As you mentionned, sensitivity 11 is close to normal value but can vary due to your developping time and developping temperature. If 11 is too much developped (too dark) then you can try higher number (usually until 14) to tell microscope that film is more sensitive , in another way use lower number for film not enough developped (too blank)
Michel RIBARDIERE TEM MANAGER JEOL (EUROPE)S.A.
-----Message d'origine----- De : zejianl-at-physics.unc.edu [mailto:zejianl-at-physics.unc.edu] Envoyé : lundi 31 mai 2004 19:22 Ā : Microscopy-at-MSA.Microscopy.Com Objet : Kodak EM4489 films sensitivity
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Dear colleagues, Do you have any experience of setting the sensitivity of EM 4489 films in TEM? I found the default value (11) in JEOL 2010F did not work very well. Your suggestions will be much appreciated.
best regards, zejian liu University of North Carolina at Chapel Hill Chapel Hill, NC 27599
Has anybody noticed a second round of changes in this film?
A while back there was a discussion about the "New Formulation" of this film and all the problems it caused with thin, patchy negatives. We finally got our new agitation procedures and exposures worked out and were getting good results again. Then, some months ago our negatives began coming out consistently too dark, so we began diluting our D-19 developer by an additional 25%, which gave us good results again.
Just last week we ran tests on MACO TEM film as a possible cheaper replacement for 4489. We processed a batch taken with three different exposure settings on our JEOL 1200EX and two agitation procedures. One procedure was our new current one of two seconds of nitrogen burst every 10 seconds, with one manual agitation every 30 seconds (lift and tilt five seconds each side), and the other was our old method of 10 seconds nitrogen burst every minute, with no manual agitation. To my surprise, the 4489 film came out beautifully using the old agitation method----no streaks or patchiness.
This leads me to think that there's a possibility that this film was reformulated yet again at some point, since nothing obvious changed in our procedures. Any similar experiences out there?
Thanks, Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 1 09:51:05 2004
First off, make sure your developer is fresh, properly diluted and at the correct temperature (68F/20C or a bit higher). I am always amazed at how some people will try to stretch old developer. Chemistry is the cheapest part of electron microscopy. Then do an exposure series as others have suggested.
Geoff
zejianl-at-physics.unc.edu wrote:
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-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 1 12:50:15 2004
We use Kodak 4489 film, and have for ages in a Philips CM10. Lately, I've noticed when I take the film out of the film box from the microscope, some of the film is not flat, but has popped up just a little on the edge of the film cassette that is open where you slide the film in. They aren't all popped up, just some. And others, if I take them out, can watch them pop up as they sit on the counter in the cassette. Possibly it's just from moisture in the air? I'm hoping they are flat in the scope and only pop up when removed from the scope, but I don't know. And have never noticed this before. Has anyone else encountered this? The images all look fine after development, I don't notice any out of focus spots on one side, but we haven't been doing any real high mag work lately.
Nancy Piatczyc
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 1 13:52:22 2004
On May 31, 2004, at 2:56 PM, Sergey Ryazantsev wrote:
} I don't remember exactly, but 0.5-07 OU (optical units, visual area of } the spectrum) should be OK, I guess (hey, EM community, correct me if } I am wrong).
Dear Sergey, We always calculated our exposure for 4489 so that we got an OD of 1.0. 4489 has a linear range to about OD 2--better than SO163--so the larger OD is still well within the linear response range and provides a really good image for prints and enlargements. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 1 14:32:55 2004
You can enhance/intensify the DAB reaction product with osmium, or nickel or nickel+cobalt before you embedd the tissue in Durcupan (or whatever). After embedding ......... I don't know. Cutting thicker sections might help a bit, also, staining with only UA (as someone else suggested) might allow the DAB to show up better. You could etch the sections with 10% hydrogen peroxide for a few minutes then try 1-2% osmium, that might give you enough contrast. Good luck.
Geoff
by way of MicroscopyListserver wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 1 16:05:57 2004
Quoting Nancy.P.Piatczyc-at-williams.edu: } We use Kodak 4489 film, and have for ages in a Philips CM10. } Lately, I've noticed when I take the film out of the film box } from the microscope, some of the film is not flat, } but has popped up just a little on the edge } of the film cassette that is open where you slide the film in. } They aren't all popped up, just some. } And others, if I take them out, can watch them pop up } as they sit on the counter in the cassette. } Possibly it's just from moisture in the air? } I'm hoping they are flat in the scope and only pop up } when removed from the scope, but I don't know. And } have never noticed this before. Has anyone else encountered this? } The images all look fine after development, } I don't notice any out of focus spots on one side, } but we haven't been doing any real high mag work lately. } } Nancy Piatczyc ================== Nancy, I have just loaded three cassettes of 4489 film and it does seem to curl more than I had remembered but not to the point where it was difficult to load.
Is there much play in the slots that the film slides into? If there is, you might consider applying a LITTLE pressure with a vise, just to reduce the gap a bit. My holders are a different type and periodically I need to open up the slots a bit when the film no longer slides into the holder.
Pat Connelly Dept of Biology Univ. of Pennsylvania psconnel-at-sas.upenn.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 2 07:58:53 2004
On behalf of the Executive Council of the Midwest Microscopy and Microanalysis Society, I would like to thank those of you who responded to my request for student poster session guidelines. The information that you provided will be very helpful to us, and we appreciate receiving so much useful feedback in such a short time.
Regards,
Elaine
Elaine Schumacher Senior Research Scientist McCrone Associates, Inc. 850 Pasquinelli Drive Westmont, IL 60559-5539 USA 630-887-7100 (tel) 630-887-7417 (fax) E-mail: eschumacher-at-mccrone.com Web Site: www.mccrone.com
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 2 11:07:32 2004
Seems like everyone makes a digital camera for light microscope applications. I would appreciate hearing any recommendations or experiences with any of the high resolution cameras available. This is for a metallurgical microscope with a c-mount and would need at least 1300x1000 resolution in color and a fast, TV like refresh rate for previewing. Any ideas?
Robert W. Skwarok Syncrude Research Center 9421-17 Ave. Edmonton, Alberta Canada, T6N-1H4 ph: 780-970-6931 fax: 780-970-6805 skwarok.robert-at-syncrude.com
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 3 00:28:21 2004
We use a Nikon DXM 1200F - excellent camera, 12 megapixel - so max pixel resolution is 3840 X 3072 and minimum 640 X 480 (10 different sizes to choose from). Image formats JPEG, BMP and TIFF. Very user friendly and everything is software driven through the PC.
I have no connection with the company - I am just a satisfied user!
Med vänliga hälsningar/With best regards
Gareth
***Come to Stockholm in 2004 to the 26th IFBLS congress*** Take a look at http://www.vardforbundet.se/ifbls2004
Gareth Morgan MPhil MSc FIBMS, Department of Laboratory Medicine (Labmed), Karolinska Institute, Karolinska University Hospital at Huddinge, F46 SE 141 86 Stockholm Sweden
OBS! Besöksadress: F-Huset, Forskningsgatan 2 F52, Rum 2.10. Laboratoriet för klinisk patologi och cytologi.
NB! Visiting address: Building F, Research Corridor 2 F52, Room 2.10. Clinical Histo- and Cytopathology Laboratory.
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 3 04:06:36 2004
The images were taken by people at Jeol in Japan. As far as I understand they introduced an image shift during exposure. (If you just shift the same image in the computer you get fringes all the way to the edge of the micrograph, which is not the case with the test images.)
} From the Young fringes Jeol claims an information limit better than 2 Angstroms, but I haven't seen Thon rings beyond about 4 Angstroms in any of the images. (I think you might have misunderstood this point: Young fringes extend up to twice as far as the outermost Thon ring, not only beyond the first zero.)
} From this discussion I got the impression that manufacturers use Young fringes because they extend further than Thon rings and somehow show the true information limit (which brings me back to my original question about the meaningfulness of the Youngfringe criterion).
} From Michael O'Keefes E-mails (offline) I understand that he distinguishes between information limit and convergence limit. Maybe that's what we are seeing here. (All these images were taken at magnifications around 50K and defocus between 1000 and 2000 nm.)
The main question for me is how to compare microscopes for the sort of microscopy we intend to do. Does the information limit really tell us what sort of resolution our reconstructions will have or is that determined by the extent of Thon rings.
I can send some images offline or put them on my homepage, if anybody wants to see them.
Philip
-----Original Message----- } From: Bill Tivol [mailto:tivol-at-caltech.edu] Sent: 28 May 2004 21:29 On May 26, 2004, at 2:30 AM, Philip Koeck wrote: } I have some test images (defocus around 1000 nm) from a FEG-TEM where } Young fringes seem to extend up to twice as far as Thon rings. Is that } usual and if so which of the two defines the information limit? } What do manufacturers mean by information limit? } Do Young fringes really show up to which resolution I can get useful } information from a micrograph? } At least in TEM of non-periodic biological specimens you normally don't } expect anything beyond the last clearly visible Thon ring. } Dear Philip, Do you generate the Young's fringes by shifting the image in the scope during the exposure, or do you use a computer to generate a shifted image and sum it with the unshifted image? In the latter case, the noise, which does not have a CTF related to the lenses, is perfectly correlated, so there will be fringes, but no rings. In the former case, the noise is independent for the shifted and unshifted parts of the image, so it will not add as much intensity to the fringes (there will be some correlation of random noise, so some intensity). The information limit is measured by the Thon rings. Whether the information from Thon rings beyond the first zero of the CTF is useful depends on the nature of the specimen, the accuracy with which the exact value of the defocus can be measured, and the processing algorithm used to compensate for the CTF. For thick, non-periodic biological specimens, the defocus varies through the specimen, the contrast is low, and the dose is low, so the S/N is low. It is still a matter of discussion whether such techniques as CTF compensation or focal series reconstruction are of benefit. Yours, Bill Tivol, PhD
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 3 08:00:31 2004
I thought I posted this a week ago, but I never saw it come up. Curious. Here goes again (TIA):
Hello Listers,
I have a colleague who has been trying to use TEM to look at isolated chloroplasts from a marine alga (Bryopsis). She has also tried looking at intact algae. The problem is that following a standard protocol (Gta fix using isolation media, Os, UA en bloc, dehyd in ethanols and PO, 'epon' embedment; and Pb and UA on-grid), the membranes along the perimeter of both the isolates as well as the supposedly intact algae appear fragmented, or missing. Mostly missing. The cytoplasmic matrix and internal membrane-bound structures such as thykaloids appear essentially normal in terms of electron density and membrane integrity, so on first inspection it would appear the membranes are disappearing following initial fixation, or, they are stubbornly invisible.
We thought the missing isolated chloroplast membrane was attributable to the isolation procedure, until the intact algae also came up with no plasma membrane.
Any comments on ways to keep the PM intact? Or is this a usual appearance for these tissues? As others have noted, the chloroplasts do remain mostly green following Os.
There was a recent thread regarding missing membranes. We plan to test the acetone dehydration idea, and we already incorporate some of the suggestions in our standard protocol such as en bloc UA, so before we embark on a voyage of discovery using additives, I thought I would toss this problem out to any marine algae experts out there. Or anyone else who has another thought to share.
Thanks, all!
Ann Lehman
++++++++++++++++++++++++++++++++++++++ Ann Hein Lehman Assistant Director, Electron Microscopy Facility Mailstop: LSC-314 Trinity College 300 Summit Street Hartford, CT 06106 v. 860-297-4289 e. ann.lehman-at-trincoll.edu f. 860-297-2538 www.trincoll.edu/~alehman
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 3 12:08:18 2004
-----Original Message----- } From: James Chalcroft Sent: Thursday, June 03, 2004 7:21 PM To: 'Lehman, Ann R'
I thought I posted this a week ago, but I never saw it come up. Curious. Here goes again (TIA):
Hello Listers,
I have a colleague who has been trying to use TEM to look at isolated chloroplasts from a marine alga (Bryopsis). She has also tried looking at intact algae. The problem is that following a standard protocol (Gta fix using isolation media, Os, UA en bloc, dehyd in ethanols and PO, 'epon' embedment; and Pb and UA on-grid), the membranes along the perimeter of both the isolates as well as the supposedly intact algae appear fragmented, or missing. Mostly missing. The cytoplasmic matrix and internal membrane-bound structures such as thykaloids appear essentially normal in terms of electron density and membrane integrity, so on first inspection it would appear the membranes are disappearing following initial fixation, or, they are stubbornly invisible.
We thought the missing isolated chloroplast membrane was attributable to the isolation procedure, until the intact algae also came up with no plasma membrane.
Any comments on ways to keep the PM intact? Or is this a usual appearance for these tissues? As others have noted, the chloroplasts do remain mostly green following Os.
There was a recent thread regarding missing membranes. We plan to test the acetone dehydration idea, and we already incorporate some of the suggestions in our standard protocol such as en bloc UA, so before we embark on a voyage of discovery using additives, I thought I would toss this problem out to any marine algae experts out there. Or anyone else who has another thought to share.
Thanks, all!
Ann Lehman
++++++++++++++++++++++++++++++++++++++ Ann Hein Lehman Assistant Director, Electron Microscopy Facility Mailstop: LSC-314 Trinity College 300 Summit Street Hartford, CT 06106 v. 860-297-4289 e. ann.lehman-at-trincoll.edu f. 860-297-2538 www.trincoll.edu/~alehman
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 3 13:22:43 2004
I was just reading a superb paper by Mobius et al (2002 J. Histochem. Cytochem. 50(1):43-55) in which they use PHEM buffer for their fixatives and immunostaining. PHEM is 60 mM PIPES, 25 mM HEPES, 2 mM MgCl2, 10 mM EGTA, pH 6.9. Can anyone comment on the rationale behind the design of this buffer? Specifically, why EGTA? In addition, why a PIPES/HEPES mixture rather than just PIPES or HEPES? Thanks, Tom
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (tauria-at-hotmail.com) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Thursday, June 3, 2004 at 11:29:54 ---------------------------------------------------------------------------
Email: tauria-at-hotmail.com Name: FRANCIS J. PRONESTI
Organization: INDEPENDENT CONSULTANT
Education: Graduate College
Location: CASTELLANA GROTTE, BARI, ITALY
Question: HELLO, I HAVE JUST PURCHASED A PHILIPS SEM 505, WHICH WAS SOLD AS SURPLUS FROM A UNIVERSITY, DUE TO UPGRADING. THIS INSTRUMENT WAS FUNCTIONING WHEN REMOVED FROM SERVICE AND APPEARS TO BE IN EXCELLENT SHAPE. UNFORTUNATELY, LIKE IN ALL THESE CIRCUNSTANCES, NO ONE NEW ANYTHING ABOUT SET UP DOCUMENTATION AND OPERATING INSTRUCTIONS. WHILE I AM NOT A COMPLETE NEOPHYTE WHEN IT COMES TO ELECTRON MICROSCOPES, I HAVE NO KNOWLEDGE NOR EXPERIENCE WITH THIS PARTICULAR MAKE AND MODEL. I HAVE CONTACTED PHILIPS AT THEIR WEBSITE REQUESTING ASSISTANCE, BUT SO FAR I HAVE HEARD NOTHING. CAN YOU DIRECT ME TO ANY SOURCE OF DOCUMENTATION FOR THIS MICROSCOPE? I WOULD BE VERY GRATEFUL. BEST REGARDS, FRANCIS J. PRONESTI (MSME.MSEE,PhD, IEEE MEMBER NO.41399650)
} The images were taken by people at Jeol in Japan. As far as I } understand } they introduced an image shift during exposure. } (If you just shift the same image in the computer you get fringes all } the way to the edge of the micrograph, which is not the case with the } test images.) } } From the Young fringes Jeol claims an information limit better than 2 } Angstroms, but I haven't seen Thon rings beyond about 4 Angstroms in } any } of the images. (I think you might have misunderstood this point: Young } fringes extend up to twice as far as the outermost Thon ring, not only } beyond the first zero.) } } From this discussion I got the impression that manufacturers use Young } fringes because they extend further than Thon rings and somehow show } the } true information limit (which brings me back to my original question } about the meaningfulness of the Youngfringe criterion). } } From Michael O'Keefes E-mails (offline) I understand that he } distinguishes between information limit and convergence limit. Maybe } that's what we are seeing here. (All these images were taken at } magnifications around 50K and defocus between 1000 and 2000 nm.) } } The main question for me is how to compare microscopes for the sort of } microscopy we intend to do. Does the information limit really tell us } what sort of resolution our reconstructions will have or is that } determined by the extent of Thon rings. } } I can send some images offline or put them on my homepage, if anybody } wants to see them. } Dear Philip, JEOL did do the test properly, but the Thon rings do not go out as far as they might. One has to find an appropriate area of the proper specimen to get good Thon rings. Two of the better specimens are Pt/Ir, which is strongly scattering and has both crystals (which give high-res Bragg spots) and amorphous parts (which give continuous scattering power), and amorphous C, which has continuous scattering power, but is more weakly scattering and, therefore, a more stringent test of performance. The acceptance tests for our scope were performed with Au on C, so there is continuous power from the C out to ~0.3 nm, and Au Bragg spots at 0.2 nm and 0.14 nm (as well as some even further out). The Thon rings do not extend as far out as the Bragg spots, primarily due to the low scattering power of C beyond 0.3 nm, but one can see segments of rings in the Bragg spots, so the envelope function has not yet become zero. Clearly, the scattering into the Bragg spots is real information, not an artifact, and it corresponds to lattice fringes visible in the image. I routinely take images of amorphous C at 2 um underfocus and count Thon rings to monitor the coherence of the FEG, and the rings extend beyond the 0.34 nm graphite peak. The scope has to be pretty well aligned and stigmated to get all the rings, and a thin, flat area of the specimen has to be found, otherwise areas at different heights will be at different defocus, and the Thon rings will be washed out. The service people were able to get many more than 30 rings--I think the record exceeds 45--but I don't remember the specimen or defocus value for which they were able to achieve this. Another potential difficulty may be the mag you chose. 50 kx may be too low if you are using a CCD, since the Nyquist limit may be a lot larger than 0.3 nm. For a given pixel size, any spatial frequency one wishes to measure must correspond to a wavelength greater than or equal to 2 pixels. Film may also present a problem at that mag; Nyquist is larger than the film grains, but your scanner may not go to high enough spatial frequency. We took many films (SO163) at that mag, and we used several scanners, and only one gave sufficient results to see Thon rings to 0.3 nm, despite the fact that all the scanners claimed 4000 dpi resolution. The specimen was Pt/Ir, and one could see the 0.2 nm fringes, even at 2 um underfocus (it took some experience to find them at this defocus), so we knew that we were getting info at high spatial frequency, but most of the scanned images (from the poor scanners) had nothing in the power spectrum at those frequencies. I'm not sure what your specimens are, so I don't know what would be acceptable performance. For focal series reconstructions of materials specimens, you will need to have information out to the spatial frequency corresponding to the resolution you want, and for something like biological single-particle analysis, where different particles are at slightly different defocus you still need information from each particle beyond the desired resolution, so that averaging the signal from a large number of particles will give acceptable S/N; however, in the first case, the resolution you need is likely to be near the info limit of the instrument, whereas, in the second case, radiation damage limits the attainable resolution to much poorer than the info limit, but the ability to obtain maximum resolution in the reconstruction depends upon how rapidly the envelope function decays. Since we do the latter, we want to be sure that we get Thon rings out to ~0.3 nm at a suitable defocus with a weakly-scattering specimen, and the info limit is less important. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 3 14:20:13 2004
} I have a colleague who has been trying to use TEM to look at isolated } chloroplasts from a marine alga (Bryopsis). She has also tried looking } at intact algae. The problem is that following a standard protocol } (Gta fix using isolation media, Os, UA en bloc, dehyd in ethanols and } PO, 'epon' embedment; and Pb and UA on-grid), the membranes along the } perimeter of both the isolates as well as the supposedly intact algae } appear fragmented, or missing. Mostly missing. The cytoplasmic matrix } and internal membrane-bound structures such as thykaloids appear } essentially normal in terms of electron density and membrane } integrity, so on first inspection it would appear the membranes are } disappearing following initial fixation, or, they are stubbornly } invisible. } } We thought the missing isolated chloroplast membrane was attributable } to the isolation procedure, until the intact algae also came up with } no plasma membrane. } } Any comments on ways to keep the PM intact? Or is this a usual } appearance for these tissues? As others have noted, the chloroplasts } do remain mostly green following Os. } } There was a recent thread regarding missing membranes. We plan to test } the acetone dehydration idea, and we already incorporate some of the } suggestions in our standard protocol such as en bloc UA, so before we } embark on a voyage of discovery using additives, I thought I would } toss this problem out to any marine algae experts out there. Or anyone } else who has another thought to share. } Dear Ann, If you have access to the appropriate equipment, you might try cryo-fixation and cryo-substitution. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 3 14:47:41 2004
Dear List, I am trying to track down a good description--including figures, if possible--of why the image shifts as a consequence of beam tilt when the image is defocussed, but not when it is focussed. I have only found a very short, and not entirely satisfactory, description in a Royal Microscopy Society handbook from 1984. I have, of course, seen references to beam-tilt-induced image shift, but not the details of the mechanism, in several EM texts. Can anyone point me in the right direction? TIA. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 3 14:52:42 2004
} I was just reading a superb paper by Mobius et al (2002 J. Histochem. } Cytochem. 50(1):43-55) in which they use PHEM buffer for their } fixatives and immunostaining. PHEM is 60 mM PIPES, 25 mM HEPES, 2 mM } MgCl2, 10 mM EGTA, pH 6.9. Can anyone comment on the rationale behind } the design of this buffer? Specifically, why EGTA? In addition, why } a PIPES/HEPES mixture rather than just PIPES or HEPES? Thanks, Tom } Dear Tom, I don't know the pKs of PIPES and HEPES off hand, but that may be a clue why both are included. EGTA is a Ca++ chelator, so it is undoubtedly there to lower or buffer the [Ca++]. Since there is quite likely to be some Ca impurity in the MgCl2, it makes sense to add EGTA, which binds Ca, but not Mg. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 3 16:59:13 2004
Not all membranes are equally reactive with osmium. Protein storage vacuoles in plant cells have a region known as the "globoid" which contains phytic acid crystals. It was thought to be a non-membrane bound region within vacuole's matrix. Conventional aldehyde and osmium fixation fails to show any sign of a membrane. When we used KMnO4 as a fixative, however, a unit membrane was present surrounding this compartment (Jiang et al., 2001 J. Cell Biol 155(6):991-1002). This type of phenomenon has been reported before. Plastid envelopes were observed to become progressively less distinct using conventional fixation in the fern Paesia but they could still be seen using KMnO4. Mitochondria membranes in the same cells were always stained so it seemed to be due to changes in plastid membrane lipids and not some trivial problem related to the osmication procedure as a whole (Bell 1983 Eur J Cell Biol 30:279-282). so maybe your membranes aren't missing - just hiding!
2:41 PM 06/03/04 -0700, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
The principle of the beam wobbler to assist focusing (originally an invention by Le Poole, patented by Philips back in the late 1940s as far as I remember) has been developed over recent years by Abraham (Bram) Koster and his group in Utrecht for application in computer-aided automatic focusing systems for 3D TEM tilt reconstructions. He should be able to help you. His publication list may be found in: http://www.bio.uu.nl/mcb/3dem/publications.html Best wishes,
Jim
-----Original Message----- } From: Bill Tivol [mailto:tivol-at-caltech.edu] Sent: Thursday, June 03, 2004 10:09 PM To: microscopy-at-msa.microscopy.com
Dear List, I am trying to track down a good description--including figures, if possible--of why the image shifts as a consequence of beam tilt when the image is defocussed, but not when it is focussed. I have only found a very short, and not entirely satisfactory, description in a Royal Microscopy Society handbook from 1984. I have, of course, seen references to beam-tilt-induced image shift, but not the details of the mechanism, in several EM texts. Can anyone point me in the right direction? TIA. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 4 03:36:50 2004
Tom, THe PHEM buffer was introduced by Manfred Schliwa and von (van?) Blerkum (OK, I probably didn't spell that one right) in the mid 80's for their ultrastructural studies of the cytoskeleton. The idea of the EGTA to remove calcium which can cause cytoskeletal degradation and supposedly the pH buffering of pipes + hepes works better than either alone. They may have reported some other reasons for mixing pipes and hepes, its been a while since I read that paper. I do think that they showed (or at least reported) that they got better results with both pipes and hepes than with either alone. The buffering business is odd because I would have thought that pipes does just fine thanks at pH 6.9. It also seems odd to me that one needs to worry about calcium causing depolymerization in the presence of aldehyde fixatives. In my own work on plant cells, we have found far better preservation of both actin and microtubules when calcium replaces the EGTA in a pipes buffer. It is true that animal microtubules are more sensitive to calcium than are plant microtubules, so it may well be needed there. Hope this helps, Tobias
} } } I was just reading a superb paper by Mobius et al (2002 J. } Histochem. Cytochem. 50(1):43-55) in which they use PHEM buffer for } their fixatives and immunostaining. PHEM is 60 mM PIPES, 25 mM } HEPES, 2 mM MgCl2, 10 mM EGTA, pH 6.9. Can anyone comment on the } rationale behind the design of this buffer? Specifically, why EGTA? } In addition, why a PIPES/HEPES mixture rather than just PIPES or } HEPES? Thanks, Tom } } } } Thomas E. Phillips, PhD } Professor of Biological Sciences } Director, Molecular Cytology Core } 3 Tucker Hall } University of Missouri } Columbia, MO 65211-7400 } } 573-882-4712 (office) } 573-882-0123 (fax) } PhillipsT-at-missouri.edu
The principle of the beam wobbler to assist focusing (originally an invention by Le Poole, patented by Philips back in the late 1940s as far as I remember) has been developed over recent years by Abraham (Bram) Koster and his group in Utrecht for application in computer-aided automatic focusing systems for 3D TEM tilt reconstructions. He should be able to help you. His publication list may be found in: http://www.bio.uu.nl/mcb/3dem/publications.html Best wishes,
Jim
-----Original Message----- } From: Bill Tivol [mailto:tivol-at-caltech.edu] Sent: Thursday, June 03, 2004 10:09 PM To: microscopy-at-msa.microscopy.com
Dear List, I am trying to track down a good description--including figures, if possible--of why the image shifts as a consequence of beam tilt when the image is defocussed, but not when it is focussed. I have only found a very short, and not entirely satisfactory, description in a Royal Microscopy Society handbook from 1984. I have, of course, seen references to beam-tilt-induced image shift, but not the details of the mechanism, in several EM texts. Can anyone point me in the right direction? TIA. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 4 03:44:12 2004
Greetings, In his comment about the "missing membranes", Jim Chalcroft wrote, in part... } } Although not directly relevant in your case, some help might come } from the discovery recently by P. Walther that cell membranes, which } are notoriously poorly contrasted after freeze substitution, are } rendered easily visible if 1 - 5% water is maintained in the } substitution fluids.
Can Jim (or anyone else) give citation to that result? It sounds extremely interesting, if a little iconoclastic considering all the wise advice given over the years to eliminate water ruthlessly from freeze substitution routines.
Yes, the following published paper refers to his discovery: Walther P, Ziegler A. (2002) Freeze Substitution of High-Pressure Frozen Samples: The Visibility of Biological Membranes is Improved when the Substitution Medium Contains Water. Journal of Microscopy 208, 3-10.
Jim
-----Original Message----- } From: Debby Sherman [mailto:dsherman-at-purdue.edu] Sent: Thursday, June 03, 2004 9:28 PM To: James Chalcroft
Hi
If it is of assistance to you I have some basic instructions in the operation of the Philips 505 that I would be happy to mail on to you? In our training courses we have and use our own operating procedures for most SEM, TEM and EDS systems
Good luck
Steve Chapman Senior Consultant Protrain Electron Microscopy Training and Consultancy World Wide Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Mob +44 (0)7711 606967 www.emcourses.com
----- Original Message ----- } From: "by way of Ask-A-Microscopist" {tauria-at-hotmail.com} To: {microscopy-at-ns.microscopy.com} Sent: Thursday, June 03, 2004 7:45 PM
Dear listers: below is a job listing that I am posting for a colleague. Please do not respond to me. Respond to the addresses supplied in the announcement. The position is for US citizens only, as this is a secure facility. Good luck.
Materials and Device Characterization Scientist/Engineer
Maxion Technologies, Inc., located near the University of Maryland, College Park campus, has a post-doctoral/contractor position open for a physicist, materials scientist or electrical engineer with experience in micro-structural, chemical, electrical and optical characterization of advanced compound semiconductor materials and devices using a scanning electron microscope and associated tools including Energy /Wavelength Dispersive X-Ray Analysis, Cathodoluminescence, Electron Beam Induced Current and Electron Backscattered Kikuchi Pattern Imaging (EBKP). The primary job responsibility will be to characterize bulk materials and thin films as well as complex device structures with the objective of shedding fundamental light on the material and device properties, thereby assisting with ongoing projects. Ph.D in Physics, Materials Science or Electrical Engineering is preferred. Candidates with MS or BS degree but with 3-5 years of relevant experience in the field will also be considered. The position is for an initial period of 1-2 years with a possibility for extension. The successful candidate will work with both Maxion and Army Research Laboratory (ARL) scientists/engineers at the ARL site in Adelphi, Maryland. US Citizenship is required. Maxion is an equal opportunity employer. Send resume to dwortman-at-maxion.com {mailto:dwortman-at-maxion.com } . For specific job related questions, please send email to prboyd-at-arl.army.mil {mailto::prboyd-at-arl.army.mil}
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 7 05:13:27 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sswaffe-at-abv.bg) from http://microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Sunday, June 6, 2004 at 04:07:47 ---------------------------------------------------------------------------
Email: sswaffe-at-abv.bg Name: Veselin
Organization: 54 st.Ivan Rilski
Education: 6-8th Grade Middle School
Location: Sofia, Bulgaria
Question: Should I use solvents when cleaning lenses from immersion oil or just otical lens tissue?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (donaldawbrey-at-texashealth.org) from http://www.microscopy.org/MicroscopyListserver/MLFormMail.html on Monday, June 7, 2004 at 07:43:28 ---------------------------------------------------------------------------
Email: donaldawbrey-at-texashealth.org Name: Donald G. Awbrey HT(ASCP) QIHC
Does anyone know of any salary information for image analysis technicians.
I happen to be a histotechnician with 15 years experience. As far as I know I may be one of very, very few people who are histotechnicians and also do IA. Most of the people who perform this task are cytology technicians.
That is my quandry.......
I am paid a histotechnician's salary and I believe that I am not being compensated properly.
If any one knows anything about IA salary please respond.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (wharton.sinkler-at-uop.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, June 7, 2004 at 07:03:05 ---------------------------------------------------------------------------
Question: I'm curious whether there are commercially available attachments for SEM which can measure micro-hardness, or crush strength (with an indenter) on particles down to the 20 micron range. I can envisualize something in which a highly tilted sample would be imaged and simultaneously impinged on by an indenter diamond or steel tip, subjected to a ramping force, then recording the force at which the crushing or a specific indentation depth occurs. I would imagine an SEM would be a suitable tool for imaging and positioning the tip.
If anyone is aware of such an attachment please inform. Similarly if you have experience with something of this nature I'd be curious to know some of the details.
Thanks, Wharton
**************************************************************** Wharton Sinkler, PhD. UOP LLC 25 E. Algonquin Rd. Des Plaines, IL 60017-5017 tel. 847-391-3878 fax. 847-391-3719 mailto Wharton.Sinkler-at-uop.com
On Jun 7, 2004, at 7:22 AM, by way of MicroscopyListserver wrote:
} Question: } I'm curious whether there are commercially available attachments for } SEM which can measure micro-hardness, or crush strength (with an } indenter) on particles down to the 20 micron range. I can envisualize } something in which a highly tilted sample would be imaged and } simultaneously impinged on by an indenter diamond or steel tip, } subjected to a ramping force, then recording the force at which the } crushing or a specific indentation depth occurs. I would imagine an } SEM would be a suitable tool for imaging and positioning the tip. } Dear Wharton, This also may be a question for the AFM people. Not being one of them, I have no idea whether an AFM tip can exert sufficient force without damaging itself, but it could probably detect the dimple afterwards. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 7 11:48:56 2004
There was an article entitled "Lens Cleaning - Best Practices Review," by C.V. Duke in the July issue of Microscopy Today. I will send you a PDF of the article in a separate email.
Ron Anderson, Editor Microscopy Today
-----Original Message----- } From: by way of Ask-A-Microscopist [mailto:sswaffe-at-abv.bg] Sent: Monday, June 07, 2004 6:40 AM To: microscopy-at-ns.microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sswaffe-at-abv.bg) from http://microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Sunday, June 6, 2004 at 04:07:47 ---------------------------------------------------------------------------
Email: sswaffe-at-abv.bg Name: Veselin
Organization: 54 st.Ivan Rilski
Education: 6-8th Grade Middle School
Location: Sofia, Bulgaria
Question: Should I use solvents when cleaning lenses from immersion oil or just otical lens tissue?
Wharton: contact a company called Ominprobe (http://www.omniprobe.com). They have something to do, I believe. They love this sort of thing.
by way of MicroscopyListserver wrote:
} } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (wharton.sinkler-at-uop.com) from } http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on } Monday, June 7, 2004 at 07:03:05 } --------------------------------------------------------------------------- } } } Email: wharton.sinkler-at-uop.com } Name: Wharton Sinkler } } Organization: UOP LLC } } Title-Subject: [Microscopy] [Filtered] MListserver: } } Question: } I'm curious whether there are commercially available attachments for } SEM which can measure micro-hardness, or crush strength (with an } indenter) on particles down to the 20 micron range. I can envisualize } something in which a highly tilted sample would be imaged and } simultaneously impinged on by an indenter diamond or steel tip, } subjected to a ramping force, then recording the force at which the } crushing or a specific indentation depth occurs. I would imagine an } SEM would be a suitable tool for imaging and positioning the tip. } } If anyone is aware of such an attachment please inform. Similarly if } you have experience with something of this nature I'd be curious to } know some of the details. } } Thanks, } Wharton } } **************************************************************** } Wharton Sinkler, PhD. } UOP LLC } 25 E. Algonquin Rd. } Des Plaines, IL 60017-5017 } tel. 847-391-3878 } fax. 847-391-3719 } mailto Wharton.Sinkler-at-uop.com } } } } --------------------------------------------------------------------------- } } }
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-995-2360 972-648-8743 (pager) SC Packaging FA Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 7 15:06:47 2004
I'm looking for spring metal clips that I will screw into one portion of a stage insert (that is being machined custom), that will hold down a slide set in this insert.
Good source anyone?
Sorry for the double post for some.
Best,
Gary
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 7 15:29:42 2004
Hi We have an old, not-worth-saving, Philips 410 TEM. If there are some small parts etc. you would like please let me know and I will see if we can send them to you. I may be able to remove the small and large screens if anyone wants those - they had been recently recoated and are in good condition. Parts are free but you must pay for shipping (fedX number?). Also I have some film holder plates (like new) from an EM300 (fits both 410 and 300 series) as well as EM300 specimen holder exchange tips.
Jon Mulholland, Director Cell Sciences Imaging Facility Beckman Center, B050 Stanford University School of Medicine Stanford, CA 94305-5301
voice 650-725-7532 fax 650-725-4951
http://taltos.stanford.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 8 06:18:53 2004
Does anyone know what the depreciation period is for a Transmission electron microscope?
Thanks,
Mike G
Michael P. Goheen Electron Microscopy Lab Dept. of Pathology & Lab Medicine Indiana University School of Medicine Tel. 317-274-7604 Fax 317-274-5346 mgoheen-at-iupui.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 8 10:07:37 2004
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail to Geology West Chester University of Pennsylvania Schmucker II Science Center, Room SS024 South Church Street and Rosedale Avenue West Chester, PA, 19383 Phone/FAX: 610-738-0437 eMail: fmonson-at-wcupa.edu CASI Page and Scheduling http://darwin.wcupa.edu/CASI/
-----Original Message----- } From: Becky Holdford [mailto:r-holdford-at-ti.com] Sent: Monday, June 07, 2004 2:41 PM To: wharton.sinkler-at-uop.com Cc: microscopy-at-ns.microscopy.com
Wharton: contact a company called Ominprobe (http://www.omniprobe.com). They have something to do, I believe. They
love this sort of thing.
by way of MicroscopyListserver wrote:
} } } ---------------------------------------------------------------------- } -------- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------ ------- } } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (wharton.sinkler-at-uop.com) from } http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on } Monday, June 7, 2004 at 07:03:05 } ------------------------------------------------------------------------ --- } } } Email: wharton.sinkler-at-uop.com } Name: Wharton Sinkler } } Organization: UOP LLC } } Title-Subject: [Microscopy] [Filtered] MListserver: } } Question: } I'm curious whether there are commercially available attachments for } SEM which can measure micro-hardness, or crush strength (with an } indenter) on particles down to the 20 micron range. I can envisualize
} something in which a highly tilted sample would be imaged and } simultaneously impinged on by an indenter diamond or steel tip, } subjected to a ramping force, then recording the force at which the } crushing or a specific indentation depth occurs. I would imagine an } SEM would be a suitable tool for imaging and positioning the tip. } } If anyone is aware of such an attachment please inform. Similarly if } you have experience with something of this nature I'd be curious to } know some of the details. } } Thanks, } Wharton } } **************************************************************** } Wharton Sinkler, PhD. } UOP LLC } 25 E. Algonquin Rd. } Des Plaines, IL 60017-5017 } tel. 847-391-3878 } fax. 847-391-3719 } mailto Wharton.Sinkler-at-uop.com } } } } ---------------------------------------------------------------------- } ----- } } }
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-995-2360 972-648-8743 (pager) SC Packaging FA Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 8 11:49:27 2004
That's a question for the accountants and the IRS.
-----Original Message----- } From: Goheen, Michael P. [mailto:mgoheen-at-iupui.edu] Sent: Tuesday, June 08, 2004 10:40 AM To: Microscopy-at-MSA.Microscopy.com
Hello,
Does anyone know what the depreciation period is for a Transmission electron microscope?
Thanks,
Mike G
Michael P. Goheen Electron Microscopy Lab Dept. of Pathology & Lab Medicine Indiana University School of Medicine Tel. 317-274-7604 Fax 317-274-5346 mgoheen-at-iupui.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 8 13:02:27 2004
The annual EMBO EM course will be held this year at the Pasteur Institute in Paris, France. Course dates are September 12 - 22 2004. More details can be found on the following web page.
We have been a large EM unit for many years and have had several TEMs (currently running 9). I cannot remember disposing of any of them less than 20 years old.
It would be different if you had only one TEM and wanted to keep up with new developments, 5 to 7 years?
Regards, Ron
On Tue, 8 Jun 2004 09:40:05 -0500 "Goheen, Michael P." {mgoheen-at-iupui.edu} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Hello, } } Does anyone know what the depreciation period is for a Transmission } electron microscope? } } Thanks, } } Mike G } } Michael P. Goheen } Electron Microscopy Lab } Dept. of Pathology & Lab Medicine } Indiana University School of Medicine } Tel. 317-274-7604 } Fax 317-274-5346 } mgoheen-at-iupui.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 http://www-em.materials.ox.ac.uk/ *********************************
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 8 15:06:12 2004
Hi, When I was at a Fortune 500 Company Microscopy Lab we depreciated all of our TEMs for 10 yrs. We had 4 TEMs. Regards, Judy Murphy
Ron Doole wrote: } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Hi, } } We have been a large EM unit for many years and have had } several TEMs (currently running 9). I cannot remember } disposing of any of them less than 20 years old. } } It would be different if you had only one TEM and wanted to } keep up with new developments, 5 to 7 years? } } Regards, } Ron } } On Tue, 8 Jun 2004 09:40:05 -0500 "Goheen, Michael P." } {mgoheen-at-iupui.edu} wrote: } } } } } } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------------------------- } } } } Hello, } } } } Does anyone know what the depreciation period is for a Transmission } } electron microscope? } } } } Thanks, } } } } Mike G } } } } Michael P. Goheen } } Electron Microscopy Lab } } Dept. of Pathology & Lab Medicine } } Indiana University School of Medicine } } Tel. 317-274-7604 } } Fax 317-274-5346 } } mgoheen-at-iupui.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 } http://www-em.materials.ox.ac.uk/ } *********************************
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 8 15:27:52 2004
I am doing basic ultramicrotomy on lowricryl embedded lung tissue. When thin sectioning the bottome of the tissue sticks to the knife causing it to crinkle. I have changed every parameter I could think of from water level to clearance angle. Does anyone have any suggestions?
Todd Hamm Oklahoma Medical Research Foundation Oklahoma City, Ok
_________________________________________________________________ FREE pop-up blocking with the new MSN Toolbar get it now! http://toolbar.msn.click-url.com/go/onm00200415ave/direct/01/
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 8 15:33:09 2004
Hi all, Does anyone on the list have experience imaging DNA after rotary shadowing and/or negative staining? I need some advice and would like to correspond off-list. thanks!
Beth ********************************************************************** Beth Richardson EM Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
Phone - (706) 542-1790 & FAX - (706) 542-1805
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) *******************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull (Aqualung)
Assuming it is the same as a SEM, you can use either accelerated or straight line depreciation. Typical depreciation period is five years. But you only get 10% the first year.
gary g.
At 07:40 AM 6/8/2004, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 8 17:04:59 2004
If there is a way to calculate resolution directly, that would be terrific. However, the only way I know is by experiment. The method is not all that exact and is not easy to perform. But if you want to compare one SEM vs. another, it will work.
What is needed is a very (very) sharp edge. I use a 30u Pt aperture Faraday cup. Center an edge at zero tilt on the display. Increase mag as high as you can reasonably get and handle (at high mag, all things become significant obstacles). Make the image range between black and white, i.e., full dynamic range. If the SEM has a waveform display mode, use that to get from top limit to bottom limit.
What you will see is:
--- \ \ \ \ \ -----
as the beam moves from the aperture body (left side) to the aperture hole (right side). Then measure the transcending horizontal distance from 90% to 10% P-P and this is your resolution figure...so to speak.
For modest resolutions of perhaps 50-100A, this method may work OK. For higher resolution SEMs, I doubt that it would work since the margin is so small. I have not tried it on SEMs with really high resolution figures.
No claim of authorship for this method, and I forget how I came across this. I probably got it from this list some years ago.
An alternate method may be to measure the "clarity" of an atomic lattice using EBSD (if you have it). Since the lattice spacing is known, how well the SEM can resolve this ought to be a method of determining resolution. But perhaps this is wild speculation on my part. I may try this later on.
gary g.
At 04:37 AM 6/8/2004, you wrote:
} hi all! } } just a simple question about SEM! } } how can I calculate a FE-SEM resolution ? does some calculation theories } exist? } how can it be verified practicaly?? } } thanx in advance... } } Sylvain Maury
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 8 17:52:47 2004
I had been active in microscopy for nearly 30 years until I was 'kicked upstairs' a few years back. At our federal department here in 'almost-Stanley-Cup-hockey-champions-land', we have some 35 microbeam instruments scattered across the country, with an average age of 13 years. In my former lab, we had a workhorse EM400T that was 20 years old, yet still useable. One of the principal reasons for that longevity was the acquisition of a CM20 FEG ~12 years ago, which picked up a lot of the load. When I was Group Leader I never hesitated to trot out the '10 year' figure, except that I used it, not in the depreciation sense, but more in the 'now outmoded' sense.
Now, here's the problem, be very careful of what figures you quote and to whom. We were audited by the Canadian federal Auditor-General last year and it was a most enlightening/frightening experience. These people (and probably accountants from Finance Depts, etc) don't know an SEM from a probe (literally, they even had side-by-side pictures of the two and claimed them as evidence of 'identical scientific instruments'). Holding an old instrument together with binder twine and scotch tape made no impresssion whatsoever. So long as an innocent tech replied that 'we use this instrument only about 10% of the time" (emergency backup, spare parts, etc), that was all they wanted to know and hung us out to dry for poor utilization of capability.
The other side of the coin is when you know you really need a current generation instrument, quote the 10 year number, but then have to explain to management why the incumbent still runs reasonably well, and perhaps even is still just fine for clients with simple requests.
A murky picture to which I have no clear answers, other than to proceed cautiously!
Tom
Dr. Tom Malis Scientist Advisor Natural Resources Canada Govt. of Canada Phone: 613-995-7358 cell: 613-371-4577 FAX: 613-947-6606 malis-at-nrcan.gc.ca
-----Original Message----- } From: Judy Murphy To: Ron Doole; home; office Cc: Goheen, Michael P.; Microscopy community Sent: 6/8/2004 4:22 PM
Hi, When I was at a Fortune 500 Company Microscopy Lab we depreciated all of our TEMs for 10 yrs. We had 4 TEMs. Regards, Judy Murphy
Ron Doole wrote: } } ------------------------------------------------------------------------ ------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------ ------- } } Hi, } } We have been a large EM unit for many years and have had } several TEMs (currently running 9). I cannot remember } disposing of any of them less than 20 years old. } } It would be different if you had only one TEM and wanted to } keep up with new developments, 5 to 7 years? } } Regards, } Ron } } On Tue, 8 Jun 2004 09:40:05 -0500 "Goheen, Michael P." } {mgoheen-at-iupui.edu} wrote: } } } } } } } ------------------------------------------------------------------------ ------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------------------ ------- } } } } Hello, } } } } Does anyone know what the depreciation period is for a Transmission } } electron microscope? } } } } Thanks, } } } } Mike G } } } } Michael P. Goheen } } Electron Microscopy Lab } } Dept. of Pathology & Lab Medicine } } Indiana University School of Medicine } } Tel. 317-274-7604 } } Fax 317-274-5346 } } mgoheen-at-iupui.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 } http://www-em.materials.ox.ac.uk/ } *********************************
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 9 02:08:40 2004
The method and period of depreciation should take into account the reason you are deprecating the equipment. If it is to properly charger the user. It should be reflect the expected usage reflect the increasing maintenance cost if the cost does increase with age. If it is for tax reasons you need to consult you tax accountants and do what is most advantageous to the bottom line.
If you choose a heavily loaded front end depreciation document your reasons very well so you don't run in the problems that Tom has and discuss it with the auditor before the fact and get him to sign off on it to stay out of that kind of problem.
Billing the cost of the equipment unfairly is second only to space wars in problems in facility meetings. Be sure that you don't run a fowl of IRS if you are running a store or the new rules on billing that came into effect a few years ago if you are in the USA.
When we increased billing on vehicles so there was money to replace them the professors that purchased the vehicles and put them in the common pool were very unhappy at paying for them twice. They continued to be upset for over a year. So finding a fair way to bill uses is important.
Gordon Gordon Couger
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward any links or information you think might be useful to others. Microscope Manual at www.science-info.org
: : : ------------------------------------------------------------------ ------------ : The Microscopy ListServer -- Sponsor: The Microscopy Society of America : To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver : On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html : ------------------------------------------------------------------ ------------- : : I had been active in microscopy for nearly 30 years until I was 'kicked : upstairs' a few years back. At our federal department here in : 'almost-Stanley-Cup-hockey-champions-land', we have some 35 microbeam : instruments scattered across the country, with an average age of 13 years. : In my former lab, we had a workhorse EM400T that was 20 years old, yet still : useable. One of the principal reasons for that longevity was the : acquisition of a CM20 FEG ~12 years ago, which picked up a lot of the load. : When I was Group Leader I never hesitated to trot out the '10 year' figure, : except that I used it, not in the depreciation sense, but more in the 'now : outmoded' sense. : : Now, here's the problem, be very careful of what figures you quote and to : whom. We were audited by the Canadian federal Auditor-General last year and : it was a most enlightening/frightening experience. These people (and : probably accountants from Finance Depts, etc) don't know an SEM from a probe : (literally, they even had side-by-side pictures of the two and claimed them : as evidence of 'identical scientific instruments'). Holding an old : instrument together with binder twine and scotch tape made no impresssion : whatsoever. So long as an innocent tech replied that 'we use this : instrument only about 10% of the time" (emergency backup, spare parts, etc), : that was all they wanted to know and hung us out to dry for poor utilization : of capability. : : The other side of the coin is when you know you really need a current : generation instrument, quote the 10 year number, but then have to explain to : management why the incumbent still runs reasonably well, and perhaps even is : still just fine for clients with simple requests. : : A murky picture to which I have no clear answers, other than to proceed : cautiously! : : Tom : : Dr. Tom Malis : Scientist Advisor : Natural Resources Canada : Govt. of Canada : Phone: 613-995-7358 : cell: 613-371-4577 : FAX: 613-947-6606 : malis-at-nrcan.gc.ca : : : -----Original Message----- : } From: Judy Murphy : To: Ron Doole; home; office : Cc: Goheen, Michael P.; Microscopy community : Sent: 6/8/2004 4:22 PM : Subject: [Microscopy] Re: Re: Depreciation period : : : : ------------------------------------------------------------------ ------ : ------ : The Microscopy ListServer -- Sponsor: The Microscopy Society of America : To Subscribe/Unsubscribe -- : http://www.msa.microscopy.com/MicroscopyListserver : On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html : ------------------------------------------------------------------ ------ : ------- : : Hi, : When I was at a Fortune 500 Company Microscopy Lab we depreciated all of : our TEMs for 10 yrs. We had 4 TEMs. : Regards, : Judy Murphy : : : : : : Ron Doole wrote: : } : } : ------------------------------------------------------------------ ------ : ------ : } The Microscopy ListServer -- Sponsor: The Microscopy Society of : America : } To Subscribe/Unsubscribe -- : http://www.msa.microscopy.com/MicroscopyListserver : } On-Line Help : http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html : } : ------------------------------------------------------------------ ------ : ------- : } : } Hi, : } : } We have been a large EM unit for many years and have had : } several TEMs (currently running 9). I cannot remember : } disposing of any of them less than 20 years old. : } : } It would be different if you had only one TEM and wanted to : } keep up with new developments, 5 to 7 years? : } : } Regards, : } Ron : } : } On Tue, 8 Jun 2004 09:40:05 -0500 "Goheen, Michael P." : } {mgoheen-at-iupui.edu} wrote: : } : } } : } } : } } : ------------------------------------------------------------------ ------ : ------ : } } The Microscopy ListServer -- Sponsor: The Microscopy Society of : America : } } To Subscribe/Unsubscribe -- : http://www.msa.microscopy.com/MicroscopyListserver : } } On-Line Help : http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html : } } : ------------------------------------------------------------------ ------ : ------- : } } : } } Hello, : } } : } } Does anyone know what the depreciation period is for a Transmission : } } electron microscope? : } } : } } Thanks, : } } : } } Mike G : } } : } } Michael P. Goheen : } } Electron Microscopy Lab : } } Dept. of Pathology & Lab Medicine : } } Indiana University School of Medicine : } } Tel. 317-274-7604 : } } Fax 317-274-5346 : } } mgoheen-at-iupui.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 : } http://www-em.materials.ox.ac.uk/ : } ********************************* : : :
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 9 02:21:40 2004
because i'm an intern in a microelectronics analysis laboratory in France, working on a HR FE-SEM, and because of that, i have to know all details and informations on SEM and methods used for...as much as possible.
The manufacturer gave a resolution of about 15A at low voltage and 9A at high voltage. i just want to know how can i verify it with theory and practice. I think i'll call the vendor...and get a copy of Goldstein (a little expensive for me :-[ )
thanx.
Sylvain
Warren E Straszheim a écrit :
} I would think that would be quite dependent on the conditions and the } sample. A program like Electron Flight Simulator might be able to estimate } a value given the appropriate parameters. Still, it may only produce the } maximum possible resolution. Actual performance will depend on your skill } in operation. } } Most vendors should be able to quote a number, but that will depend on the } sample. Maybe than can suggest the maximum possible resolution for your } sample, but maybe not. } } Why do you ask? } } Warren } } At 06:37 AM 6/8/2004, you wrote: } } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------------------------- } } } } hi all! } } } } just a simple question about SEM! } } } } how can I calculate a FE-SEM resolution ? does some calculation theories } } exist? } } how can it be verified practicaly?? } } } } thanx in advance... } } } } Sylvain Maury
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 9 04:05:30 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (caneiro-at-cab.cnea.gov.ar) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, June 8, 2004 at 14:19:23 ---------------------------------------------------------------------------
Email: caneiro-at-cab.cnea.gov.ar Name: Alberto Caneiro
Organization: Instituto Balseiro
Title-Subject: [Microscopy] [Filtered] CRT for EDAX PV9900
Question: We have a PV9900 EDAX EDS system. We are having problems with the monitor (CRT) of this equipment. We are looking for a replacement for it. We are willing to purchase it at a reasonable price from whoever can supply it.
For measurement of the spatial resolution have a look at D. C. Joy: "SMART - a program to measure SEM resolution and imaging performance" Journal of Microscopy, vol. 208 (2002) 24-34.
-----Original Message----- } From: sylvain maury [mailto:sylvain.maury-at-thalesgroup.com] Sent: 8. juni 2004 13:37 To: Microscopy community
Hello listers,
We have in our TEM storeroom several LKB knifemakers which have not been used for some years since everyone here prefers the comfort and edge longevity of diamond knives for ultrathin sectioning. Recently I received advertising material referring to an add-on device called a GUM-symparter which apparently converts the LKB knifemaker into a wondrous instrument by applying the "newest principles of equilibrium weighting" to produce optimal triangular glass knives. Could this be a useful acquisition for labs like ours? I would be interested to hear from any lab that has bought and used this attachment; in particular as to whether it has lived up to expectations. Best wishes,
Jim
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 9 06:47:30 2004
When I worked for a major US rubber company instrument depreciation was used more as a tool for determining department budgets then a tool for up grading. While I remain blissfully ignorant of the booking/tax conditions of management, we were often told that long depreciation such as 10 years on a GC helped increase the department budget, even if the GC was just a shell with the detector, column and ports stripped off of it. We transferred a SEM to another department (they had a EDS unit but no SEM, we had a SEM but no EDS) and for one year we had extra money because we no longer had a depreciating SEM to payoff. The following year we had less money because we no longer had that SEM. If this sounds confusing, it is. If I remember correctly we depreciated everything for 10 years and sometime (?) ran a second 10 years.
More to the point, the problem with long depreciation appears to be the inability to purchase new equipment while an old and now obsolete or non-functioning instrument still has appreciable value due to the extended depreciation. I've seen instruments, which existed, only as nameplates and owner's manuals in a desk drawer, still listed on the equipment roster because the depreciation value was too high to be dispose of.
The second most limiting parameter is being forced to answer the replacement question: "What can't you do today that you could do yesterday?" This question is more of a political and cultural limitation and a good topic for another day.
Frank Karl
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 9 08:00:21 2004
How long did you polymerize the blocks and at what temperature. Usually I polymerize the Lowicryl with UV light at -20C for 2-3 days and then an additional day or two at room temperature (this room temp polymerization does not affect any of the immunoEM labeling sensitivity). The resin is much more cuttable.
Also, if your diamond knife is a little dull try another one or if the edge is not really clean sometimes the sections cling to whatever film is on that knife edge from previous cutting sessions.
good luck
Karen Bentley
Karen L. Bentley, M.S.(previously Jensen) Associate Scientist & Project Manager Electron Microscope Research Core University of Rochester Medical Center Rochester, NY 14642 585-275-1954
-----Original Message----- } From: todd hamm [mailto:ripbnowell-at-hotmail.com] Sent: Tuesday, June 08, 2004 4:47 PM To: Microscopy-at-MSA.Microscopy.Com
I am doing basic ultramicrotomy on lowricryl embedded lung tissue. When thin
sectioning the bottome of the tissue sticks to the knife causing it to crinkle. I have changed every parameter I could think of from water level to
clearance angle. Does anyone have any suggestions?
Todd Hamm Oklahoma Medical Research Foundation Oklahoma City, Ok
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From MicroscopyL-request-at-ns.microscopy.com Wed Jun 9 08:46:52 2004
I am not very happy with my current supplier of secondary gold immunoreagents. I would appreciate input regarding potential new suppliers, as well as general comments about, say, the shelf-life, stability, sensitivity of your favorite immunoreagent brand.
Thanks--Peter
_______________________________ Peter Rohloff, PhD Laboratory of Molecular Parasitology Medical Scholars Program University of Illinois at Urbana-Champaign
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 9 10:34:34 2004
Does anyone know how CT numbers are mapped to the pixel values in a DICOM image?
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From MicroscopyL-request-at-ns.microscopy.com Wed Jun 9 11:34:04 2004
Which Lowacryl resin are you using? This makes a very big difference in sectioning characteristics.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
On 6/9/04 8:18 AM, "Bentley, Karen" {Karen_Jensen-at-URMC.Rochester.edu} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } Todd: } } How long did you polymerize the blocks and at what temperature. Usually I } polymerize the Lowicryl with UV light at -20C for 2-3 days and then an } additional day or two at room temperature (this room temp polymerization } does not affect any of the immunoEM labeling sensitivity). The resin is } much more cuttable. } } Also, if your diamond knife is a little dull try another one or if the edge } is not really clean sometimes the sections cling to whatever film is on that } knife edge from previous cutting sessions. } } good luck } } Karen Bentley } } Karen L. Bentley, M.S.(previously Jensen) } Associate Scientist & Project Manager } Electron Microscope Research Core } University of Rochester Medical Center } Rochester, NY 14642 } 585-275-1954 } } } -----Original Message----- } } From: todd hamm [mailto:ripbnowell-at-hotmail.com] } Sent: Tuesday, June 08, 2004 4:47 PM } To: Microscopy-at-MSA.Microscopy.Com } Subject: [Microscopy] Sectioning Lowricryl } } } } } ---------------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } --- } } I am doing basic ultramicrotomy on lowricryl embedded lung tissue. When thin } } sectioning the bottome of the tissue sticks to the knife causing it to } crinkle. I have changed every parameter I could think of from water level to } } clearance angle. Does anyone have any suggestions? } } Todd Hamm } Oklahoma Medical Research Foundation } Oklahoma City, Ok } } _________________________________________________________________ } FREE pop-up blocking with the new MSN Toolbar - get it now! } http://toolbar.msn.click-url.com/go/onm00200415ave/direct/01/ } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 9 12:54:15 2004
In a message dated 6/9/04 2:03:46 PM, tommy91779-at-hotmail.com writes:
} Does anyone know how CT numbers are mapped to the pixel values in a DICOM } image?
The DICOM standard describes in detail the greyscale standard display function that presents normalized rendering of image values. It is heavily based on the non-linear response of human vision to luminance changes. You can download the standard and much other information on DICOM formats in pdf format from {www.dclunie.com} . There is even a tool to import DICOM files into Photoshop.
John Russ
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 9 13:00:08 2004
I have an MT2C, actually, but I have a problem with too much laxity in the binoch head. I am trying to get this old microtome into usable shape. I was fnally able to free the up and down part of the arm to raise and lower the binoch (it's a bit stiff but can now be adjusted).
Now I have too much wiggle in the bit that allows the binoch head to move up and down (similar to a nodding motion). I somehow need to increase the tension or friction or something so the binoch head hold steady now matter what the pivot. Now it just keeps nodding off, you know, resting it;s little head on the fluorescent light. Like a new baby this thing can't hold it head up.
Any suggestions? I'm trying to take the joint apart and figure it out, but I'm stumped as to how to increase the tension/pressure whatever it is.
On a similar theme-- Does anybody know where I can get a reconditioned/refurbished ultramicrotome? I have an old LKB that gives all the users a little jolt every time they complete the circuit with their bodies by touching the microtome and the controller box. We are tired of having sectioning be a shocking experience and we'd like to replace it. I'd like something newer than the LKB or MT2C, something like an Ultracut E. Any ideas?
Nodding off in Norfolk,
Paula :-)
Paula Sicurello EM Lab 106 Mills Godwin Sciences Building Old Dominion University Norfolk, VA 23529 USA 757-683-3822
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 9 13:24:32 2004
Peter, Immunogold reagents are a peculiar lot. The characteristics of the gold particle itself is dependant on which reducing agent is used. Unfortunately, the most common method involves the reduction of chloroauric acid with citrate-tannic acid. There is a stochiometric relationship between the ratio of the chemicals used and diameter of the gold particle produced. This results in tremendous affinity for cytoskeletal components. Tannic acid is a mordant often used to preserve microtubules. However, if the reducing agent is sodium borohydride, one can circumvent this affinity. Jansen used to be the only company that advertised that their gold was produced by the reduction of chloroauric acid by sodium borohydride. I believe they were purchased by Amersham, who treats this procedure as a trade secret. In addition, it is paramount that one uses teleostan fish gelatin as a stabilizer instead of BSA. Amersham also sells a immunogold quality teleostan fish gelatin. (Reference: Birrel, et al, "Pitfalls of Immunogold Labeling," Journal of Histo. & Cyto., 35(8): 843-53 (1987). I have no interest in any of these companies, except as a consumer.
At 10:09 AM 6/9/2004 -0500, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Wed Jun 9 13:54:39 2004
When labeling e.g. lowicryl sections using indirect immunogold (primary antibody - secondary antibody conjugated to gold), is there a consensus for the typical (or maximal, that may be easier) distance between the antigen and the particle? I do understand this will never be a discrete number, I would like to have an idea what people think.
Thanks,
Michael Jarnik
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 9 16:13:09 2004
Hello Jim Leica used to utilize this principle in their knifemaker (which is actually originated from LKB). The idea is that you made squares not from the end of strip (as in LKB), but cutting glass strip in half every time until you got perfect squares. Leica has sort of rail with stops for the glass strip. The position of stops are: half of the glass strip length, 1/4, 1/8 and so on. Last stop is at the length for square. I could assume somebody made similar device for old LKB knifemaker. I don't think it would convert your old knifemaker into brand new machine: quality of knifes depends not only from way of glass breaking but from preciseness of mechanics. Old machine would not perform like new even breaking glass in new way. From another hand, it'll definitely improved the quality of the knifes. I have to tell you (no financial interest) Leica's knifemaker made fantastic knifes even with 10 mm glass. I used to use glass knifes for teaching purpose and diamond for science. Sergey
At 11:49 AM 6/9/2004 +0200, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Thu Jun 10 06:33:40 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (viviana.torres-at-gmail.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, June 9, 2004 at 18:56:28 ---------------------------------------------------------------------------
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (pblanken-at-wisc.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, June 9, 2004 at 16:01:04 ---------------------------------------------------------------------------
Email: pblanken-at-wisc.edu Name: Peter Blankenheim
Question: I'm driving to the Savannah Convention from Madison Wi, and I'm looking for someone to share the car, possibly driving, and possibly lodging. 50 yr old male.
Many people will probably respond to your query. Whenever I fix small numbers of cells, etc., the easiest method is to spin them down in 2% agar (after fixation, even through the Osmium stage)in small eppendorf tubes. You then can remove the agar from the tube, cut off the tip with the cells, whatever, and continue to process like a piece of tissue. Avoids constant centrifugation and loss of material with each spin.
Peggy Sherwood Lab Associate, Photopathology Wellman Laboratories of Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
-----Original Message----- } From: viviana.torres-at-gmail.com [mailto:viviana.torres-at-gmail.com] Sent: Thursday, June 10, 2004 8:00 AM To: microscopy-at-ns.microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (viviana.torres-at-gmail.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, June 9, 2004 at 18:56:28 ---------------------------------------------------------------------------
I have a an excellent book entitled "Electron Microscopic Immuncytochemistry" edited by Julia Polak and John Priestley, Oxford University Press, 1991. They state on page 74 of Chapter 3 entitled "Post-embedding Electron Microscopic Immunocytochemistry":
"The minimum theorectical lateral resolution (i.e. distance between the centre of the gold particle and the epitope recognized by the primary antibody) that can be achieved with the protein-A gold technique is about 16 nm, using a colloidal gold particle of 3 nm. Using the immunoglobulin-gold technique with a gold particle of 10 nm the lateral resolution is approximately 21 nm. It is clear that the bigger the gold particle, the less satisfactory is the lateral resolution of the technique."
Hope this answers your question.
Karen Bentley
Karen L. Bentley, M.S. Associate Scientist & Manager Electron Microscope Research Core University of Rochester Medical Center Rochester, NY 14642 585-275-1954
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 10 12:48:58 2004
The statment in the Polak & Priestley book makes an assumption that when the specimen is dried, the antibodies and gold probe fall to one side (presumably in a random direction).
As there is no evidence to support the statement, other than it was published in a book, may I propose the possibility that the gold particles have an equally good chance (or better) of drying on top of the antibodies. This would mean that the lateral resolution in practice is much better than predicted by what is expected theoretically. The resolution in three dimensions may not be so good in that the gold will end up above the antigen but this should not concern us when we examine the sections in the TEM
The theoretical resolution predicted in the Polak and Priestley book, assumes the gold particles will be bound to the far end of the antibody molecule, when in fact the particles are more likely to bind closer to the hinge region (Baschong & Wrigley 1989 small colloidal gold conjugated to FAB fragments or to immunoglobulin G as high resolution labels for electron microscopy. A technical overview. J. Electron Microsc. Tech. 14:313-323), thus reducing the theorectical resolution predicted by Polak & Priestley.
Anyone who performs immuno-EM on a routine basis will tell you that gold particles are very often much closer to the antigen site than could be expected if the gold was "falling off the pile of protein". Quite often the gold is located in precisely the location predicted.
A good demonstration of how precise immunolabeling can be is seen in the study by Sodeik et al (1997 Microtuble-mediated transport of incoming Herpes simplex virus 1 capsids to the nucleus. J. Cell Biol 136:1007-1021). An antibody to dynein (which attaches the capsid to microtubules) labels exactly 40 - 50 nm away from the virus, where the antigen is predicted to be. If the immunolabeling were to be as poor resolution as predicted by theorectical considerations, the technicnique could not be used for such high resolution studies as it has been applied.
Obviously we need more experimentation, and thus more data, to continue this discussion and to provide Michael with the answer he wants.
Regards,
Paul Webster.
Paul Webster, Ph.D. House Ear Institute 2100 West Third Street Los Angeles CA 90057 phone (213) 273 8026 fax (213) 413 6739 email: pwebster-at-hei.org
} ---------- } From: Bentley, Karen } Sent: Thursday, June 10, 2004 7:20 AM } To: 'microscopy-at-msa.microscopy.com' } Subject: [Microscopy] Gold Distance from Antigen } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Michael: } } I have a an excellent book entitled "Electron Microscopic } Immuncytochemistry" edited by Julia Polak and John Priestley, Oxford } University Press, 1991. They state on page 74 of Chapter 3 entitled } "Post-embedding Electron Microscopic Immunocytochemistry": } } "The minimum theorectical lateral resolution (i.e. distance between the } centre of the gold particle and the epitope recognized by the primary } antibody) that can be achieved with the protein-A gold technique is about 16 } nm, using a colloidal gold particle of 3 nm. Using the immunoglobulin-gold } technique with a gold particle of 10 nm the lateral resolution is } approximately 21 nm. It is clear that the bigger the gold particle, the } less satisfactory is the lateral resolution of the technique." } } Hope this answers your question. } } Karen Bentley } } Karen L. Bentley, M.S. } Associate Scientist & Manager} } Electron Microscope Research Core } University of Rochester Medical Center } Rochester, NY 14642 } 585-275-1954 } } } }
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 10 16:12:30 2004
Does anyone know if special precautions are required before inserting a high-tension cable back into the oil tank of a TEM. We have a TEM that was shipped with the transformer separate from the instrument. I want to finish installing the instrument, but wanted to inquire as to whether there were any special precautions to follow. Thanks in advance, Robert
Dr. Robert Kyle Pope Indiana University South Bend Department of Biology 1700 Mishawaka Avenue South Bend, IN 46615-1408 Email: ropope-at-iusb.edu Fax: 574-237-6589 Tele: 574-237-4411
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 10 16:57:34 2004
Roth (1982 The protein A-gold technique..... pp 107-133 in Techniques in Immunocytochemistry vol. 1, eds GR Bullock and P Perutz) also discusses size of antibodies, and estimates that they are about 8 nm in diameter. So if both primary and secondary antibodies are lying flat on your section, there's a maximum ~16 nm between antigen and gold particle.
As Paul notes, this is a maximum distance, the gold can be stacked directly on top of the antigen. In immunoFESEM work on microtubules, for example (Vesk et al. 1994 Imaging of plant microtubules with high resolution scanning electron microscopy. Protoplasma 182: 71-74), using primary anti-tubulin and gold-labelled secondary, you see a single or occasionally a double blob on the microtubule with the gold on top. The tissue has been dried, of course, so I guess the antibodies and gold are more likely to appear this way. cheers, Rosemary
Dr. Rosemary White rosemary.white-at-csiro.au Microscopy Centre ph. 61-2-6246 5475 CSIRO Plant Industry mob. 61-0402 835 973 GPO Box 1600 fax. 61-2-6246 5000 Canberra, ACT 2601 Australia
} From: "Webster, Paul" {PWebster-at-hei.org} } Date: Thu, 10 Jun 2004 11:09:55 -0700 } To: "MSA listserver submission (E-mail)" {Microscopy-at-MSA.Microscopy.Com} } Subject: [Microscopy] RE: Gold Distance from Antigen } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } Hi Karen, } } The statment in the Polak & Priestley book makes an assumption that when the } specimen is dried, the antibodies and gold probe fall to one side (presumably } in a random direction). } } As there is no evidence to support the statement, other than it was published } in a book, may I propose the possibility that the gold particles have an } equally good chance (or better) of drying on top of the antibodies. This would } mean that the lateral resolution in practice is much better than predicted by } what is expected theoretically. The resolution in three dimensions may not be } so good in that the gold will end up above the antigen but this should not } concern us when we examine the sections in the TEM } } The theoretical resolution predicted in the Polak and Priestley book, assumes } the gold particles will be bound to the far end of the antibody molecule, when } in fact the particles are more likely to bind closer to the hinge region } (Baschong & Wrigley 1989 small colloidal gold conjugated to FAB fragments or } to immunoglobulin G as high resolution labels for electron microscopy. A } technical overview. J. Electron Microsc. Tech. 14:313-323), thus reducing the } theorectical resolution predicted by Polak & Priestley. } } Anyone who performs immuno-EM on a routine basis will tell you that gold } particles are very often much closer to the antigen site than could be } expected if the gold was "falling off the pile of protein". Quite often the } gold is located in precisely the location predicted. } } A good demonstration of how precise immunolabeling can be is seen in the study } by Sodeik et al (1997 Microtuble-mediated transport of incoming Herpes simplex } virus 1 capsids to the nucleus. J. Cell Biol 136:1007-1021). An antibody to } dynein (which attaches the capsid to microtubules) labels exactly 40 - 50 nm } away from the virus, where the antigen is predicted to be. If the } immunolabeling were to be as poor resolution as predicted by theorectical } considerations, the technicnique could not be used for such high resolution } studies as it has been applied. } } Obviously we need more experimentation, and thus more data, to continue this } discussion and to provide Michael with the answer he wants. } } Regards, } } Paul Webster. } } } } } } } } Paul Webster, Ph.D. } House Ear Institute } 2100 West Third Street } Los Angeles } CA 90057 } phone (213) 273 8026 } fax (213) 413 6739 } email: pwebster-at-hei.org } } } } ---------- } } From: Bentley, Karen } } Sent: Thursday, June 10, 2004 7:20 AM } } To: 'microscopy-at-msa.microscopy.com' } } Subject: [Microscopy] Gold Distance from Antigen } } } } } } } } -----------------------------------------------------------------------------} } - } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ----------------------------------------------------------------------------- } } -- } } } } Michael: } } } } I have a an excellent book entitled "Electron Microscopic } } Immuncytochemistry" edited by Julia Polak and John Priestley, Oxford } } University Press, 1991. They state on page 74 of Chapter 3 entitled } } "Post-embedding Electron Microscopic Immunocytochemistry": } } } } "The minimum theorectical lateral resolution (i.e. distance between the } } centre of the gold particle and the epitope recognized by the primary } } antibody) that can be achieved with the protein-A gold technique is about 16 } } nm, using a colloidal gold particle of 3 nm. Using the immunoglobulin-gold } } technique with a gold particle of 10 nm the lateral resolution is } } approximately 21 nm. It is clear that the bigger the gold particle, the } } less satisfactory is the lateral resolution of the technique." } } } } Hope this answers your question. } } } } Karen Bentley } } } } Karen L. Bentley, M.S. } } Associate Scientist & Manager} } } Electron Microscope Research Core } } University of Rochester Medical Center } } Rochester, NY 14642 } } 585-275-1954 } } } } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 10 17:37:42 2004
This message invites you to contribute to the next on a series of CD-ROM published by our laboratory. To date, we have distributed over 50,000 copies of 9 CD-ROMs --- 8 in cytometry and 1 in imaging and microscopy. The second volume of our imaging and microscopy CD-ROM with 5000 copy distribution is imminent. We have 2-3 weeks to complete this next electronic publication. You may see the others at http://www.cyto.purdue.edu/flowcyt/cdroms/newcd6.htm
We invite you contribute your unpublished materials - images, movies, protocols, lectures, etc. You retain all copyrights to all materials published. This is an opportunity to publish quality materials in a well recognized series of CD-ROMs that will receive worldwide distribution.
This particular CD-ROM will be distributed initially at the MSA meeting in Savannah, and then at a number of meetings around the world. As with all of the Purdue CD-ROMs, it is free to those attending meetings, or if obtained from one of the sponsoring companies. The disc is produced by our staff at Purdue University Cytometry Laboratories and is done so as part of our "Cytomics Initiatives in Education" program.
You may contribute an entire section on your work, a small section describing an image or series of images, animations or videos or other materials. Book reviews, manual evaluations, protocols, technical support information, software routines, macros and instrument setup materials are welcome and useful. You must verify that you own the copyright by submitting the form at
http://tinyurl.com/3bmgn
and provide permission to us to place this material onto the CD-ROM. You retain all ownership rights. If the materials are already published, you may require permission of the publisher to place the materials onto this CD-ROM by using the form which is found at
http://tinyurl.com/2sw6o
All of the above information is somewhere on our website starting here http://www.cyto.purdue.edu/flowcyt/cdroms/newcd6.htm
Time is short, but this means that the materials published are very recent. You have 21 days left to contribute!!
You may submit materials directly to us at cd2-at-flowcyt.cyto.purdue.edu or to my personal email address. Instructions for FTP of large files are on the webpage above.
If you are a company and wish to participate in sponsorship which is still available, please contact us directly.
Kind Regards
J. Paul Robinson & Bartek Rajwa Purdue University Cytometry Laboratories
------------------ J.Paul Robinson, PhD Professor of Immunopharmacology Professor of Biomedical Engineering
Bartlomiej Rajwa, PhD Purdue University Cytometry Laboratories Hansen Research Bld, B050 West Lafayette, 47907 IN tel. (765) 494 0757
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 11 02:02:52 2004
Lets see if I can help? I assume this is an oil filled tank, its heavy?
Keep the air in the room as still as possible and make sure that there is the minimum amount of dust in the air. Make sure the cable end is absolutely clean and clean round the top of the high voltage tank before opening the tank itself. Use a cleaning media/cloth that does not leave a dust behind. Some instruments have an insulating grease on the connection, these cables have a taper fit.
The cable end will probably have a locating pin so that it only fits in one position. Gently lower it into place and try to feel it settling in the contact area, do not force it!
Once in place clamp or bolt it down and you are ready to go! Run the high voltage up slowly to give time for any trapped air to be flushed out.
Good luck
Steve Chapman Senior Consultant Protrain Electron Microscopy Training and Consultancy World Wide Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Mob +44 (0)7711 606967 www.emcourses.com
----- Original Message ----- } From: "Dr. Robert K. Pope" {ropope-at-iusb.edu} To: {Microscopy-at-MSA.Microscopy.Com} Sent: Thursday, June 10, 2004 10:30 PM
Hi Listers,
I just wanted to thank all who replied to my question with a nod of my head ;-).
I was trying all the suggestions and nothing was working. I left the bits alone for a night came back put all the screws and widgets back together. Withthe exception of putting the tightening screw on the top instead of the bottom all was the same--except this time it worked.
Now I can go back and thick section Wah, oh, I mean Yippee!
Thanks again all!
No more nodding off in Norolk,
Paula :-)
Paula Sicurello EM Lab 106 Mills Godwin Sciences Building Old Dominion University Norfolk, VA 23529 USA 757-683-3822
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 11 10:25:31 2004
We are trying to label silica particles with rhodamine B-ITC. The first time we tried using a standard bead labeling protocol it seemed to work, but now we can't repeat it. The thing I find wierd is that the pellet of particles is clearly pink by eye, so I would think that would mean they are labeled. Yet in the microscope, very few are fluorescent. I suppose it could be the small number of labeled ones generating the color, but I wondered if anyone had any alternative ideas as to why they might not look fluorescent even though colored. Thanks- Dave --
Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 91 N. Eagleville Rd. U-3125 Storrs, CT 06269-3125 knecht-at-uconn.edu 860-486-2200 860-486-4331 (fax) home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 11 10:43:30 2004
My lab is looking into purchasing a new coater for our FESEM sample preparation. We have been extremely satisfied with our current system, a BOC Edwards Xenosput (Dynavac), for Pt coating. Unfortunately Edwards no longer sells this particular technology. At present we are considering an Osmium coater or Ion beam system. I would appreciate frank opinions from experienced users of these, or other, viable candidates for this type of coating. Specifically, we are interested in sample through put, routine maintenance, overall reliability, cost of consumables, limitations on sample size and ease of use. Thanks in advance, jr
FYI Generally speaking we operate at {2kv and magnifications up to ~150K.
John A. Robson Boehringer Ingelheim Pharmaceuticals, Inc. PO Box 368 900 Ridgebury Rd Ridgefield, CT 06877
BAL-TEC AG and Boeckeler Instruments / RMC Products, Inc. Cordially invite you to the First U.S. Users' Group Meeting for High Pressure Freezing during the M&M Meeting in Savannah, Georgia. Current users and potential users are invited to the meeting.
When: Sunday, August 1, 2004, 12:00 noon - 5:00 pm.
Where: Grand Ballroom F, Westin Savannah Harbor Golf Resort and Spa, Savannah, Georgia.
A buffet lunch will be provided.
Presenters are being invited from current users of the HPM 010 High Pressure Freezer.
Attendance is limited to 50 persons, so please RSVP at your earliest convenience.
If you have questions, wish to RSVP, or have already made travel arrangements, please contact Boeckeler Instruments for assistance at 800.552.2262 or 502.745.0001 and ask for Ms. Kim Megaw. You can also e-mail Kim at { {kim-at-boeckeler.com} } .
Thank you. We look forward to seeing you in Savannah!
Bob Chiovetti, Ph.D. Senior Product Specialist RMC Products Boeckeler Instruments, Inc.
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 11 13:09:46 2004
We have been experiencing a problem using Aclar for flatembedding lately: After polimerization, it is very difficult to peel it off the tissue. Often it rips out pieces of tissue as it comes off. This problem started when we opened a new order from EMS, while samples with the old Aclar batch (and processed parallelly with the new batch), worked fine. We tried new orders from EMS and Ted Pella... without much improvement. Aclar also feels more rigid and thicker than we used in the past, even though it is the same catalog number and thickness. We use EMbed 812, polimerized for 1-2 days at 60 degrees C.
Has anybody experienced a similar problem? Can you recommend alternative material for flatembedding?
Alev
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Alev Erisir University of Virginia Department of Psychology 102 Gilmer Hall, PO Box 400400 Charlottesville, VA 22904-4400
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 11 14:26:41 2004
We're making metal plates to raise the Olympus IX71 microscope stage by 12mm to accommodate a piezo focusing device.
Does anybody know the screw size for holding the stage in place so that we can order the appropriate replacement screws?
Thanks. ____________________________________________________________________________ Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 Fax: 430-8996 URL: http://www.aecom.yu.edu/aif/
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 11 15:38:24 2004
We need to put an extension ring 12 mm high between the objective and the nosepiece.
Does anybody out there have such rings or know where we could purchase them?
An alternative would be to have the machine shop here make a few. Would somebody please post the thread size in case we have to do this.
Thanks!
____________________________________________________________________________ Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 Fax: 430-8996 URL: http://www.aecom.yu.edu/aif/
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 11 16:28:13 2004
} We are trying to label silica particles with rhodamine B-ITC. The } first time we tried using a standard bead labeling protocol it seemed } to work, but now we can't repeat it. The thing I find wierd is that } the pellet of particles is clearly pink by eye, so I would think that } would mean they are labeled. Yet in the microscope, very few are } fluorescent. I suppose it could be the small number of labeled ones } generating the color, but I wondered if anyone had any alternative } ideas as to why they might not look fluorescent even though colored. } Thanks-
Dear David, Are you sure that all the particles have exactly the same composition? One reason that a dye can absorb light--to generate color--but not fluoresce is if there are non-radiative decay processes, which in general are strongly affected by the environment of the dye. So the dye molecules could be bound to some of the particles in one way, where they fluoresce, and in another way on others of the particles (if some particles have a different surface environment, for example) where they do not. In either case, there would likely be absorption of light, although the spectrum could be shifted, so the particles would appear colored. Not only is this possible if there are two different compositions, surface treatments, etc. for the particles, but some subtle effects where different crystal faces predominate on the surfaces of some of the particles will also affect fluorescence. I have had no experience either with rhodomine B-ITC or silica particles, so if the real experts disagree, go with their advice. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Sun Jun 13 00:30:03 2004
Alev Erisir wrote: ============================================================================ ============ We have been experiencing a problem using Aclar for flatembedding lately: After polimerization, it is very difficult to peel it off the tissue. Often it rips out pieces of tissue as it comes off. This problem started when we opened a new order from EMS, while samples with the old Aclar batch (and processed parallelly with the new batch), worked fine. We tried new orders from EMS and Ted Pella... without much improvement. Aclar also feels more rigid and thicker than we used in the past, even though it is the same catalog number and thickness. We use EMbed 812, polimerized for 1-2 days at 60 degrees C.
Has anybody experienced a similar problem? Can you recommend alternative material for flatembedding? ============================================================================ ============= We have offered Aclar film, specifically Aclar 33c, since it seemed to be the one with the most attractive properties for applications in microscopy and histology. It is not "sticky" either before or after resin curing. See URL http://www.2spi.com/catalog/photo/aclar-film.shtml for additional information.
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 ######################## ============================================
From MicroscopyL-request-at-ns.microscopy.com Sun Jun 13 10:07:02 2004
The 30th Annual Nikon 2004 Small World Photomicrography Competition
Celebrating its 30th year, the annual Nikon International Small World Competition was founded in 1975 to recognize excellence in photography through the microscope. The competition's reputation has grown throughout the years and is regarded as the leading forum for recognizing the beauty and complexity of life as seen through the light microscope. The 2004 deadline for entries is Wednesday June 30, 2004.
Astonishing displays of color and remarkable subject matter are among the highlights in the annual Nikon International Small World Photomicrography contest. These award-winning photographs merge the techniques of scientific inquiry with aesthetic beauty to create vibrant and dynamic images reflective of the intriguing world of microscopy. Hailed as the worldâs leading forum for photographic excellence in the art and science of photomicrography, images submitted to the contest capture the mysterious and often unseen universe viewed through a light microscope.
Microscopists from all areas are encouraged to enter the contest and can upload their images from www.nikonsmallworld.com . Submissions may include up to three 35mm photographs or digital images of any subject matter viewed using a light microscopy technique. The first and second of twenty prizewinners receive a selection of Nikon products and equipment worth $3,000 and $2,000 respectively. Use of a Nikon microscope or camera is not required. Participants can enter online by uploading their images or sending in 35mm transparencies to Nikon's headquarters in Melville, New York.
The deadline for entries is June 30, 2004.
For complete contest information and to enter online, go to www.nikonsmallworld.com . You can also call Nikon at 631-547-8569.
} From Stan at Nikon
From MicroscopyL-request-at-ns.microscopy.com Sun Jun 13 20:19:16 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (charles-at-3dcircuits.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, June 11, 2004 at 12:51:05 ---------------------------------------------------------------------------
Question: We are attempting to restore the "image calibration file" on a Noran Voyager (V3.7 software). The loss of this file occurred after the Voyager 3.7 software was installed (from scratch) on a new computer. We do not have a backup or copy of this file, however, we do have a printout of the calibration constants from the previous setup. According to Thermo, this file is entitled "wima.column1.cal", and cannot be easily "created" except by the field engineer. Does anyone know how to create this file, or could a correct calibration file be created with a "template" and filling in the calibration constants that we have in hardcopy.
I have a cressington turbo 208 carbon coater with the metal evaporator accessory.
I use it regularly for carbon coating, but am only just starting to use it for metal coating.
I would like to use gold wire, but am not sure how thick the wire should be, or the length of gold wire I require to obtain a suitable coat.
I have tried using 4cm of 0.25mm gold wire for 20 seconds at 20mA, but am not seeing a 'charge-free' coat. (I don't want to coat too thickly but am not sure how much gold per sample I should be using.)
Can anyone give me some ideas? Does anyone who has this coater already have some settings I could try?
thanks
Samantha Taylor TDG Experimental Officer Alcoa World Alumina XRD, Microscopy and Thermal Analysis Phone:(08) 9410 3588 Fax: (08) 9410 3167 Email: Samantha.Taylor-at-Alcoa.com.au
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 14 05:51:52 2004
The Ask-the-Experts session at the M&M 2004 meeting in Savannah takes on a different format this year to provide programming closer to the time of the annual Microscopy and Microanalysis (M&M) meeting, and to better address the needs of the meeting attendees.
Rather than listing topics in the Call for Papers, which must be produced nearly a year before the meeting, an on-line Form is now available for you to suggest topics for this informal session.
You can find a link to this form in the News section of the Meeting Home Page. :
http://mm2004.microscopy.org
This link will allow meeting participants to submit questions, and/or as appropriate suggest name(s) of experts to whom the question might be directed.
The Ask-the- Experts organizers will then seek to schedule a time to address topics of significant interest during the meeting.
The schedule of topics will be announced and posted to the meeting website a couple of weeks before the early registration deadline, so that potential M&M 2004 attendees can schedule in the topics of interest to them. Please plan to participate both in the topic selection and the sessions, so that we can gauge whether this change of format is beneficial!
Ask-The-Experts Organizers.
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 14 07:39:27 2004
Our SEM (JEOL 6360 LV w/tungsten filament) is likely being relocated along with the rest of our lab to a second-floor location (the room is on a concrete slab) in the general vicinity of a mainframe computer, an elevator, a men's room and a large outside air conditioning unit. I have asked to give an expert opinion on the feasibility of this move from a noise (electromagnetic and vibration) standpoint. The proposed room is ~ 40 feet from the mainframe (as the crow flies, across the hall with multiple walls in between), ~ 40 feet from the elevator (same side of the hallway, four walls between), next door to the men's room (but the stalls are on the opposite wall) and ~ 20 feet from the outside A/C unit. The scope is on a JEOL-supplied air table, and I typically image at less than 5,000x magnification (usually no higher than 1,000x). My samples are predominantly organic in nature (cloths, polymers, hair).
I have proposed to have the room assessed for the various types of noise (~$3,500), but have been greeted with resistance by management, who expected about $300 (and have no expertise in microscopy). So I ask those on the ListServer who have encounted a similar noise concern for an SEM lab relocation, should I expect to have issues with electromagnetic or vibrational noise due to any of the mainframe, elevator, men's room or A/C unit (or even second-floor on concrete location)?
I thank you in advance for any assistance.
Kirk
Brian (Kirk) Kirkmeyer, Ph.D. Research Scientist, Microscopy International Flavors and Fragrances 1515 State Highway 36 Union Beach, NJ 07735-3542 732-335-2426 / 732-335-2350 FAX brian.kirkmeyer-at-iff.com
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 14 09:06:31 2004
Once upon a time our sem was moved to a second floor location adjacent to the MTS Lab (fatigue/tensile testing lab). That lasted a month of so before management was convinced that the system was nearly useless in the new location. Images had the "jiggles" at a few thousand x and higher. The lab was re-relocated back.
You can take your chances or measure the environmental conditions. You *may* have sufficient EMI isolation, given the distances. Vibration isolation is very dependent on the building construction and source levels and more difficult to guess/estimate.
} From your description, there are no high current wiring or switchboxes nearby. I would be most concerned about mechanical vibrations with EMI concerns secondary.
The question is: How expensive is a vibration survey vs. moving the sem and finding it will not do your work?
Woody
Woody White BWXT Services: http://www.bwxt.com/bwxt.html My Site: http://woody.white.home.att.net
-----Original Message----- } From: Brian.Kirkmeyer-at-iff.com [mailto:Brian.Kirkmeyer-at-iff.com] Sent: Monday, June 14, 2004 9:04 AM To: Microscopy-at-MSA.Microscopy.Com
Our SEM (JEOL 6360 LV w/tungsten filament) is likely being relocated along with the rest of our lab to a second-floor location (the room is on a concrete slab) in the general vicinity of a mainframe computer, an elevator, a men's room and a large outside air conditioning unit. I have asked to give an expert opinion on the feasibility of this move from a noise (electromagnetic and vibration) standpoint. The proposed room is ~ 40 feet from the mainframe (as the crow flies, across the hall with multiple walls in between), ~ 40 feet from the elevator (same side of the hallway, four walls between), next door to the men's room (but the stalls are on the opposite wall) and ~ 20 feet from the outside A/C unit. The scope is on a JEOL-supplied air table, and I typically image at less than 5,000x magnification (usually no higher than 1,000x). My samples are predominantly organic in nature (cloths, polymers, hair).
I have proposed to have the room assessed for the various types of noise (~$3,500), but have been greeted with resistance by management, who expected about $300 (and have no expertise in microscopy). So I ask those on the ListServer who have encounted a similar noise concern for an SEM lab relocation, should I expect to have issues with electromagnetic or vibrational noise due to any of the mainframe, elevator, men's room or A/C unit (or even second-floor on concrete location)?
I thank you in advance for any assistance.
Kirk
Brian (Kirk) Kirkmeyer, Ph.D. Research Scientist, Microscopy International Flavors and Fragrances 1515 State Highway 36 Union Beach, NJ 07735-3542 732-335-2426 / 732-335-2350 FAX brian.kirkmeyer-at-iff.com
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 14 10:00:54 2004
I have a 305FE Schottky field emitter gun that is decommissioned. It is eight months old and was working fine when pulled off. I'm toying with the idea of documenting the reverse engineering of this gun, unless someone can make good use of it. I also have the separate FE support box that contains the ion pump power supplies. This is probably just useful as a boat anchor.
It seems interesting to see what is inside a current production thermal FE gun. There would be pix and dimensions on my Web site if I do this.
gary g.
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 14 10:34:00 2004
I am writing today to try and answer a question with the help of knowledgeable experts in the field of microscopy. I am a MBA graduate student here at Texas A&M University working with them in collaboration and development of new and existing patents. Recently, I was tasked with trying to determine whether there is currently in the market a device which can measure the forces BETWEEN cells in the pico newton range. From the responses of industry and academic contacts so far it is evident that there are quite a number of companies which can measure the forces between cells and OTHER objects such as those to measure cell adhesion and cell surface tension by way of optical tweezers and AFM's. I have also found many devices (such as those from Cell Robotics) which can easily manipulate or surgically cut away portions of cellular material. But I have as of yet found no device which is used for the explicit measurement of forces between cells in the tens of pico newtons.
I really wish that I could be more specific in my explanation of what it the customer is seeking. It is my belief that because of possible patent disclosure I am not allowed to have a wider scope for the intended use of this device.
I would gladly welcome any responses through either the list server or to the phone number below at NASA. Please remember that I am not an expert in the field of microscopy and therefore simplicity in explanation is requested because of my ignorance. Thank you in advance for taking the time to read this and respond. Gig 'em!
Ryan M Lee NASA Technology Transfer Center Texas Engineering Extension System Texas A&M University System 979-862-8603
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 14 13:11:29 2004
The 2 small fluorescent tubes, which are situated just above where the knife and boat sit, both "blew" last week. The person using the ultramicrotome at the time told me that the bulbs were no longer working. I installed 2 new tubes today, and still can't get overhead illumination. I am now suspecting a blown fuse might be the cause, but I don't know where to look for this fuse - there is not much explanation in the manual about these overhead lights.
Could anyone please point me in the right direction?
Thanks in advance for any help.
Regards,
Paula.
Paula M. Allan-Wojtas Research Scientist - Food Microstructure/chercheur scientique - microstructure des aliments Food Safety and Quality team/ Salubrité et qualité des aliments Agriculture and Agri-Food Canada/Agriculture et Agroalimentaire Canada Telephone/Téléphone: 902-679-5566 Facsimile/Télécopieur: 902-679-2311 32 Main Street/ 32, rue Main Kentville, Nova Scotia/ Kentville (Nouvelle-Écosse) B4N 1J5
allanwojtasp-at-agr.gc.ca
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 14 13:11:07 2004
Does anyone out there have the service manual/circuit diagrams for a Cambridge Stereoscan 90 SEM, or can you tell me where I can purchase them? I'd also just like to know who else has this microscope. I have some spares (e.g., turbo pump, camera system) available for barter. I also have documentation for a Stereoscan 150 if anybody wants it.
Paul Grover Dept. of Biological Sciences Purdue University
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 14 13:33:55 2004
Woody, Last year I moved one FESEM and three TEMs into a new lab. I did the site check with a $200 EMF meter/probe, available from several vendors on-line, and about 20mls of clean mercury in a well sealed 60ml poly bottle (wider is better). The required EMF meter readings are available in the equipment installation manual. I placed the bottle of mercury on the floor where the EM was to go, removed the cap (very carefully) and peeked down through the bottle neck at the surface of the liquid. With no vibrations I would see a smooth mirror. If rings are visible in the mercury's surface than the floor is vibrating. Low EMF and no 'jiggles' here. All of my scopes are running at spec.
Disclaimer: I am not responsible for your results or any mercury spills due to operator error. There are many variables to consider. Vibrations may vary over the course of time (trains, intermittent local equipment operation). Audio noise and air flow patterns can not be detected using the above methods.
Good luck. Jim
} From: "White, Woody N." {nwwhite-at-bwxt.com} } To: "'Brian.Kirkmeyer-at-iff.com'" {Brian.Kirkmeyer-at-iff.com} , } Microscopy-at-msa.microscopy.com } Subject: [Microscopy] RE: SEM site assessment } Date: Mon, 14 Jun 2004 09:25:42 -0500 } MIME-Version: 1.0 } X-UConn-MailScanner-Information: Contact UConn Help Desk 860-486-4357 for } more information. } X-UConn-MailScanner: Found to be clean } X-UConn-MailScanner-SpamCheck: } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
James S. Romanow The University of Connecticut Physiology and Neurobiology Department Electron Microscopy Facility Unit 3242 BSP Building, Room G06 91 North Eagleville Road Storrs, CT 06269-3242
Kirk, Last year I moved one FESEM and three TEMs into a new lab. I did the site check with a $200 EMF meter/probe, available from several vendors on-line, and about 20mls of clean mercury in a well sealed 60ml poly bottle (wider is better). The required EMF meter readings are available in the equipment installation manual. I placed the bottle of mercury on the floor where the EM was to go, removed the cap (very carefully) and peeked down through the bottle neck at the surface of the liquid. With no vibrations I would see a smooth mirror. If rings are visible in the mercury's surface than the floor is vibrating. Low EMF and no 'jiggles' here. All of my scopes are running at spec.
Disclaimer: I am not responsible for your results or any mercury spills due to operator error.
Good luck. Jim
} From: "White, Woody N." {nwwhite-at-bwxt.com} } To: "'Brian.Kirkmeyer-at-iff.com'" {Brian.Kirkmeyer-at-iff.com} , } Microscopy-at-msa.microscopy.com } Subject: [Microscopy] RE: SEM site assessment } Date: Mon, 14 Jun 2004 09:25:42 -0500 } MIME-Version: 1.0 } X-UConn-MailScanner-Information: Contact UConn Help Desk 860-486-4357 for } more information. } X-UConn-MailScanner: Found to be clean } X-UConn-MailScanner-SpamCheck: } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
James S. Romanow The University of Connecticut Physiology and Neurobiology Department Electron Microscopy Facility Unit 3242 BSP Building, Room G06 91 North Eagleville Road Storrs, CT 06269-3242
We're loosing our Hitachi 3500 SEM due to a split in the EM facility here at Clemson University. Is there anything out there available, comparable on a used basis. We're looking for equipment and a price. Thanks in advance for any information.
Darryl Krueger, RA BioScience Dept. Clemson University
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 14 15:39:17 2004
For cheap vibration detection, I've used a large (40 cm diameter) dish of water, placed on whatever surface you're testing, no a/c on, turn overhead lights off, shine a lamp onto the water surface at a low angle from the side while standing perfectly still and observe the reflections on adjacent wall - this is quite sensitive - can pick up vibrations from folk walking down the hall outside. But perhaps a denser liquid, like mercury, picks up a different vibration spectrum, or responds differently?
I used this demonstration to get a proper anti-vibration table for one of our light microscopes....
cheers, Rosemary
Dr. Rosemary White rosemary.white-at-csiro.au Microscopy Centre ph. 61-2-6246 5475 CSIRO Plant Industry mob. 61-0402 835 973 GPO Box 1600 fax. 61-2-6246 5000 Canberra, ACT 2601 Australia
} From: Jim Romanow {bsgphy3-at-uconnvm.uconn.edu} } Date: Mon, 14 Jun 2004 14:55:49 -0400 } To: microscopy-at-SPARC5.microscopy.com } Subject: [Microscopy] RE: SEM site assessment } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } Woody, } Last year I moved one FESEM and three TEMs into a new lab. I did the site } check with a $200 EMF meter/probe, available from several vendors on-line, } and about 20mls of clean mercury in a well sealed 60ml poly bottle (wider } is better). The required EMF meter readings are available in the equipment } installation manual. I placed the bottle of mercury on the floor where the } EM was to go, removed the cap (very carefully) and peeked down through the } bottle neck at the surface of the liquid. With no vibrations I would see a } smooth mirror. If rings are visible in the mercury's surface than the floor } is vibrating. Low EMF and no 'jiggles' here. All of my scopes are running } at spec. } } Disclaimer: } I am not responsible for your results or any mercury spills due to operator } error. There are many variables to consider. Vibrations may vary over the } course of time (trains, intermittent local equipment operation). Audio } noise and air flow patterns can not be detected using the above methods. } } Good luck. } Jim } } } } } From: "White, Woody N." {nwwhite-at-bwxt.com} } } To: "'Brian.Kirkmeyer-at-iff.com'" {Brian.Kirkmeyer-at-iff.com} , } } Microscopy-at-msa.microscopy.com } } Subject: [Microscopy] RE: SEM site assessment } } Date: Mon, 14 Jun 2004 09:25:42 -0500 } } MIME-Version: 1.0 } } X-UConn-MailScanner-Information: Contact UConn Help Desk 860-486-4357 for } } more information. } } X-UConn-MailScanner: Found to be clean } } X-UConn-MailScanner-SpamCheck: } } } } } } } } -----------------------------------------------------------------------------} } - } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ----------------------------------------------------------------------------- } } -- } } } } Once upon a time our sem was moved to a second floor location adjacent to } } the MTS Lab (fatigue/tensile testing lab). That lasted a month of so before } } management was convinced that the system was nearly useless in the new } } location. Images had the "jiggles" at a few thousand x and higher. The lab } } was re-relocated back. } } } } You can take your chances or measure the environmental conditions. You } } *may* have sufficient EMI isolation, given the distances. Vibration } } isolation is very dependent on the building construction and source levels } } and more difficult to guess/estimate. } } } } } From your description, there are no high current wiring or switchboxes } } nearby. I would be most concerned about mechanical vibrations with EMI } } concerns secondary. } } } } The question is: How expensive is a vibration survey vs. moving the sem and } } finding it will not do your work? } } } } Woody } } } } } } } } Woody White } } BWXT Services: } } http://www.bwxt.com/bwxt.html } } My Site: } } http://woody.white.home.att.net } } } } -----Original Message----- } } } From: Brian.Kirkmeyer-at-iff.com [mailto:Brian.Kirkmeyer-at-iff.com] } } Sent: Monday, June 14, 2004 9:04 AM } } To: Microscopy-at-MSA.Microscopy.Com } } Subject: [Microscopy] SEM site assessment } } } } } } } } ---------------------------------------------------------------------------- } } -- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ---------------------------------------------------------------------------- } } --- } } } } Our SEM (JEOL 6360 LV w/tungsten filament) is likely being relocated along } } with the rest of our lab to a second-floor location (the room is on a } } concrete slab) in the general vicinity of a mainframe computer, an } } elevator, a men's room and a large outside air conditioning unit. I have } } asked to give an expert opinion on the feasibility of this move from a } } noise (electromagnetic and vibration) standpoint. The proposed room is ~ } } 40 feet from the mainframe (as the crow flies, across the hall with } } multiple walls in between), ~ 40 feet from the elevator (same side of the } } hallway, four walls between), next door to the men's room (but the stalls } } are on the opposite wall) and ~ 20 feet from the outside A/C unit. The } } scope is on a JEOL-supplied air table, and I typically image at less than } } 5,000x magnification (usually no higher than 1,000x). My samples are } } predominantly organic in nature (cloths, polymers, hair). } } } } I have proposed to have the room assessed for the various types of noise } } (~$3,500), but have been greeted with resistance by management, who } } expected about $300 (and have no expertise in microscopy). So I ask those } } on the ListServer who have encounted a similar noise concern for an SEM } } lab relocation, should I expect to have issues with electromagnetic or } } vibrational noise due to any of the mainframe, elevator, men's room or A/C } } unit (or even second-floor on concrete location)? } } } } I thank you in advance for any assistance. } } } } Kirk } } } } Brian (Kirk) Kirkmeyer, Ph.D. } } Research Scientist, Microscopy } } International Flavors and Fragrances } } 1515 State Highway 36 } } Union Beach, NJ 07735-3542 } } 732-335-2426 / 732-335-2350 FAX } } brian.kirkmeyer-at-iff.com } } } } James S. Romanow } The University of Connecticut } Physiology and Neurobiology Department } Electron Microscopy Facility } Unit 3242 } BSP Building, Room G06 } 91 North Eagleville Road } Storrs, CT 06269-3242 } } 860 486-2914 voice } 860 486-6369 fax } james.romanow-at-uconn.edu } } } }
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 14 21:23:27 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (muchphoto-at-Earthlink.net) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Monday, June 14, 2004 at 14:11:43 ---------------------------------------------------------------------------
Email: muchphoto-at-Earthlink.net Name: Michael Reese Much
Education: Undergraduate College
Location: Bethlehem, PA USA
Question: I am photographing protozoa and am looking for a way to slow them down or even kill them for digital photomicroscopy. I am using methyl cellulose to slow them down but I'd like to find a way to incapacitate them without their cellular structure disintegrating.
muchphoto-at-Earthlink.net (by way of Ask-A-Microscopist) wrote: } } } ------------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America} To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver} On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html} -------------------------------------------------------------------------------} } Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (muchphoto-at-Earthlink.net) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Monday, June 14, 2004 at 14:11:43} ---------------------------------------------------------------------------} } Email: muchphoto-at-Earthlink.net } Name: Michael Reese Much} } Education: Undergraduate College} } Location: Bethlehem, PA USA} } Question: I am photographing protozoa and am looking for a way to slow them down or even kill them for digital photomicroscopy. I am using methyl cellulose to slow them down but I'd like to find a way to incapacitate them without their cellular structure disintegrating.} } ---------------------------------------------------------------------------Dear Michael Much: What kind of microscopy are you using for digital photomicroscopy of protozoa? Your post doesn't specify what kind, but I'm presuming that you're using light microscopy in conjunction with digital photography. I'm hardly an expert on protozoa in general, but I did some work on ciliates while I was working on my M.A. thesis about 4 years ago. Methyl cellulose should indeed slow down protozoa (and definitely on ciliates), but apparently it's affecting the cell's cellular structure. I'm not sure what other chemicals could slow down protozoa other than methyl cellulose, but I do know of various methods in light microscopy that can kill protozoa. For example, if you're working in a lab that uses traditional electron microscopy techniques, then you might have access to toxic but effective chemicals like osmium tetroxide (OsO4). The vapors from that chemical! are usually sufficient to kill protozoa. Alternatively, you could pursue using a small amount of, say, glutaraldehyde or some other kind of fixative. I'm guessing that adding a small amount, and then diluting it alot by repeated washes might result in a clean enough slide for photomicrography. Good luck with your assignment. Nelson Conti
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From MicroscopyL-request-at-ns.microscopy.com Tue Jun 15 03:31:00 2004
Below are some articles from Miscape that address slowing down protozoa and vital staining of live protozoa. Many of the classic formula for slowing various wee creatures call for narcotics that we considered difficult to get. I have found that most are not if you know a doctor well enough that he trusts you to use the substance for what you say you are going to use it for and don't live in an area the US Drug Enforcement is making an example of at the time. The last time I needed chloroform I got a prescription from a veterinarian. My doctor had no problems with a small bottle of coral hydrate already diluted and if I wanted cocaine I would talk to my eye doctor and as for an amount so small that it would be worthless as a stimulant and dilute it on the spot. A compounding pharmacy will have cocaine as well but when it gets to class 3 narcotics that have to be written on the DEA pad most doctors are pretty careful not write any that look odd.
CO2 bubbled in the media slows and kills many organisms. Physical compression in thick media also helps. If you can't find a compressing cell I have been looking for and excuse to make some for my own use and this might get me off high center.
Matching the chemical to the desired action generaly relaxation at death or as the processes slow is what is desired and sometimes can be achieved by using weaker slower acting solution of the chemical. If you can find a proven protocol and the chemical it is far better to use it than to water time developing you own.
Far better to walk on the shoulders of giants than to wander then wilderness alone doing the same work over and over again.
On slowing and killing protozoa http://www.microscopy-uk.org.uk/mag/indexmag.html?http://www.microscopy-uk.org.uk/mag/artmay02/rhmicrotech.html
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward any links or information you think might be useful to others. Microscope Manual at www.science-info.org
----- Original Message ----- } From: "by way of Ask-A-Microscopist" {muchphoto-at-Earthlink.net} To: {microscopy-at-microscopy.com} Sent: Monday, June 14, 2004 9:46 PM
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (R.H.Olley-at-reading.ac.uk) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, June 14, 2004 at 12:51:52 ---------------------------------------------------------------------------
Email: R.H.Olley-at-reading.ac.uk Name: Robert H. Olley
Organization: Reading University
Title-Subject: [Microscopy] [Filtered] Shadow direction in EM and planetary exploration
Question: I have recently seen some planetary exploration photographs which remind me of features seen in polymers under SEM / TEM. If you look at the following two
Don't you think they've got the shadow direction wrongly oriented? The Phoebe craters look like tents and the caldera appears to be sticking up. Not the sort of mistake an electron microscopist would make!?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (R.H.Olley-at-reading.ac.uk) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, June 15, 2004 at 03:05:19 ---------------------------------------------------------------------------
Email: R.H.Olley-at-reading.ac.uk Name: Robert H. Olley
Organization: Reading University
Title-Subject: [Microscopy] [Filtered] Slowing down protozoa
Question: to: Michael Reese Much
I remember my biology teacher telling us that one used menthol to kill amoeba. Most agents are so aggressive that the amoeba withdraws its pseudopods and goes into a lump, but menthol kills the amoeba so gently that "it doesn't know what's happening to it". That way, one can get static amoeba with pseudopods extended.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (romano-at-mpie.de) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, June 15, 2004 at 03:30:45 ---------------------------------------------------------------------------
Question: Dear Sir or Madam, Currently I am trying to use SEM and LOM to see the structure of crustacean shells. I was looking on the net for information related to preparation of the samples. I was wondering if it would be possible to get some information related to this. Could you please give me any hints about how to prepare the samples?, or where should I look to get a deeper knowledge of biological tissues?. Could you recommend some literature related to it?. Thank you very much in advance. Best regards.
Patricia Romano
Dr. Patricia Romano Triguero Max-Planck-Institut fuer Eisenforschung Abteilung - Mikrostrukturphysik und Umformtechnik Max-Planck-Str. 1 40237 Duesseldorf Deutschland
Tel +49 (0)211-6792-663 Fax +49 (0)211-6792-333 Email romano-at-mpie.de
Getting ready for new SEM, I find the following odd items if someone can use them:
2ea FEI 13423 LaB6 cathodes, appear to be new 1ea FEI 12629 LaB6 cathode 1ea FEI 18570 Schottky FE cathode, appears new 1ea Denka M1-EM {100} 90 degrees 15uR LaB6 cathode, new 1ea box with 5 large Mo apertures 13-1 200234, new 10ea Topcon filaments AW030WET(?) new in box Strange looking LeMont Scientific Quad solid state BSE preamp with diodes. Square 4-hole mounting flange with O-ring. Diode plate swings out and back manually.
If anyone knows what these are for and can use them, let me know. Otherwise, they go to trash.
gary g.
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 15 10:36:42 2004
Rosemary, Back in 1980 I would put the Hg in a 10 cm petri dish to check for vibration. It 'got the job done' but I like your idea better.
Regards, Jim
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 15 12:52:04 2004
Unfortunately for images like these too often interpretation is complicated. But the outline of shadows tells us that we have impressions, not plateaus. For example, smaller round caldera on the top has a long sharp shadow that definitely looks like shadow from the rim and not shadow formed by the rounded top of the plateau. If you rotate this image 90 degrees left, so that brightest rim will be on the bottom, you'll get image more familiar for our eyes (illuminated from the top) and caldera definitely will be seen as depression with smaller calderas as even deeper depressions.
Vladimir
} -----Original Message----- } From: by way of MicroscopyListserver [mailto:R.H.Olley-at-reading.ac.uk] } Sent: Tuesday, June 15, 2004 8:08 AM } To: microscopy-at-microscopy.com } Subject: [Microscopy] viaWWW: Shadow direction in EM and } planetary exploration } } } } } -------------------------------------------------------------- } ---------------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ----------------- } } Below is the result of your feedback form (NJZFM-ultra-55). } It was submitted by (R.H.Olley-at-reading.ac.uk) from } http://microscopy.com/MicroscopyListserver/MLFormMail.html on } Monday, June 14, 2004 at 12:51:52 } -------------------------------------------------------------- } ------------- } } Email: R.H.Olley-at-reading.ac.uk } Name: Robert H. Olley } } Organization: Reading University } } Title-Subject: [Microscopy] [Filtered] Shadow direction in EM } and planetary exploration } } Question: I have recently seen some planetary exploration } photographs which remind me of features seen in polymers } under SEM / TEM. If you look at the following two } } (a) Olympus Mons Caldera on Mars: } } http://nssdc.gsfc.nasa.gov/imgcat/html/object_page/vo1_890a68.html } } (b) Craters on Phoebe from Cassini: } } http://antwrp.gsfc.nasa.gov/apod/ap040614.html } } Don't you think they've got the shadow direction wrongly } oriented? The Phoebe craters look like tents and the caldera } appears to be sticking up. Not the sort of mistake an } electron microscopist would make!? } } * * * * * * * * * * * * * * * * * * } } -------------------------------------------------------------- } ------------- } }
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 15 13:54:20 2004
On Jun 15, 2004, at 6:07 AM, by way of MicroscopyListserver wrote:
} Title-Subject: [Microscopy] [Filtered] Shadow direction in EM and } planetary exploration } } Question: I have recently seen some planetary exploration photographs } which remind me of features seen in polymers under SEM / TEM. If you } look at the following two } } (a) Olympus Mons Caldera on Mars: } } http://nssdc.gsfc.nasa.gov/imgcat/html/object_page/vo1_890a68.html } } (b) Craters on Phoebe from Cassini: } } http://antwrp.gsfc.nasa.gov/apod/ap040614.html } } Don't you think they've got the shadow direction wrongly oriented? } The Phoebe craters look like tents and the caldera appears to be } sticking up. Not the sort of mistake an electron microscopist would } make!? } Dear Robert, You are quite correct that the direction of the shadow changes the perception of height so that depressions appear raised and mounds appear as craters; this is a fairly well-known visual phenomenon. The advantage of a laptop is that one can hold the screen upside down and see the proper perspective. As to whether "they've got the shadow direction wrongly oriented", one disadvantage of ultramacroscopy is that the lighting conditions are fixed, and, until we can figure out how either to move the sun or provide a new one, we have to take what is given. Of course, the images could be rotated by 180 degrees before they're posted, but that would interchange the north and south poles, which goes against astronomical convention. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 15 14:21:17 2004
Don't know anything about the company......but perhaps you might look into SEMTech Solutions. They offer service for AMRAY SEMs, and also offer Re-manufactured Pre-owned Scanning Electron Microscopes.
http://www.semtechsolutions.com/
Kelly A. Ramos Metallurgical Engineer / Supervisor Argo-Tech Materials Laboratories 23555 Euclid Avenue Cleveland, OH 44117 216-692-5904 or 216-692-5446 (fax) 216-692-5816 http://www.atclabs.com
darryl krueger {dkruege-at-CLEMS To: Microscopy-at-MSA.Microscopy.com ON.EDU} cc: Subject: [Microscopy] Looking to buy SEM 06/14/04 03:30 PM
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Listees,
We're loosing our Hitachi 3500 SEM due to a split in the EM facility here at Clemson University. Is there anything out there available, comparable on a used basis. We're looking for equipment and a price. Thanks in advance for any information.
Darryl Krueger, RA BioScience Dept. Clemson University
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 15 15:06:43 2004
Two bags of Pella 16292 100ea 15mmx10mm specimen mounts and four Pella 1666 mount grabbing tweezers. If these fit your SEM and you can use them, first $15 for UPS shipping gets all.
gary g.
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 15 20:53:05 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (muchphoto-at-Earthlink.net) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, June 15, 2004 at 10:50:50 ---------------------------------------------------------------------------
Email: muchphoto-at-Earthlink.net Name: Michael Much
Education: Undergraduate College
Location: Bethlehem, PA, USA
Question: Thanks to everybody for your replies regarding slowing down protozoa. The diversity of answers was quite interesting. I really liked the link to vital staining of protozoa--something I have been experimenting with. I called my veterinarian regarding the suggestion of lidocaine and she felt it would most likely destroy any cellular organisms. Some of you asked what type of scope I'm using--it's a light microscope--a trinocular and I have fabricated my own camera mounts for the third tube to accept film cameras, digital cameras and video cameras. I am not a student but rather an amateur microscopist who is doing his own experimentation in shooting microscopic invertebrates.
To researchers out there in cyberspace. We are very intereted in localising types of collagen by immunocytochemistry on paraffin or resin sections of human tympanic membranes. If anyone has any suggestions on where to buy collagen antibodies that will work on the above please contact us.
Many thanks
Terry
Dr Terry Robertson (PhD) Senior Research Fellow and Acting Manager School of Surgery and Pathology First Floor M Block, QE2 Medical Centre Mailbag M504 University of Western Australia Nedlands, Australia 6907 Phone 61 8 93462935 Fax 61 8 93462891 Mobile 0403025440 email terryr-at-cyllene.uwa.edu.au
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 16 06:30:10 2004
Thank you to everyone who provided me with input! It has been a ton of help.
Kirk
Brian (Kirk) Kirkmeyer, Ph.D. Research Scientist, Microscopy International Flavors and Fragrances 1515 State Highway 36 Union Beach, NJ 07735-3542 732-335-2426 / 732-335-2350 FAX brian.kirkmeyer-at-iff.com
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 16 10:16:45 2004
If you haven't already taken a look at the new online microscopy journal www.modernmicroscopy.com, we suggest that you do, as it is becoming a great microscopy resource. This peer reviewed journal features articles, reviews, and tutorials about microscopy by some of the most experienced microscopists in the field, and access is free. As the main sponsor of the site, The McCrone Group staff has contributed a number of articles to get the site started. We are actively looking for new topics and ideas from the entire microscopy community. Submission information can be found on the site, and contributions are always welcome. The success of the site is determined by the participation from and the benefits provided to the microscopy community.
Thank you, on behalf of the McCrone Group,
Elaine Schumacher Senior Research Scientist McCrone Associates, Inc. 850 Pasquinelli Drive Westmont, IL 60559-5539 USA 630-887-7100 (tel) 630-887-7417 (fax) E-mail: eschumacher-at-mccrone.com Web Site: www.mccrone.com
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 16 11:26:24 2004
Join your colleagues this October in Cork, Ireland for the Conference on Food Structure and Food Quality. Sponsored by the Food Structure & Functionality Forum Division and the European Section of the AOCS.
This three-day symposium will hold a comprehensive technical program in addition to three intensive short courses.
The conference will also feature the winner of the 2004 Food Structure & Functionality Division Lifetime Achievement Award, Professor Anne-Marie Hermansson, SIK, The Swedish Institute for Food and Biotechnology, Gothenburg, Sweden.
Organizers are still accepting abstracts for oral and poster presentations through 30 June. Visit : http://www.aocs.org/meetings/fsq/call.asp for more information, including online registration and the complete technical program.
Register by 3 September to receive the early registration rate.
Paula M. Allan-Wojtas, Past Chair, Food Structure and Functionality Forum, a division of AOCS.
Research Scientist - Food Microstructure/chercheur scientique - microstructure des aliments Food Safety and Quality team/ Salubrité et qualité des aliments Agriculture and Agri-Food Canada/Agriculture et Agroalimentaire Canada Telephone/Téléphone: 902-679-5566 Facsimile/Télécopieur: 902-679-2311 32 Main Street/ 32, rue Main Kentville, Nova Scotia/ Kentville (Nouvelle-Écosse) B4N 1J5
allanwojtasp-at-agr.gc.ca
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 16 12:11:22 2004
Contributions about FIB are certainly welcome. Articles related to any aspect of optical and electron microscopy, related techniques and instrumentation will be considered for publication. We invite you to take a look at the site, and help us to expand range of subjects that you find there.
Regards,
Elaine
Elaine Schumacher Senior Research Scientist McCrone Associates, Inc. 850 Pasquinelli Drive Westmont, IL 60559-5539 USA 630-887-7100 (tel) 630-887-7417 (fax) E-mail: eschumacher-at-mccrone.com Web Site: www.mccrone.com
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 16 15:39:20 2004
A recent question has come up around our labs, and that is, "What effect do wireless laptop computers employing 802.11 B\A\G, Wireless Access Points, Cell Phones, Cordless Phones and other hand held wireless deices have on the performance of SEMs, TEMs, EELS spectrometers, and EDX Spectrometers". We are going to try to formulate some test methods to evaluate this, but I was hoping that maybe someone out there in the Microscopy World has had some experience with this subject, or has some Words of Wisdom on the subject.
John Mardinly Intel
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 16 17:33:17 2004
BAL-TEC AG and Boeckeler Instruments/RMC Products cordially invite you to the Bal-Tec Tutorial on SEM-Controlled Broad Ion Beam Sample Preparation, presented by Dr. Wolfgang Gruenewald.
(6/16/04 14:01) Mardinly, John {john.mardinly-at-intel.com} wrote:
} A recent question has come up around our labs, and that is, "What effect } do wireless laptop computers employing 802.11 B\A\G, Wireless Access } Points, Cell Phones, Cordless Phones and other hand held wireless deices } have on the performance of SEMs, TEMs, EELS spectrometers, and EDX } Spectrometers". We are going to try to formulate some test methods to } evaluate this, but I was hoping that maybe someone out there in the } Microscopy World has had some experience with this subject, or has some } Words of Wisdom on the subject. } } John Mardinly } Intel
John,
802.11 uses the 2.4 GHz band. If there is an effect, I'd expect you'd be able to detect it with a Fourier transform on the resulting image (much like you can find 60 Hz noise from the E field of power lines). If you were trying to design an experiment, I'd use a directional WiFi antenna at a known angle across the sample. My sense of the orders of magnitude makes me think that the SEM would not show any effect except at perhaps the highest possible magnifications, but the math on that is pretty easy to work out.
Cell phones use different bands, depending on the carrier and the service type. Those numbers are available online, but if memory serves. Most phones use 900, 1800 or 1900MHz bands. I think that US cell phones primarily use 900 and 1900 MHz.
Brent
-- Brent Neal, Ph.D. Reindeer Graphics, Inc. brent-at-reindeergraphics.com
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 17 05:55:21 2004
FYI, Tutorial Announcement at the M&M 2004 meeting are done by the Education Committee. A single announcement is made annually. I appreciate your enthusiasm to let people know about your tutorial, however, can you in the future please coordinate with me and/or the education committee with respect to Education Committee functions. It is preferrable to do a one time posting rather than lots of individual ones.
Thanks in advance
Nestor
At 6:55 PM -0400 6/16/04, RCHIOVETTI-at-aol.com wrote: } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 17 07:53:15 2004
I've received a reply to yesterday's posting indicating that there's a problem with getting past the main page of the modernmicroscopy.com site and accessing the articles. I tried it myself this morning both through the link that I provided and through Internet Explorer, and found the site to be locked up at present. I checked with one of our IT staff, and found that this has been an intermittent problem since the site was set up. The problem is with the internet service provided, and they haven't been able to resolve it yet, but I've been told that it seems to clear up on its own, and the site becomes accessible again. I had replies yesterday that indicated that people had been able to use the link in my posting to get to the journal articles.
Of the many times that I've gone to the journal in the past several months, this is the first time that I've encountered this problem. I guess it's Murphy's Law in action, to have the glitch occur just when I send out an invitation to the listserver community to visit the site! I'm sorry for any frustration that this may have caused, and urge you to try again at a later time. I'll check the site periodically myself. If you continue to have access problems, please let me know.
Regards,
Elaine
Elaine Schumacher Senior Research Scientist McCrone Associates, Inc. 850 Pasquinelli Drive Westmont, IL 60559-5539 USA 630-887-7100 (tel) 630-887-7417 (fax) E-mail: eschumacher-at-mccrone.com Web Site: www.mccrone.com
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 17 09:21:22 2004
Your submission of other meetings will be very appreciated (submission form is accessible from the page mentioned, the direct link is http://www.petr.isibrno.cz/microscopy/PMRform.php).
Petr Schauer
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 17 10:16:17 2004
A customer of mine is trying to view osteoblasts grown on a peptide layer on titanium disks. In both her conventional SEM and my ESEM, she has seen patches of cellular adhesive on the peptide layer, but the cells are missing (she knows from other procedures that the cells have been growing well on the disks).
Do you have any suggestions for specimen preparation (other than glutaraldehyde fixation) that would keep the osteoblasts intact for SEM?
Thanks!
Leslie Eibest SEM lab Dept. of Biology Duke University
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 17 11:34:39 2004
We have started long-term live cell experiments (} 24 hours) and although I have followed basic instructions learned over the years, there are still problems with our setup.
If anyone is interested in the details, we have a fair-sized plexiglass incubator housing the microscope stage into which I have put a water-filled flask through which CO2 is bubbled, we have a heater stage, a blower providing warm air and an extra container of water for humidity. Still, after several hours, when the temperature should have stabilized, we see drift in focus, condensation on the inside of the lid of the cell culture dish and inadequate humidity. CO2 is not regulated accurately.
Does anyone know of any webiste or other literature - with lots of pictures - to see how others have designed and used live cell setups.
Thank you in advance Judy
Judy Trogadis Bio-Imaging Coordinator St. Michael's Hospital, 7Queen 30 Bond St. Toronto, ON M5B 1W8 Canada ph: 416-864-6060 x6337 pager: 416-685-9219 fax: 416-864-6043 trogadisj-at-smh.toronto.on.ca
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 17 11:48:22 2004
I am not sure how you would measure a GHz signal on an image. Even if you acquire a 2kx2k image in 1 second, the sample frequency is a few MHz. That means, that the highest frequency you can measure is of the same order (Nyquist). If you can't measure it, it will not show up in FFTs either.
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Brent Neal [mailto:brent-at-reindeergraphics.com] Sent: Wednesday, June 16, 2004 17:42 To: Mardinly, John Cc: Microscopy-at-MSA.Microscopy.Com
All items have new homes.
Thanks for the replies.
gary g.
At 08:51 AM 6/15/2004, I wrote:
} Getting ready for new SEM, I find the following } odd items if someone can use them: } } 2ea FEI 13423 LaB6 cathodes, appear to be new } 1ea FEI 12629 LaB6 cathode } 1ea FEI 18570 Schottky FE cathode, appears new } 1ea Denka M1-EM {100} 90 degrees 15uR LaB6 cathode, new } 1ea box with 5 large Mo apertures 13-1 200234, new } 10ea Topcon filaments AW030WET(?) new in box } Strange looking LeMont Scientific Quad solid state BSE } preamp with diodes. Square 4-hole mounting flange } with O-ring. Diode plate swings out and back manually. } } If anyone knows what these are for and can use them, } let me know. Otherwise, they go to trash. } } gary g. }
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 17 12:52:48 2004
} More odd items. } } Two bags of Pella 16292 100ea 15mmx10mm } specimen mounts and four Pella 1666 } mount grabbing tweezers. If these } fit your SEM and you can use them, } first $15 for UPS shipping gets all. } } gary g. }
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 17 13:08:13 2004
Thanks to all for your responses and suggestions regarding the current problem in accessing the ModernMicroscopy site. We appreciate the feedback, and are working with our internet service provider to correct the problem. I'll post an update when the issues are resolved, and again, I apologize for the frustration that this may have caused to anyone trying to access the site.
Regards,
Elaine
Elaine Schumacher Senior Research Scientist McCrone Associates, Inc. 850 Pasquinelli Drive Westmont, IL 60559-5539 USA 630-887-7100 (tel) 630-887-7417 (fax) E-mail: eschumacher-at-mccrone.com Web Site: www.mccrone.com
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 17 13:21:54 2004
Has anyone out there looked at shark (OK, dogfish) vertebrae in an SEM? A colleague from down the hall in the fisheries department has some vertebrae (sectioned and unsectioned) that he'd like to look at in order to count the rings (kind of like the rings in a tree stump). The rings are just too close together to clearly differentiate in a binocular scope, but I was thinking in order for them to show up in an SEM there has to be a density or textural difference between "ring" and "inter-ring" areas. Would etching of sectioned ones with something help? (These vertebrae are not true bone of course, but a sort of calcified cartilage, so I suppose an acid might do something usefull......) I'm open to all ideas......
Frank
F.C. Thomas FThomas-at-NRCan.gc.ca, 902-426-4635, facsimile 902-426-6152 GSC Atlantic Natural Resources Canada, Bedford Institute of Oceanography, P.O. Box 1006, Dartmouth, Nova Scotia B2Y 4A2 Ressources naturelles Canada, l'Institut Oceanographique du Bedford, B.P. 1006, Dartmouth, (Nouvelle-Ecosse) B2Y 4A2 Government of Canada/Gouvernement du Canada
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 17 13:49:48 2004
I don't think that the GHz signal is going to affect the electron beam inside the column, it's all about skin depth. The most common way for disturbances of this kind is through cables between equipment, power lines and tracks on circuit boards. Any non linear electronics will create overtones, and if you have a signal in your system it will also create modulation products that is added as noise. Since all electronic circuits are nonlinear it will add noise, then it is up to the system design to compensate and filter out any noise. If you modulate (turning on and off) the GHz signal with a slower frequency you should get a detectable signal in the picture of more or less sharp features. Since most of the devices you named in the first letter is using time division multiplexing this should be simple to set up. In the most basic way you could use a cellular phone and just make a call. The duty cycle in a cellular system is in the order of 1/8th active and have a repetition frequency of around a couple of hundred hertz. For the exact numbers it depends on the phone system used and local settings. But it shouldn't be hard to find out in a web search or a call to the phone company. If you adjust the scanning speed you could probably get a striped picture as the signal is adding to the noise level in your system.
For a quick demo of the effect try putting your cellular phone near your computer monitor and make a call to see the effects. You don't see the GHz signal but the added noise is affecting circuits and you will see the time division.
I have never done any measurements like this since I just got our TEM working after a year of repairs and renovation. Now it's grounded again while we build a closed cooling system. Last week we got an EDS so we have to put some work into it again and get the system up and running. http://www.home.neab.net/gandalf/EM-lab/TEM100CX/index.htm
What I have done is worked with micro wave equipment and also designed electronic equipment with radios.
Btw, it's fully possible to sample a GHz signal at a couple of MHz. You can't recreate the original signal (that's Nyquist) but you will get a downconversion in the frequency band (folding distortion). This is a trick used in software frontends in the mobile phone system in the base stations. But any right designed A/D sampler in a system should have a filter in front of it to remove any signal above Nyquist, that's why the signal shouldn't appear directly in the picture.
As a final disclaimer I'm not going to take any responsibility if you damage your equipment. The signal levels from a mobile could be really high and affect the electronics, especially older equipment not designed for the wireless age. Start with a good distance between the phone and the equipment before getting closer.
Regards, Göran
Mike Bode wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 17 13:50:04 2004
(6/17/04 11:07) Mike Bode {mb-at-soft-imaging.com} wrote:
} Brent, } } I am not sure how you would measure a GHz signal on an image. Even if you } acquire a 2kx2k image in 1 second, the sample frequency is a few MHz. That } means, that the highest frequency you can measure is of the same order } (Nyquist). If you can't measure it, it will not show up in FFTs either. } } mike
That was the point I was making exactly, and what I meant when I said "the math is straightforward."
Brent
-- Brent Neal, Ph.D. Reindeer Graphics, Inc. brent-at-reindeergraphics.com
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 17 14:09:25 2004
Lets be careful what we play with at home. http://www.biomedcentral.com/news/20040617/04
Gordon Gordon Couger gcc-at-couger.com
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward any links or information you think might be useful to others. Microscope Manual at www.science-info.org
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 17 14:38:48 2004
The lockup problem with modernmicroscopy.com has been resolved, and the site is now accessible. We're not sure at this point if a high request volume resulting from my initial posting was the cause of the problem, but we'll continue to monitor the situation and work toward resolution of any ongoing issues. Thanks again to all for your patience and your feedback. As I said in my original message, please consider submitting articles for publication. We look forward to your contributions.
Regards,
Elaine
Elaine Schumacher Senior Research Scientist McCrone Associates, Inc. 850 Pasquinelli Drive Westmont, IL 60559-5539 USA 630-887-7100 (tel) 630-887-7417 (fax) E-mail: eschumacher-at-mccrone.com Web Site: www.mccrone.com
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 17 15:02:35 2004
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} A customer of mine is trying to view osteoblasts grown on a peptide layer on } titanium disks. In both her conventional SEM and my ESEM, she has seen } patches of cellular adhesive on the peptide layer, but the cells are missing } (she knows from other procedures that the cells have been growing } well on the disks). } } Do you have any suggestions for specimen preparation } (other than glutaraldehyde fixation) that would keep the osteoblasts } intact for SEM? } } Leslie Eibest } SEM lab, Dept. of Biology, Duke University =========== Leslie, Please let us know if the cells were Critical Point Dried, HMDS, etc.
There are some factors that apply to all preparations and some not. If the cell side of the disc comes in contact with other surfaces the cells can be mechanically knocked off. Were the discs secured in a holder? There may be expansion/contraction factors in play because a metal was used instead of glass or a plastic as a carrier for the peptide and cells.
Has any TEM shown the contact between the peptide and the osteoblasts? Do they stuck together well or only in small patches? I assume that a cross-section work-up was done to establish the fact that the cells do adhere well to the peptide.
Pat Connelly Univ. of Pennsylvania, Biology Dept.
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 17 16:30:39 2004
Dear Goran, I remember when Hitachi brought a FETEM to the Seattle ICEM meeting, the highest resolution was affected when the security guards activated their walkie-talkies. It just blurred the lattice they were showing a little. I think this is a much lower frequency than the GHz that cordless phones use, but certainly the highest resolution may be affected by radio interference affecting the electronics. Regards, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "Göran Axelsson" {axelsson-at-acc.umu.se} To: {Microscopy-at-MSA.Microscopy.Com} Sent: Thursday, June 17, 2004 12:16 PM
Howdie!
I don't think that the GHz signal is going to affect the electron beam inside the column, it's all about skin depth. The most common way for disturbances of this kind is through cables between equipment, power lines and tracks on circuit boards. Any non linear electronics will create overtones, and if you have a signal in your system it will also create modulation products that is added as noise. Since all electronic circuits are nonlinear it will add noise, then it is up to the system design to compensate and filter out any noise. If you modulate (turning on and off) the GHz signal with a slower frequency you should get a detectable signal in the picture of more or less sharp features. Since most of the devices you named in the first letter is using time division multiplexing this should be simple to set up. In the most basic way you could use a cellular phone and just make a call. The duty cycle in a cellular system is in the order of 1/8th active and have a repetition frequency of around a couple of hundred hertz. For the exact numbers it depends on the phone system used and local settings. But it shouldn't be hard to find out in a web search or a call to the phone company. If you adjust the scanning speed you could probably get a striped picture as the signal is adding to the noise level in your system.
For a quick demo of the effect try putting your cellular phone near your computer monitor and make a call to see the effects. You don't see the GHz signal but the added noise is affecting circuits and you will see the time division.
I have never done any measurements like this since I just got our TEM working after a year of repairs and renovation. Now it's grounded again while we build a closed cooling system. Last week we got an EDS so we have to put some work into it again and get the system up and running. http://www.home.neab.net/gandalf/EM-lab/TEM100CX/index.htm
What I have done is worked with micro wave equipment and also designed electronic equipment with radios.
Btw, it's fully possible to sample a GHz signal at a couple of MHz. You can't recreate the original signal (that's Nyquist) but you will get a downconversion in the frequency band (folding distortion). This is a trick used in software frontends in the mobile phone system in the base stations. But any right designed A/D sampler in a system should have a filter in front of it to remove any signal above Nyquist, that's why the signal shouldn't appear directly in the picture.
As a final disclaimer I'm not going to take any responsibility if you damage your equipment. The signal levels from a mobile could be really high and affect the electronics, especially older equipment not designed for the wireless age. Start with a good distance between the phone and the equipment before getting closer.
Regards, Göran
Mike Bode wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 17 20:41:44 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (dorit-at-burnham.org) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, June 17, 2004 at 12:12:40 ---------------------------------------------------------------------------
Email: dorit-at-burnham.org Name: Dorit Hanein
Organization: The Burnham Institute
Title-Subject: [Microscopy] [Filtered] Position Open: Electron Microscopist Research Assistance
Question: Position Open: Electron Microscopist Research Assistance, to start August, 2004. (http://www.burnham-inst.org).
Requirements include working knowledge of high resolution TEM and a BA/BS or equivalent experience in biological or material science, or bioengineering. Responsibilities include negative stain sample preparation and electron microscopy, vitreous ice sample preparation and electron cryo-microscopy, some microscope maintenance and standard biochemical analyses.
Application should include a statement of career goals and addresses for three references. Salary is competitive and commensurate with experience. EOE.
Send applications to: Dr. Dorit Hanein, e-mail dorit-at-burnham.org
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Wall1-at-LLNL.GOV) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, June 17, 2004 at 13:12:50 ---------------------------------------------------------------------------
Email: Wall1-at-LLNL.GOV Name: Mark A. Wall
Organization: Lawrence Livermore National Laboratory
Question: The Characterization group in the Chemistry and Materials Science Directorate at LLNL, Livermore, CA, has an opening for a PhD. level candidate. The selected candidate will be responsible for the daily operations of a new dual-beam FIB equiped with EDS, OIM, CL, lift-out, injections gases, STEM, etc. The selected candidate will supervise one dedicated technican who will maintain and operate the instrument. In addition, the selected candidate will be responsible for working with numerous staff scientists for the application of FIB technology as well as developing dual-beam experimentation, applications, etc. as required for numerous materials programs at LLNL. Experience with hands-on FIB techniques is required. Experience with TEM and other materials characterzation techniques is highly desired.
Applications shall be submitted on-line at http://jobs.llnl.gov/prod_index.html
The job posting number is 001906.
Please refer questions to Mark A. Wall -at- wall1-at-LLNL.GOV
Frank, Cartilage is nearly all protein. Have you considered tagging it with fluorescent antibodies? There may be one available for use in a non-shark ( bovine? human?) cartilage imaging application that would bind well enough to shark. Some of genetic machinery (and the proteins that result) which allowed early the chordates to flourish remains essentially unchanged. Perhaps that optical scope will still be the quick and easy solution.
Bob Carter 2000 Bayshore Road Lopez Island, WA 98261
-----Original Message----- } From: Thomas, Frank [mailto:FThomas-at-nrcan.gc.ca] Sent: Thursday, June 17, 2004 11:44 AM To: Microscopy-at-MSA.Microscopy.Com
Listers -
Has anyone out there looked at shark (OK, dogfish) vertebrae in an SEM? A colleague from down the hall in the fisheries department has some vertebrae (sectioned and unsectioned) that he'd like to look at in order to count the rings (kind of like the rings in a tree stump). The rings are just too close together to clearly differentiate in a binocular scope, but I was thinking in order for them to show up in an SEM there has to be a density or textural difference between "ring" and "inter-ring" areas. Would etching of sectioned ones with something help? (These vertebrae are not true bone of course, but a sort of calcified cartilage, so I suppose an acid might do something usefull......) I'm open to all ideas......
Frank
F.C. Thomas FThomas-at-NRCan.gc.ca, 902-426-4635, facsimile 902-426-6152 GSC Atlantic Natural Resources Canada, Bedford Institute of Oceanography, P.O. Box 1006, Dartmouth, Nova Scotia B2Y 4A2 Ressources naturelles Canada, l'Institut Oceanographique du Bedford, B.P. 1006, Dartmouth, (Nouvelle-Ecosse) B2Y 4A2 Government of Canada/Gouvernement du Canada
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 18 07:32:13 2004
} =========== } Leslie, } Please let us know if the cells were Critical Point Dried, HMDS, etc.
For her conventional SEM workup, the cells were dehydrated in ethanol, then critical point dried and sputter coated. For ESEM work in my lab, we put disks with cells directly in the scope, and viewed them wet at about 5 Torr and 3 C.
} There are some factors that apply to all preparations and some not. } If the cell side of the disc comes in contact with other surfaces } the cells can be mechanically knocked off. Were the discs secured in a holder?
For ESEM work, since we didn't do any processing, I don't think the cells contacted any other surfaces.
} There may be expansion/contraction factors in play because a metal was } used instead of glass or a plastic as a carrier for the peptide and cells.
That could be, since the samples were chilled.
} Has any TEM shown the contact between the peptide and the osteoblasts? } Do they stuck together well or only in small patches? I assume that a } cross-section work-up was done to establish the fact that the cells do } adhere well to the peptide.
She hasn't done any TEM that I'm aware of, but has been able to recover growing cells from the disks. The fact that we could see where the cells used to be attached gave us the impression that they had adhered to the peptide layer. We don't know about cell distribution on the disk - that's what she was trying to determine with the SEM.
Thanks for your help!
Leslie
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 18 07:55:39 2004
i used to do a lot of work on bone/implant interface and osseointegration including culture studies. i would grow long-term (2-6 months) osteogenic cells (released from fetal mouse calvaria) on commercially pure titanium and titanium alloy dics. i viewed specimens that i ran through CPD after glut fixation. partly, i was looking for neural "contaminant" cells that were growing on top of the osteogenic cells and they were fairly easy to see. the osteo cell lawn is very difficult to see since the cells attach and flatten out much like epithelial or fibroblast cells and form a thin layer on the surface. this was true even before confluency and this was on uncoated titanium surfaces. adding a protein coating will make the visualization even more difficult as i found using various extracellular matrix protein coatings. if i used fine sandpaper to rough up the surface, i could easily see the surface groves "through" the cell layer. in short, don't expect the cells to look distinct especially after several days in culture since they lay down their own matrix and adhesion proteins and are pretty much part of the surface.
hope this helps.
tbudd
Pat Connelly wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } A customer of mine is trying to view osteoblasts grown on a peptide layer on } } titanium disks. In both her conventional SEM and my ESEM, she has seen } } patches of cellular adhesive on the peptide layer, but the cells are missing } } (she knows from other procedures that the cells have been growing } } well on the disks). } } } } Do you have any suggestions for specimen preparation } } (other than glutaraldehyde fixation) that would keep the osteoblasts } } intact for SEM? } } } } Leslie Eibest } } SEM lab, Dept. of Biology, Duke University } =========== } Leslie, } Please let us know if the cells were Critical Point Dried, HMDS, etc. } } There are some factors that apply to all preparations and some not. } If the cell side of the disc comes in contact with other surfaces } the cells can be mechanically knocked off. Were the discs secured in a holder? } There may be expansion/contraction factors in play because a metal was } used instead of glass or a plastic as a carrier for the peptide and cells. } } Has any TEM shown the contact between the peptide and the osteoblasts? } Do they stuck together well or only in small patches? I assume that a } cross-section work-up was done to establish the fact that the cells do } adhere well to the peptide. } } Pat Connelly } Univ. of Pennsylvania, Biology Dept.
-- Dr. T. Budd Professor and Chair of Biology St. Lawrence University Canton, NY 13617 Phone = 315-229-5640 Fax = 315-229-7429 E-mail = tbudd-at-stlawu.edu
This message is made of 100% recycled electrons!
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 18 08:10:34 2004
If you are using a cell phone for this test, be aware that the output power the antenna provides is a function of what the cell base station tells it to put out and that's a function of the power the base station receives. You will unlikely have constant output power from a cell phone. If this is a test you really need to do, you should simulate the RF with a generator and the modulation scheme you wish to use, i.e. CDMA, TDMA, GSM etc, You will also need an antenna and power meter/spectrum analyzer to really know what power level you have at the intended target, EM column/electronics?
Peter Tomic Agere Systems
-----Original Message----- } From: Brent Neal [mailto:brent-at-reindeergraphics.com] Sent: Wednesday, June 16, 2004 7:42 PM To: Mardinly, John Cc: Microscopy-at-MSA.Microscopy.Com
(6/16/04 14:01) Mardinly, John {john.mardinly-at-intel.com} wrote:
} A recent question has come up around our labs, and that is, "What } effect do wireless laptop computers employing 802.11 B\A\G, Wireless } Access Points, Cell Phones, Cordless Phones and other hand held } wireless deices have on the performance of SEMs, TEMs, EELS } spectrometers, and EDX Spectrometers". We are going to try to formulate
} some test methods to evaluate this, but I was hoping that maybe someone
} out there in the Microscopy World has had some experience with this } subject, or has some Words of Wisdom on the subject. } } John Mardinly } Intel
John,
802.11 uses the 2.4 GHz band. If there is an effect, I'd expect you'd be able to detect it with a Fourier transform on the resulting image (much like you can find 60 Hz noise from the E field of power lines). If you were trying to design an experiment, I'd use a directional WiFi antenna at a known angle across the sample. My sense of the orders of magnitude makes me think that the SEM would not show any effect except at perhaps the highest possible magnifications, but the math on that is pretty easy to work out.
Cell phones use different bands, depending on the carrier and the service type. Those numbers are available online, but if memory serves. Most phones use 900, 1800 or 1900MHz bands. I think that US cell phones primarily use 900 and 1900 MHz.
Brent
-- Brent Neal, Ph.D. Reindeer Graphics, Inc. brent-at-reindeergraphics.com
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 18 11:40:25 2004
Cartilage has a lot of glycosamionglycans (hyaluronic acid and various chondroitin sulfates) to go along with the proteins, cells and fibers of various types. All of these components vary in propotion to the type of cartilage, its location and (I would suspect) the species. An LM stain like Alcian Blue or colloidal iron might be of use
Bob Carter wrote:
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-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
From MicroscopyL-request-at-ns.microscopy.com Sat Jun 19 07:35:13 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (k.bustamante-at-surrey.ac.uk) from http://microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Friday, June 18, 2004 at 14:09:33 ---------------------------------------------------------------------------
Email: k.bustamante-at-surrey.ac.uk Name: Karla Bustamatne
Organization: University of Surrey
Education: Graduate College
Location: Surrey, UK
Question: HI. I had taken histological samples on the locust ventral cord. I am taking neural recordings using penetrating electrodes. I was hoping to take the recordings and then remove the probe and be able to find track where the probe was positioned. I was unable to find any. Therefore i left one of the probes inside the tissue and proceed with the histology procedure. I found that close to the region where the probe was teh staining is poor. I was wondering if the probe which is make of silicon oxide and gold electrodes could have had an effect on the staining. I used bodian, cresyl fast violet and haemotoxily and eosin staining methods. I am not an experience histologist.Actually my main degree is in electronics so apologize if i am asking something obvious.
HPH (Hello, and Please Help.) I have been trying to make formvar support grids by methods that have always produced good results in years past. Now, however, the formvar refuses to leave the glass slides as I lower them into a container of water. Has anyone experienced this? Can you give ne a clue? My procedure now and in the past is to dip clean slides in a solution (w/v) of 0.25% formvar in dichloroethane, air dry, then scrape bottom and sides with a razor blade. What do you think? Humidity? Phase of the moon? Bad hands? Please advise. WJP
From MicroscopyL-request-at-ns.microscopy.com Sun Jun 20 01:55:32 2004
HHH (Hello, Here's help), Yes, it is probably humidity, phase of the moon, or the pollen. What I have found helps ever so much with Formvar is to take a kimwipe (a brand of tissue paper--probably lens paper would work too) and rub it vigorously over the surface of the slide and then dip into the Formvar and cast as usual. I guess this charges the surface with a little static electricity, but whatever the mechanism this seems to work a treat.
Happy casting, Tobias
} } } HPH (Hello, and Please Help.) I have been trying to make formvar } support grids by methods that have always produced good results in } years past. Now, however, the formvar refuses to leave the glass } slides as I lower them into a container of water. Has anyone } experienced this? Can you give ne a clue? My procedure now and in } the past is to dip clean slides in a solution (w/v) of 0.25% formvar } in dichloroethane, air dry, then scrape bottom and sides with a } razor blade. What do you think? Humidity? Phase of the moon? Bad } hands? Please advise. WJP
After scraping an outline of the film to be cast with a clean razor blade, I breathe on the coated slide and the vapor of my breath makes the film look frosty. Then I quickly dip the slide at a 45 degree angle into a bowl of water and the film floats off. If you don't want to blow on the slide, I have held the slide over a beaker of hot water (recently boiled) and the water vapor likewise makes the film on the slide look frosty. As an aside, I no longer use Formvar or Parlodian, but prefer Butvar which, in my hands, makes very stable films even during the humidity of summer in Iowa. I don't even bother to carbon coat my films which I routinely cast on 1X2 copper or nickel slot grids.
Dean Abel Biological Sciences 143 BB University of Iowa Iowa City IA 52242-1324
At 09:15 PM 6/19/2004 -0500, you wrote: } HPH (Hello, and Please Help.) I have been trying to make formvar support } grids by methods that have always produced good results in years past. } Now, however, the formvar refuses to leave the glass slides as I lower } them into a container of water. Has anyone experienced this? Can you give } ne a clue? My procedure now and in the past is to dip clean slides in a } solution (w/v) of 0.25% formvar in dichloroethane, air dry, then scrape } bottom and sides with a razor blade. What do you think? Humidity? Phase of } the moon? Bad hands? Please advise. WJP
From MicroscopyL-request-at-ns.microscopy.com Sun Jun 20 14:58:19 2004
I am working on a project to examine the optical crystallography of fossil bones and teeth in thin section. There are abundant publications with SEM images of fossil teeth (particularly enamel) that have been etched to show the orientation of the apatite crystallites. Similar studies of bone materials appear to be sparse (!). Since I'm a bit out of my area here, could anyone on the list direct me to a site, or publications, that might supply me with some SEM images of fossil (or recent) bone that show the microstructure in detail? Thanks for any help in this area.
Henry Barwood Associate Professor of Science, Earth Science Department of Math and Physics MSCX 312G Troy State University Troy, Alabama 36082 hbarwood-at-troyst.edu
From MicroscopyL-request-at-ns.microscopy.com Sun Jun 20 22:18:02 2004
The problem is not one that can be blamed on, or cured with black magic, but simply to do with the glass surface interacting with the formvar film. Did you by any chance change the make of the slides you are using? Or perhaps you changed the way you clean the glass.
I use glass slides from VWR that are cleaned superficially by dipping in alcohol and then wiping clean with lens paper. The coating and floating always works if I keep things the same.
If I leave the slides in the alcohol for too long, the film reamins stuck to the glass. If I wipe with anything other than the lens tissue, the film sticks. If I use other glass slides, the film sticks. If I clean any glass surface too well (e.g. in chromic acid) the film always sticks. Interestingly, when I try to grow cells on super-cleaned glass coverslips, the cells avoid the glass surface completely.
I am always surprized when rational scientists who meet a problem with a technique are willing to blame it on some supernatural event. Look more closely at the details and the mystery will be explained (i.e. do some experiments).
I once had a problem with Lowicryl not polymerizing. Instead of blaming anyone or any thing, we figured out the problem. It turns out that plastic tubes with orange tops can be fabricated in two sorts of plastic. We had been using polypropylene tubes but accidentally switched to polystyrene without really noticing. Mix Lowicryl in polystyrene tubes and it will not polymerize! No magic there.
The answers are out there!
Regards,
Paul Webster.
-----Original Message----- } From: Perreault,William [mailto:william.j.perreault-at-lawrence.edu] Sent: Sat 6/19/2004 7:15 PM To: Microscopy-at-msa.microscopy.com Cc: william.j.perreault-at-lawrence.edu
HPH (Hello, and Please Help.) I have been trying to make formvar support grids by methods that have always produced good results in years past. Now, however, the formvar refuses to leave the glass slides as I lower them into a container of water. Has anyone experienced this? Can you give ne a clue? My procedure now and in the past is to dip clean slides in a solution (w/v) of 0.25% formvar in dichloroethane, air dry, then scrape bottom and sides with a razor blade. What do you think? Humidity? Phase of the moon? Bad hands? Please advise. WJP
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 21 02:49:35 2004
......... clean the glass slide under warm tap water with a bristle brush and curd soap (no soap with smelling substances etc) rinse only shortly with destilled or deionised water, let it dry at the air and polish it with an absolute clean linnen cloth. then go on as you always did but breath "slowly and heavily " on the slide before immersing it extremely slowly at an 45 degree angle into the water. works with all glass slides (don't use superfrost slides for this special purpose).
peter heimann
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 21 06:57:02 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (flechedorfr-at-netscape.net) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, June 21, 2004 at 05:59:47 ---------------------------------------------------------------------------
Question: Hello everyone I am searching some internet courses on transmission electron microscopy. Can anyone please provide me to on-line websites? Thanks in advance
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (akoorts-at-medic.up.ac.za) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Monday, June 21, 2004 at 01:13:51 ---------------------------------------------------------------------------
Question: IN SITU HYBRIDIZATION for electron microscopy on core bone marrow biopsies. I need all the help I can get. First, the hybridization solution - damages the sections, I have found that two hours at 37 degrees celcius is perhaps the limit for hybridization incubation time but I get no labeling. I have tried 4 hours and 12 hours - perhaps a little labeling but the sections are damaged, difficult to distinguish the cells. I have used a probe concentration of 600 ng/ml - is this high or low? The probe that I use is about 250 bp long, PCR digoxygenin labeled. The hybridization solution contains 50% formamide, is this perhaps too stringent? Pretreatment of the sections - 10 ug/ml. Any other suggestions for pretreatment? Regards Alida
I had a similar sample last year - The researcher was looking at otoliths from fish 'ears' and wanted to count the rings to age his sample - DIC in a metallurgical (reflection) microscope worked well - growth rings were quite visible, well differentiated and easy to count. Dogfish vertebrate are much bigger than the otoliths though. Your colleague might have to slice them thinner and perhaps cut into quarters to properly fit the stage of the 'scope
Richard Harris Technical Manager Imaging and Analysis Department of Biology University of Western Ontario London Ontario CANADA Ph. 519-661-2111 ext. 86780 Fax 519-661-3935
-----Original Message----- } From: Thomas, Frank [mailto:FThomas-at-nrcan.gc.ca] Sent: Thursday, June 17, 2004 2:44 PM To: Microscopy-at-MSA.Microscopy.Com
Listers -
Has anyone out there looked at shark (OK, dogfish) vertebrae in an SEM? A colleague from down the hall in the fisheries department has some vertebrae (sectioned and unsectioned) that he'd like to look at in order to count the rings (kind of like the rings in a tree stump). The rings are just too close together to clearly differentiate in a binocular scope, but I was thinking in order for them to show up in an SEM there has to be a density or textural difference between "ring" and "inter-ring" areas. Would etching of sectioned ones with something help? (These vertebrae are not true bone of course, but a sort of calcified cartilage, so I suppose an acid might do something usefull......) I'm open to all ideas......
Frank
F.C. Thomas FThomas-at-NRCan.gc.ca, 902-426-4635, facsimile 902-426-6152 GSC Atlantic Natural Resources Canada, Bedford Institute of Oceanography, P.O. Box 1006, Dartmouth, Nova Scotia B2Y 4A2 Ressources naturelles Canada, l'Institut Oceanographique du Bedford, B.P. 1006, Dartmouth, (Nouvelle-Ecosse) B2Y 4A2 Government of Canada/Gouvernement du Canada
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 21 09:24:24 2004
Hi William, Our lab has experienced this problem in the past, too. We use slides from Corning Glassworks and found when we switched from using double frosted slides to plain, non-frosted slides, the Formvar stuck relentlessly to the slides. Switching back to double frosted slides cured the problem. Shannan
Shannan Little Research Technician Electron Microscopy/ Image Analysis Agriculture & Agri-Food Canada Lethbridge, Alberta http://res2.agr.ca/lethbridge/emia/index_e.htm
-----Original Message----- } From: Perreault,William [mailto:william.j.perreault-at-lawrence.edu] Sent: Saturday, June 19, 2004 8:16 PM To: Microscopy-at-msa.microscopy.com Cc: william.j.perreault-at-lawrence.edu
HPH (Hello, and Please Help.) I have been trying to make formvar support grids by methods that have always produced good results in years past. Now, however, the formvar refuses to leave the glass slides as I lower them into a container of water. Has anyone experienced this? Can you give ne a clue? My procedure now and in the past is to dip clean slides in a solution (w/v) of 0.25% formvar in dichloroethane, air dry, then scrape bottom and sides with a razor blade. What do you think? Humidity? Phase of the moon? Bad hands? Please advise. WJP
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 21 11:17:25 2004
Project MICRO is MSA's educational outreach program. The MICRO pages on the MSA website (URL below) have just been updated (THANK YOU, Nestor!) and are definitely worth a visit. The bibliography of books, videos, CD-ROMs, and web links now has a powerful search engine, which makes it easy to find items. And the quotes about microscopy are always fun...
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/ Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 21 14:05:03 2004
These organisms, or spores, were found in large masses on the surface of cat feces, which had been in a moist environment for some time. The individual spores appear brown under a microscope, but the masses of spores are bright red. The spores are about 5 microns in diameter, and all the same size and distinctive shape. I considered the possibility that they might be a myxomycete, but have not been able to find any with spores resembling these. If anybody knows what species this is, or even is sure of what class of organisms it belongs to, please let me know. The spores can be seen at: http://www.msu.edu/~common/Unknown.jpg. Thanks.
Ralph Common
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 21 15:44:58 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mark.nathanson-at-umdnj.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, June 21, 2004 at 12:54:59 ---------------------------------------------------------------------------
Email: mark.nathanson-at-umdnj.edu Name: Mark Nathanson
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jean-ross-at-uiowa.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, June 21, 2004 at 08:58:37 ---------------------------------------------------------------------------
Email: jean-ross-at-uiowa.edu Name: Jean Ross
Organization: Central Microscopy Research Facility, Univ. of Iowa
I would like the input of anyone who is a user of the Lynx EM processor. We have been having some mixed results in TEM processing of various tissues, resulting in soft blocks.
I have observed that the dehydrants are evaporating so that by the time the tissues reach those stations they are half their original volume. We have the unit in a fume hood. How do others handle possible evaporation and water uptake issues? We generally follow our normal protocols that we use for hand processing. Any tips or suggestions would be welcome.
On Jun 19, 2004, at 7:15 PM, Perreault,William wrote:
} Now, however, the formvar refuses to leave the glass slides as I } lower them into a container of water. Has anyone experienced this? Can } you give ne a clue?
Dear William, I had similar problems, not with plain formvar, but with holey formvar, and the solution was to put a layer of Apiazon L onto the slide. Two ways that worked were to put a small dab on the slide and spread it with a finger and thumb on opposite sides of the slide or to dissolve the Apiazon in a hydrocarbon solvent and dip the slide as one would into the formvar. The former method has advantages for holey films in that it causes the holes to line up in the direction that one rubbed one's finger; the latter method should give a more uniform film--although I never measured this--so is more suitable for plain formvar films. I also dissolves 0.25 g of Alconox in 1 L water, heated the solution to 50 C and used the warm solution to float off the formvar. Good luck. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 21 19:44:41 2004
We are interested in electropolishing away some electroless nickel in order to do TEM on some thin gold palladium plating on the electroless nickel. Does anyone know of a good electrolyte for electropolishing the electroless nickel? Thanks.
John Mardinly Intel
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 22 01:21:48 2004
Ralph, They look like diatoms to me. Do you ever see anything that look like hyphae in the cultures?
Just an off the cuff guess though, I don't know.
TB
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If your dehydrants are evaporating to half their volumn; the vials are not sealing correctly to the "Vial Sealing Assembly" which is attached to the processor lid. The only time vials are open to the air and subject to evaporation is when the processor is changing stations or the vial is being used. Whether or not the processor is in the fume hood really is not in play because the lid on the Lynx should be closed during process.
Chances are the "Seal Assembly" is defective due to age and exposure to solvents & resins. Other possibilities exsist but this is the most common cause.
Best Regards,
Al Coritz Electron Microscopy Sciences 215-412-8400 www.emsdiasum.com
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 22 11:40:19 2004
If you teach microscopy (at any educational level!) you'll be interested in an article that has just appeared in the online journal Cell Biology Education: "Microscopy images as interactive tools in cell modeling and cell biology education", by a group in Brazil. You'll find it at http://www.cellbioed.org/articles/vol3no2/article.cfm?articleID=104 -- 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/ Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 22 13:30:12 2004
John, I don't know if this will work for you, but for my Ph.D. work, I electropolished Ni emitters with a solution of HCl at room temperature using 5-10 V ac. I think that the solution was about 10%. The trick for Ni was that it didn't work well with fresh solution. What I did was put a pinch of NiCl in the solution to get the right slightly green color and it worked very nicely. I assume that this will work for you and probably be inert to the Au-Pd, but I can't be certain. I will check my dissertation to see if I have the recipe right. It has been a few years since then!
-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, June 21, 2004 9:10 PM To: microscopy-at-msa.microscopy.com Cc: Sim, Kian Sin
We are interested in electropolishing away some electroless nickel in order to do TEM on some thin gold palladium plating on the electroless nickel. Does anyone know of a good electrolyte for electropolishing the electroless nickel? Thanks.
John Mardinly Intel
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 22 17:13:48 2004
Is there a simple test available for testing deionized water quality? The building in which I work has a couple of different water filtration systems. One is the millique system, while the other is a 'homemade' distillation system. I want to know if there is a diference in water quality between the two. I have used a simple water tester for a microbiology class I took a couple of years ago. It had a grayish colored grid pattern that estimates the number of microbes present in the water. You add some water to the unit and let it set for a set time period. After the requisite time period, you check the grid pattern to see if anything grew. This device may/may not be appropriate for what I am looking for. Any suggestion will be appreciated. Sara Goldston
__________________________________ Do you Yahoo!? Yahoo! Mail is new and improved - Check it out! http://promotions.yahoo.com/new_mail
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 22 18:49:57 2004
} Is there a simple test available for testing deionized } water quality? The building in which I work has a } couple of different water filtration systems. One is } the millique system, while the other is a 'homemade' } distillation system. I want to know if there is a } diference in water quality between the two. I have } used a simple water tester for a microbiology class I } took a couple of years ago. It had a grayish colored } grid pattern that estimates the number of microbes } present in the water. You add some water to the unit } and let it set for a set time period. After the } requisite time period, you check the grid pattern to } see if anything grew. This device may/may not be } appropriate for what I am looking for. Any suggestion } will be appreciated. } Dear Sara, There are three concerns for water purity. The crudest is whether anything is growing in it; since not a lot will grow in a saturated NaCl solution, you can see that the microbe test isn't very stringent. The presence of ions increases the conductivity, so having a conductance meter will tell you how free of ions your water is. There can be non-ionic solutes, however, that do not increase the conductivity, so, while conductivity is the usual measure of water purity, there can still be dissolved material. In fact, the output of ion exchange resins--used in many water-purification systems--can contain non-ionic material generated by the resin itself, so, for really pure water, the output of the millique system can be distilled. There may even then be impurities, but deionized, distilled water is usually adequate. I don't know any test better than conductivity for water purity. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 22 22:30:07 2004
I have some nanotubes which show structures in CCl4 or chlorofrom. I wanted to scan them in TEM, What kind of film can I use apart from Formvar or colloidin, Carbon coating also did not help. Any suggestions
Shashi Singh CCMB Hyderabad INDIA
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From MicroscopyL-request-at-ns.microscopy.com Wed Jun 23 09:22:18 2004
Is there a way to make SEM pictures 3D? I know that tilting is involved, what degree? Is it better to colorize and use the red/blue glasses or is just tilting and using the stereo glasses best?
Any pointers about making a 3D pix would be greatly appreciated.
Tilting at windmills covered in algae,
Paula :-)
Paula Sicurello EM Lab 106 Mills Godwin Sciences Building Old Dominion University Norfolk, VA 23529 USA 757-683-3822
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 23 10:06:34 2004
You might try Tek-Net in Lakewood, NJ. 732-905-5530.
by way of MicroscopyListserver wrote:
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-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 23 10:24:20 2004
We want to put a simple high resolution camera on our Zeiss Axiophot and are considering a Nikon Coolpix 5000 or something similar. However, before purchasing one, we would really like to see one attached to a computer in use. This is for a multi-user facility and we want to see how simple it really is to use and to what extent it can take low-light images.
Is there anyone in NYC or lower Westchester who has such a set up and a few minutes to show me?
Thanks!
____________________________________________________________________________ Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 Fax: 430-8996 URL: http://www.aecom.yu.edu/aif/
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 23 11:05:33 2004
Ah, at last, a subject where I can contribute. Yes. 3D SEM images are quite possible.
I know that tilting is involved, what degree?
Do not tilt. You simply want horizontal displacement. If you tilt you will introduce perspective errors.
} Any pointers about making a 3D pix would be greatly appreciated.
See if you can contact Dee Breger. She published a book of 3D SEM photos ("Through the Electronic Looking Glass: 3D Images from a Scanning Electron Microscope," Cygnus Graphic, 1995). They were anaglyphs. Since SEM photos are by nature monochromatic, an anaglyph is a very suitable means of reproducing the image. If you plan on making false color images then you may want to consider a more sophisticated method.
I believe Ms. Breger is now at Drexel with an email deebre-at-coe.drexel.edu.
Good luck,
Bruce Girrell
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 23 11:26:13 2004
Here's the method that I've been taught. Pick an area and take the photo normally Tilt the subject/sample 6 degrees. If you're doing very high mag make sure you can get pack to the exact original area. We use a program called PCI Quartz which lets us process it under Stereo Pair. Pick the color, match the photos. When your satisfied with the overlay and have saved it rotate the photo 70 degrees.
Linda S. McCorkle Ohio Aerospace Institute Materials Division, Polymers Branch NASA John H. Glenn Research Center at Lewis Field 21000 Brookpark Road Cleveland, OH 44135
Please spread the word to interested qualified persons. (Please don't respond to me, I have nothing to do with this position)
Histology/Electron Microscopy position
Children's Hospital and Health Center in San Diego has an immediate opening for an Electron Microscopist with some histology experience. For more details regarding salary, job responsibilties and job requirements please contact Eric A. Breisch at ebreisch-at-chsd.org or Anne Peters at apeters-at-chsd.org. Interested candidates may also apply online at the Children's Hospital web site. (http://www.chsd.org)
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 23 11:49:57 2004
The typical value of tilt difference for stereo shots is 6 degrees.
Although not critical, it has been my experience that the optimum value can vary slightly, depending on magnification, the specimen elevation variation, and the tilt starting value.
Good depth of focus helps keep the images sharp. This translates to a small final aperture and a long working distance.
I almost exclusively use two images and a viewer. I ran across a web site that offers a number of viewers. Mine is (similar to) the F71. Works great, but pricy. I have no info about the site other than the address. http://www.ascscientific.com/stereos.html
While the anaglyph method (red/blue) works, the perception (for me) was never as crisp and dramatic as the two image/viewer method. Perhaps a function of my less than perfect red/green vision...
With practice, some can perceive stereo without a viewer. I have no trouble doing that, but the effect for me is less than with a good viewer.
To try without viewer: Use ~4x5 inch images held at "reading distance" and separated by about the width of an image. Cross your eyes (hope no one hits you in head :), defocus, and a third image should form in the empty space between the two actual images. Concentrating on the center pseudo-image, bring the eyes into focus and relax the crossed eyes. Voila, Stereo!
One more thing... Be sure to position the images correctly. Transposing the images can invert the perceived "ups & downs". In most cases the tilt of the "average" surface can be observed. "Down hill" should be in the same direction as the actual tilt.
Have fun! Woody
Woody White BWXT Services: http://www.bwxt.com/bwxt.html My Site: http://woody.white.home.att.net
-----Original Message----- } From: Paula Sicurello [mailto:PSicurel-at-odu.edu] Sent: Wednesday, June 23, 2004 10:48 AM To: microscopy-at-msa.microscopy.com
Hi listers,
Is there a way to make SEM pictures 3D? I know that tilting is involved, what degree? Is it better to colorize and use the red/blue glasses or is just tilting and using the stereo glasses best?
Any pointers about making a 3D pix would be greatly appreciated.
Tilting at windmills covered in algae,
Paula :-)
Paula Sicurello EM Lab 106 Mills Godwin Sciences Building Old Dominion University Norfolk, VA 23529 USA 757-683-3822
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 23 12:15:10 2004
John, according to ASTM E407, one of the less hazardous etchants (polishes at high currents) you can use for Ni and high Ni alloys (EN would fall into this category) is made up with 70%v of H3PO4 and the balance (30%v) water. Use electrolytic at 5-10 V for etching. So I would presume, that for polishing use at } = 10 V. I haven't tried this one and precautions are: USE IN HOOK. DO NOT STORE. Mix (make-up) fresh.
Happy Polishing, Ed Komarnicki Sr. Analytical Chemist MacDermid Inc.
To {microscopy-at-msa.microscopy.com} cc "Sim, Kian Sin" {kian.sin.sim-at-intel.com} Subject Electropolishing ELectroless nickel
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We are interested in electropolishing away some electroless nickel in order to do TEM on some thin gold palladium plating on the electroless nickel. Does anyone know of a good electrolyte for electropolishing the electroless nickel? Thanks.
John Mardinly Intel
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 23 12:22:30 2004
the technique you are referring to is called "Stereo imaging" or similar (see for example http://www.soft-imaging.com/rd/english/405.htm). Basically you acquire 2 images with a specified tilt difference. What is best depends on what you want to do with the images.
Display on a computer monitor: If you don't have a special monitor that allows the use of polarized glasses, you have two options:
1) create an anaglyphic image (red/blue or green/blue), and use the appropriate glasses. This will give you a good 3D display on the monitor, although the image appears yellowish.
2) Switch between the two stereo images repeatedly. If done correctly, it will also provide a sense of depth without using any glasses, but the image is not fixed (seems to be oscillating).
The stereo angle for these types of images should be in the range of 6-10 degrees. If you go much beyond that, the brain cannot fuse the two images anymore. This also depends on the 3-dimensionality of the sample.
For printing you can go the anaglyphic route (see 1 above), or you can mount two prints at appropriate distances and use special 3D glasses for looking at the prints. These glasses make sure that each eye sees only the print that it is supposed to see, and you get again a 3D impression, which can also be in color. The anaglyphic images have the advantage that they are independent of distance from the print. The stereo glasses require a specified geometric arrangement, which includes the distance from the prints.
Finally, you can also use the stereo images to calculate a 3D surface profile of your sample (see again the link above). Once you have done that, you can use 3D display software to rotate and look at the 3D object and actually measure in 3 dimensions.
In all cases you need to "adjust" the images, i.e., pick one point that is at the identical position on both images. If you don't do that, there may be a lateral shift between the images that can destroy the 3D appearance.
Hope that explains a few things.
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Paula Sicurello [mailto:PSicurel-at-odu.edu] Sent: Wednesday, June 23, 2004 08:48 To: microscopy-at-msa.microscopy.com
Hi listers,
Is there a way to make SEM pictures 3D? I know that tilting is involved, what degree? Is it better to colorize and use the red/blue glasses or is just tilting and using the stereo glasses best?
Any pointers about making a 3D pix would be greatly appreciated.
Tilting at windmills covered in algae,
Paula :-)
Paula Sicurello EM Lab 106 Mills Godwin Sciences Building Old Dominion University Norfolk, VA 23529 USA 757-683-3822
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 23 13:02:09 2004
Sorry Bruce, but tilting is crucial to making an anaglyph (or enabling the viewer to see a 3D image when using red/green or red/blue glasses) of SEM images. In short what you are doing is mimicking what your two eyes do when you look at an object. Your eyes are tilted at slight angles relative to each other. The Image seen thorough both eyes are combined in your brain to produce an image with 3D information. If you only have one eye than you cannot see in 3D unless shadows help fool your brain in interpreting the image.
You can do the same thing with an SEM image. You need to take two images with one tilted relative to the other. The amount of tilt depends on the degree of roughness of the sample which is in turn somewhat related to magnification, but usually 7 degrees is a good starting point. What is important is to keep the center of the image stable for both images. This is easily done by putting a piece of transparent film (such as from a clear sheet protector) on the viewing screen and using a grease pencil to mark relevant features near the center of the image. Then move the stage to line up the second image.
Any adjustment in focus should be done by adjusting working distance. If you use your focus knob you will be changing the strength of your final (objective) lens which in turn affects the rotational path of the electrons in the beam.
Modern microscopes have software to easily overlay the images to create a red/blue or red/green image which, when viewed thorough the appropriate glasses, will give you your 3D image. You can also take the two images and combine them in PhotoShop, as well as other programs, to make the final anaglyph.
If you need more complete instructions then E-mail me. I can send the article that appeared in Microscopy Today, Vol #99-1 (Jan. 1999). The book by Bozzola & Russell also contains a discussion of making stereo images.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
On 6/23/04 11:33 AM, "Bruce Girrell" {bigirrell-at-microlinetc.com} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- --} - } } Paula Sicurello asked } } } Is there a way to make SEM pictures 3D? } } Ah, at last, a subject where I can contribute. Yes. 3D SEM images are quite } possible. } } I know that tilting is involved, what degree? } } Do not tilt. You simply want horizontal displacement. If you tilt you will } introduce perspective errors. } } } Any pointers about making a 3D pix would be greatly appreciated. } } See if you can contact Dee Breger. She published a book of 3D SEM photos } ("Through the Electronic Looking Glass: 3D Images from a Scanning Electron } Microscope," Cygnus Graphic, 1995). They were anaglyphs. Since SEM photos } are by nature monochromatic, an anaglyph is a very suitable means of } reproducing the image. If you plan on making false color images then you may } want to consider a more sophisticated method. } } I believe Ms. Breger is now at Drexel with an email deebre-at-coe.drexel.edu. } } Good luck, } } Bruce Girrell } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 23 13:24:24 2004
Tilt angle of between 5 and 7 degrees gives best results. Large tilt angles may give excessive impression of depth.
Vital ingredients of success are to register the two images carefully, and to use height (z) adjustment only to refocus.
Also turn off any raster rotation.
Take first picture.
Mark a prominent central feature at approx mean height (z) on screen using a water soluble OHP pen, and return the feature to that position after tilting.
After tilting focus using specimen height adjustment only. (Do not focus using focus controls, which change image magnification).
Take second picture.
In most SEMs tilt in the horizontal image axis, so the images must be rotated 90 degrees for viewing. If you do this Clockwise then the left eye image (coloured red in anaglyph images) is the one with more tilt, and the right eye image (coloured green or cyan) image is the one with less tilt.
To make a cyan/red anaglyph Open both images in Photoshop (or similar) Change the right eye (cyan) image's Mode to RGB Select All of the Left eye (red) image, Copy and paste it into the Red colour channel of the Right eye image. Save as ...
View with cyan red glasses.
Chris
Quoting Paula Sicurello {PSicurel-at-odu.edu} :
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } } } } Hi listers, } } Is there a way to make SEM pictures 3D? I know that tilting is involved, } what degree? Is it better to colorize and use the red/blue glasses or is } just tilting and using the stereo glasses best? } } Any pointers about making a 3D pix would be greatly appreciated. } } Tilting at windmills covered in algae, } } Paula :-) } } Paula Sicurello } EM Lab } 106 Mills Godwin Sciences Building } Old Dominion University } Norfolk, VA 23529 } USA } 757-683-3822 } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 23 15:05:25 2004
This may be a long shot, but here goes. We are in dire need of an external computer control kit for a JEOL JSM-5800 SEM,(MP-87010 (EXT)). If anyone has one that they can part with, or have a retired 5800, please contact me by phone or email. Thanks in advance. Folks on this list server are great - Thanks Nestor.
Joseph
Joseph M. Oparowski Center for Materials Science - Consulting and Failure Analysis Bose Corporation Joseph_Oparowski-at-bose.com The Mountain, M/S 415 Phone: (508) 766-1371 Framingham, MA 01701-9168 Fax: (508) 766-1313
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 23 15:11:25 2004
} } Paula Sicurello asked } } } Is there a way to make SEM pictures 3D? } } Ah, at last, a subject where I can contribute. Yes. 3D SEM } images are quite possible. } } I know that tilting is involved, what degree? } } Do not tilt. You simply want horizontal displacement. If you } tilt you will introduce perspective errors.
Unfortunately horizontal displacemen works only at really low mags. Tilting involves no errors in calculation and for human eye at tilt angle 6 degrees there will be nothing unusual in stereo images. For pretty flat surfaces I used tilt angles as high as 16 degrees without any problems with observation (with tilt plus-minus 8 degrees).
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 23 16:24:54 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (ssamuelsson-at-eyetk.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, June 23, 2004 at 16:08:14 ---------------------------------------------------------------------------
Email: ssamuelsson-at-eyetk.com Name: Steven Samuelsson
Organization: Eyetech Pharmaceuticals, Inc
Title-Subject: [Microscopy] [Filtered] MListserver: Imaging Specialist and Postdoc positions available
Question: Eyetech Pharmaceuticals Inc., located in Woburn, MA., is recruiting for a biological light microscopy/IA associate. This talented individual will provide hands-on support to facility investigators. Required expertise includes quantitative image analysis, computer science, light/laser microscopy. We are also recruiting for a postdoctoral fellow to investigate issues and mechanisms of retinal vascular leakage and therapies using morphology-based approaches. Minimum requirements include competence with light and electron microscopy instruments and advanced research techniques in cell and membrane biology. It is not necessary to have a background in eye research. Eyetech is an equal opportunity employer. US citizenship/permanent residency visa for USA required, as is fluency in English.
Please respond with a CV and career objective statement to:
Steven J. Samuelsson, Ph.D. Head: Morphology; Cell Biologist Eyetech Pharmaceuticals, Inc. Eyetech Research Center 42 Cummings Park Woburn, MA. 01801 Tel: 781 938-3937 x314 Fax: 781 935-9083 ssamuelsson-at-eyetk.com www.eyetk.com
Dear friends, in order to check some possible FRET between FITC and ALEXA 555, when using bleaching of acceptor as method, we found that 543 nm bleached FITC as well. Did you ever experience such a behaviour? Alby
------------------------------------------------------------------------ --------------------------- Alberto Diaspro, Department of Physics, University of Genoa Via Dodecaneso 33, 16146 Genova, Italy facsimile +39-010314218 - voice +39-0103536426/480/309 URL: http://www.lambs.it http://www.wiley.com/WileyCDA/WileyTitle/productCd-0471409200.html ------------------------------------------------------------------------ --------------------------
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 24 13:46:07 2004
Here is the July 2004 Microscopy Today table of contents. I will close the subscription list for this issue on Tuesday 29 June 2004. Microscopists in North America and MSA members anywhere can subscribe for free. Anyone else may subscribe for US$35 per year (to PARTIALLY cover postage). All subscriptions at http://www.microscopy-today.com Thank you.
Carmichael Right at Your Fingertips
Alden& Galvis On The Trail of the Fathers: The Serendipitous Santos
Sedgewick Advantages of Adobe Photoshop Elements 2.0 over the full version of Photoshop
Boyes & Gai ETEM Issues and Opportunities for Dynamic In-situ Experiments
Diebold Microscopical Evaluation of Glass Delamination in Pharmaceutical Vials: A Look at Three Different Vial Manufacturers
Clarke Chromatic Aberration in Digital Photomicrographs from Microscopes Requiring Compensating Eyepieces
Brooker, et al. The Structural and Chemical Analyser - A New Analytical Technique for SEM
Hi All: I have been asked to check into the following by one of our personnel:
For the JEOL 200CX, how much total power does TEM supposed to draw at 200KV without the ASID? Our system draws 15A (210V) at 200KV with HT on. With beam current at 150uA, it draws roughly 17A. Among 15A, 8A is for all the electronics and vacuum pump, and 7A is for lens. Is this normal?
For the Tracor TN5520, we are looking for a computer interface (PC board, software, OR the original Tracor hardware) that will enable us to generate spectrum. The closer to free the better. Any donations would be gladly accepted.
Thanks, Michael Coviello UT Arlington
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 24 14:10:36 2004
Stereo image of the sample? http://biology.berkeley.edu/EML/stereo.html
Tilting can be from anywhere from 2 - 8 degrees or so. It depends on the sample. If the sample has a lot of protrusions which stick out, stay with a small tilt angle. Gordon
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
On Wed, 23 Jun 2004, Paula Sicurello wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } } } } Hi listers, } } Is there a way to make SEM pictures 3D? I know that tilting is involved, } what degree? Is it better to colorize and use the red/blue glasses or is } just tilting and using the stereo glasses best? } } Any pointers about making a 3D pix would be greatly appreciated. } } Tilting at windmills covered in algae, } } Paula :-) } } Paula Sicurello } EM Lab } 106 Mills Godwin Sciences Building } Old Dominion University } Norfolk, VA 23529 } USA } 757-683-3822 } }
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 24 14:21:49 2004
I have a 80 x 1 column vector of numbers that represents a PSF for a micro CT scanner. These numbers approximately follow a Gaussian distribution. I wish to expand this distribution into an multiple dimensional matrix, 80 x 80 for example, for processing through deconvolution. Does any one have any idea how I would go about doing this???
Thank you in advance,
Thomas Sadowski Southern Connecticut State University
_________________________________________________________________ FREE pop-up blocking with the new MSN Toolbar get it now! http://toolbar.msn.click-url.com/go/onm00200415ave/direct/01/
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 24 15:39:16 2004
Hi all, One of the researchers here has contacted me about doing an EM autoradiography experiment using 14C isotope. I believe he wants to look at the uptake and fate of carbon from labeled nanotubes that are presented to cell mono layers. I have read that 90% of the 14C radiation stays within 20um of its entry point in the emulsion. Does anyone know what the resolution can be for this technique and in particular the 14C isotope?
Greg
-- ================================================================== Greg Strout Electron Microscopist, University of Oklahoma WWW Virtual Library for Microscopy: http://www.ou.edu/research/electron/www-vl/ e-mail: gstrout-at-ou.edu (405)325-4391 Opinions expressed herein are mine and not necessarily those of the University of Oklahoma ==================================================================
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 24 16:16:42 2004
Hi all, One of the researchers here has contacted me about doing an EM autoradiography experiment using 14C isotope. I believe he wants to look at the uptake and fate of carbon from labeled nanotubes that are presented to cell mono layers. I have read that 90% of the 14C radiation stays within 20um of its entry point in the emulsion. Does anyone know what the resolution can be for this technique and in particular the 14C isotope?
Greg
-- ================================================================== Greg Strout Electron Microscopist, University of Oklahoma WWW Virtual Library for Microscopy: http://www.ou.edu/research/electron/www-vl/ e-mail: gstrout-at-ou.edu (405)325-4391 Opinions expressed herein are mine and not necessarily those of the University of Oklahoma ==================================================================
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 24 17:13:02 2004
Workshop Announcement University of California at Santa Barbara
The Department of Molecular, Cellular, and Developmental Biology and the Neuroscience Research Institute are sponsoring an advance course on light microscopy. This 5-day workshop will be offered from September 13 through September 17, 2004 and will consist of lectures and laboratory exercises that will run from 9 am to approximately 5 pm each day. The seminar/workshop will be intensive enabling participants to develop theoretical and hands-on expertise with light microscopes. Attendees will interact closely with the instructors while using modern research grade microscopes, cameras, and computers. The seminars and laboratories will cover basic optical theory and how it pertains to increasing contrast (signal to noise ratio) in biological samples. Fundamental techniques such as fluorescence, phase contrast, Nomarski differential interference contrast, and darkfield imaging will be taught and attendees will use microscopes equipped with these optical enhancement accessories. In addition, the theory and practice of electronic image acquisition (analog and digital) will be discussed and attendees will work with low-light cameras, digital image processing computers, and morphometric programs. There are five research grade microscopes, five electronic imaging cameras, two computer workstations, and one confocal microscope. With a maximum enrollment of 10 students, there will be ample opportunity to work with all of the microscopes and cameras. For those so interested, intensive hands-on instruction and guidance on the confocal microscope will be provided.
For a fuller description of the workshop, please check the web address below. Enrollment forms can be completed online and this workshop provides an opportunity to have a working-vacation in Santa Barbara, California.
(6/23/04 11:50) Michael Cammer {cammer-at-aecom.yu.edu} wrote:
} ------------------------------------------------------------------------------- } } We want to put a simple high resolution camera on our Zeiss Axiophot and } are considering a Nikon Coolpix 5000 or something similar. However, before } purchasing one, we would really like to see one attached to a computer in } use. This is for a multi-user facility and we want to see how simple it } really is to use and to what extent it can take low-light images. } } Is there anyone in NYC or lower Westchester who has such a set up and a few } minutes to show me?
Michael,
I would be very careful about using a cheap digital camera. Most especially, I would recommend against the specific model you have mentioned (Nikon Coolpix 5000) for one simple reason: there is no raw output. The only scientifically useful data format that the Coolpix 5000 outputs is TIFF, and even that is not very useful since the TIFFs are only 8 bit. Of course, you'll remember from the Computer-Assisted Image Analysis course at NCSU that JPEG is worse than useless. (You probably won't remember me - I was the young guy in the back running the computers.) The specifications of many cameras can be easily found from sites such as DPReview {http://www.dpreview.com} . If you really want to use a 'prosumer' camera, I'd start looking at specs there first.
You've probably paid a good deal of money from your scope. You shouldn't cripple your instrument by putting a POS camera on it. This is especially the case since you've already thrown up a warning flag by talking about low light images. Most consumer or 'prosumer' digital cameras deal with low light conditions extremly poorly - you'll get noisy images.
Brent
-- Brent Neal, Ph.D. Reindeer Graphics, Inc. brent-at-reindeergraphics.com
From MicroscopyL-request-at-ns.microscopy.com Thu Jun 24 21:17:46 2004
Not that I know anything much about this stuff, but the Canon Powershot G5 is listed on that link you gave as having RAW as the uncompressed file format.
Does this make it OK?
cheers
rtch
Date sent: Thu, 24 Jun 2004 21:05:42 -0400 } From: Brent Neal {brent-at-reindeergraphics.com}
: : (6/23/04 11:50) Michael Cammer {cammer-at-aecom.yu.edu} wrote: : : : } ------------------------------------------------------------------- ------------ : } : } We want to put a simple high resolution camera on our Zeiss Axiophot and : } are considering a Nikon Coolpix 5000 or something similar. However, before : } purchasing one, we would really like to see one attached to a computer in : } use. This is for a multi-user facility and we want to see how simple it : } really is to use and to what extent it can take low-light images. : } : } Is there anyone in NYC or lower Westchester who has such a set up and a few : } minutes to show me? : : Michael,
There is no way to connect the CoolPix camera directly to the computer you have to connect the camera to the USB port and power cycle the camera or remove the flash card and read it. Nor is there an easy way to control the camera with a computer. There is some soft ware out there that does control the camera but it is not very good. Also test the camera for artifacts. I have heard that some of the newer Coolpix camera don't have the problems that 995 and early 4500 and 5000 cameras had I document here http://www.couger.com/microscope/shootout/shootout.html.
If you can't get a camera free of ring artifacts a used Coolpix 950 for $100 to $175 is a lot less frustrating to use and won't have the problems that antialiasing software causes in the later models that infuriates some users and doesn't bother others much at all. Most users find the 950 is good enough for kind of pictures you will get from any of the consumer cameras all things considered.
No consumer camera gives you the kind of imaging you need for professional work. They all process the image before recording it and work poorly in low light conditions. That can be fixed with software ticks but cameras designed for microscope use will give a lot better account for them selves and should fit in the work flow much better. They don't cost that much more and the cost will be quickly repaid by the ware and tear on your nerves of who ever is using it every day.
Good luck Gordon Gordon Couger gcc-at-couger.com
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward any links or information you think might be useful to others. Microscope Manual at www.science-info.org
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 25 04:17:58 2004
} I would be very careful about using a cheap digital camera. Most } especially, I would recommend against the specific model you have } mentioned (Nikon Coolpix 5000) for one simple reason: there is no } raw output. ...
This is not true ... raw output from the CP5000 is possible with a firmware update that's been available for a couple of years. I.E., our CP5000 has raw output. I will agree with your concerns regarding low light acquisition, but the original post asked that he see it for himself, as he should. I'd imagine he'd have little problem in the NYC area ... with help from his peers ... sorry I cannot help.
cheerios ... shAf :o) Avalon Peninsula, Newfoundland www.micro-investigations.com (in progress)
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 25 05:58:49 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (longli_tem-at-hotmail.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, June 24, 2004 at 08:57:38 ---------------------------------------------------------------------------
Email: longli_tem-at-hotmail.com Name: Long Li
Organization: University of Pittsburgh
Title-Subject: [Microscopy] [Filtered] MListserver: A post-doctoral position is available
Question: Dear All:
A post-doctoral position is available starting immediately on the characterization of nanoparticles by advanced scanning transmission electron microscopy methods, such as quantitative Z-contrast imaging, electron energy loss spectroscopy, as well as high resolution electron microscopy. Experience with VG-STEM and Z-contrast imaging is specially appreciated. This is part of a collaborative program with University of Illinois at Urbana-Champaign and Brookhaven National Labs, to predict and determine the 3-dimensional supported structure of nanoparticles utilized in heterogeneous catalysis either as model systems or for water purification for fundamental understanding of nano-catalytic reactions.
To apply for this position, please send a resume with the names and contact information for three references, or to obtain more information, please contact:
Professor Judith Yang 848 Benedum Hall Materials Science & Engineering Dept. University of Pittsburgh Pittsburgh, PA 15261
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Title-Subject: [Microscopy] [Filtered] MListserver: EDS Detector available for donation
Question: We have an older Oxford EDS detector from a decomissioned ESEM 2020 WITHOUT the computer/electronics that's available for donation (qualified group). It's model is 6807 10mm2 ATW1 with a retrograde angle of incline - specific to ESEM 2020. In working order when we pulled it off the tool ~ 2 years ago. Let me know offline if you are interested in it. Would need to pay shipping. Thanks.
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Email: juan-at-nanostellar.com Name: Juan
Title-Subject: [Microscopy] [Filtered] software for nano-particle shape analysis
Question: Dear all,
We need to do shape-resolved nano-particle size analysis. For examples, TEM images show triangles, squares, pentagons..., and we would like to know if there is any software which can automatically recognize the shape and give the size of them.
Hi for all ! I have big problem with this operation BINXFER in the qbasic for quantimet 570 color,
I have binary image, in the window "field measurement" we have a good data for count, when we have program in qbasic "test-c. qba" we need use image analysis comand BINXFER with function "paralysis" to obtain a good data for count I have problem with this function; you see now this program; 10 rem feature count test-c.qba 20 rem calibrate - pixel=1 30 cls 40 clearplane -1 50 mframe 31 61 450 450 60 iframe 0 0 512 512 70 display 0 0 3 6 80 loadbin 1 '1.bin' 90 binxfer 1 0 2 5 0 0 0 rem problem with paralysed not working !! 100 measfield 2 110 rfieldres fieldarray(1) 120 area=fieldarray(1) 130 perimeter=fieldarray(2) 140 count=fieldarray(5) 150 rem postext 5 10 160 print "area - " area 170 print "perimeter - " perimeter 180 print "count - " count 190 stop 200 end
when in the line 100 i give 100 measfield 1 program computing count for all end !!!
How use this function BINXFER correct to obtain good data mesurements for count ?
best regards
Chris Hübner
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 25 09:58:18 2004
Here's a question that goes back into the dim past. I need to make a small front surface mirror. I have the blank glass, and I thought that I would use my old em training and evaporate some chromium chips onto the surface, in a Denton Vacuum Evaporator, model DV-515. However, I can't find tungsten baskets anywhere in the lab. We do have lots of carbon rods, and I am wondering if any of you have ever used such rods to evaporate the chromium. I've done it with Pt, of course.
Thanks,
Joel
Joel B. Sheffield, Ph.D. Biology Department, Temple University 1900 North 12th Street Philadelphia, PA 19122 jbs-at-temple.edu (215) 204 8839, fax (215) 204 0486 http://astro.temple.edu/~jbs
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 25 11:32:38 2004
Has anyone done, or has anyone seen done, microwave enhanced immunohistochemistry on cell cultures using alkaline phosphatase secondary antibody. So far I have been able to find only preliminary treatment with microwave for antigen retrieval. Any info will be greatly appreciated.
Greg Erdos
Gregory W. Erdos Ph.D. Assistant Director, Biotechnology Program Scientific Director, Electron Microscopy P.O. Box 118525 217 Carr Hall University of Florida Gainesville, FL 32611 gwe-at-ufl.edu 352-392-1295 fax- 352-846-0251
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 25 12:53:32 2004
I was asked to use ethylene glycol for TEM specimen preparation: as a dehydration fluid, a fixative, in a boat of diamond knife, and a solvent for stain. Any comments and advise are greatly apprciated.
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
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Email: lbustillos-at-amalab.com Name: Luis Bustillos
Question: We are having a puzzling problem with our JEOL 100CX II. It shuts itself off sometimes after 1-2 hours but sometimes it will run a day or two before it will eventually shut itself off. We have checked all that we could think of that could be causing the problem. For example, the water circulation, air pressure, heating plates, fore pump sensors, exchanged the whole vacuum board, checked the input voltage but to no avail. I would really appreciate it if anybody has any suggestions.
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Email: satyam-at-iopb.res.in Name: P. V. Satyam
Organization: Institute of Physics, Bhubaneswar, India
Title-Subject: [Microscopy] [Filtered] MListserver: Thermostat for JEOL 2010 TEM
Question: Hello friends, I need a help in finding out the following thermostat to get the for our recently purchased TEM machine (JEOL 2010, UHR).
THERMOSTAT PART NO. - 440001765 50'C OFF AND 35'C ON USED IN LENS PS CKT. BOARD.
At present I am in Japan. I will be going back to my Institute in India, where our machine requires this thermostat (on July 3rd). But, I can't purchase directly from JEOL due to their policies. We need the above item as soon as possible. I need to go back and import from them eventhough it is a small item!
I plan to visit Akihabara, Tokyo in another couple of days for this purpose. Please suggest.
I am willing to pay for these,If some one wants to ship them. We need very urgently.
What are the other sources where I can get the above thermostat?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (stephanie.l.mccracken-at-medtronic.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, June 25, 2004 at 14:23:57 ---------------------------------------------------------------------------
Question: Is anyone aware of contract labs that perform BeCu Cross-sections? I am interested in contracting this to avoid having to deal with the Health and Safety Issues.
On Jun 25, 2004, at 11:19 AM, Dusevich, Vladimir wrote:
} I was asked to use ethylene glycol for TEM specimen preparation: } as a dehydration fluid, a fixative, in a boat of diamond knife, and } a solvent for stain. Any comments and advise are greatly } apprciated. } Dear Vladimir, I wouldn't use ethylene glycol either for a dehydrating agent--it is completely miscible with water, so it wouldn't extract it--or a fixative--it preserves many proteins as do other polyalcohols, e.g. sucrose. On the other hand, it might work well as a fluid in the boat of a diamond knife--I've never used it that way, but sections should float and not be damaged--and it could be a good solvent for some stains, although it is quite hydrophyllic. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Fri Jun 25 17:02:37 2004
Luis, I would suggest you check the freon presure for EHT.
Long Li ____________________________________________________________________________ _________ Materials Science and Engineering Department University of Pittsburgh 3700 O'Hara St., 848 Benedum Hall Pittsburgh, PA 15261
----- Original Message ----- } From: "by way of MicroscopyListserver" {lbustillos-at-amalab.com} To: {microscopy-at-microscopy.com} Sent: Friday, June 25, 2004 2:31 PM
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (innap-at-savion.huji.ac.il) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Sunday, June 27, 2004 at 03:02:29 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] [Filtered] MListserver: software for simulation of CBED
Question: Dear All,
I am trying to use Convergent Beam Electron Diffraction for precise measurement of lattice parameter. Do you know any (either commercial or free) software which construct CBED patterns (high order Laue zones patterns)? I would also deeply appreciate any recommendation regarding Software for constucting crystalline shapes (3D polygons) based on know Miller indexes. Best, Inna
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (saram-at-duke.edu) from http://microscopy.org/MicroscopyListserver/MLFormMail.html on Sunday, June 27, 2004 at 09:37:05 ---------------------------------------------------------------------------
Email: saram-at-duke.edu Name: Sara E. Miller
Organization: Duke
Title-Subject: [Microscopy] [Filtered] MListserver: Electron Microscopy Technologist Job
Question: Electron Microscopy Technologist Job
I have an electron microscopy technologist position open. We do all TEM--a mixture of negative staining and thin sectioning; 75% is clinical (diagnosing viral diseases by EM plus surgical pathology EM) and 25% is research. We have 5 EM tech positions that rotate and perform various duties.
Requirements for the position include a bachelorĮs degree and electron microscopy training (a course or experience) or an associateĮs degree in EM. I am looking for someone who is proficient in cutting thin sections and has experience running an electron microscope. I would like to have someone who has done some negative staining and examination of viruses.
I am looking for a person who enjoys challenging and interesting cases, is dedicated to accuracy, and is willing occasionally to put in extra effort in return for appropriate compensation. I am particularly looking for someone who can manage several jobs at once while having a good time sharing camaraderie with the rest of us, i.e., is not high strung. I know this special person exists, since there are 4 lovely folks remaining (3 guys and 1 gal) with these same qualifications. I will be happy to answer any questions you have by phone or email.
Durham, NC, is a pleasant place in which to live and work--3 hrs from the mountains or the beach, and has more entertainment than you could want with 3 major universities and their arts programs in close proximity. Durham is one of the apexes of a triangle formed by it, Raleigh, and Chapel Hill, each only 20-30 min apart. In the center of the triangle are numerous businesses that comprise the Research Triangle Park, a huge research center with close ties to the various universities in the 3 cities. Duke Medical Center is composed of a medical school and a tertiary care hospital. Many basic and applied research projects are underway. The medical center is adjacent to the undergraduate campus where it has been rumored that basketball competition is fierce.
If you are interested, please contact me directly, off line. Sincerely,
Sara E. Miller, Ph. D. Director, Electron Microscopy Laboratory Department of Pathology, Box 3712 Duke University Medical Center Durham NC 27710 Phone: (919) 684-3452 Fax: (919) 684-3265 saram-at-duke.edu
On my JEOL 840A I have used RS #228-2636 Industrial Thermostat, much cheaper than the JEOL type although a different shape (I hold it in place with a spring that wraps around the diff pump). This one opens at 50 deg, closes at 35 deg.
But I prefer to use the type which needs to be manually reset, to avoid temperature recycling ---- if there's a problem which overheats the DP I want to know about it.
RS # 228-2513 is their lowest, it opens at 70 deg.
However, you have to watch out that you don't invalidate any guarantees by using non-JEOL parts.
cheers
rtch
Question: Hello friends, I need a help in finding out the following thermostat to get the for our recently purchased TEM machine (JEOL 2010, UHR).
THERMOSTAT PART NO. - 440001765 50'C OFF AND 35'C ON USED IN LENS PS CKT. BOARD.
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
From MicroscopyL-request-at-ns.microscopy.com Sun Jun 27 17:26:48 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (subramss-at-email.uc.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Sunday, June 27, 2004 at 14:45:04 ---------------------------------------------------------------------------
Question: I am trying to find out more about using Methane as an imaging gas in the ESEM and would appreciate the input of any of you who may have used alternate gases for imaging in the ESEM.
Regards,
Jimble
Srinivas Subramaniam Advanced Materials Characterization Center University of Cincinnati
Hello Inna, At the recent school on CBED http://www.zmn.tu-ilmenau.de/cat74.html this topic was covered in detailes. The program mbfit from Dr. K.Tsuda allows advanced dynamical (Bloch waves) calculation of CBED patterns, and it was free at least for school participants. There was a program as well (one of many, I believe) called Winkiku for kynematical calculations. We also demonstrated there a multislice based code for CBED calculation including HOLZ lines, however it is not yet ready for external users. But if you are facing the problems of lines splitting and symmetry breaks on you patterns, multislice is the only way to go and we are opened for cooperation. There was also a program based on modified Hough transformation for precise HOLZ line position measurement, which you may find usefull and which is freeware. You may contact Dr Ute Kaiser at ute.kaiser-at-tu-ilmenau.de to get one.
Hope this helps Andrey
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (innap-at-savion.huji.ac.il) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Sunday, June 27, 2004 at 03:02:29 } --------------------------------------------------------------------------- } } Email: innap-at-savion.huji.ac.il } Name: Inna Popov } } Organization: The Hebrew University of Jerusalem } } Title-Subject: [Microscopy] [Filtered] MListserver: software for simulation of CBED } } Question: Dear All, } } I am trying to use Convergent Beam Electron Diffraction for precise measurement of lattice parameter. Do you know any (either commercial or free) software which construct CBED patterns (high order Laue zones patterns)? } I would also deeply appreciate any recommendation regarding Software for constucting crystalline shapes (3D polygons) based on know Miller indexes. } Best, } Inna }
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 28 02:32:23 2004
I am not sure that vacuum evaporation of chromium from carbon is a good idea. I have never heard of that being done, even for such "macro" applications as mirror coating. Have you considered using aluminium metal instead for your front-surfaced mirror? It might be easier in your case. For that you would not need a tungsten basket, but simply a tungsten wire with a v-notch kink in it. Perhaps you can find (or make) some of these? The aluminium can be simply a small area of household foil which you can hang over the v-kink and melt to a blob under vacuum beforehand, although even this may be unnecessary. To prevent larger contaminant particles from falling on the mirror surface you might be wise to support it somehow so that it is not directly under the evaporation source. Good luck with your evaporation!
Jim
-----Original Message----- } From: Joel Sheffield [mailto:jbs-at-temple.edu] Sent: Friday, June 25, 2004 5:26 PM To: Microscopy-at-MSA.Microscopy.Com
Hi,
Here's a question that goes back into the dim past. I need to make a small front surface mirror. I have the blank glass, and I thought that I would use my old em training and evaporate some chromium chips onto the surface, in a Denton Vacuum Evaporator, model DV-515. However, I can't find tungsten baskets anywhere in the lab. We do have lots of carbon rods, and I am wondering if any of you have ever used such rods to evaporate the chromium. I've done it with Pt, of course.
Thanks,
Joel
Joel B. Sheffield, Ph.D. Biology Department, Temple University 1900 North 12th Street Philadelphia, PA 19122 jbs-at-temple.edu (215) 204 8839, fax (215) 204 0486 http://astro.temple.edu/~jbs
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 28 06:21:32 2004
I'm afraid I can't help much with your question: in our ESEM we've ever only used water vapour, or HV mode. But I'm curious - is there some particular reason why you want to use methane? Would it have an advantage over water vapour, or are you especially interested in observing a particular reaction in a methane environment? Also, of course, there's the small matter of a chamber full of methane being exposed to a spark from, say, a stage drive motor. I suppose once you had a partial pressure of only a couple torr of methane in there it would be safe enough, but I'm wondering about the early evacuation stages, when there could still be enough oxygen in the chamber to get a sudden and highly unpleasant reaction going. (Maybe I'm a bit paranoid - here in Nova Scotia we lost 26 coal miners in a methane explosion a few years back). Anyway, good luck, but be careful with that stuff......
F.C. Thomas FThomas-at-NRCan.gc.ca, 902-426-4635, facsimile 902-426-6152 GSC Atlantic Natural Resources Canada, Bedford Institute of Oceanography, P.O. Box 1006, Dartmouth, Nova Scotia B2Y 4A2 Ressources naturelles Canada, l'Institut Oceanographique du Bedford, B.P. 1006, Dartmouth, (Nouvelle-Ecosse) B2Y 4A2 Government of Canada/Gouvernement du Canada
-----Original Message----- } From: subramss-at-email.uc.edu [mailto:subramss-at-email.uc.edu] Sent: Sunday, June 27, 2004 7:57 PM To: microscopy-at-ns.microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (subramss-at-email.uc.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Sunday, June 27, 2004 at 14:45:04 ---------------------------------------------------------------------------
Question: I am trying to find out more about using Methane as an imaging gas in the ESEM and would appreciate the input of any of you who may have used alternate gases for imaging in the ESEM.
Regards,
Jimble
Srinivas Subramaniam Advanced Materials Characterization Center University of Cincinnati
IF THIS ISN'T COMMERCIAL ADVERTISING THEN WHAT IS, THIS KIND OF REPLY SHOULD BE OFF LINE, WE'RE SURE HOPE IT WAS INADVERTANT, NEVER THE LESS IT IS ADVERTISING. ----- Original Message ----- } From: {ramos-at-argo-tech.com} To: {Microscopy-at-MSA.Microscopy.com} Sent: Tuesday, June 15, 2004 2:41 PM
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (tem_iopb-at-iopb.res.in) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, June 28, 2004 at 02:14:54 ---------------------------------------------------------------------------
Email: tem_iopb-at-iopb.res.in Name: P. V. Satyam
Organization: Institute of Physics
Title-Subject: [Microscopy] [Filtered] MListserver: Thermostat : JEOL Help
Question: Dear Friends,
I posted the following help message few days back: } } Question: Hello friends, } } I need a help in finding out the following thermostat to get the } } for our recently purchased TEM machine (JEOL 2010, UHR). } } } } THERMOSTAT } } PART NO. - 440001765 } } 50'C OFF AND 35'C ON } } USED IN LENS PS CKT. BOARD. } }
This message is to thank you all who gave their suggestions. Also, I would like to thank Mr. Takebe, Regional Sale Manager in JEOL, Japan who has going all the way to help me in getting the component. He has promised me to hand deliver the components at me personally (at present I am near Tokyo). Initially I was rather not that confident; But this gesture from JEOL to help when it is badly needed is highly appreciative from my side. I was worried that it will take more time to get it if I go back India and place an order and hence posted for faster ways to cricumvent and find some new methods to solve the problems in using thermostat.
Please note that this is just my experience which I am sharing. No commercial interests are involved in this.
Inna, I invite you to try the online CBED simulation routines at: http://emaps.mrl.uiuc.edu. This site provides an online interface to a collection of diffraction and imaging simulation routines written by Prof. Zuo of the University of Illinois through a web based user interface written by Dr. Jim Mabon. Both kinematic and full Bloch wave simulations of CBED patterns can be performed. For more information, please see: http://cmm.mrl.uiuc.edu/EMAPS/WebEmapsInfo.htm
Cheers,
Ray D. Twesten, PhD. Center for Microanalysis of Materials Seitz Materials Research Laboratory 104 S. Goodwin Ave., Urbana, IL 61801 +1 217 244-6177 (Fax -2278)
-----Original Message----- } From: by way of MicroscopyListserver [mailto:innap-at-savion.cc.huji.ac.il] Sent: Sunday, June 27, 2004 10:04 AM To: microscopy-at-microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (innap-at-savion.huji.ac.il) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Sunday, June 27, 2004 at 03:02:29 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] [Filtered] MListserver: software for simulation of CBED
Question: Dear All,
I am trying to use Convergent Beam Electron Diffraction for precise measurement of lattice parameter. Do you know any (either commercial or free) software which construct CBED patterns (high order Laue zones patterns)? I would also deeply appreciate any recommendation regarding Software for constucting crystalline shapes (3D polygons) based on know Miller indexes. Best, Inna
Frank, It's the dilution of methane and oxygen you have to worry about (Excluding burning metals, fluorine gas and so forth) The ratio called explosive limits determine the hazard. Too much methane or too little methane no explosion. On the plus side, methane is slightly lighter than air, so as you vent the system you would not have to worry about heavy vapors collecting in low spots creating suffocation traps and explosive concentrations.
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Jimble -
I'm afraid I can't help much with your question: in our ESEM we've ever only used water vapour, or HV mode. But I'm curious - is there some particular reason why you want to use methane? Would it have an advantage over water vapour, or are you especially interested in observing a particular reaction in a methane environment? Also, of course, there's the small matter of a chamber full of methane being exposed to a spark from, say, a stage drive motor. I suppose once you had a partial pressure of only a couple torr of methane in there it would be safe enough, but I'm wondering about the early evacuation stages, when there could still be enough oxygen in the chamber to get a sudden and highly unpleasant reaction going. (Maybe I'm a bit paranoid - here in Nova Scotia we lost 26 coal miners in a methane explosion a few years back). Anyway, good luck, but be careful with that stuff......
F.C. Thomas FThomas-at-NRCan.gc.ca, 902-426-4635, facsimile 902-426-6152 GSC Atlantic Natural Resources Canada, Bedford Institute of Oceanography, P.O. Box 1006, Dartmouth, Nova Scotia B2Y 4A2 Ressources naturelles Canada, l'Institut Oceanographique du Bedford, B.P. 1006, Dartmouth, (Nouvelle-Ecosse) B2Y 4A2 Government of Canada/Gouvernement du Canada
-----Original Message----- } From: subramss-at-email.uc.edu [mailto:subramss-at-email.uc.edu] Sent: Sunday, June 27, 2004 7:57 PM To: microscopy-at-ns.microscopy.com
---
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (subramss-at-email.uc.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Sunday, June 27, 2004 at 14:45:04 ---------------------------------------------------------------------------
Question: I am trying to find out more about using Methane as an imaging gas in the ESEM and would appreciate the input of any of you who may have used alternate gases for imaging in the ESEM.
Regards,
Jimble
Srinivas Subramaniam Advanced Materials Characterization Center University of Cincinnati
I DON'T THINK THAT THIS IS ADVERTISING. THE CLEMSON UNIVERSITY PERSON ASKED FOR HELP IN LOCATING A REPLACEMENT SEM. RAMOS OFFERED A SUGGESTED SOURCE. ADVERTISEMENT WOULD BE IF THE ACTUAL SOURCE POSTED TO THE LIST DIRECTLY. THEY DID NOT. USERS, INCLUDING MYSELF, ARE ALWAYS SEEKING SOURCES. NO ONE CAN KNOW ALL OF THEM.
IF IT IS ADVERTISING, I'M SURE NESTOR WOULD LET US AND THE POSTER KNOW. OR NESTOR WOULD NOT HAVE ALLOWED THE POSTING.
GARY G.
At 06:24 AM 6/28/2004, you wrote:
} IF THIS ISN'T COMMERCIAL ADVERTISING } THEN WHAT IS, THIS KIND OF REPLY } SHOULD BE OFF LINE, WE'RE SURE HOPE IT } WAS INADVERTANT, NEVER THE LESS IT } IS ADVERTISING. } ----- Original Message ----- } } From: {ramos-at-argo-tech.com} } To: {Microscopy-at-MSA.Microscopy.com} } Sent: Tuesday, June 15, 2004 2:41 PM } Subject: [Microscopy] Re: Looking to buy SEM } } } } } } } } } Darryl, } } } } } } Don't know anything about the company......but perhaps you might look into } } SEMTech Solutions. They offer service for AMRAY SEMs, and also offer } } Re-manufactured Pre-owned Scanning Electron Microscopes. } } } } http://www.semtechsolutions.com/ } } } } } } Kelly A. Ramos } } Metallurgical Engineer / Supervisor } } Argo-Tech Materials Laboratories } } 23555 Euclid Avenue } } Cleveland, OH 44117 } } 216-692-5904 or 216-692-5446 } } (fax) 216-692-5816 } } http://www.atclabs.com } } } } } } } } darryl krueger } } {dkruege-at-CLEMS To: } Microscopy-at-MSA.Microscopy.com } } ON.EDU} cc: } } Subject: [Microscopy] } Looking to buy SEM } } 06/14/04 03:30 } } PM } } } } Dear Listees, } } } } We're loosing our Hitachi 3500 SEM due to a split in the EM facility here } } at Clemson University. Is there anything out there available, comparable } } on a used basis. We're looking for equipment and a price. Thanks in } } advance for any information. } } } } Darryl Krueger, RA } } BioScience Dept. } } Clemson University } } } } } } } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 28 09:01:30 2004
} ... But I'm curious - is there some particular reason } why you want to use methane? Would it have an advantage over } water vapour, or are you especially interested in observing a } particular reaction in a methane environment?
It is curious to note the fact methane is used as a quenching gas in ionization-type detectors (e.g., x-ray argon gas flow detectors). The methane molecule redily supplies the ionized argon cation with the electron it needs for being immediately available for ionization again. I don't know the physical chemistry of it, ... but, in a context of specimen charging, the phenomenon does beg the question Jimble asks.
} Name: Jimble } } Organization: University of Cincinnati } } Title-Subject: [Microscopy] [Filtered] ESEM imaging gases: Methane } } Question: I am trying to find out more about using Methane as an } imaging gas in the ESEM and would appreciate the input of any of you } who may have used alternate gases for imaging in the ESEM. } } Regards, } } Jimble } } Srinivas Subramaniam } Advanced Materials Characterization Center } University of Cincinnati } } ------------------------------------------------------------------ } --------- } } }
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 28 09:27:40 2004
A post-doctoral position is available starting immediately on the characterization of nanoparticles by advanced scanning transmission electron microscopy methods, such as quantitative Z-contrast imaging, electron energy loss spectroscopy, as well as high resolution electron microscopy. Experience with VG-STEM and Z-contrast imaging is specially appreciated. This is part of a collaborative program with University of Illinois at Urbana-Champaign and Brookhaven National Labs, to predict and determine the 3-dimensional supported structure of nanoparticles utilized in heterogeneous catalysis either as model systems or for water purification for fundamental understanding of nano-catalytic reactions.
To apply for this position, please send a resume with the names and contact information for three references, or to obtain more information, please contact:
Professor Judith Yang 848 Benedum Hall Materials Science & Engineering Dept. University of Pittsburgh Pittsburgh, PA 15261
jyang-at-engr.pitt.edu (412) 624-8613
_________________________________________________________________ MSN Toolbar provides one-click access to Hotmail from any Web page FREE download! http://toolbar.msn.click-url.com/go/onm00200413ave/direct/01/
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 28 09:32:46 2004
Here I am at my new job and I get to clean out the old chemicals. I've already found 17 year old ether, an unmarked bottle of who know what and various other old chemicals.
My question today regards the propylene oxide. Does it age? Can it still be used as an intermediate solvent if it's 24 (bottle dated 9/1980) or 12 (bottle dated 9/1992) years old?
If not I will discard it, if it just ages like a fine wine we'll use it.
I've looked at the MSDS's and can't find anything about age, stability and usefulness.
So if you know the answer to my question, please let me know.
Is it a tasty wine or a nasty vinegar?
Cheers,
Paula :-)
Paula Sicurello EM Lab 106 Mills Godwin Sciences Building Old Dominion University Norfolk, VA 23529 USA 757-683-3822
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 28 09:26:06 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mark.nathanson-at-umdnj.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, June 28, 2004 at 09:07:49 ---------------------------------------------------------------------------
Email: mark.nathanson-at-umdnj.edu Name: Mark Nathanson
Question: I am trying to recondition an LKB Nova Ultramicrotome, but we have no documentation. Is there anyone in the Phila-New York area with an instruction maual or repair manuals that thyey would be willing to share?
At the partial pressure you are talking about I would not worry about possible explosion unless the methane build up in any cold traps or in the pumping system. Purging the chamber should mean that there is pretty much just methane in the chamber and therefore no oxygen for it to combine with!
-- John Mansfield PhD MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 USA Phone: (734) 936-3352 FAX (734) 763-2282 Cell. Phone: (734) 834-3913 (Leaving a phone message at 936-3352 is preferable to 834-3913) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42° 16' 48" Long. 83° 43' 48" AIM: thejfmjfm
Home address: 4304 Spring Lake Boulevard Ann Arbor MI 48108-9657 Phone (734) 994-3096
On Jun 28, 2004, at 7:50 AM, Thomas, Frank wrote:
} } } ----------------------------------------------------------------------- } ------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } -------- } } Jimble - } } I'm afraid I can't help much with your question: in our ESEM we've } ever only used water vapour, or HV mode. But I'm curious - is there } some } particular reason why you want to use methane? Would it have an } advantage } over water vapour, or are you especially interested in observing a } particular reaction in a methane environment? } Also, of course, there's the small matter of a chamber full of } methane being exposed to a spark from, say, a stage drive motor. I } suppose } once you had a partial pressure of only a couple torr of methane in } there it } would be safe enough, but I'm wondering about the early evacuation } stages, } when there could still be enough oxygen in the chamber to get a sudden } and } highly unpleasant reaction going. (Maybe I'm a bit paranoid - here in } Nova } Scotia we lost 26 coal miners in a methane explosion a few years back). } Anyway, good luck, but be careful with that stuff...... } } F.C. Thomas } FThomas-at-NRCan.gc.ca, 902-426-4635, facsimile 902-426-6152 } GSC Atlantic } Natural Resources Canada, Bedford Institute of Oceanography, } P.O. Box 1006, Dartmouth, Nova Scotia B2Y 4A2 } Ressources naturelles Canada, l'Institut Oceanographique du Bedford, } B.P. } 1006, Dartmouth, (Nouvelle-Ecosse) } B2Y 4A2 } Government of Canada/Gouvernement du Canada } } } -----Original Message----- } } From: subramss-at-email.uc.edu [mailto:subramss-at-email.uc.edu] } Sent: Sunday, June 27, 2004 7:57 PM } To: microscopy-at-ns.microscopy.com } Subject: [Microscopy] viaWWW: ESEM imaging gases: Methane } } } } } ----------------------------------------------------------------------- } ----- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } ----- } --- } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (subramss-at-email.uc.edu) from } http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on } Sunday, June 27, 2004 at 14:45:04 } ----------------------------------------------------------------------- } ---- } } Email: subramss-at-email.uc.edu } Name: Jimble } } Organization: University of Cincinnati } } Title-Subject: [Microscopy] [Filtered] ESEM imaging gases: Methane } } Question: I am trying to find out more about using Methane as an } imaging gas in the ESEM and would appreciate the input of any of you } who may have used alternate gases for imaging in the ESEM. } } Regards, } } Jimble } } Srinivas Subramaniam } Advanced Materials Characterization Center } University of Cincinnati } } ----------------------------------------------------------------------- } ---- }
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 28 09:39:27 2004
Dear All: A post-doctoral position is available starting immediately on the characterization of nanoparticles by advanced scanning transmission electron microscopy methods, such as quantitative Z-contrast imaging, electron energy loss spectroscopy, as well as high resolution electron microscopy. Experience with VG-STEM and Z-contrast imaging is specially appreciated. This is part of a collaborative program with University of Illinois at Urbana-Champaign and Brookhaven National Labs, to predict and determine the 3-dimensional supported structure of nanoparticles utilized in heterogeneous catalysis either as model systems or for water purification for fundamental understanding of nano-catalytic reactions. To apply for this position, please send a resume with the names and contact information for three references, or to obtain more information, please contact: Professor Judith Yang 848 Benedum Hall Materials Science & Engineering Dept. University of Pittsburgh Pittsburgh, PA 15261 jyang-at-engr.pitt.edu (412) 624-8613
_________________________________________________________________ Make the most of your family vacation with tips from the MSN Family Travel Guide! http://dollar.msn.com
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 28 09:56:12 2004
A post-doctoral position is available starting immediately on the characterization of nanoparticles by advanced scanning transmission electron microscopy methods, such as quantitative Z-contrast imaging, electron energy loss spectroscopy, as well as high resolution electron microscopy. Experience with VG-STEM and Z-contrast imaging is specially appreciated. This is part of a collaborative program with University of Illinois at Urbana-Champaign and Brookhaven National Labs, to predict and determine the 3-dimensional supported structure of nanoparticles utilized in heterogeneous catalysis either as model systems or for water purification for fundamental understanding of nano-catalytic reactions.
To apply for this position, please send a resume with the names and contact information for three references, or to obtain more information, please contact:
Professor Judith Yang 848 Benedum Hall Materials Science & Engineering Dept. University of Pittsburgh Pittsburgh, PA 15261
jyang-at-engr.pitt.edu (412) 624-8613
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 28 11:12:51 2004
Hello, We have just sent an Electroscan E3 ESEM to UC Berkeley's Salvage. UC Berkeley people have first crack at it, then other UC people, and then finally other organizations.
It was in a functioning state when decommissioned. You'll need a rotary pump, and water chiller to make it fully functional I think.
For information about obtaining it, you'll need to contact UC Berkeley's salvage department at:
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
On Fri, 25 Jun 2004, jiahe xu wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Free or low price. I only need parts.Any information } would be great help. } Thanks. } } Tim } } } __________________________________________________ } Do You Yahoo!? } Tired of spam? Yahoo! Mail has the best spam protection around } http://mail.yahoo.com }
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 28 14:03:39 2004
Some reagents, unlike wine, undergo minimal change with time. Low MW solvents (ethanol, acetone, propylene oxide) should be OK if they are unopened and sealed tightly. About the only concern would be evaporative loss which would lead to cooling and possible condensation of water in the solvent. In that case, pass them along to someone who could use them for cleaning purposes, etc.
We have even successfully used unopened, original Epon (yes, a truly fine, 30 yr old vintage stored in a dark and cool environment) with no problems. However, the additional components (DDSA, NMA, BDMA were fresh reagents).
JB
} Here I am at my new job and I get to clean out the old chemicals. I've } already found 17 year old ether, an unmarked bottle of who know what and } various other old chemicals. } } My question today regards the propylene oxide. Does it age? Can it still } be used as an intermediate solvent if it's 24 (bottle dated 9/1980) or 12 } (bottle dated 9/1992) years old? } } If not I will discard it, if it just ages like a fine wine we'll use it. } } I've looked at the MSDS's and can't find anything about age, stability and } usefulness. } } So if you know the answer to my question, please let me know. } } Is it a tasty wine or a nasty vinegar? } } } Cheers, } } Paula :-) } } Paula Sicurello } EM Lab } 106 Mills Godwin Sciences Building } Old Dominion University } Norfolk, VA 23529 } USA } 757-683-3822
-- ############################################################## 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 Email: bozzola-at-siu.edu Web: http://www.siu.edu/~image/ ##############################################################
From MicroscopyL-request-at-ns.microscopy.com Mon Jun 28 15:38:18 2004
Kelly said he didn't know anything about the company, didn't he?
Mentioning a company name isn't advertising.
Let's not get so PC that we can't make recommendations to each other on the list.
cheers
rtch
ps Kelly also was polite in that he said who he is and where he's from, and that he didn't shout
} From: "JERRY SMITH" {jsmit51-at-tampabay.rr.com} To: {ramos-at-argo-tech.com} , {Microscopy-at-MSA.Microscopy.com}
Hi,
For those interested in microscopy education and/or lattice fringe imaging, we've put together a collection of web-explorable models* for general access. The lion's share of those on the main menu are for prospective nanoworld explorers to "do detective work" with, e.g. to measure sizes, angles, perhaps even Fourier analyze, and then practice reporting on their results.
The visibilitymaps link, via "More here", leads to a collection of research-relevant but similarly interactive 3D maps** of visible lattice fringe geometry for various specimen thicknesses and microscope resolutions ala Ultramicroscopy 94 (2003) 245 and subsequent MSA papers***. We'll eventually set up through webMathematica to host user-requested plots.
Meanwhile, if you're contemplating analysis of fringe images from a particular crystal type, feel free to drop me a line (off the listserver, specifying effective scope resolution and crystal thickness as well) to see if we can manage to post the model you'd like on request.
I am in the final evaluation process for the purchase of new optical brightfield microscopes . They will be used for general biology, embryology, and histology.
Can anyone give me their opinion off the server about the Zeiss Axiostar Plus and the Nikon E200? Thank you for your assistance.
Ken _______________________________________ Kenneth L. Tiekotter, Adjunct Professor The University of Portland Department of Biology 5000 N Willamette Blvd. Portland, OR 97203 USA
Tel.: 503.493.8861
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 29 05:31:55 2004
Paula, I hope you're very careful with the old ether. As I understand it, the stuff ages and becomes VERY unstable. I heard about a 5 gallon can that was found in Woods Hole, MA. Required a bomb squad and resulted in a big fine for the lab head.
Ken Converse owner Quality Images third party SEM service Delta, PA
Paula Sicurello wrote:
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From MicroscopyL-request-at-ns.microscopy.com Tue Jun 29 05:49:19 2004
Lou, I haven't seen any replies to your question, so I'll give it a shot.
I only work on SEMs, so I'm not familiar with the plumbing on the 100CX, but on the JSM-840 and 6400, JEOL splits the water supply and sends one feed the the Optics console and the other feed to the Electronics console. When they exit their respective cooling loops they are recombined so there is no way to tell if one or the other has a restriction. A single flow meter may say the you have the correct flow, but half of the system may be starved. This is sometimes caused by the quick connect fittings getting old and the internal check valves closing while the coupler is in the correct position.
What I have done on several systems is to put a flowmeter on each feed and run half the water through each. If a blockage occurs on one leg, it will be apparent on the respective flowmeter. One quick solution to the blockage is to remove the check valve from the offending connector until you can get a new connector.
Ken Converse owner Quality Images third party SEM service Delta, PA
by way of MicroscopyListserver wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 29 06:44:58 2004
we need for an rather old Philips EM 420 the two-piece plate holders. The format is 9.2cm x 11.2cm (eq. 3.6" x 4.4"). The insert is for the 8cm x 10cm (3.2" x 3.9") negative format. Please let me know if you know anybody who has some of this holders in stock or wants to get rid of his used ones.
Please contact me offline. Thank you in advance.
Andreas
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 29 06:52:37 2004
Thanks to all who expressed concern over my 17 year old ether. It was in a metal can and I called our Safety people as soon as I saw the age. The safety person said that ether forms less peroxides in metal than in glass-which is a good thing because I was moving that can all over the place before I saw what it was. Then I had to move it again to try and find a date on the can (the only label was an expiration date).
Let this be a reminder to all to put "date received" labels on all bottles and cans of reagents and chemicals. Later down the line someone might need to now the age of the stuff after inheriting it.
Once again Thanks for the concern-my ether did not go BOOM. Yippee!
I'll probably find some old really wet picric acid next.
Back to finding more old chemicals,
Paula :-)
Paula Sicurello EM Lab 106 Mills Godwin Sciences Building Old Dominion University Norfolk, VA 23529 USA 757-683-3822
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 29 07:50:11 2004
I had a similar problem with my old 100CXII. It's been a while, but I can tell you a couple of things that went wrong towards the end. I seem to remember that my mysterious shut-down was related to a big breaker in the HT unit. If memory serves, this breaker is the main power switch for the HT and is located on the left side facing the front of the unit. I replaced it and the shut-downs stopped. My assumption is that over time the contacts weakened and it would begin to get hot, then kick out. Another problem was that the pneumatic valve blocks located under the left console began to develop slow leaks. I fixed a couple of them, but eventually they all began to fail. Replacement parts arrive 6 months after being ordered as they had to come on special order from Japan, but by then I had replaced the microscope. Hope this helps
Dr Robert Simmons Program Director Biological Imaging Core Laboratory Georgia State University Atlanta, GA 30302-4010
404-651-3138 404-651-2509 FAX
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 29 08:27:20 2004
My job and EM lab were eliminated. My servicemen knew that my lab was being eliminated, if I still wanted to work in EM, and knew the condition of my TEM, SEM, image analyzer, and microtome. They knew I had them rebuild the whole vacuum system within the last two years on the TEM and the care it got over the years. Servicemen can be a networking resource for you. Here's why.
Servicemen know who's lab is being closed or down sized, who's thinking of upgrading their equipment, and who is actually upgrading. Even if the lab also has some other manufacturer's equipment, they are usually told what is happening with that total lab setup. Just ask them if they can help you in your search.
Contact the salesmen with any manufacturer. If a customer knows the salesman has a buyer for his used equipment being replaced, then the salesman can use that to close a deal on a new piece of equipment from him. The salesman wins, you win, the seller wins, and the manufacturer wins. Be sure to ask the serviceman what the condition of the equipment is and for some service reports for the last two years or so. Ask if the equipment was serviced by factory people under a factory contract or under insurance that used factory people.
Also, some industrial companies will donate their used equipment to a university. If the thing is not any good, it will usually be dumpstered. If it is still good equipment, they will try to donate it and take a tax write-off. This 'free stuff' might be a bit too old for you. In that case, you have to pay for something newer.
If a company does not want to fund an SEM lab anymore, you can make an offer to take it off their hands as a donation. In exchange, you can offer to let them use their old scope for a specified number of hours using their own microscopist. This was done at a local university near me through the serviceman. He got them together with a company I didn't even know had a microscope.
There are a lot of options out there in addition to buying an SEM on the open market. Hopefully, someone will read this and contact you about a donation or a good SEM you can buy used.
Paul Beauregard Senior Research Associate
} From: darryl krueger {dkruege-at-CLEMSON.EDU} To: Microscopy-at-MSA.Microscopy.com cc:
} Research Position in Characterization of Boron Carbide } } } } The primary aim of the project is to understand the root cause of } the dramatic failure of B4C at pressures near 20GPa. The project } will deal with carrying out thorough characterization of existing } Boron Carbides and derivatives using analytical techniques such as } high resolution transmission electron microscopy with electron } energy loss spectroscopy, Raman spectroscopy, Nuclear Magnetic } Resonance etc. in order to correlate the elastic modulus and } hardness to the microstructure. In addition, the collapse of the } crystal structure will be investigated theoretically using first } principle calculations. Specifically, we will design a stable B4C } crystal based on crystallographic and spectroscopic data available } in the literature. The changes in the crystal structure will then be } monitored while being subjected to various stresses below and above } the threshold impact pressure of B4C. The calculations will provide } a link from the atomistic level to the micro-structural level. The } theoretical study could shed light on whether the poor ballistic } performance above the threshold impact pressure is an intrinsic } property of B4C. } } } } The successful candidate should have a strong background in } characterization of materials using transmission electron } microscopy. A thorough understanding of microstructure and } crystallography of ceramic materials is essential. Applicants are } requested to contact Prof. Manish Chhowalla, Rutgers University, } Ceramic and Materials Engineering, 607 Taylor Road, Piscataway, NJ } 08854. Email: } {mailto:manish1-at-rci.rutgers.edu} manish1-at-rci.rutgers.edu. Phone: } 732-445-5619
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 29 10:08:22 2004
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I agree with Robert. Our 100CX developed leaks in the gang of air pressure valves next to the HT tank. The JEOL service technician was able to rebuild them and that fixed the problem.
Randall Massey Operations Manager UW-Electron Microscope Facility 1300 University Ave. Madison, WI 53706 voice (608)262-2993 Fax (608)262-7306
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 29 10:17:23 2004
For any IC folks out there, I'm wondering if anyone can explain what I believe is highly amorphous ILD at vias in single and dual damascene ICs? Relevant pix can be seen at:
http://www.microtechnics.com/ild1.jpg
http://www.microtechnics.com/ild2.jpg
Picture 2 shows cratering in the open areas but both pix show massive opening at the via. I attribute this to the ILD being more amorphous at the via. But why? Or is something else going on?
Etching is wet chemistry. Mag is about 60KX.
Thanks for any inputs.
gary g.
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 29 10:20:42 2004
Thanks to all who expressed concern over my 17 year old ether. It was in a metal can and I called our Safety people as soon as I saw the age. The safety person said that ether forms less peroxides in metal than in glass-which is a good thing because I was moving that can all over the place before I saw what it was. Then I had to move it again to try and find a date on the can (the only label was an expiration date).
Let this be a reminder to all to put "date received" labels on all bottles and cans of reagents and chemicals. Later down the line someone might need to now the age of the stuff after inheriting it.
Once again Thanks for the concern-my ether did not go BOOM. Yippee!
I'll probably find some old really wet picric acid next.
Back to finding more old chemicals,
Paula :-)
Paula Sicurello EM Lab 106 Mills Godwin Sciences Building Old Dominion University Norfolk, VA 23529 USA 757-683-3822
__________________________________ Do you Yahoo!? Yahoo! Mail - 50x more storage than other providers! http://promotions.yahoo.com/new_mail
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 29 10:28:33 2004
Call me please. I am used to be a service engineer for JEOL.
Tim 301-405-5244
--- Long Li {longli_tem-at-hotmail.com} wrote: } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Luis, } I would suggest you check the freon presure for EHT. } } } } } } Long Li } ____________________________________________________________________________ } _________ } Materials Science and Engineering Department } University of Pittsburgh } 3700 O'Hara St., 848 Benedum Hall } Pittsburgh, PA 15261 } } Tel: (412) 624-9753 } FAX: (412) 624-8069 } e-mail: Lil2-at-pitt.edu } longli_tem-at-hotmail.com } } } } } ----- Original Message ----- } } From: "by way of MicroscopyListserver" } {lbustillos-at-amalab.com} } To: {microscopy-at-microscopy.com} } Sent: Friday, June 25, 2004 2:31 PM } Subject: [Microscopy] viaWWW: Microscope keeps } shutting itself off. } } } } } } } } } -------------------------------------------------------------------------- } ---- } } The Microscopy ListServer -- Sponsor: The } Microscopy Society of America } } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -------------------------------------------------------------------------- } ----- } } } } Below is the result of your feedback form } (NJZFM-ultra-55). It was } submitted by (lbustillos-at-amalab.com) from } http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html } on } Friday, June 25, 2004 at 08:44:42 } } } -------------------------------------------------------------------------- } - } } } } Email: lbustillos-at-amalab.com } } Name: Luis Bustillos } } } } Title-Subject: [Microscopy] [Filtered] } MListserver: Microscope keeps } shutting itself off. } } } } Question: We are having a puzzling problem with } our JEOL 100CX II. It } shuts itself off sometimes after 1-2 hours but } sometimes it will run a day } or two before it will eventually shut itself off. } We have checked all that } we could think of that could be causing the problem. } For example, the water } circulation, air pressure, heating plates, fore pump } sensors, exchanged the } whole vacuum board, checked the input voltage but to } no avail. I would } really appreciate it if anybody has any suggestions. } } } } Lou } } } } } -------------------------------------------------------------------------- } - } } } } } }
__________________________________ Do you Yahoo!? Yahoo! Mail is new and improved - Check it out! http://promotions.yahoo.com/new_mail
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 29 10:41:28 2004
We will be studying changes in mitochondrial morphology after drug application to cell monolayers. I have reviewed the listings in the archives from few years ago regarding effects of osmolarity and choices of buffer and fixative on muscle mitochondria. I would think that mitochondria in cell monolayers would be particularly sensitive to these factors.
I am condidering fixing the monolayers in 2.5 % glutaraldehyde in serum free medium in an effort to maintain physiological conditions up to the moment of fixation. Then, what about rinses before going into osmium?
Can anyone offer any pointers regarding preservation of mitochondrial morphology in cell monolayers?
Thanks, Dotty
Dorothy Sorenson Microscopy and Image Analysis Laboratory Department of Cell and Developmental Biology University of Michigan Medical School 4643 Medical Science Building II 1301 Catherine Ann Arbor, MI 48109-0616
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 29 12:43:05 2004
When cells are cryo-fixed (metal mirror or high pressure freezing), the mitochondria can have a very different morphology than after chemical fixation. I remember an ancient paper from the 80's which looked at the morphology of isolated mitochondria cryo-fixed before and after exposure to an uncoupling agent. the uncoupled cryo-fixed mitochondria had an appearance similar to chemically fixed ones while the coupled ones were more compact and therefore denser. sorry I can't remember the actual citation. but the bottom line is that I think any attempt to see subtle drug effects on mitochondrial morphology after chemical fixation is suspect.
At 12:11 PM 06/29/04 -0400, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
I've been attempting to make gold-conjugated antibodies and I've had varying degrees of success. I'm currently working with 1.4nm Nanogold, 2nm and 5nm colloidal gold from various vendors. I believe each conjugate is made correctly, but my methods for purifying and concentrating the 1.4nm and 2nm products are not adequate. Depending upon the conjugate, the samples are unrecoverable from the centricon or ultracentrifuge tube.
I'm figuring that my centrifuge conditions are not appropriate and that the Nanogold may be binding irreversibly to the centricon membrane. The protocols supplied by the colloidal gold vendors do not give sufficient details and my efforts to extrapolate the speed at which to spin the 2nm conjugates have not worked as of yet. I would appreciate any input or experience that one could share. Feel free to contact me using the email address below.
Thanks in advance, Henry
-- Henry C. Grisé grise-at-bio.fsu.edu Biology Graduate Student Florida State University Tallahassee, FL 32306
From MicroscopyL-request-at-ns.microscopy.com Tue Jun 29 16:06:25 2004
I agree with Tom on this one. How will you know if all of the mitochondria in one cell are doing the same thing at the same time (and thus equally sensitive to the drug and giving you a valid measurement of its effects) ? You won't! I think subtle changes would be better tracked via biochemistry. Huge changes might be another matter but even then I think you will need to do some stereology to validate what ever you see.
Geoff
Tom Phillips wrote:
} When cells are cryo-fixed (metal mirror or high pressure freezing), } the mitochondria can have a very different morphology than after } chemical fixation. I remember an ancient paper from the 80's which } looked at the morphology of isolated mitochondria cryo-fixed before } and after exposure to an uncoupling agent. the uncoupled cryo-fixed } mitochondria had an appearance similar to chemically fixed ones while } the coupled ones were more compact and therefore denser. sorry I } can't remember the actual citation. but the bottom line is that I } think any attempt to see subtle drug effects on mitochondrial } morphology after chemical fixation is suspect. } } At 12:11 PM 06/29/04 -0400, you wrote: } } } ------------------------------------------------------------------------------ } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------------------------- } } } } } } Dear listers, } } } } We will be studying changes in mitochondrial morphology after drug } } application to cell monolayers. I have reviewed the listings in the } } archives from few years ago regarding effects of osmolarity and } } choices of buffer and fixative on muscle mitochondria. I would think } } that mitochondria in cell monolayers would be particularly sensitive } } to these factors. } } } } I am condidering fixing the monolayers in 2.5 % glutaraldehyde in } } serum free medium in an effort to maintain physiological conditions } } up to the moment of fixation. Then, what about rinses before going } } into osmium? } } } } Can anyone offer any pointers regarding preservation of mitochondrial } } morphology in cell monolayers? } } } } Thanks, } } Dotty } } } } Dorothy Sorenson } } Microscopy and Image Analysis Laboratory } } Department of Cell and Developmental Biology } } University of Michigan Medical School } } 4643 Medical Science Building II } } 1301 Catherine } } Ann Arbor, MI 48109-0616 } } } Thomas E. Phillips, PhD } Professor of Biological Sciences } Director, Molecular Cytology Core } 3 Tucker Hall } University of Missouri } Columbia, MO 65211-7400 } } 573-882-4712 (office) } 573-882-0123 (fax) } PhillipsT-at-missouri.edu } } }
-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 30 02:23:30 2004
Hi, we are working up an animal model of allergic rhinitis and currently favour the guinea-pig. Conventional toluidine blue works great on other species but hardly demonstrates them at all in the g-p. The best I've acheived is with a Luna's method, does any-one have any good working methods tinctorial or IHC (we have not found an antibody to work yet)? It has been suggested that I do some TEM but am loathe to go there as I suspect the mast cell ultrastructure is also different to other species (and hence the lack of histochemistry). This is making life extremely difficult as the degranulation of mast cells is crucial
Histopathology Group Asthma and Allergy Disease Biology ri- CEDD. GlaxoSmithKline Medicines Research Centre, Gunnelswood Road, STEVENAGE, Hertfordshire. SG1 2NY tel. +44 (0)1438 764119 fax. +44 (0)1438 764782 email. gillian.2.brown-at-gsk.com
You could try periodic acid Schiff as a general starting point and then try Alcian Blue (pH 2.5 and pH1.0) and also aldehyde fuchsin. Have you tried an antibody to mast cell tryptase.
Gareth Morgan MPhil MSc FIBMS, Department of Laboratory Medicine (Labmed), Karolinska Institute, Karolinska University Hospital at Huddinge, F46 SE 141 86 Stockholm Sweden
OBS! Besöksadress: F-Huset, Forskningsgatan 2 F52, Rum 2.10. Laboratoriet för klinisk patologi och cytologi.
NB! Visiting address: Building F, Research Corridor 2 F52, Room 2.10. Clinical Histo- and Cytopathology Laboratory.
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 30 08:36:21 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rpowell-at-nanoprobes.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, June 29, 2004 at 18:13:44 ---------------------------------------------------------------------------
Email: rpowell-at-nanoprobes.com Name: Rick Powell
Organization: Nanoprobes, Incorporated
Title-Subject: [Microscopy] [Filtered] MListserver: Re: Gold conjugated IgG and Fab'
Question: (Vendor reply)
Hello Henry:
We manufacture NanogoldÆ conjugates, and often begin the purification of IgG and Fab' fragments by Centricon membrane centrifugation. The conditions we usually use are a 30,000 MW cutoff membrane ("Centricon-30"); the conjugate is dissolved in reaction buffer, usually phosphate-buffered saline at pH 6.5 or 7.5 depending on whcih type of Nanogold you used (one to a few mL). We usually use a 6,000 g filtration for 20 minutes, then invert, and extract into the caps at 3,000 g for two minutes.
Some preps are more "sticky" than others; if this is a problem (you can often tell because the liquid does not filter through the membrane as quickly as it should), then you can try vortexing the tubes before inverting them, or adding a few drops of dimethy lsulfoxide (DMSO) then vortexing (make sure that total DMSO stays below 20% of what's left above the membrane) - DMSO is highly effective for dissolving Nanogold.
One consideration: we don't recommend doing this as a purification method (i.e. multiple times), but only to concentrate the samples before liquid chromatography, which is our principal method for separation. We recommend gel filtration liquid chromatography to separate the Nanogold conjugate from unbound excess Nanogold (Superose-12 is a good resin for this).
The best method for separating colloidal gold conjugates is widely held to be density gradient ultracentrifugation over a 10 to 30% sucrose or glycerol gradient. For 5 nm colloidal gold, the literature procedure (see, for example, Slot and Geuze, Immunolabeling for EM, 1984) consists of an initial pelleting for 45 mins at 60,000 to 125,000 g; the loose part of the pellet is then resuspended and recentrifuged over a 10 to 30% gradient of sucrose or glycerol in 1% BSA-Tris buffer, and centrifuged again for 45 minutes at 125,000 g (41,000 rpm on a Beckman SW41 rotor), usually at 4C. The good stuff (the right size and not aggregated) is usually about one-third to half-way down. You can then remove the glycerol by dialysis if necessary.
We usually can't pellet Nanogold, although we've noticed that it occasionally precipitates rather than pellets - for this or 2 nm gold you will probably need to use a very high-g centrifuge such as a Beckman Airfuge compressed-air centrifuge.
Research Diagnostics, Inc. have a thorough set of instructions for preparing colloidal gold conjugates:
http://www.researchd.com/gold/gold8.htm
We include a section on our web site which discusses what Nanogold is, how labeling works, and how to optimize the labeling reaction and the separation of conjugates:
http://www.nanoprobes.com/LGuide.html
We would also recommend double-checking the conjugation reaction - particularly the pH and (in the case of Nanogold) the stoichiometry, or ratio of the amounts of Nanogold to protein.
Hope this helps,
Rick Powell Nanoprobes, Incorporated rpowell-at-nanoprobes.com www.nanoprobes.com
I've been attempting to make gold-conjugated antibodies and I've had varying degrees of success. I'm currently working with 1.4nm Nanogold, 2nm and 5nm colloidal gold from various vendors. I believe each conjugate is made correctly, but my methods for purifying and concentrating the 1.4nm and 2nm products are not adequate. Depending upon the conjugate, the samples are unrecoverable from the centricon or ultracentrifuge tube.
I'm figuring that my centrifuge conditions are not appropriate and that the Nanogold may be binding irreversibly to the centricon membrane. The protocols supplied by the colloidal gold vendors do not give sufficient details and my efforts to extrapolate the speed at which to spin the 2nm conjugates have not worked as of yet. I would appreciate any input or experience that one could share. Feel free to contact me using the email address below.
Thanks in advance, Henry
-- Henry C. GrisČ grise-at-bio.fsu.edu Biology Graduate Student Florida State University Tallahassee, FL 32306
Second try. We're trying to cut a couple of Indium/Thallium alloys: 78 at. % In/22 at. % Tl and 18 at. % In/82 at. % Tl. We're actually trying to produce a good face for examination in a reflecting metallographic (light) microscope, not produce sections. A good SEM specimen would be nice. Grain structure at the LM level is desired. The alloy is very soft, however, and isn't cooperating. Tried: Breaking in LN2. It bent. Cutting with EMD. Pitted the face. A diamond saw. Smeared the face. Lapping. Embedded polishing compound in the face. Electro-jet polishing, but the solutions tried didn't work. I don't know what solutions were tried, though, any hints? We're trying glass-knife microtomy, but I suspect we'll have smearing problems. We don't have a materials science diamond knife, nor can we buy one. We're looking for a laser cutter or high-pressure water jet cutter. Any other ideas? TIA
Phil -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 30 11:17:19 2004
You have a challenge, no doubt! I have no experience with your particular alloy, but have had limited success with soft metals using typical metallographic grind/polish methods, followed by a long session in a vibratory polisher with sub-micron alumina (C) or colloidal silica. Use a light load, embedments *are* a problem...
Regards, Woody ------------------
Woody White BWXT Services: http://www.bwxt.com/bwxt.html My Site: http://woody.white.home.att.net
-----Original Message----- } From: Philip Oshel [mailto:peoshel-at-wisc.edu] Sent: Wednesday, June 30, 2004 11:09 AM To: Microscopy-at-microscopy.com
Listers,
Second try. We're trying to cut a couple of Indium/Thallium alloys: 78 at. % In/22 at. % Tl and 18 at. % In/82 at. % Tl. We're actually trying to produce a good face for examination in a reflecting metallographic (light) microscope, not produce sections. A good SEM specimen would be nice. Grain structure at the LM level is desired. The alloy is very soft, however, and isn't cooperating. Tried: Breaking in LN2. It bent. Cutting with EMD. Pitted the face. A diamond saw. Smeared the face. Lapping. Embedded polishing compound in the face. Electro-jet polishing, but the solutions tried didn't work. I don't know what solutions were tried, though, any hints? We're trying glass-knife microtomy, but I suspect we'll have smearing problems. We don't have a materials science diamond knife, nor can we buy one. We're looking for a laser cutter or high-pressure water jet cutter. Any other ideas? TIA
Phil -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 30 11:19:45 2004
One of the best methods for cutting soft materials is the wire saw. There are 2 ways to use the wire saw - with diamond coated wire or with an uncoated wire and an abrasive slurry. The method with an abrasive slurry would be best for your application. I understand that you had problems with embedding of abrasive grains during lapping, but the wire sawing process is forcing the abrasive against the area you are cutting away as opposed to towards the surface that will remain as in a lapping process.
The wire saw will not smear the material as happens with a diamond wheel saw. If you'd like, I'd be happy to cut a sample for you in our lab. Please contact me off-line to make arrangements.
We also have a number of relevant application notes that you can download from our website which deal with the wire saw as well as our other materials processing equipment. Go to www.southbaytech.com and navigate to "Applications Support". There are also links to FAQs and Application Notes from the product pages. Type "Wire Saw" in the search box at the upper right corner of the page to see our listing of 3 wire saws.
DISCLAIMER: South Bay Technology has been producing wire saws for more than 40 years and, therefore, has a vested interest in promoting their use.
Best regards-
David
-- David Henriks Vice President
South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673 USA
TEL: +1-949-492-2600 Toll-free in the USA: +1-800-728-2233 FAX: +1-949-492-1499
email: henriks-at-southbaytech.com
Celebrating 40 years of providing Materials Processing Solutions for Metallogaphy, Crystallography and Electron Microscopy.
Please visit us online at www.southbaytech.com.
The information contained in this message and any attachments is privileged and confidential. This message is intended for the individual or entity addressed. If you are not the intended recipient, please do not read, copy or disclose this communication. Notify the sender of the mistake by calling +1-949-492-2600 and delete this message from your system.
Philip Oshel wrote:
} } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } Listers, } } Second try. } We're trying to cut a couple of Indium/Thallium alloys: 78 at. % In/22 } at. % Tl and 18 at. % In/82 at. % Tl. } We're actually trying to produce a good face for examination in a } reflecting metallographic (light) microscope, not produce sections. A } good SEM specimen would be nice. Grain structure at the LM level is } desired. The alloy is very soft, however, and isn't cooperating. } Tried: } Breaking in LN2. It bent. } Cutting with EMD. Pitted the face. } A diamond saw. Smeared the face. } Lapping. Embedded polishing compound in the face. } Electro-jet polishing, but the solutions tried didn't work. I don't } know what solutions were tried, though, any hints? } We're trying glass-knife microtomy, but I suspect we'll have smearing } problems. We don't have a materials science diamond knife, nor can we } buy one. } We're looking for a laser cutter or high-pressure water jet cutter. } Any other ideas? TIA } } Phil
-- David Henriks Vice President
South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673 USA
TEL: +1-949-492-2600 Toll-free in the USA: +1-800-728-2233 FAX: +1-949-492-1499
email: henriks-at-southbaytech.com
Celebrating 40 years of providing Materials Processing Solutions for Metallogaphy, Crystallography and Electron Microscopy.
Please visit us online at www.southbaytech.com.
The information contained in this message and any attachments is privileged and confidential. This message is intended for the individual or entity addressed. If you are not the intended recipient, please do not read, copy or disclose this communication. Notify the sender of the mistake by calling +1-949-492-2600 and delete this message from your system.
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 30 12:19:46 2004
} Second try. } We're trying to cut a couple of Indium/Thallium alloys: 78 at. % In/22 } at. % Tl and 18 at. % In/82 at. % Tl. } We're actually trying to produce a good face for examination in a } reflecting metallographic (light) microscope, not produce sections. A } good SEM specimen would be nice. Grain structure at the LM level is } desired. The alloy is very soft, however, and isn't cooperating. } Tried: } Breaking in LN2. It bent. } Cutting with EMD. Pitted the face. } A diamond saw. Smeared the face. } Lapping. Embedded polishing compound in the face. } Electro-jet polishing, but the solutions tried didn't work. I don't } know what solutions were tried, though, any hints? } We're trying glass-knife microtomy, but I suspect we'll have smearing } problems. We don't have a materials science diamond knife, nor can we } buy one. } We're looking for a laser cutter or high-pressure water jet cutter. } Any other ideas? TIA } Dear Phil, Could you prepare a suitable face by pulling a "pre-scored" rod? By this I mean to take a fairly thick cylinder of the alloy, cut into it to make a fairly short length of very thin material (diameter to be determined by what size face you need), then apply a short, intense pull to separate the rod at its thinnest point, like breaking silly putty. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 30 12:57:05 2004
does anyone have a foot switch and/or specimen holders for a leica 2800 frigocut-E cryomicrotome in your spare parts inventory?? would be much appreciated as we set up our latest "donation" to our lab.
thanks
tbudd
-- Dr. T. Budd Professor and Chair of Biology St. Lawrence University Canton, NY 13617 Phone = 315-229-5640 Fax = 315-229-7429 E-mail = tbudd-at-stlawu.edu
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From MicroscopyL-request-at-ns.microscopy.com Wed Jun 30 13:52:57 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (parthara-at-ucmail.uc.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, June 30, 2004 at 09:48:30 ---------------------------------------------------------------------------
We are using permanox chamber slides (Nunc) to grow keratinocytes, infect them with viruses and then perform immunofluorescence. We are observing that under the microscope the cells do not look very clear. Nunc says that glass slides are better then permanox for immunofluorescence, as the permanox has minimal fluorescence and that the nature of the plastic is that it has a slight cloudiness to it. I have several chambers with cells treated and fixed and ready to be stained. Is there any way to be able to use these chambers and not have to redo the entire experiment?
Thanks for any suggestions Ranjani Research Associate University of Cincinnati
We are looking for a darkfield system for an Olympus BH-2, used is OK. Please contact me offline if you have any information. Thanks in advance,
Martín J. Ramírez Divisiķn Aracnología Museo Argentino de Ciencias Naturales Av. Angel Gallardo 470 C1405DJR Buenos Aires Argentina tel +54 11 4982-8370 fax +54 11 4982-4494 ramirez-at-macn.gov.ar
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 30 14:53:44 2004
Phil, To me it looks like your best bet is to cut it with whatever and then polish it with polishing film to avoid embedding particles in the sample. You will soil the film very fast and will need more fresh film perhaps, but you will expose the substructure of the sample. The water jet sounds promising, but it might smear the face too. Polishing in LN2? Wet etching? This is one brain buster..
Philip Oshel wrote:
} } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } Listers, } } Second try. } We're trying to cut a couple of Indium/Thallium alloys: 78 at. % In/22 } at. % Tl and 18 at. % In/82 at. % Tl. } We're actually trying to produce a good face for examination in a } reflecting metallographic (light) microscope, not produce sections. A } good SEM specimen would be nice. Grain structure at the LM level is } desired. The alloy is very soft, however, and isn't cooperating. } Tried: } Breaking in LN2. It bent. } Cutting with EMD. Pitted the face. } A diamond saw. Smeared the face. } Lapping. Embedded polishing compound in the face. } Electro-jet polishing, but the solutions tried didn't work. I don't } know what solutions were tried, though, any hints? } We're trying glass-knife microtomy, but I suspect we'll have smearing } problems. We don't have a materials science diamond knife, nor can we } buy one. } We're looking for a laser cutter or high-pressure water jet cutter. } Any other ideas? TIA } } Phil
-- Milen Shishkov Wellman Center for Photomedicine Massachusetts General Hospital Harvard Medical School
Address: MGH BAR 712 50 Blossom St. Boston, MA 02114 Phone: (617) 726 1589 Fax: (617) 726 4103
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 30 16:18:18 2004
How are you looking at the slides? Have you removed the chamber and coverslipped the cells or are you trying to look through the plastic slide? If so, that is your problem. In addition, plastic can absorb a lot of UV light so that can attenuate the signal. You might want to consider putting small round (12 mm diameter) glass cover glasses into 24 well trays. Seed your cells, infect them, and then mount cell side facing down onto a glass slide. this works well. good luck. tom
At 02:22 PM 06/30/04 -0500, you wrote:
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Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
This is probably an FCC alloy with that much In in it and it makes sense that it smears and will not fracture at LN2 temp, it does not have a ductile-brittle transition. As you are finding out, soft materials are a problem to polish. Here are some suggestions:
1) Find an FIB and dry cut a small sample. No problems and you may even get enhanced optical imaging with playing with ion imaging the cut sample. Your sample size will be extremely small but still accessible unless the feature sizes are bigger than the sample size.
2) You don't say what the treatment of this alloy is. Are you interested in the phases and microstructure after some processing or deformation or what? If it is cast structure, why not recast it on a very smooth surface and then chemically etch the smooth surface when you remove it from that surface? You might try glass, mica, or something similar. That surface should be as clean as possible prior to casting the sample on it.
3) South Bay Technology also has a chemical wire saw system that drips an acid onto the wire cutting through the sample. I have never seen this system work, but I have seen it in their brochure. I would take Dave Henriks up on his offer of wire cutting the sample using a smooth wire and the boron carbide slurry mixture made with glycerin and water.
4) Use one of the other treatments that you have already tried followed by vibratome polishing. If you used EDM, you could put a coating of epoxy on surface to fill in the pits, cure it and then polish. Periodically redo the epoxy. You will have to chemically finish the sample to remove the "smeared" material. You could also try the vibratome route. You could also use a low angle ion mill to ion polish the surface after the mechanical cutting and polishing steps. On an EDM sample you might have to have in there for a while, but you would succeed. Also an option is to low angle ion mill the wire saw cut sample.
5) Check with Buehler, Struers, and other companies who have polishing equipment and who publish metallographic "how to" books. You want to look for examples of copper, lead, or solder alloys to see how they handle the smearing problem.
-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)
Philip Oshel wrote:
} } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } Listers, } } Second try. } We're trying to cut a couple of Indium/Thallium alloys: 78 at. % In/22 } at. % Tl and 18 at. % In/82 at. % Tl. } We're actually trying to produce a good face for examination in a } reflecting metallographic (light) microscope, not produce sections. A } good SEM specimen would be nice. Grain structure at the LM level is } desired. The alloy is very soft, however, and isn't cooperating. } Tried: } Breaking in LN2. It bent. } Cutting with EMD. Pitted the face. } A diamond saw. Smeared the face. } Lapping. Embedded polishing compound in the face. } Electro-jet polishing, but the solutions tried didn't work. I don't } know what solutions were tried, though, any hints? } We're trying glass-knife microtomy, but I suspect we'll have smearing } problems. We don't have a materials science diamond knife, nor can we } buy one. } We're looking for a laser cutter or high-pressure water jet cutter. } Any other ideas? TIA } } Phil
-- Milen Shishkov Wellman Center for Photomedicine Massachusetts General Hospital Harvard Medical School
Address: MGH BAR 712 50 Blossom St. Boston, MA 02114 Phone: (617) 726 1589 Fax: (617) 726 4103
From MicroscopyL-request-at-ns.microscopy.com Wed Jun 30 21:08:50 2004
Ranjani, I agree with Tom that growing the cells in glass cover slips in well plates get rid of most of such problems. Other possibility is you can use, coverslip bottom chambers. If you wish you can get them from http://www.nuncbrand.com/page.asp?ID=236&lang=GB If you wish to save the current experiment where cells already grown and infected with virus in the Permanox chambers you can remove the chamber completely including the small silicon base which holds the plastic chamber on to the slides and coverslip (glass) them with suitable antifade mountant, then find an inverted fluorescence microscope and view them from through the cover slip side. I hope this helps. Shiv
Mayandi Sivaguru, Ph.D., Associate Director Molecular Cytology Core Facility Molecular Biology Program 2, Tucker Hall University of Missouri Columbia, MO 65211-7400 sivagurum-at-missouri.edu
Voice: 1-573-882-4895 Fax: 1-573-882-0123
www.biotech.missouri.edu/mcc/
At 02:22 PM 6/30/04, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Sat Jul 3 14:50:26 2004
Karl, please, don't vent the column, and don't do anything with the roughing tank.
Assuming the pump is working properly (pulls proper vacuum, does not make funny noises and smoke, doesn't leak oil), and does not need cleaning:
1) Depress vacuum ON button (right control panel), hold it down for a minute or two, let the pump run and warm up. 2) Turn off TEM by pressing mains OFF button on left control panel. 3) Swing power supply cabinet open.
Quick oil change.
4) Disconnect pump intake hose and exhaust hose if present. 5) Carefully lift pump, rotate it so that drain plug is facing back of the TEM, and elevate pump by placing it upon a piece of wood (4x4 works well), a brick, etc. Avoid stretching pump power cable. If cable is too short, unplug it from power supply after un-screwing it's connector lock. Make sure pump is slightly tilted - drain plug down, motor-up. 6) Use oil pan (auto parts store), unscrew oil drain plug from the pump, drain oil, replace plug. 7) Make sure pump position is horizontal. Fill pump with fresh oil. Slowly, don't over-fill. Watch the sight glass.
Very quick-n-dirty oil change
4) Have a shallow dish and aluminum foil, plenty of paper towels, inspection mirror. 5) Bend aluminum foil as a U-channel, put it between drain plug and shallow dish. 6) Carefully unscrew drain plug, but don't remove it completely. Hold it at the drain hole and control oil flow, keep oil from spilling over U-channel. Drain oil, replace plug. 7) Fill pump with fresh oil. Slowly, don't over-fill. Watch the sight glass with inspection mirror (you can't see it directly unless pump is rotated).
Very quick-n-dirty oil change is not recommended as a standard practice. Old oil does not drain completely, and some oil will be spilled.
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax (678)467-0012 mobile www.sia-cam.com
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----- Original Message ----- } From: {Frank.Karl-at-degussa.com} To: {microscopy-at-msa.microscopy.com} Sent: Thursday, July 01, 2004 2:00 PM
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