: Is this normal? If not, what is wrong? How can I } avoid this problem? } } Thanks } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } } } }
First I would like to thank everyone for their help and suggestions = with my problem of "dirty grids" or as I call it (technical term) ka ka = on the sections.
I know it has been a while since I asked this question, but I have not = been able to go through and try out all the suggestions sent to me .
The solvent I am using is a mixture of 50% Methanol and 50% ddH20 Staining was done on dental wax for 10 minutes.
For lead staining it was done for 5 minutes with sodium hydroxide = pellets in a covered petri dish.
Everything was filtered through .45 micron filters.
Solutions I was given for my staining artifact were:
1. Change filter size to .20 microns. 2. Use parafilm instead of dental wax for staining. 3. Shorten my staining time from 10 minutes to 5 minutes. 4. Change suppliers of UA stain 5. Try a new batch of stain.
Solutions I tried.
1. Used parafilm instead of Dental wax 2. used .20 micron filters 3. mixed up a new batch of stain.
By changing all these criteria I fixed the staining problem that occurred a few weeks ago.
Now I get about 90-95% clean grids. Sometimes there is a little schmutz around, but not like occurred before.
Thanks to everyone for their suggestions
Eric A. Rosen UCLA Medical Center Electron Microscopy Lab Dept. Pathology and Lab Medicine Los Angeles, CA 90095 310-825-5737
I haven't had trouble with clays but certainly with gypsum (CaSO4 2H2O). I imagine a very hydrous clay would cause similar problems achieving vacuum. You could dehydrate the sample in a (vacuum) oven, but then of course you're looking at a different clay from the one you started with. Some variety of Environmental or Cryo-SEM might be required, depending on your needs.
Matt
Matthew J. Lynn Center for Advanced Microscopy University of Miami (305)284-4736 mlynn-at-miami.edu
On Friday, July 27, 2001 8:03 AM, Alun [SMTP:alun-at-deepsouth.co.nz] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Has anyone had any experience with SEM photographing minerals, } or had a problem in obtaining a vacuum in the case of clay type minerals. } } Bernard. } }
I don't think that filtering achieves much since that will only eliminate the boulders; 0.5um rocks are 5mm at only 10,000x mag. Use 2x distilled water.
UA in methanol goes off much faster than UA in water and that may be your major problem. Made up in water I trust UA for one month, when refrigerated and mostly kept in the dark. Made up in methanol, I would trust that solution only for a day or two.
The second problem is over-staining. Ten or even twenty minutes for aqueous UA is normal, but methanolic UA may be over-staining after 2 minutes or so.
I like your technical term "schmutz" - for the uninitiated, schmutzig is German for "dirty" and schmutz means dirt or just muck. I vote that "schmutz" should be official jargon in microscopy. Another little hint: We all have noticed that a grid when applied to a drop of staining solution floats to the top. So does all the schmutz. It makes sense to apply a grid to the side of a drop, that way any top floating schmutz is not picked up but pushed away. With aqueous UA and lead stain it makes sense to only stain one side of the section. Methanolic stains may not support the grid and the grid must be placed into the drop. This increased the chance of contamination and also causes more intense staining. More intense staining may sound desirable, but there are some negatives. This contributes to over-staining, also, some stains only penetrate part of a section and so give the effect of a thinner section and sharper image. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Saturday, July 28, 2001 4:28 AM, Eric [SMTP:biology-at-ucla.edu] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } To the Microscopy List, } } } First I would like to thank everyone for their help and suggestions = } with my problem of "dirty grids" or as I call it (technical term) ka ka = } on the sections. } } I know it has been a while since I asked this question, but I have not = } been able to go through and try out all the suggestions sent to me . } } } The solvent I am using is a mixture of 50% Methanol and 50% ddH20 } Staining was done on dental wax for 10 minutes. } } For lead staining it was done for 5 minutes with sodium hydroxide = } pellets in a covered petri dish. } } Everything was filtered through .45 micron filters. } } } Solutions I was given for my staining artifact were: } } 1. Change filter size to .20 microns. } 2. Use parafilm instead of dental wax for staining. } 3. Shorten my staining time from 10 minutes to 5 minutes. } 4. Change suppliers of UA stain } 5. Try a new batch of stain. } } } Solutions I tried. } } 1. Used parafilm instead of Dental wax } 2. used .20 micron filters } 3. mixed up a new batch of stain. } } By changing all these criteria I fixed the staining problem that } occurred a few weeks ago. } } Now I get about 90-95% clean grids. Sometimes there is a little } schmutz around, but not like occurred before. } } Thanks to everyone for their suggestions } } Eric A. Rosen } UCLA Medical Center } Electron Microscopy Lab } Dept. Pathology and Lab Medicine } Los Angeles, CA 90095 } 310-825-5737 } }
I know that this server and most universities and other large institutions are virus protected, but many readers will be interested to learn that my usual tally of (eliminated) virus messages is about 2 per week. During the last ten days this has blown out 26. My server scans for and eliminates the virus part of a message and we have our own anti virus system. Just now, a lot of people out there are badly affected by the TROJ_SIRCAM.A virus and its variants. All of these message come in with different headers and all these files terminate with these extensions: exe, com, pif, bat, 1nk, scr. I received a couple of virus messages from SE Asian Universities.
Incidentally: That amazing Nigerian scam was supposedly squashed. Hmmm, I used to get one or two messages weekly, now I get ten or more from them and some clones. Trouble is that emails are too cheap to send and since no legislator is tough on spaming, we all must waste our time (and some fools part with their money). 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
I was wondering if anyone could offer some advice regarding my 2010F gun.
I periodically measure beam current using the small viewing screen after aligning the microscope in a reproducible configuration (A1 and A2 set at specific values, 50micron condenser aperture, CBD mode 1nm alpha 9, 1MX magnification with beam focused using Brightness knob). Over the last several months I have observed that the size of the spot is approximately 4 times larger than when the microscope was commissioned in early 1999. The FWHM of the probe was measured yesterday to be 2.2nm. The emission timer reads 19,200 hrs.
Obviously our analytical capabilities are compromised by the gun's current condition but we can still perform the majority of our work.
Is the probe size likely to continue to get larger?
Is our emitter about to expire completely?
What are the chances of the gun lasting another 6 to 12 months without the probe deteriorating dramatically?
Thanks for any advice you can offer, cheers
Mark Blackford Materials Division, ANSTO PMB 1, Menai, N.S.W., 2234 Australia
Phone 61 2 9717 3027 Fax 61 2 9543 7179
Disclaimer: The views expressed in this E-mail message do not necessarily represent the official views of ANSTO from which this message was conveyed.
We are seeking to buy an operational 100CX objective lens power supply. Please contact me off line at the address below if you have one for sale.
Thanks in advance.
Owen
Owen P. Mills Electron Optics Facility Engineer Michigan Technological University Department of Materials Science and Engineering Rm 512 M&M Building Houghton, Mi 49931 Ph: 906-487-2002 FAX: 906-487-2934 Email: opmills-at-mtu.edu URL: http://www.mm.mtu.edu/~opmills/index.html
Hello fellow microscopists, has anyone out there know how I can get the free trial version ACDSee 16 bit installation software suitable for Win 3.11 operating system? I tried getting it from their web site but it appears they have discontinued it -only programs suitable for Win 95 and higher.I know that until recently this free down loadable version was available on their website. I have tried contacting their helpline, but so far to no avail. I'd be very grateful to anyone who could contact me off-line with information where I can get it. Or perhaps does anyone know of something very similar -allowing very quick viewing of images in file manager of 3.11? Regards Martin Roe
Hello fellow microscopists, has anyone out there know how I can get the free trial version ACDSee 16 bit installation software suitable for Win 3.11 operating system? I tried getting it from their web site but it appears they have discontinued it -only programs suitable for Win 95 and higher.I know that until recently this free down loadable version was available on their website. I have tried contacting their helpline, but so far to no avail. I'd be very grateful to anyone who could contact me off-line with information where I can get it. Or perhaps does anyone know of something very similar -allowing very quick viewing of images in file manager of 3.11? Regards Martin Roe
I read your message just after carrying out my monthly emitter checks on our 3000F at 20,386 hours. We also hope to predict the degradation in emitter performance and plan a service replacement for a few minutes before the tip fails:-) Well the're not cheap are they?
I have a slightly different set up; I adjust A1 to get 200uA emission and record similar data to you. I also record the beam current on a picoammeter. I don't measure the spot size accurately but I note the magnification required to fill the centre spot on the small screen. It's a subjective test but the result has been either 1, 1.2 or 1.5M times at every test and the spot does not show any trend towards getting larger.
What I have noticed is that the spot size is very dependent on the objective lens setting (now standardised) and beam tilt (rotation centre). If there is any objective astigmatism then it is 'corrected' by the condenser stigmators giving a larger spot.
If you do measure the current on the small screen you may find that it alters with both probe size and position on the screen (different absorption at different positions).
Make sure you are at DV=0 (or -14 in our case to allow for the GIF voltage offset) and that the column is always fully aligned in the mode you use (in our case CBD, alpha 9, spot 0.4nm and smallest C aperture).
I know that Sheffield University were monitoring their 2010F emitter but I don't think that they got any warning of a tip failure before their's was replaced.
Maybe others have better information for you.
Regards, Ron
On Mon, 30 Jul 2001 17:19:26 +1000 Mark G BLACKFORD {mgb-at-ansto.gov.au} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear All, } } I was wondering if anyone could offer some advice regarding my 2010F gun. } } I periodically measure beam current using the small viewing screen after } aligning the microscope in a reproducible configuration (A1 and A2 set at } specific values, 50micron condenser aperture, CBD mode 1nm alpha 9, 1MX } magnification with beam focused using Brightness knob). Over the last } several months I have observed that the size of the spot is approximately 4 } times larger than when the microscope was commissioned in early 1999. The } FWHM of the probe was measured yesterday to be 2.2nm. The emission timer } reads 19,200 hrs. } } Obviously our analytical capabilities are compromised by the gun's current } condition but we can still perform the majority of our work. } } Is the probe size likely to continue to get larger? } } Is our emitter about to expire completely? } } What are the chances of the gun lasting another 6 to 12 months without the } probe deteriorating dramatically? } } Thanks for any advice you can offer, cheers } } } Mark Blackford } Materials Division, ANSTO } PMB 1, } Menai, N.S.W., 2234 } Australia } } Phone 61 2 9717 3027 } Fax 61 2 9543 7179 } } Disclaimer: } The views expressed in this E-mail message do not necessarily represent the } official views of ANSTO from which this message was conveyed. }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
I am considering options for a CCD on a 35mm port or in the PEELS compatible location of a Gatan PEELS. Any input, including from vendors, would be useful. This would be for HREM applications.
------------------------------------------------------- Laurence Marks Department of Materials Science and Engineering & Center for Transportation Nanotechnology Northwestern University Evanston, IL 60201, USA Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu http://www.ctn.northwestern.edu ------------------------------------------------------- The Other Nanotubes http://focus.aps.org/open/st12.html Boron Nitride Nanotubes http://pubs.acs.org/cen/topstory/7912/7912notw1.html
Are you aware that in some german dialects "Schmutz" does means other things that dirt? And accidentally some people have family names as Schmutz and even worst they happen to be quite good microscopists using this server as well. May be some latin name or older "dead" language name would help? Or why not leva is as dirt? Just a suggestion. svetla -- Svetla Stoilova-McPhie, PhD The Burnham Institute, 10901 N. Torrey Pines Road, La Jolla, CA 92037 tel: 858 646 3100 ext.3484, fax: 858 646 3196
Hello everyone, All of my previous experience in preparing thick and thin sections for LM and TEM, respectively, is with epon. Should I expect similar behavior with araldite when sectioning with a diamond knife and staining with toluidine blue or aqueous UA and Reynolds lead citrate? I would appreciate any tips or references. Thanks.
Donald Gantz Dept. Physiology & Biophysics Boston University School of Medicine Boston, Massachusetts 02118 Email: Gantz-at-Biophysics.bumc.bu.edu Phone: 617-638-4017
The Microscopy Society of America is seeking an Archivist to manage the historical documents of our Society. This position is appointed by MSA Council for a period of three years.
We are looking for someone who is experienced in the storage and cataloging of (primarily) paper documents. Although this is a voluntary position (non-paying), the Society would provide funding for expendibles and day-to-day operations (and possibly a part-time, student-worker assistant).
If interested (or if you know of a qualified person), please contact me at the M&M 2001 meeting in Long Beach next week or at the following address:
-- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ##############################################################
We have a Hitachi S-650 with two 160VP pumps that need to be replaced. I have called Hitachi about information on the pumps, but they are having some difficulty in locating any specs on them. Searching the Internet did not give any information either.
The pumps plug into what appears to be a standard 3-prong socket and the pumps have information on the housing for 110 V, so I'm thinking that the power requirements are fairly standard. What I don't know is what flow rate, et cetera, to get for the replacement pumps.
Does anyone know where I could locate the pump specs or could you recommend some pumps that would work? And, in case you haven't figured it out already, I'm not an expert on vacuum systems, so I may be going about this the wrong way by looking at specs that really don't matter. If someone could let me know what parameters are the important things to look for, I'd be grateful.
Postdoctoral Research Associate in Quantitative Electron Microscopy of Nano-Structures
Applications are invited for a postdoctoral research associate position at the Dept. of Materials Science and Engineering, University of Illinois-Urbana/Champaign. The position involves developing electron diffraction and spectroscopy techniques for quantitative characterization of the electronic and structural properties of nano-structures, using the state of art electron microscopy facilities (JEOL 2010F-GIF and VG HB501-STEM) at the Center for Microanalysis of Materials. The successful candidate should have strong experience in advanced materials electron microscopy and diffraction techniques. Experience with STEM, energy-filtered imaging and diffraction, and electron energy-loss spectroscopy is desired, but not required. Preference will be given to candidates with strong experimental aptitude. A PhD. in Physics, Materials Science or related field is required. The position is funded by DOE and is initially for one year with possibility for renewal up to 3.5 years. Applicants should send a curriculum vitae, including a list of publications and research experience, and the names of three references to: Prof. J.M. Zuo, Dept. of Materials Science and Engineering, University of Illinois at Urbana and Champaign, 1304 W Green Street, Urbana, IL 61801; 217-333-2736 (fax); (217) 244-6504 (phone); jianzuo-at-uiuc.edu (email).
Application materials should be received before Oct. 1, 2001 for full consideration.
Dear Mary, As you've been told, NIH Image will do this, saving the timelapse images as a stack held in RAM. However, Object Image, the extended version of NIH Image (avalilable from simon.bio.uva.nl), will do it with a means to create a similar control window to add integration or averaging, and to save the image series to hard drive. I will send you the macro for Object Image directly.
Regards, Glen
Mary Tyler wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 Folks, } I'm in the process of setting up a system for doing time-lapse video } microscopy and would love to get information from others on what set-ups } work best for them. I'm interested in people's experience with time-lapse } VCRs and digital capture. I'm recording in color using a three-chip CCD } video camera. We have a Scion capture card on a Mac G4. I will be } recording over 24-hour periods. } In particular, has anyone been using NIH Image or Imagej to do time-lapse } in color? } } I am new to this listserv, so my apologies if I'm raking over old } territory. I did do a search of the listserv archives and didn't find any } postings in this subject. Many thanks, Mary } =============================================== } Mary_Tyler-at-umit.maine.edu or mtyler-at-maine.edu } } Mary S. Tyler } Professor of Zoology } Dept. of Biological Sciences } University of Maine } Orono, ME 04469-5751 } FAX: 207-581-2537
Hello, I am working with apoptoic cells in plant tissues. I am using Evan's Blue as a test of viability and of apoptosis. Can anyone tell me what Evan's blue (EB) stains in the cell? I know that it can not enter a cell with an intact membrane but when it enters a damaged or dead cell what does the EB actually bind to? Ann
They are pretty good in locating info and fixing all sorts of items. They do mech, ion and diffusion pump rebuilds. Very good service and high quality.
gary g.
At 12:41 PM 7/30/2001, you wrote:
} We have a Hitachi S-650 with two 160VP pumps that need to be replaced. I } have called Hitachi about information on the pumps, but they are having some } difficulty in locating any specs on them. Searching the Internet did not } give any information either. } } The pumps plug into what appears to be a standard 3-prong socket and the } pumps have information on the housing for 110 V, so I'm thinking that the } power requirements are fairly standard. What I don't know is what flow } rate, et cetera, to get for the replacement pumps. } } Does anyone know where I could locate the pump specs or could you recommend } some pumps that would work? And, in case you haven't figured it out } already, I'm not an expert on vacuum systems, so I may be going about this } the wrong way by looking at specs that really don't matter. If someone } could let me know what parameters are the important things to look for, I'd } be grateful. } } Thanks, } } -Norman Kay } AME Dept. } The Univ. of Arizona
Once again the MSA Education Committee is organizing Exhibitor Demonstrations on Tuesday, August 7 from 5:00 to 6:00pm in the Exhibit Hall. These mini-seminars or tutorial demonstrations are held in the booths of the participating companies after the Hall is closed to non-participants at 5:00 pm.
Signup sheets with titles and descriptions are at the MSA Education table in the MSA Mega Booth. When you sign up you will be issued a ticket, which you will need to renter the Hall after it is closed. You need to sign up no later than noon on Tuesday. The number of attendees is limited, so visit the MSA Education table soon, since the demonstrations get filled up quickly!
Here's a list of participating Exhibitors and titles:
Asylum Research Force Measurements 101 - Featuring the New, Low Noise BioLever
Carl Zeiss, Inc., Microscopy & Imaging Systems Zeiss LSM 5 PASCAL Personal Confocal Microscope Features and Capabilities.
Digital Instruments/Veeco Metrology Group Recent Advances in Throughput, Resolution, Sensitivity, and Functionality with Atomic Force/Scanning Probe Microscopy
Evex Analytical Video SEM
FEI Company High Resolution STEM Imaging on a TEM
Gatan, Inc. Fast Mapping with Hi-Speed EELS Spectrum Imaging
Hamamatsu Photonic Systems Hamamatsu Introduces IEEE1394
Hitachi Scientific Instruments Hitachi's NEW WxB Filter System on the S-4700 FESEM
IATIA Group Optical Phase Microscopy with QPm: Quantitative Imaging and Conventional Visualization Analogs
IXRF Systems, Inc. SEM-EDS Integration: The Future of Microanalysis!
Micro Photonics, Inc. Non-Destructive 3D Microscopy Using X-Ray Microtomography
Molecular Imaging PicoSPM and In Situ Imaging Under Controlled Environments
Olympus America, Inc. Motorized Microscopy - Making Your Life easier
Polaroid Corp. Polaroid's Digital Microscope Camera and Simple PCIp Software
Princeton Gamma Tech, Inc. "What is That PTS I Hear So Much About?"
Q3DM, Inc. Automating High Resolution Microscopy: Addressing the Challenges of High Throughput Fluorescence Microscopy for Cell-Based Assays
Quesant Instruments Corp. Quesant Instruments New Metrology System
Soft Imaging System Corp. Digital Imaging on Your TEM: Everything You Wanted to Know But Were Afraid to Ask.
South Bay Technology, Inc. #1-Sneak Preview of New Techniques and Tool Sets for the D500i DIMPLER #2-Ion Beam Sputter Deposition & Etching (IBS/e) #3-Dedicated Low Energy Ion Milling
Ted Pella, Inc. New Methods of Microwave-Assisted Processing for Electron Microscopy
TSL (EDAX booth) Phase Identification and TEM Tool
See you in Long Beach!
Aloha, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
We are hoping someone out there, preferably within Australia, may have an ultramicrotome within their facility that has become redundant or obsolete (preferably an Ultracut E, Ultracut s or a Reichert OMU 3). If so, please reply ASAP.
We plan to buy CCD cameras for our two TEMs, JEM-2000 EXII and JEM2010F. The models on our final (almost) list are Gatan 692 (1K by 1K) for 2000Ex and 795.201.2 (2K by 2K) for 2010F
We use 2000 EX for routine IC failure analysis and 2010F for HR and EDS. Both CCDs are quite expensive. We will appreciate any user experience on both modles or any suggestions on lower cost ones.
You will be responsible for operating a transmission electron microscope (TEM), scanning electron microscope (SEM), taking and processing images. Responsibilities include collecting, processing, embedding, and sectioning tissues in support of electron microscopy services. You will keep records in compliance with SOP's and GLP's and maintain laboratory instruments in accordance with SOP's, troubleshooting equipment problems as needed. You will process samples in a timely manner according to SOP's. You will maintain and keep the laboratory clean and organized and ensure that instrumentation is cleaned, calibrated, and properly maintained according to facility SOP's. You will ensure that laboratory computer equipment operates smoothly and work with computer support to trouble-shoot network and software problems. You may also act as Pathology monitor for assigned Drug Safety studies. You will tabulate and report Imaging data to the laboratory supervisor and work with the laboratory supervisor to ensure the accuracy and completeness of all Pathology data presented in the final report to the FDA and sent to archives. You will assist in writing SOP's and appropriate forms to ensure compliance in all GLP studies and to maintain records for all studies that are non-GLP or in the spirit of GLP. You may also assist investigators with (SEM) and digital imaging needs. You will perform additional departmental and facility support functions such as (but not limited to) inventory, scheduling, or purchasing.
A Bachelor's degree is required in Biology, Biochemistry, or Chemistry or a related field with 7 years relevant experience or a Master's degree with 5 years relevant experience. 5-7 years experience working in transmission and scanning electron microscopy, in an industrial, hospital, or highly specialized academic setting is also required. Candidate should have strong communication and computer skills.
David W. Pereira Wyeth-Ayerst Laboratories 641 Ridge Road Chazy, NY 12921 Tel. (518) 846 6464 Fax. (518) 846 6345 email pereird-at-war.wyeth.com
Evans Blue will spontaneously react with albumenoids to form a covalent linkage. Under those circumstances, the dye becomes fluorescent (red).
Evans Blue has been used to tag albumen (1:1). Then it is dialyzed to remove unbound dye. Finally the tagged albumen is injected into an animal vascular system. It is used this way to test for sites of extravasation following injury or as an injection to test for blood volume. It is NOT known to be hazardous (I believe).
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 CASI Home Page: email: fmonson-at-wcupa.edu Schedule: http://bio.wcupa.edu/cgi-bin/webreserve.acgi Please call before visiting.
} ---------- } From: Ann P. Ketter } Sent: Monday, July 30, 2001 8:17 PM } To: listserver microscopy } Subject: LM- what does evan's blue stain? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello, } I am working with apoptoic cells in plant tissues. I am using Evan's } Blue as a test of viability and of apoptosis. Can anyone tell me what } Evan's blue (EB) stains in the cell? I know that it can not enter a } cell with an intact membrane but when it enters a damaged or dead cell } what does the EB actually bind to? } Ann } } }
Wyeth-Ayerst Research, the pharmaceutical division of American Home Products, is seeking an experienced and responsible microscopy technician at its Drug Safety facility in Chazy NY. This is a new, state of the art facility located just minutes from Lake Champlain and the Adirondack Mountains. Although located in rural setting it is only an hours drive to Montreal, Quebec and Burlington Vermont. You will be responsible for operating a transmission electron microscope (TEM), scanning electron microscope (SEM), taking and processing images. Responsibilities include collecting, processing, embedding, and sectioning tissues in support of electron microscopy services. You will keep records in compliance with SOP's and GLP's and maintain laboratory instruments in accordance with SOP's, troubleshooting equipment problems as needed. You will process samples in a timely manner according to SOP's. You will maintain and keep the laboratory clean and organized and ensure that instrumentation is cleaned, calibrated, and properly maintained according to facility SOP's. You will ensure that laboratory computer equipment operates smoothly and work with computer support to trouble-shoot network and software problems. You may also act as Pathology monitor for assigned Drug Safety studies. You will tabulate and report Imaging data to the laboratory supervisor and work with the laboratory supervisor to ensure the accuracy and completeness of all Pathology data presented in the final report to the FDA and sent to archives. You will assist in writing SOP's and appropriate forms to ensure compliance in all GLP studies and to maintain records for all studies that are non-GLP or in the spirit of GLP. You may also assist investigators with (SEM) and digital imaging needs. You will perform additional departmental and facility support functions such as (but not limited to) inventory, scheduling, or purchasing.
A Bachelor's degree is required in Biology, Biochemistry, or Chemistry or a related field with 7 years relevant experience or a Master's degree with 5 years relevant experience. 5-7 years experience working in transmission and scanning electron microscopy, in an industrial, hospital, or highly specialized academic setting is also required. Candidate should have strong communication and computer skills. Interested individuals should send their resumes, including a description of their expertise and references to:
David W. Pereira Wyeth-Ayerst Laboratories 641 Ridge Road Chazy, NY 12921 Tel. (518) 846 6464 Fax. (518) 846 6345 email pereird-at-war.wyeth.com
At 2:24 PM -0400 7/30/01, Don Gantz wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
************** Hi Donald,
Much would depend on the formulation of araldite used. Like Epon, et al, it can be harder or softer if the ratios of components is varied. My experience is that, keeping relative hardness in mind, it behaves pretty much like epon. I'm sure others, with more experience working with it will have more info on this.
Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
I expect that no two vacuum engineers would agree on the ideal pump size for a particular system: to wit the tiny pumps used by JEOL and the huge twin pumps used on most Hitachi scopes. Mechanical (also called rotary and fore pumps) are purchased largely on three figures: ultimate vacuum, pumping speed / volume (expressed as a volume over a time) and price. A small pump is cheaper to buy and uses less electricity, they tend to be quieter running (depending on make), larger pumps are the converse in properties but they pump down faster especially with new film in a TEM and when evacuating a larger space rather then a small airlock. In either case you must opt for a two-stage pump. Direct drive types are less trouble since belts break more frequently than couplings. Couplings become noisy and so warn the user that they need attention, but belts break with no warning. (It pays to inspect belts every three months or so, cracks on the inside of a belt indicated that a replacement is needed). Belt driven pumps run a little slower and live a little longer. Having said that, I would buy a direct drive pump one size larger than the reasonable minimum required. I think that 4cubic meters/ hour is a reasonable minimum for most EMs and so a 6 might be a good size. A 2 would work, but encourage an awful lot of long coffee breaks and a ten is a bit of overkill. I don't know how the Hitachi twin pumps share the work load, but it may not be easy to combine the pumping circuits, so you may need to retain two somewhat smaller pumps (2x 3 or 4 m3/h). Over the years I have used pumps of numerous makes and while they all work and most are quiet reliable, I would get a Leybold or an Edwards because they seemed to run quietest and achieved, for a given size pump the best ultimate vacuum. But these things may have changed in recent times. Perhaps other listers have an opinions on pumps performance. However, I should also declare my bias: ProSciTech is an agent (in Australasia) for Edwards pumps. Happily I have predilections for Edwards. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Tuesday, July 31, 2001 5:42 AM, SEM Machine [SMTP:SEM-at-ACATC.AME.Arizona.edu] wrote: } } } We have a Hitachi S-650 with two 160VP pumps that need to be replaced. I } have called Hitachi about information on the pumps, but they are having some } difficulty in locating any specs on them. Searching the Internet did not } give any information either. } } The pumps plug into what appears to be a standard 3-prong socket and the } pumps have information on the housing for 110 V, so I'm thinking that the } power requirements are fairly standard. What I don't know is what flow } rate, et cetera, to get for the replacement pumps. } } Does anyone know where I could locate the pump specs or could you recommend } some pumps that would work? And, in case you haven't figured it out } already, I'm not an expert on vacuum systems, so I may be going about this } the wrong way by looking at specs that really don't matter. If someone } could let me know what parameters are the important things to look for, I'd } be grateful. } } Thanks, } } -Norman Kay } AME Dept. } The Univ. of Arizona } } } }
There is no micro user onboard our research ship, and you are inundating our inmarsat connection with your email list. Please take care of this as soon as possible please,
Thank you,
Jeff Turmelle Sys Admin, R/V Maurice Ewing Lamon-Doherty Earth Observatory of Columbia University jefft-at-ldeo.columbia.edu
Question: I want to know the exact meaning of terms defocus,underfocus,focus,infocus and overfocus how they are classified for a particular object.
I would appreciate receiving reply in this regard
Dear Mullapudi Srinvas, For an ideal lens, all the electrons leaving a point on a specimen will be brought together in a point in a plane--the focal plane. When the lens currents are adjusted so that this plane is coincident with the detection system (film, CCD, etc.), the image will be in exact focus. (N.b., focus is a property of the image and is the same regardless of the particular object, although different points in thick objects can have different focal planes.) The distance between the focal plane and the detector plane is the amount of defocus--positive for overfocus (focal plane above the detector plane) and negative for underfocus. If you look at how information from the specimen is conveyed to the image as a function of spatial frequency--i.e., how the large and small features appear--you will find that there is a curve which defines the relationship between the amplitude of the wave of electrons leaving the specimen and that of the wave incident on the detector. This curve is the contrast transfer function (CTF), and it is of the same sign for frequencies lower than a certain value, then it changes sign and oscillates at higher frequencies. The amount of defocus which produces a CTF extending to the largest frequency before it changes sign gives the condition called Schertzer (sp?) focus. This condition results in images in which easily interpretable information extends to the highest resolution possible. Images are often taken somewhat underfocus, since that condition causes interference fringes to occur at edges due to the sign reversal of the CTF and enhances their visibility. This is often useful for images of biological specimens. I hope this answers your questions. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
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The Microscopy Lab at KU is getting a new SEM at the beginning of October. Consequently our current SEM, a Hitachi S-570 needs to be moved out before that time. If anyone needs an older SEM please contact me. Thank you Bruce Bruce Cutler, Ph.D. Director, Microscopy & Electronic Imaging Laboratory University of Kansas
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id NAA27436 for dist-Microscopy; Tue, 31 Jul 2001 13:32:17 -0500 (CDT) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id NAA27433 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Tue, 31 Jul 2001 13:31:47 -0500 (CDT) Received: from mail2.lsil.com (mail2.lsil.com [147.145.40.22]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id NAA27426 for {Microscopy-at-sparc5.microscopy.com} ; Tue, 31 Jul 2001 13:31:34 -0500 (CDT) Received: from mhbs.lsil.com (mhbs [147.145.31.100]) by mail2.lsil.com (8.9.3+Sun/8.9.1) with ESMTP id LAA03065; Tue, 31 Jul 2001 11:28:44 -0700 (PDT) Received: from [172.21.35.125] by mhbs.lsil.com with ESMTP; Tue, 31 Jul 2001 11:28:02 -0700 Received: by exccolorado.co.lsil.com with Internet Mail Service (5.5.2650.21) id {P0GAJR2S} ; Tue, 31 Jul 2001 12:22:29 -0600 Message-Id: {BC3CE0375D29D51199A400508B66CA6C367E5C-at-exc-snake.co.lsil.com} Microscopy-at-sparc5.microscopy.com
Lee,
There is a substance called choline hydroxide (Sigma Aldrich) that has 10,000:1 selectivity (Si:SiO2). Dilute 25 ml. choline with 65 ml of DI water at 90 degrees C (must be maintained accurately). Depending on the thickness of your poly, it should take 1 - 5 minutes to remove the poly and keep the GOX intact. Hope this helps. Hey, if you know Yih-Shung Lin and he is still there at TSMC tell him that John Staman said "Hi".
Regards,
John Staman, LSI Logic
-----Original Message----- } From: "TCLEEG-at-tsmc.com.tw"-at-sparc5.microscopy.com [mailto:"TCLEEG-at-tsmc.com.tw"-at-sparc5.microscopy.com] Sent: Tuesday, July 31, 2001 6:35 AM To: Microscopy-at-sparc5.microscopy.com
I am in search of SEM services for small metallic medical products in the Minneapolis or St. Paul area in Minnesota. If there are any suggestions, I would appreciate them. Thank you. Dan May IntraTherapeutics dmay-at-iti-med.com {mailto:dmay-at-iti-med.com}
Dear Norman, The Cutevac 160VP is a 160 l/min direct-drive rotary pump. It plugs into any 110v socket. Any direct-drive single or double stage vacuum pump with the same capacity should work. BTW, what are you doing with the old pumps? I have seven of them running in my lab and the oldest have been running continuously for 19.5 years. I love them, fix them myself when they leak oil and would love to get some more. At 12:41 PM 7/30/01 -0700, you wrote: } } We have a Hitachi S-650 with two 160VP pumps that need to be replaced. I } have called Hitachi about information on the pumps, but they are having some } difficulty in locating any specs on them. Searching the Internet did not } give any information either. } } The pumps plug into what appears to be a standard 3-prong socket and the } pumps have information on the housing for 110 V, so I'm thinking that the } power requirements are fairly standard. What I don't know is what flow } rate, et cetera, to get for the replacement pumps. } } Does anyone know where I could locate the pump specs or could you recommend } some pumps that would work? And, in case you haven't figured it out } already, I'm not an expert on vacuum systems, so I may be going about this } the wrong way by looking at specs that really don't matter. If someone } could let me know what parameters are the important things to look for, I'd } be grateful. } } Thanks, } } -Norman Kay } AME Dept. } The Univ. of Arizona } Regards adn good luck, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society 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 } } We plan to buy CCD cameras for our two TEMs, JEM-2000 EXII and } JEM2010F. The models on our final (almost) list are } Gatan 692 (1K by 1K) for 2000Ex and 795.201.2 (2K by 2K) for 2010F } } We use 2000 EX for routine IC failure analysis and 2010F for HR and } EDS. Both CCDs are quite expensive. We will appreciate any } user experience on both modles or any suggestions on lower cost ones. } } Regards, } Tan-Chen Lee
Dear Tan Chen Lee!
It would be useful to contact You directly to avoid direct advertisement on this mail list. Please send Your e-mail adress to the adress specified below.
My experience is that belt-drive pumps last a lot longer than direct, I have a couple of JEOL ones that have run more-or-less continuously for almost 30 years, with annual oil changes, frequent belt inspection and replacement as Jim says, and one replacement of the vane springs! Direct-drive ones don't seem to be able to do anything like that.
They are also quieter, particularly if the belts are sprayed occasionally with belt dressing.
Gimme belt-drives every time.
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
I don't remember the procedure, but you may remove PERCOLL after purification by centrifugation at some particular acceleration (g). It's clearly stated in the PERCOLL Instruction Manual. If you need real data, I may check. I was using negatively stained mitochondria for TEM after PERCOLL purification and did not noticed any PERCOLL particles in the preparation, so it's not a problem if you remove PERCOLL properly after purification.
Sergey.
At 06:54 PM 7/26/01 -0600, you wrote: } I have a question about using Percoll for TEM - someone told me just last } week that you shouldn't use Percoll if you'll be doing thin sections, } since it is a silica (can damage knives) - has anyone used it and either } seen or not seen gunk in their sections? Scratches &/or damaged a knife? } Sea creatures or other beach denizens? (Sorry, I'm tired and couldn't } resist) } } I'm not planning to use it any time soon, but hearing about it again makes } me curious about using it to prep for EM. I've seen nasty sucrose crystals } a few times with samples from sucrose gradients, but they are mostly an } annoyance. } } Thanks! } } Tamara } } } On Thu, 26 Jul 2001, Sergey Ryazantsev wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } It's better to use PERCOLL solution. It's isoosmotic and you could } } separate cells from microsomal fraction or viruses etc. You may also } } separate different cell types by this technique. PERCOLL was manufactured } } by Pharmacia, which is a part of Amersham now, I believe. Good } luck. Sergey } } } } At 11:06 AM 7/26/01 -0700, you wrote: } } |--------------------------------------------------| } Tamara Howard } Department of Cell Biology and Physiology } University of New Mexico - Health Sciences Center } Albuquerque, NM 87131 } thoward-at-unm.edu } |--------------------------------------------------|
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Many thanks to everyone who replied. I now have some idea of where I need to go from here.
To answer a few of the questions that came up, the problem was that before the SEM was donated to us, I believe it was not serviced for a while nor was it handled very gently during transit. I just started working with the unit and recently found out that when it was delivered, it was left at the wrong building & sat on the loading dock for a few days - during monsoon season. So, needless to say, this unit has had a rough life. We've had it serviced & the Hitachi tech said that the pumps would most likely need to be replaced soon, but I should have questioned more into why she thought that. The pump failed a little over 3 months after the service, so I guess she was on to something. I'm also learning as I go, since there isn't anyone here to teach me, which I'm sure has contributed to the early demise of the pump.
What happened to the pump was that one morning when I came in, there was a puddle of oil around the pump, so something obviously gave out during the night (all those years of schooling have taught me a thing or two...). Of the two pumps, the one that died was the "stronger" one, since we already knew that the other pump was a bit weaker. I called Hitachi & they told me that they didn't have a rebuild kit, per se, but that they could send a list of parts - no guarantees on if they were still available, though. Based on my past experience with trying to find parts for our SEM, I'm pretty sure very few of the parts are still available. I did give the pump to our machine shop, which has done a bit of work on vacuum pumps, and they rebuilt it as best they could, but it wasn't enough, since I cannot get a high vacuum now.
I should mention, though, that a Hitachi tech did contact me after I submitted the initial post & was very helpful on getting me some information on pumps that could replace our unit. Based on what he said, and also from replies here, it appears that we need something that can do about 9 cfm. I also plan on contacting a few of the companies suggested here on costs to rebuild the pumps, since they may be salvageable.
I am trying to stain some formaldehyde preserved (no osmium) fungal hyphae so I can visualize them after embedding in LRWhite.
Any suggestions? Tryphan blue works until you transfer the hyphae to LRWhite, which extracts all color. I have oriented the hyphae in small blocks of agar, but the small size of the hyphae makes it difficult to see them if I stain the agar, say with Sudan black as was once suggested years ago.
We have purchased Gatan Model 600 Dual Ion Mill for use in the preparation of foils of geological material for TEM applications. Since this unit was purchased used from a government lab it was delivered without technical support and with incomplete documentation, leaving many unanswered questions. We are having problems maintaining high vacuum (the diffusion pump shuts off) and cannot find the source of the problem. Help from anyone familiar with this device would be greatly appreciated.
We are attempting to thin quartz arenites and have spent the majority of our time (when the unit is operational) in an effort to optimize the thinning process. Any input in that regard would also be welcome.
Thanks in advance for any information you can provide.
Kevin Macke Bowling Green State University Department of Geology Bowling Green, OH Email: macke-at-bgnet.bgsu.edu
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Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (dlongerbeam-at-captusnetworks.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, July 31, 2001 at 19:24:09 ---------------------------------------------------------------------------
Email: dlongerbeam-at-captusnetworks.com Name: Donald Longerbeam
Organization: none
Education: Graduate College
Location: Roseville Ca.
Question: I was trained as a molecular biologist and did work with various microscopes over the years. However I have been out of the field for 25 years. I now have a son who I wish to introduce to the microscopic world. Who is making a good and reasonable microscope at this time. Olympus and Zeiss are certainly good - but they are too expensive. I am willing to spend up to $1000 apiece for a good binocular dissecting microscope and a good general higher power scope - . Thanks. Donald Longerbeam.
My usual golf partners won't be able to make it to M&M this year. Brian Cook and I are looking for two others to pair up with.
Thanks, Lucille Giannuzzi
******************************************************************* Lucille A. Giannuzzi, Ph.D.
Associate Professor, Mechanical, Materials, and Aerospace Engineering Director, UCF/Cirent Materials Characterization Facility University of Central Florida 12443 Research Parkway, Suite 305 Orlando, FL 32826 email lag-at-mail.ucf.edu phone (407) 882-1500 fax (407) 275-4321 -------------------------------------------------------------------- "Good judgement comes from experience.
Experience comes from making bad judgement."
Mark Twain ********************************************************************
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