-Obviously an advantage for many bacteria species, but a nightmare for the microscopist who wants to count them. For several reasons we want to split the aggregates of bacteria into single cells before counting. We have tried mild detergent treatment and ultrasound though with limited success. The samples are initially taken, as filter samples in working atmospheres were bacteria could be airborne, e.g. farm work. Afterwards the bacteria are washed off the filter, resuspended, stained (AO), refiltered on black pc-filter, mounted and counted. Any suggestions for a treatment, which will de-aggregate the bacteria, are most welcome.
Asbjorn Skogstad National Inst. of Occup. Health, Oslo, Norway asbjorn.skogstad-at-stami.no
That's why I tried to bring in a useful element to it by mentioning, a little tongue in cheek, my humidity vs formvar films question. But thanks to the silly weather thread I had some useful and interesting responses. May I be forgiven for saying that there's a silver lining..............
Tony Bruton University of Natal South Africa
} } } "Chuck Butterick" {cbutte-at-ameripol.com} 02/01/00 04:06AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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Hi,
Are we like kids sitting in a sandbox and squabbling? Yes weather, no weather! He said it first! She started it! I don't want weather on the listserver. I want to read about weather in the boondocks. Who cares about you, anyway? Sort of funny when you think of it that way, isn't it?
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Does anyone have the geometry settings for a Kevex Quantum detector on a JEOL 2000FX? I am using DTSA and need the values. I would like to have the sample to detector distance and the azimuthal angle. I am using +45 for the azimuthal angle and 90 for the detector angle. Are these correct? I know that the takeoff angle is 70. -Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Hi all. I have just joined this list and am posting without the usual "lurking time" due to time constraints - please forgive if recently covered. I am also a novice in all microscopy techniques, especially fluorescence.
I am trying to detect GFP-containing cells in liver tissue and having some problems.
1. I can't determine the spectra for the filter blocks I am using (on a Nikon Eclipse TE300 with TE-FM Epifluorescence attachment). The blocks are marked "UV", "B-2" (blue from source, green in field) and "G" (green from source, red in field). I have e-mailed Nikon and to be fair they've only had a few days but I'm under some pressure. No help from the manual. I have been using B-2 for GFP.
2. I get quite a lot of background fluorescence even with frozen sections (fixed in neutral buffered formalin). Can anyone suggest a way to reduce this? (eg any extra filters?)
3. I have read conflicting opinions on fixation. Most previous work has used thick sections (50 microns cut eg with Vibratome, ? to avoid having to embed tissue blocks). GFP is interfered with by acetone (and probably other organic solvents) but I would have thought that after fixation with formalin it should be stabilized and resistant to the xylene/alcohol used in paraffin embedding. I have seen fluorescence retained after this treatment but perhaps it can be improved.
4. For those with liver fluoro experience - on "UV" setting I see bright fluorescence which photobleaches. I believe I am looking at retinoids in stellate cells, although the bleaching is incomplete and a bit slower than I have seen before. Can anyone confirm this?
Apologies for long post. Any help much appreciated.
David Lockwood University of Qld Dept of Surgery PA Hospital Brisbane Australia
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A colleague would like your responses to these questions. I believe that I say something on the first question about a year ago and apologize for asking again.
} Here's what I/we need to know. } } 1) Are Microstar diamond knives equal to Diatome. We have an offer that } will save us about $1200 on three knives ( one 3mm ultra with 45 degree } angle, one 3mm ultra with 35 degree angle, one 10 mm histo knife). } } } 2) Is the power controller feature on the Pelco Microwave Tissue processor } (model #3451) worth it? (costs about $1300 additional compared to the } Pelco 3450 without the power controller). I plan to do immunolocalization } work, but am not sure this power controller feature is necessary or worth } the cost. } } Casey } } } Thanks for your time } Marty Reed Equipment Technician Biology Department Humboldt State University Arcata CA 95521 707-826-3234 707-826-3201 FAX mmr7001-at-axe.humboldt.edu
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Hi! Could anybody tell me what would be the sale price for a Hitachi H600? Thanks Dorota
I don't use a CM series microscope on a regular basis, but what is important is that you set it up the same way every time.
You can not make the back focal plane of the objective lens coincident with the objective aperture. The back focal plane is well above the location of the objective aperture plane.
There are two ways that I use to set up diffraction patterns reproducibly depending on whether I am using CBED or SAD. Both are set up after the sample has been made eucentric and the image focussed.
CBED: This method can be done for both CBED and SAD. The shadow of the condenser aperture defines the diameter of the diffraction disk. When the intermediate lens is adjusted properly, the edges of the diffraction disks will be in focus. You are grabbing the back focal plane for your diffraction pattern in the projector lens system. You will note that all of the HOLZ lines (if you can see them) are the sharpest at this condition. If you have a highly polycrystalline sample with continuous diffraction rings, this method is difficult to do.
SAD. Spread the beam with the condenser all the way. (clockwise in the CM-12 will go to more parallel beam faster than CCW -I think.) Then focus the spot to the smallest that you can. You can take a really long exposure or cheat a little and put some intensity back into the pattern with the condenser lens. You will note that the objective aperture is not focused in this method.
The most important thing to remember is to make the sample eucentric and focus before during either of these methods and to do the diffraction focussing consistently from sample to sample. -Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies." --
} -----Original Message----- } From: Valerie Leppert [mailto:vjleppert-at-ucdavis.edu] } Sent: Monday, January 31, 2000 6:44 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Question on Philips CM-12 SAD Alignment } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Hi there; } } We have a new (to us) Philips CM-12 TEM in our lab and are } wondering how } to get the intermediate lens focused on the diffraction aperture (for } making the first image plane and the diffraction aperture coincident } prior to obtaining a SAED pattern). It doesn't seem to be } covered in the } manual. } } Any help would be appreciated as this is a completely new } microscope to } us. } } Thanks, } Valerie Leppert } }
I received Simon Watkins' note about the microscopy class in Maine and wonder if anyone knows about similar courses closer to California in the near future? We've got a number of technicians working on fluorescence microscopy in our lab, and I think it would be great to have some more formal training for us!
Sincerely, Laurie Wallin UCSD Department of Anesthesiology 9500 Gilman Drive, 0629, La Jolla, CA 92093
We are disposing of our old Philips 410 transmission electron microscope. It is in pretty good shape but is not fully functioning. At minimum it needs its ion getter reconditioned. Does anyone know of someone who might want to buy it and fix it up or use it for parts? -Robert
____________________________________
Robert S. Dotson, Ph.D., Laboratory Supervisor, Microscopy
Coordinated Instrumentation Facility 605 Lindy Boggs Building Tulane University 6823 St. Charles Avenue New Orleans, LA 70118-5698
I am thankful to those who took this weather thread and imparted some information that I can really use. The retired professor that taught me how to release forvar films had no idea why sometimes it worked and sometimes it didn't. Now at least I have a couple of likely variables to check.
Thanks! Chris Best Mol. Biol. Juniata College Huntingdon, PA 16652
PS - I can't help but be amused that even on a forum for scientists, the petty &/or silly items get the most responses. Tell the truth, do you stay up at night watching Jerry Springer? (For heaven sake, don't answer that!)
I am afraid that I do not agree with Scott Walck about back focal planes. I do not have a CM 12 in the lab here. So I can not check that I am not confusing the CM 12 with other Philips/FEI instruments. However, when Scott says “You can not make the back focal plane of the objective lens coincident with the objective aperture. The back focal plane is well above the location of the objective aperture plane.” he is wrong. On all microscopes except the very few which have very small pole-piece gaps for high resolution, the objective aperture should coincide with the back focal plane.
There is an easy and accurate way to find the true back focal plane on a CM 12 (or any other microscope with an immersion lens). Use a crystalline sample, go to convergent-beam diffraction then use the diffraction focus to make the Kikuchi lines as sharp as you can. That is the back focal plane.
If the microscope is set up properly, the image of the objective aperture should be sharply in focus at nearly the same setting of the diffraction focus. If the diffraction focus to give a sharp image of the objective aperture is very different from the diffraction focus to make the Kikuchi lines sharp, get your service engineer to reset the height of the objective aperture until they agree.
If the sample is at the correct eucentric height, a selected-area diffraction pattern in the true back focal plane will have sharp spots for a C2 setting almost but not quite to the maximum (almost fully clockwise). Again, if it does not, ask the service engineer to fix it.
Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvannia 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
Alwyn, We have a CM-12 at our other facility and I will check it out in a day or so.
I thought that I did and it worked like our JEOL 2000FX. In the 2000FX, because the lens is highly excited, there are 3 cross overs in the objective lens after the sample. That means the true back focal plane is inside the objective lens and it is not possible to put the aperture at that plane. In the 2000FX, I have focused the CBED pattern and have found the objective aperture not in focus. I have also found that the two methods that I outlined do not agree with the camera constants. The 2000FX has a condenser mini lens to make the beam parallel, but the highly excited lens allows the small probes. Since the CM-12 can form the small probes, I thought that the lens system was working in a similar manner.
I will try it out unless someone beats me to it. How about it CM owners?
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
} -----Original Message----- } From: Alwyn Eades [mailto:jae5-at-lehigh.edu] } Sent: Tuesday, February 01, 2000 3:02 PM } To: microscopy-at-sparc5.microscopy.com } Subject: SAD and the back focal plane } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } } I am afraid that I do not agree with Scott Walck about back } focal planes. } I do not have a CM 12 in the lab here. So I can not check } that I am not } confusing the CM 12 with other Philips/FEI instruments. } However, when } Scott says "You can not make the back focal plane of the } objective lens } coincident with the objective aperture. The back focal plane } is well above } the location of the objective aperture plane." he is wrong. On all } microscopes except the very few which have very small } pole-piece gaps for } high resolution, the objective aperture should coincide with } the back focal } plane. } } There is an easy and accurate way to find the true back focal } plane on a CM } 12 (or any other microscope with an immersion lens). Use a } crystalline } sample, go to convergent-beam diffraction then use the } diffraction focus to } make the Kikuchi lines as sharp as you can. That is the } back focal plane. } } If the microscope is set up properly, the image of the } objective aperture } should be sharply in focus at nearly the same setting of the } diffraction } focus. If the diffraction focus to give a sharp image of } the objective } aperture is very different from the diffraction focus to make } the Kikuchi } lines sharp, get your service engineer to reset the height of } the objective } aperture until they agree. } } If the sample is at the correct eucentric height, a selected-area } diffraction pattern in the true back focal plane will have } sharp spots for } a C2 setting almost but not quite to the maximum (almost } fully clockwise). } Again, if it does not, ask the service engineer to fix it. } } } } Alwyn Eades } Department of Materials Science and Engineering } Lehigh University } 5 East Packer Avenue } Bethlehem } Pennsylvannia 18015-3195 } Phone 610 758 4231 } Fax 610 758 4244 } jae5-at-lehigh.edu } }
Jordi: For Reichert repair and parts I would suggest Helmut Patzig of MOC (Microscopical Optical Consulting Inc.), phone (914) 268-6450 or e-mail MOCLeica-at-Aol.com Helmut is known to carry spare parts for Reichert and LKB ultramicrotomes. I have no vested interest in MOC other than as a satisfied customer. If he cannot help, you could ask him what your Reichert transformer voltage output should be and using a voltage meter adjust the voltage of a variable voltage transformer to the correct amount. Make sure to incorporate a "stop" on the dial so the voltage can't be accidentally moved above the correct amount. Henry
Henry Eichelberger, Manager Electron Microscopy Facility Department of Biological Sciences Binghamton University Binghamton, NY 13902-6000
-----Original Message----- ------------------------------------------------------ On Mon, 24 Jan 2000, Marti, Jordi wrote:
} The power supply in our cryo microtome is having problems which might be } related to the transformer. I was told by the service engineer that the } transformer is no longer supported by Reichert. Does any one know where I } can get a replacement ? } } Thanks } } Jordi Marti
I apologize in advance if this posting offends anyone. However there are a good many people who use silver membranes in their work and to have the supply from the worlds only manufacturer (to my knowledge) come to a halt, has the potential of being highly disruptive at least to some programs: ================================================= Osmonics has announced the closing of our Phoenix, AZ manufacturing facility effective 1 May 2000. Since our silver membranes are manufactured in this facility, Osmonics has decided to end production of this membrane because of declining sales and the very expensive costs associated with moving the mfg. . plant to another location. All orders placed before 1 March, 2000 will be honored and filled. ================================================== We plan to make a "last buy" before the cut off date of March 1, 2000. I would advise anyone depending on these silver membranes for their work to take stock of their future requirements because after the cut off date, sales will be possible only from remaining stocks.
For those who do run out, and are in a bind, we are prepared help them find alternative filtration media, of which there are indeed some alternatives for some applications.
Chuck
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Look for us! ######################## WWW: http://www.spi.cc ######################## ============================================
I have someone here who wants to freeze and cut cryostat sections of marine larvae (5 microns?) for immuno/light microscopy. The specimens are 100-200 microns in length.
1. What would be the best way of handling these small items? 2. What would be the best support/medium e.g TissueTek? 3. What about cryoprotection? I am familiar with 2.3 molar sucrose for EM. 4. Any general tips for immuno (protein/amino peptidases)?
Thanks - Keith _______________________ Keith Ryan (Dr) Marine Biological Association Citadel Hill Plymouth Devon PL1 2PB England
Like the other respondees I am not a CM12 user (where are they?), however, as an ex CM12 user of some 10 years ago I think that I remember the alignment that Valerie is asking about. In the basic alignment of the machine there was a step during which the SAD aperture was focussed. I think that it was in the `service calibrations' page. This may have changed in later software revisions.
The alignment of this microscope was usually carried out by the engineer when the instrument was installed. They would set up the objective lens current and adjust the specimen goniometer height to ensure that when it was at the eucentric position it was correctly in focus. Following this a complete column alignment was carried out including focussing the SAD aperture. The manufacturers then assumed that the operator would set up the specimen to the eucentric height, it should focus in the same position and the aperture should be in focus.
If the aperture is not in focus when in the SA range (as noted next to the mag readout) then check you are correctly at the eucentric position, if you are then either it was not aligned properly after installation or something has changed. NOTE: DO NOT ADJUST THE SERVICE ALIGNMENTS UNLESS YOU KNOW WHAT YOU ARE DOING AS YOU CAN LOSE THE INSTRUMENT ALIGNMENTS.
Good luck, Ron
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
I got a tip for finding the condenser lens settings required for parallel illumination from Angus Kirkland (Cambridge).
Make sure you are in microprobe mode and set up for SAED and the specimen is out of the way. Spread the beam, put in the SA Aperture, go to diffraction and then sweep the SA aperture (SAA) from side to side and minimise the deflection of the diffraction spot by tweaking the illumination control (C2 or second condenser lens). A little bit of thought will convince you that anything other than a parallel beam will give you a side to side motion (tilt) when the SAA is swept.
I have found it useful to keep a table of C2 condenser lens current settings for each spot size (C1 excitation) in microprobe mode. Setting the C2 lens for parallel illumination and adjusting the diffraction focus to get the sharpest spot will then give you near perfect SAD conditions.
Anyone contemplating electron holography for example, will have to start from the parallel illumination to get the best spatial coherence for their holograms.
******************************************************** Dr Jonathan Barnard
Analytical Materials Physics The Angstrom Laboratory, Uppsala University P O Box 534, SE-751 21 Uppsala, Sweden Phone: +46-(0)18-4716838 Fax: +46-(0)18-500131 http://www.angstrom.uu.se/analytical/home.html
I am working on fluorescent imaging of thin sections cut from samples embedded in TEM embedding resin, and am having a significant problem with resin fluorescence. Does anyone know of an embedding resin that doesn't autofluoresce?
Matt Plantinga Amway Corporation matt_plantinga-at-amway.com
We have a Zeiss Axiophot microscope and a Dvorak-Stotler Chamber that we would like to assemble into a temperature-controlled environment for digital live-cell imaging. I have several questions about the best way to combine these elements:
i) Can anyone recommend a heating stage compatible with both the microscope and the chamber?
ii) Would the best position for a temperature sensor be on the top (near the objective?) or on the bottom of the chamber, or both (two sensors)?
iii) Is it better to heat the media/fluids in their containers, or to have an in-line heater ( brand recommendations?)
iv) Are there any fuid flow or temperature considerations specific to this chamber that need to be addressed (gravity vs. pump feed, shear forces, heating stage redundant, etc.)?
v) Are there any problems related to objective lens mag/NA that need to be overcome when using this D- S apparatus?
Any references, anecdotes, or words of wisdom would be appreciated. Dealers, if you have a product that you think I could use, please don't hesitate to contact me. Thank you in advance. {FontFamily} {param} BrushScript BT {/param} {bigger}
{nofill} Regards,
Andrew Ochalski, Microscopy Technician, Dept. of Biology, University of Ottawa Room 108, Gendron Bldg. 130 Louis Pasteur, Ottawa, Ontario CANADA K1N 6N5 613-562-5800 x6343 FAX: 613-562 5486
I would like to get some information from some of the top companies on microscopes with photographic and possibly fluourescent capabilities. Please email me or call 313-993-4195 Thank you
Not really a reply but another question!! Anybody know of a similar course a little further east - England (Great Britain) to be exact !!? Thanks in Advance,
Baz
Barry Shaw Senior Imaging Technician Molecular & Cell Biology E Floor School Of Biomedical Sciences University of Nottingham Medical School NG7 2UH
We are a company that has specialized on extracting 3D information from stereoscopic SEM images. Currently we try to figure out the potential applications and the future prospects of such techniques for the microscopy community.
What we do exactly is that we take two images from a sample on the SEM from slightly different tilt angles and use digital image processing
to compute a 3D elevation model. From there you can then perform various analysis steps like extraction of 3D profiles, determination of surface roughness etc.
My questions now are:
1.) How would you judge the importance of obtaining 3D data from a SEM image? 2.) What would be the major requirements for such a data set to be useful (like density, accuracy, number of false alarms, etc.)? 3.) Is it of importance to you that you get the 3D data and the image data together and not separated (like it is the case with AFM)? 4.) How would you judge the future prospects and influence of such a technique on your personal field of work? 5.) Do you generally trust the measurement of 3D data via stereoscopic images? 6.) Do you already have experience with stereoscopic depth measurments and what are your conclusions?
Best regards, Manfred Prantl R&D Alicona GmbH, Germany www.alicona.com
Irene Piscopo of FEI/Philips probably can answer your question. I'm copying this message to her although if she's in the "field" a response may take a few days. Also, Max Otten of FEI/Philips is another good source for that type of information.
We have a CM-120 so we're aware of your problems in deciphering the operator's manual. Good luck!
Bruce F. Ingber, Biologist USDA-ARS, SRRC 1100 Robert E. Lee Blvd. New Orleans, LA 70124
-----Original Message----- } From: Valerie Leppert [mailto:vjleppert-at-ucdavis.edu] Sent: Monday, January 31, 2000 5:44 PM To: Microscopy-at-sparc5.Microscopy.Com
I can't speak about Philips CM12's, but the back focal plane of the objective coincides with the objective aperture in the CM30 here.
---------------------------------------------------------------------- Russell E. Cook, Ph.D. Electron Microscopy Center for Materials Research Argonne National Laboratory Materials Science Division, Building 212 9700 South Cass Avenue Argonne, IL 60439-4838 (630)252-7194 FAX: (630)252-4289 cook-at-horus.msd.anl.gov
Hello all: Valerie Leppart, you can reach me at 203-853-3256. I cannot call you since there is no phone number to contact. Irene Piscopo
The CM 12 microscope has four mag ranges: LM, M, SA, Mh
As long as you are eucentric and using the SA range the image plane and the diffraction aperture plane will be in focus. (It is done automatically by the instrument.) This will give you accurate SAD down to 1um. The objective mag in the plane of the diffraction aperture is approxmately 27X. If you wish to obtain diffraction from areas smaller than one micron, use uD, or uuD. If you call me I will discuss these methods with you and send you detailed instructions on using these methods.
Irene Piscopo
} -----Original Message----- } From: Ingber, Bruce F. [SMTP:bingber-at-commserver.srrc.usda.gov] } Sent: Wednesday, February 02, 2000 10:14 AM } To: Microscopy-at-MSA. Microscopy. com (E-mail) } Cc: Piscopo Irene (CS) (E-mail); Max T. Otten (E-mail) } Subject: FW: Question on Philips CM-12 SAD Alignment } } Irene Piscopo of FEI/Philips probably can answer your question. I'm } copying } this message to her although if she's in the "field" a response may take a } few days. Also, Max Otten of FEI/Philips is another good source for that } type of information. } } We have a CM-120 so we're aware of your problems in deciphering the } operator's manual. Good luck! } } Bruce F. Ingber, Biologist } USDA-ARS, SRRC } 1100 Robert E. Lee Blvd. } New Orleans, LA 70124 } } (504) 286-4270 phone } (504) 286-4419 fax } bingber-at-nola.srrc.usda.gov } } -----Original Message----- } From: Valerie Leppert [mailto:vjleppert-at-ucdavis.edu] } Sent: Monday, January 31, 2000 5:44 PM } To: Microscopy-at-sparc5.Microscopy.Com } Subject: Question on Philips CM-12 SAD Alignment } } ------------------------------------------------------------------------ } The Microscopy ListServer-Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe-Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi there; } We have a new (to us) Philips CM-12 TEM in our lab and are wondering how } to } get the intermediate lens focused on the diffraction aperture (for making } the first image plane and the diffraction aperture coincident prior to } obtaining a SAED pattern). It doesn't seem to be covered in the manual. } Any help would be appreciated as this is a completely new microscope to } us. } Thanks, } Valerie Leppert
I don't have the reference right in front of me, but I believe that Gonzalez and Wintz say that
1. The eye can respond to an intensity range of ~10^10 in light intensity, but this is the adaptive response. The perceived brightness is a log (some will argue power law) function of the incident intensity.
2. In any one point in an image, the eye can distinguish at most ~20-30 gray levels... But... in a complex image you need at least 100 gray levels for the eye to see it as smooth. In other words, the eye seems to adapt as it scans the image.
Cheers, Henk
At 02:15 PM 2/2/00 -0600, rgriffin-at-eng.uab.edu"-at-sparc5.Microscopy.Com wrote:
} How many gray levels can the human eye distinguish? We've found several } references that disagree. } If anyone knows of a reference - that would be best. } } } Robin Griffin } UAB
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://web.ceof.ohio-state.edu Murphy's Law: "If anything can go wrong, it will." Commentary: "Murphy was an optimist."
Thanks for your reply and everyone else's reply regarding the SAD alignment. There seem to be a lot of different opinions about how this done!
We did set the sample at the eucentric and did notice that the SAD aperture is not quite in focus when in image mode. Of course this will cause the diffraction information to come from an area that is displaced with respect to the aperture position.
I am accustomed to being able to independently focus the intermediate lens on the SAD aperture, and then bring the image into focus using the objective lens - making the image plane and the SAD aperture coincident. This is on a much older Philips model and a much older Hitachi model.
However, on the CM-12, there doesn't seem to be a way for the user to do this in normal operation. I've tried a few of the suggestions (haven't worked my way through them all yet!) with no luck yet.
It does seem that this adjustment has been grouped under the category of "service alignment" in newer models.
We have a sony DXC-760 MD ccd camera that we have mounted to a standard camera lens with a bayonet adapter. Unfortunately, we have borrowed the adapter and have to return it. Is there a standard size (i.e. Nikon, Minolta, Canon) lens adapter that works with this camera mount? We can't find a part number or description for the adapter, and the book doesn't describe the size. We have already ordered and returned one that didn't fit.
Thanks in advance.
Karl Hagglund Proctor and Gamble Food and Beverage Analytical Microbiology
Fellow Listers, Does anyone know of a (preferrably free) software package which simulates ray paths, including the diffraction mode, in the TEM? We would like to use such a package to teach physics students a bit of optics.
Thanks, Eric
---------------------- Dr Eric Lachowski Department of Chemistry University of Aberdeen Meston Walk Old Aberdeen AB24 3UE Scotland +44 (0)1224 272934 fax 272921 e.lachowski-at-abdn.ac.uk
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Reply to: RE: LSCM Need help with non-fluorescing resin Dear Matt,
Try any of the immunocytochemical resins (Lowicryl, LR White or Gold, Unicryl etc). When specific antibodies are used on embedded tissue, they produce very beautiful fluorescent patterns that look like line drawings.
Paul Webster
Matt_Plantinga-at-amway.com wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I am working on fluorescent imaging of thin sections cut from samples } embedded in TEM embedding resin, and am having a significant problem with } resin fluorescence. Does anyone know of an embedding resin that doesn't } autofluoresce? } } Matt Plantinga } Amway Corporation } matt_plantinga-at-amway.com } } } } } RFC822 header } ----------------------------------- } } Received: from ultra5.microscopy.com [206.69.208.10] by mailhouse.hei.org } with ESMTP } (SMTPD32-5.05) id A77A9A2E007C; Wed, 02 Feb 2000 16:10:34 -0800 } Received: (from daemon-at-localhost) } by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id HAA02782 } for dist-Microscopy; Wed, 2 Feb 2000 07:22:38 -0600 (CST) } Received: from no_more_spam.com (sparc5.microscopy.com [206.69.208.10]) } by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id HAA02778 } for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Wed, 2 Feb 2000 } 07:21:41 -0600 (CST) } From: "Matt_Plantinga-at-amway.com"-at-sparc5.Microscopy.Com } Received: from mail1.mailrouter.net ([167.23.241.35]) } by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id HAA02771 } for {Microscopy-at-sparc5.microscopy.com} ; Wed, 2 Feb 2000 07:21:30 -0600 (CST) } Received: from lnot21.mailrouter.net ([10.10.16.153]) } by mail1.mailrouter.net (Pro-8.9.3/Pro-8.9.3) with ESMTP id IAA16955 } for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 2 Feb 2000 08:11:43 -0500 (EST) } Subject: LSCM Need help with non-fluorescing resin } To: Microscopy-at-sparc5.microscopy.com } Date: Wed, 2 Feb 2000 08:15:48 -0500 } Message-ID: {OF23F7A00C.431C958F-ON85256879.00486259-at-mailrouter.net} } X-MIMETrack: Serialize by Router on LNOT21/ANet(Release 5.0.2 |November 4, } 1999) at 02/02/2000 08:16:41 AM } MIME-Version: 1.0 } Content-type: text/plain; charset=us-ascii } Errors-to: Microscopy-request-at-sparc5.Microscopy.Com } X-RCPT-TO: {pwebster-at-hei.org} } X-UIDL: 242867701 } Status: U } Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm
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In a message dated 2/2/00 4:00:26 PM, rgriffin-at-eng.uab.edu-at-sparc5.Microscopy.Com writes:
} How many gray levels can the human eye distinguish? We've found several } } references that disagree. } } If anyone knows of a reference - that would be best.
Most of the books on image processing say the same thing (probably all copied from each other and other non-prime sources), but I don't know of an original reference based on real research. You can find the same ideas in books like Frisby "Seeing: illusion, brain and mind", Oxford, 1980; Marr "Vision", Freeman, 1982; and Rock "Perception", Freeman, 1984 which deal with the human visual system not in the context of computer graphics. These are probably closer to the original research.
The basic idea is that it takes about a 2-2.5% change in absolute brightness (i.e. a ratio of 1.025) to be just detectable, and only if it is a fairly sudden change spatially (or temporally for that matter). That is where the 20-30 distinct brightness levels over the full range of brightness that can be seen at one time (i.e. without accommodations like varying pupil size) comes from. The requirement that an entire scene have about 100 levels to avoid banding is more controversial, based on the idea that the eye can perform some accommodation as it traverses from one part of a scene to another. This probably doesn't apply when (e.g.) looking through a microscope, so the 20-30 number would be more applicable. It also ignores color, which complicates things further.
} From what is my understanding of optics - this will not give you parallel illumination at the specimen plane (if this what you meant?). By tweaking C2 you are moving the crossover plane (which is actually the diffraction plane). You can in the same way make the spot not to move by changing the diffraction focus. For each setting of C2 you can find the crossover by changing the diff. focus. Try this - spread the beam, go to diffraction, focus by diffraction focus for smallest spot, go back to imaging mode put the SAA, go to diffraction and try the procedure you described. I think the spot will not move. Ofcourse if you expand the beam too much you can no longer produce small diffraction spot (without the use of SAA) because you go out of paraxial mode. The diffraction plane is always the crossover plane not the theoretical back focal plane of the objective. Its z-position changes with the change of the C2 excitement. The spatial coherency of the electron waves is very high (actually the electrons are flying one by one through the microscope). The limiting factors are the source size, the illumination angle and the energy spread.
Please correct me if I'm wrong ... I'm still "green" in electron microscopy .. I'll be happy to learn from experienced users.
Best wishes,
Rado
--------------------------------------------------------------------- Radostin Danev Laboratory of Ultrastructure Research National Institute for Physiological Sciences Myodaiji-cho, Okazaki 444-8585, JAPAN e-mail: rado-at-nips.ac.jp --------------------------------------------------------------------- ----- Original Message ----- } From: Jonathan Barnard {Jonathan.Barnard-at-Angstrom.UU.SE} To: {microscopy-at-sparc5.microscopy.com} Sent: Wednesday, February 02, 2000 7:20 PM
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We are making preparations for experiments on determination of the chemical composition of semiconductor oxides, formed locally. The size of the modified area should be about 1 micron, and we are not sure yet if it is possible to make mentioned chemical analysis with Auger microprobe at all.
Another problem is that our newly came Auger spectroscope JAMP-7810 has no English manual with it. And believe me, it is barely possible to read that in Japanese!
We would be very grateful for your suggestions on both my questions: - how to measure the best chemical bonding on the spot with 1x1 sq. micrometer dimensions - where to get the manual (or copy of it) from. Remark: Mails to USA and Japanese representatives of JEOL gave no result!
Best regards. Dmitri
__________________________________________ Dmitri V. Sokolov, Doctor Course student Research Center for Interface Quantum Electronics, Hokkaido University, North 13, West 8, Kitaku, Sapporo 060, Hokkaido, Japan Phone 81-11-706-7174 Fax 81-11-716-6004 http://www.geocities.com/SiliconValley/Campus/1314 AOL Instant Messenger FalconDot ICQ 9418072 __________________________________________
Dear listers, I have started a research project on ammonium -bearing mineral (zeolites), and now I'm trying to quantify N using electron microprobe analysis. My problems of dealing with N are:
a) The thickness of the coating (in the literateur = approximately 150 A¡). Does anyone have information on the model of coater to making these films? Does anyone have experience with preparation of such samples?
b) Obtaining appropriate standards. (I have Si3N4, but it contains too much N (=39.96%) towards my samples (5%). Does anyone known any good nitrogen standards for such a sample ?
Any other suggestions about quantitative nitrogen analysis would be greatly appreciated !! Thanks for your time.
My best regards
Eugenia
Eugenia Marchi Universitˆ degli Studi di Modena Dip. Scienze della Terra P.le S.Eufemia n¡.19 41100 Modena (Italy) Tel. +39-059-417289 Fax. +39-059-417399 e-mail: bengi-at-unimo.it
It's unfortunate that this manufacturer is stopping production. Aren't there other manufacturers?
Dennis B. Barr (dennbarr-at-eastman.com) Physical Chemistry Research Laboratory Physical & Analytical Chemistry Research Division Eastman Chemical Company Kingsport, TN 37662-5150
B-150B, R-132E, (423) 229-2188
} -----Original Message----- } From: Garber, Charles A. [SMTP:cgarber-at-2spi.com] } Sent: Wednesday, February 02, 2000 2:37 AM } To: MICROSCOPY BB } Subject: Silver membranes being discontinued } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } Hello, } } I apologize in advance if this posting offends anyone. However there are a } good many people who use silver membranes in their work and to have the } supply from the worlds only manufacturer (to my knowledge) come to a halt, } has the potential of being highly disruptive at least to some programs: } ================================================= } Osmonics has announced the closing of our Phoenix, AZ manufacturing } facility } effective 1 May 2000. Since our silver membranes are manufactured in this } facility, Osmonics has decided to end production of this membrane because } of } declining sales and the very expensive costs associated with moving the } mfg. } .. plant to another location. All orders placed before 1 March, 2000 will } be } honored and filled. } ================================================== } We plan to make a "last buy" before the cut off date of March 1, 2000. I } would advise anyone depending on these silver membranes for their work to } take stock of their future requirements because after the cut off date, } sales will be possible only from remaining stocks. } } For those who do run out, and are in a bind, we are prepared help them } find } alternative filtration media, of which there are indeed some alternatives } for some applications. } } Chuck } } ============================================ } } Charles A. Garber, Ph. D. Ph: 1-610-436-5400 } President 1-800-2424-SPI } SPI SUPPLIES FAX: 1-610-436-5755 } PO BOX 656 e-mail:cgarber-at-2spi.com } West Chester, PA 19381-0656 USA } Cust.Service: spi2spi-at-2spi.com } } Look for us! } ######################## } WWW: http://www.spi.cc } ######################## } ============================================
Yes, I agree with John. A better question than "how many gray levels can one see" is probably "how many gray levels can one see under what circumstances". The answer is probably very different for a static image and a dynamic image, with edges or without, in low light conditions or bright light conditions.
The human eye (and brain) has had Millions of years to evolve and develop some rather sophisticated algorithms to deal with different conditions and the answer is probably also very complex. For example, our vision is extremely good at detecting movement. On a completely static image this tends to depress differences, while on a dynamic image it tends to enhance differences (we automatically focus on the changes).
Maybe you can test that yourself:
take the same image on a computer at different bit depths (=levels of gray). Then randomly put them on the screen (without watching the change) and try to decide, which image it is. Then do the same but watch the changes and try to decide how many gray levels you need before the changeover becomes invisible. You could do this with a "noise" image, an image that shows some object, and a smooth gray wedge. I for one would be interested to hear about the results.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
} ---------- } From: "DrJohnRuss-at-aol.com"-at-sparc5.Microscopy.Com[SMTP:"DRJOHNRUSS-at-AOL.COM"-at-SPA RC5.MICROSCOPY.COM] } Sent: Wednesday, February 02, 2000 5:56:04 PM } To: Microscopy-at-sparc5.Microscopy.Com; rgriffin-at-eng.uab.edu } Subject: Re: Gray level } Auto forwarded by a Rule } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
In a message dated 2/2/00 4:00:26 PM, rgriffin-at-eng.uab.edu-at-sparc5.Microscopy.Com writes:
} How many gray levels can the human eye distinguish? We've found several } } references that disagree. } } If anyone knows of a reference - that would be best.
Most of the books on image processing say the same thing (probably all copied from each other and other non-prime sources), but I don't know of an original reference based on real research. You can find the same ideas in books like Frisby "Seeing: illusion, brain and mind", Oxford, 1980; Marr "Vision",
Freeman, 1982; and Rock "Perception", Freeman, 1984 which deal with the
human visual system not in the context of computer graphics. These are probably closer to the original research.
The basic idea is that it takes about a 2-2.5% change in absolute brightness (i.e. a ratio of 1.025) to be just detectable, and only if it is a fairly sudden change spatially (or temporally for that matter). That is where the 20-30 distinct brightness levels over the full range of brightness that can be seen at one time (i.e. without accommodations like varying pupil size) comes from. The requirement that an entire scene have about 100 levels to avoid banding is more controversial, based on the idea that the eye can perform some accommodation as it traverses from one part of a scene to another. This probably doesn't apply when (e.g.) looking through a microscope, so the 20-30 number would be more applicable. It also ignores color, which complicates things further.
Most industrial cameras use C-mounts. C-mount to camera lens adapters are available from Edmund Industrial Optics (http://www.edmundoptics.com phone:800-363-1992) on page 165 of their 2000 catalog. They have mounts for several types of cameras, and list for $62 each. If your connecting the camera to a 'scope, Edmund sells a relay lens for $240 which mounts the camera to a 23mm eyepiece tube. For connecting to trinocular and CCD ports on a scope, Diagnostic Instruments sells relay lenses for many types of 'scopes (http://www.diaginc.com) for $299.
Michael Mohn Assistant Professor of Materials Technology Monroe County Community College Phone: 734-384-4122 Fax: 734-242-9711 http://www.monroe.cc.mi.us/mmohn
} } } "Karl Hagglund-KW" by way of Nestor J. Zaluzec {hagglund.kw-at-pg.com} 02/02 6:06 PM } } } We have a sony DXC-760 MD ccd camera that we have mounted to a standard camera lens with a bayonet adapter. Unfortunately, we have borrowed the adapter and have to return it. Is there a standard size (i.e. Nikon, Minolta, Canon) lens adapter that works with this camera mount? We can't find a part number or description for the adapter, and the book doesn't describe the size. We have already ordered and returned one that didn't fit.
Thanks in advance.
Karl Hagglund Proctor and Gamble Food and Beverage Analytical Microbiology
Are you equating brightness levels with grey levels here? I was told something like this years ago when we were buying our first digital B&W camera, when I asked the vendor about the competitors thousand plus grey levels vs his 256 grey levels and was told that the eye could not distinguish greater levels. Now, four or five years later everyone is pitching 10, 12 and 16? bit cameras, and yet the images from these cameras do look better. Is it the bits and increased grey levels? I know that if your doing image analysis at the pixel level that more bits are to your advantage but what about just photgraphic quality?
Sorry, but I think we are getting a little confused here.
1) The diffraction plane is (by definition) the objective lens focal length below the sample - it is the focus of parallel directions coming from the sample. The focal length (and parameters such as Cs, Cc) change rapidly with the objective lens current so an eucentric position makes life simple but is not necessarily the best condition to use. (For HREM you generaly want to have the objective lens stronger and reduce Cs.)
2) In general C2 does not change the focal length at all. You can have some effect from coupling of the magnetic fields, but all C2 is supposed to do is change the convergence angle.
3) The objective aperature is (approximately) in the back-focal plane, given that it is finite and height adjustments are only done coarsely (for a specific value of the objective current only).
4) The selected area aperature is (approximately) in the same plane as the object for the same reasons as above. Remember that there are positional errors in SA diffraction due to Cs (see Hirsch et al for instance).
5) You should focus Kikuchi-lines, not go for the smallest spot, since these are real object in the diffraction pattern. What you will see is then representative of you incident beam, sometimes a Gaussian range of directions.
6) The simplest way to get more parallel illumination in general is to reduce the condensor aperture size.
7) All bets are off if you have a FEG. Rather than having incoherent illumination things like the Cs of the prefield (above the sample) and postfield (below) add coherently making it much more complicated. For instance, the illumination angle varies across the field of view. While this is also there with LaB6, it is a weak effect (except for certain classes of diffracted beams, e.g. those with a screw-axis). With a FEG you can get a "spot" pattern many ways, and most of these will have large aberrations (e.g. pincushion).
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University fax: (847) 491-7820 mailto:l-marks-at-nwu.edu http://www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
Eugenia, the microprobe is not the best tool for the analysis of ammonia in zeolites because ammonia is very volatile in the electron beam. If your zeolite is phase pure it would be far better to dissolve it and do a classical chemical analysis.
regards, Eric On Thu, 3 Feb 2000 14:54:20 +0100 bengi {bengi-at-unimo.it} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } Dear listers, } I have started a research project on ammonium -bearing mineral (zeolites), } and now I'm trying to quantify N using electron microprobe analysis. My } problems of dealing with N are: } } a) The thickness of the coating (in the literateur = approximately 150 A°). } Does anyone have information on the model of coater to making these films? } Does anyone have experience with preparation of such } samples? } } b) Obtaining appropriate standards. (I have Si3N4, but it contains too much } N (=39.96%) towards my samples (5%). Does anyone known any good nitrogen } standards for such a sample ? } } Any other suggestions about quantitative nitrogen analysis would be greatly } appreciated !! } Thanks for your time. } } My best regards } } Eugenia } } } Eugenia Marchi } Università degli Studi di Modena } Dip. Scienze della Terra } P.le S.Eufemia n°.19 } 41100 Modena (Italy) } Tel. +39-059-417289 } Fax. +39-059-417399 } e-mail: bengi-at-unimo.it } } }
---------------------- Dr Eric Lachowski Department of Chemistry University of Aberdeen Meston Walk Old Aberdeen AB24 3UE Scotland +44 (0)1224 272934 fax 272921 e.lachowski-at-abdn.ac.uk
I need somebody to repair a Hitachi SEM S-2700. Please contact me individually not the list. Thanks for help, -- L.Paul Bédard, ing. Ph.D. Resp. Lab. Géochimie | Lab. Manager Sciences Appliquées ; Université du Québec à Chicoutimi Canada Tél. : 418/545-5011 x 2276 | Alt. E-mail : pbedard-at-saglac.qc.ca {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} La période humaine, depuis la création d'Adam jusqu'à nos jours, n'est qu'une moisissure dans l'histoire de notre planète JCK Laflamme 1886
Let me offer my guess that the improved appearance of the images is be due largely to the improved signal-to-noise ratio in the newer cameras.
Of course there is also something to be gained in the dynamic range by the extra bits of resolution. They are much more forgiving and allow substantial changes in contrast and brightness without sacrificing the information contained in the data.
Warren S.
At 09:39 AM 2/3/2000 -0600, you wrote:
} Are you equating brightness levels with grey levels here? } I was told something like this years ago when we were buying our first } digital B&W camera, when I asked the vendor about the competitors thousand } plus grey levels vs his 256 grey levels and was told that the eye could not } distinguish greater levels. Now, four or five years later everyone is } pitching 10, 12 and 16? bit cameras, and yet the images from these cameras } do look better. Is it the bits and increased grey levels? I know that if } your doing image analysis at the pixel level that more bits are to your } advantage but what about just photgraphic quality? } } Rick Vaughn
Dear Colleagues, Has anyone used gold enhancement reagents from Nanoprobes, Inc. (Stonybrook, NY - USA) as an alternative to silver enhancement for enlarging 1 nm immunogold probes?
Advertisements state that gold enhancement has lower background and is compatible with osmium. Can anyone verify this from personal experience?
In addition to these advantages, I am hoping that milder pH conditions will be less damaging to ultrastructure in cultured neurons (using a preembedment protocol).
} The diffraction plane is always the crossover plane not the theoretical back } focal plane of the objective. Its z-position changes with the change of the } C2 excitement.
Your statement above is not wrong, but isn't the standard TEM viewpoint (or terminology). Yes, the plane at which the spot is sharpest (the in focus plane for the demagnified filament image) is the crossover plane. However to generally call this the "diffraction plane" is confusing. For example in the case of CBED, the crossover plane has moved (up or down, depending on whether C2 was initially under or overfocused) to the specimen plane. This doesn't make the specimen plane the "diffraction plane". Rather, the "diffraction plane" is now considered the in-focus plane for the C2 aperture. This is at the objective lens back-focal plane - the plane at which points on the specimen are magnified to infinity and where the position variable corresponds to the illumination angle. In collecting an SADP you can correct for slightly non-parallel illumination by correcting diffraction focus slightly off of the true back focal plane, but this should be only slight.
What was proposed in the original mail was a technique of determining how parallel the incident beam is, and of improving things by tweaking C2. If the beam isn't very parallel, you'll see the diffraction spots shift when you move the SA aperture. This is because the incident beam inclination depends on the position on the sample.
I would think the "tweak" to C2 for obtaining more parallel illumination would always in the direction of greater beam spread (unless I am misunderstanding something?).
Regards, Wharton ++++++++++++++++++++++++++++++++++++++++++++++++++ Wharton Sinkler Argonne National Laboratory West P. O. Box 2528 Idaho Falls, ID 83403
} Rado } } --------------------------------------------------------------------- } Radostin Danev } Laboratory of Ultrastructure Research } National Institute for Physiological Sciences } Myodaiji-cho, Okazaki 444-8585, JAPAN } e-mail: rado-at-nips.ac.jp } --------------------------------------------------------------------- } ----- Original Message ----- } } From: Jonathan Barnard {Jonathan.Barnard-at-Angstrom.UU.SE} } To: {microscopy-at-sparc5.microscopy.com} } Sent: Wednesday, February 02, 2000 7:20 PM } Subject: TEM:finding the parallel beam } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } I got a tip for finding the condenser lens settings required for parallel } } illumination from Angus Kirkland (Cambridge). } } } } Make sure you are in microprobe mode and set up for SAED and the specimen } } is out of the way. Spread the beam, put in the SA Aperture, go to } } diffraction and then sweep the SA aperture (SAA) from side to side and } } minimise the deflection of the diffraction spot by tweaking the } } illumination control (C2 or second condenser lens). A little bit of } thought } } will convince you that anything other than a parallel beam will give you a } } side to side motion (tilt) when the SAA is swept. } } } } I have found it useful to keep a table of C2 condenser lens current } } settings for each spot size (C1 excitation) in microprobe mode. Setting } the } } C2 lens for parallel illumination and adjusting the diffraction focus to } } get the sharpest spot will then give you near perfect SAD conditions. } } } } Anyone contemplating electron holography for example, will have to start } } from the parallel illumination to get the best spatial coherence for their } } holograms. } } } } ******************************************************** } } Dr Jonathan Barnard } } } } Analytical Materials Physics } } The Angstrom Laboratory, Uppsala University } } P O Box 534, SE-751 21 Uppsala, Sweden } } Phone: +46-(0)18-4716838 Fax: +46-(0)18-500131 } } http://www.angstrom.uu.se/analytical/home.html } } } } ******************************************************** } } } } }
You can avoid your concern with C coating by coating your standards and unknowns at the same time. You should use the minimum coating thickness--just enough to prevent beam charging.
However, as stated below by Dr. Lachowski, ammonia volatility would be a big problem, especially since you will need help from high beam intensities and/or long counting times to get enough N counts to give decent precision. The expected intensity measured with a 10kV beam from a zeolite nominally of analcime composition (NaAlSi2O6.H2O) with 5 wt% N would be about 2.5% that of the intensity from Si3N4. High N intensity is difficult to achieve even with the best spectrometer and crystal designs. And to add insult to injury, the addition of the C coating absorbs even more of the N x-rays from the sample. The mass absorption coefficient for N K-alpha emission with a C absorber is huge (greater than 23,000).
Plus, you should check for possible N peak shape differences between Si3N4 (or other standards) and your ammonium-bearing zeolite.
All in all, it is a very difficult analytical problem.
Good luck!
Carl
At 4:24 PM +0000 2/3/00, Dr Eric Lachowski wrote: } Eugenia, } the microprobe is not the best tool for the analysis of } ammonia in zeolites because ammonia is very volatile in the } electron beam. If your zeolite is phase pure it would be } far better to dissolve it and do a classical chemical } analysis. } } regards, } Eric } On Thu, 3 Feb 2000 14:54:20 +0100 bengi {bengi-at-unimo.it} } wrote: } } } } } Dear listers, } } I have started a research project on ammonium -bearing mineral (zeolites), } } and now I'm trying to quantify N using electron microprobe analysis. My } } problems of dealing with N are: } } } } a) The thickness of the coating (in the literateur = } approximately 150 A°). } } Does anyone have information on the model of coater to making these films? } } Does anyone have experience with preparation of such } } samples? } } } } b) Obtaining appropriate standards. (I have Si3N4, but it contains too much } } N (=39.96%) towards my samples (5%). Does anyone known any good nitrogen } } standards for such a sample ? } } } } Any other suggestions about quantitative nitrogen analysis would be greatly } } appreciated !! } } Thanks for your time. } } } } My best regards } } } } Eugenia } } } } } } Eugenia Marchi } } Università degli Studi di Modena } } Dip. Scienze della Terra } } P.le S.Eufemia n°.19 } } 41100 Modena (Italy) } } Tel. +39-059-417289 } } Fax. +39-059-417399 } } e-mail: bengi-at-unimo.it } } } } } } } } ---------------------- } Dr Eric Lachowski } Department of Chemistry } University of Aberdeen } Meston Walk } Old Aberdeen AB24 3UE } Scotland } +44 (0)1224 272934 fax 272921 } e.lachowski-at-abdn.ac.uk
====================================== Carl Henderson Electron Microbeam Analysis Laboratory University of Michigan 2501 C.C. Little Bldg. Ann Arbor, MI 48109-1063 USA (734) 936-1550 FAX (734) 763-4690 ======================================
Does anyone out there have experience with the Technoorg-Linda IV3H ion beam thinner. We have their instruction manual but it has been a long time since we had a training session and are having some problem with the set up. Does anyone have a revised instruction manual or perhaps some tips on the alignment procedure of the guns. (please please please please).
Thanks Paul Nolan
P.S.. It is cold in Canada..everyday...all year. hehe
Now I'm really confused. I used to think I understood this somewhat.
1. In the JEOL 2010F FEG (the "all bets are off" case in your item#7), changing the illumination spread by changing C2 profoundly moves the "diffraction plane" relative to the objective aperture. [There are actually two objective apertures at different heights in this microscope, but that's beside the point]. By diffraction plane, I mean the plane below the objective lens where the diffraction spots are in sharp focus. In other words (assuming fixed OL and condenser minilens excitations), in this microscope there is only one C2 value that allows the diffraction spot pattern and OL aperture to be in focus simultaneously. If you focus on the OL aperture (with the intermediate lens), the spot pattern can be focused at only this one C2 setting. At any other C2 setting, you can focus the diffraction pattern with the intermediate lens ("diffraction focus") but the OL aperture will then be defocused. Changing the condenser minilens excitation changes the diffraction plane (relative to the OL aperture) to a different C2 setting. This is actually quite a useful feature of the microscope, e.g., for getting very intense focused diffraction patterns, but a different story.
2. The reason for choosing to focus the diffraction spots rather than the K-lines is --to put it simply-- that's where the intensity is. In amplitude contrast imaging (conventional brightfield and darkfield imaging), the intensity contribution to the image is limited by the OL aperture. It's really important to have the defining aperture and diffraction spot pattern in focus simultaneously. In addition, if I aim to measure diffraction spot patterns there is little incentive to focus the K-lines instead of the spots.
3. Textbooks such as Hirsch et al. often discuss the positional error in SA diffraction due to Cs. What readers often miss is the consequence of the fact that the error depends strongly on the diffraction angle. If measurements are confined to the low-index (low-angle) reflections, the selecting area can be reduced (to 200 nm or so at 200 kV) without big positional errors. Maybe that's an example of differences in experimental and theoretical viewpoints.
4. Reducing the condensor aperture size will give less illumination, but it won't give parallel illumination. It will change the size of the diffraction spots, but not their focus.
5. I'm not quite sure why the illumination spread has such a strong effect on the apparent position of the diffraction plane relative to the OL aperture, but suspect it arises from the action of the strong condensor-objective in this microscope. I would also question that it has much to do with coherent/incoherent beams in a FEG or with prefield/postfield Cs effects.
Larry
Larry Thomas Pacific Northwest National Laboratory MSIN P8-16 P.O. Box 999 Richland, WA 99352 Phone: (509)372-0793 Fax: (509)376-6308 Email: mailto: Larry.Thomas-at-pnl.gov
---------- From: L. D. Marks Reply To: L-marks-at-nwu.edu Sent: Thursday, February 3, 2000 8:06 AM To: Radostin Danev Cc: MSA listserver Subject: Re: TEM:finding the parallel beam
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html -----------------------------------------------------------------------.
Sorry, but I think we are getting a little confused here.
1) The diffraction plane is (by definition) the objective lens focal length below the sample - it is the focus of parallel directions coming from the sample. The focal length (and parameters such as Cs, Cc) change rapidly with the objective lens current so an eucentric position makes life simple but is not necessarily the best condition to use. (For HREM you generaly want to have the objective lens stronger and reduce Cs.)
2) In general C2 does not change the focal length at all. You can have some effect from coupling of the magnetic fields, but all C2 is supposed to do is change the convergence angle.
3) The objective aperature is (approximately) in the back-focal plane, given that it is finite and height adjustments are only done coarsely (for a specific value of the objective current only).
4) The selected area aperature is (approximately) in the same plane as the object for the same reasons as above. Remember that there are positional errors in SA diffraction due to Cs (see Hirsch et al for instance).
5) You should focus Kikuchi-lines, not go for the smallest spot, since these are real object in the diffraction pattern. What you will see is then representative of you incident beam, sometimes a Gaussian range of directions.
6) The simplest way to get more parallel illumination in general is to reduce the condensor aperture size.
7) All bets are off if you have a FEG. Rather than having incoherent illumination things like the Cs of the prefield (above the sample) and postfield (below) add coherently making it much more complicated. For instance, the illumination angle varies across the field of view. While this is also there with LaB6, it is a weak effect (except for certain classes of diffracted beams, e.g. those with a screw-axis). With a FEG you can get a "spot" pattern many ways, and most of these will have large aberrations (e.g. pincushion).
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University fax: (847) 491-7820 mailto:l-marks-at-nwu.edu http://www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
South Bay Technology does have quite a bit of experience with the TL IV3 Ion Mill and we are happy to offer our assistance. We market the IV3 system throughout the world for Technoorg-Linda and offer technical support as well. I will send you the most up to date IV3 manual that we have produced and will include by separate e-mail the section on the gun alignment.
I will follow up with you to be sure that you have all of the information you need.
David Henriks Vice President TEL: 800-728-2233 (toll free in the USA) South Bay Technology, Inc. +1-949-492-2600 1120 Via Callejon FAX: +1-949-492-1499 San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by INTERNET:"Paul.Nolan-at-Alcan.Com"-at-sparc5.Microscopy.Com } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Does anyone out there have experience with the Technoorg-Linda IV3H ion beam thinner. We have their instruction manual but it has been a long time since we had a training session and are having some problem with the set up. Does anyone have a revised instruction manual or perhaps some tips on the alignment procedure of the guns. (please please please please).
Thanks Paul Nolan
P.S.. It is cold in Canada..everyday...all year. hehe {
} Laurie, } } Now I'm really confused. I used to think I understood this somewhat. } } 1. In the JEOL 2010F FEG (the "all bets are off" case in your item#7), } changing the illumination spread by changing C2 profoundly moves the } "diffraction plane" relative to the objective aperture. [There are actually two } objective apertures at different heights in this microscope, but that's beside } the point]. By diffraction plane, I mean the plane below the objective lens } where the diffraction spots are in sharp focus. In other words (assuming fixed } OL and condenser minilens excitations), in this microscope there is only one C2 } value that allows the diffraction spot pattern and OL aperture to be in focus } simultaneously. If you focus on the OL aperture (with the intermediate lens), } the spot pattern can be focused at only this one C2 setting. At any other C2 } setting, you can focus the diffraction pattern with the intermediate lens } ("diffraction focus") but the OL aperture will then be defocused. Changing the } condenser minilens excitation changes the diffraction plane (relative to the OL } aperture) to a different C2 setting. This is actually quite a useful feature of } the microscope, e.g., for getting very intense focused diffraction patterns, but } a different story. } OK, the bets off case. In a FEG you have a somewhat coherent range of incident directions. The "diffraction pattern" is therefore something which is effected by the coherent aberrations of the microscope, unlike the case with incoherent illumination. Consider the case with focussed illumination, when the diffraction pattern shows an image of the condensor aperture. In a FEG As a you can "focus" this (coherent) image to a small spot by going out of focus in a true sense for the diffraction pattern. This gives you a spot-like pattern, but you will also see severe distortions at higher angles. With incoherent illumination you cannot do this and going out of focus (in the diffraction pattern) will give in general a blurred image of the condensor aperture - life is simple. What you need to do is focus "real diffraction" features, e.g. Kikuchi lines, then not play with the diffraction focus at all. As you change C2 the size of the spots will grow or shrink, this is fine.
} 2. The reason for choosing to focus the diffraction spots rather than } the K-lines is --to put it simply-- that's where the intensity is. In amplitude } contrast imaging (conventional brightfield and darkfield imaging), the intensity } contribution to the image is limited by the OL aperture. It's really important } to have the defining aperture and diffraction spot pattern in focus } simultaneously. In addition, if I aim to measure diffraction spot patterns } there is little incentive to focus the K-lines instead of the spots. } Yes, you can always get a "spot" pattern with a FEG, and it may be prettier. However, beware the distortions which make doing measurements from it very dangerous.
} 3. Textbooks such as Hirsch et al. often discuss the positional error } in SA diffraction due to Cs. What readers often miss is the consequence of the } fact that the error depends strongly on the diffraction angle. If measurements } are confined to the low-index (low-angle) reflections, the selecting area can be } reduced (to 200 nm or so at 200 kV) without big positional errors. Maybe that's } an example of differences in experimental and theoretical viewpoints. } Hirsch et al is just a good reference to read, about this as well as many other things!
} 4. Reducing the condensor aperture size will give less illumination, } but it won't give parallel illumination. It will change the size of the } diffraction spots, but not their focus. } It makes it closer to parallel. Of course, with really parallel illumination there is no intensity!
} 5. I'm not quite sure why the illumination spread has such a strong } effect on the apparent position of the diffraction plane relative to the OL } aperture, but suspect it arises from the action of the strong } condensor-objective in this microscope. I would also question that it has much } to do with coherent/incoherent beams in a FEG or with prefield/postfield Cs } effects. } Sorry, I don't think that is correct. I would suggested that people look at the distortions with a standard sample and see how bad they can be.
} Larry } } } Larry Thomas } Pacific Northwest National Laboratory } MSIN P8-16 } P.O. Box 999 } Richland, WA 99352 } Phone: (509)372-0793 Fax: (509)376-6308 } Email: mailto: Larry.Thomas-at-pnl.gov } } } } ---------- } From: L. D. Marks } Reply To: L-marks-at-nwu.edu } Sent: Thursday, February 3, 2000 8:06 AM } To: Radostin Danev } Cc: MSA listserver } Subject: Re: TEM:finding the parallel beam } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Sorry, but I think we are getting a little confused here. } } 1) The diffraction plane is (by definition) the objective lens } focal length below the sample - it is the focus of parallel } directions coming from the sample. The focal length (and } parameters such as Cs, Cc) change rapidly with the objective lens } current so an eucentric position makes life simple but is not } necessarily the best condition to use. (For HREM you generaly } want to have the objective lens stronger and reduce Cs.) } } 2) In general C2 does not change the focal length at all. You } can have some effect from coupling of the magnetic fields, but } all C2 is supposed to do is change the convergence angle. } } 3) The objective aperature is (approximately) in the back-focal } plane, given that it is finite and height adjustments are only } done coarsely (for a specific value of the objective current } only). } } 4) The selected area aperature is (approximately) in the same } plane as the object for the same reasons as above. Remember that } there are positional errors in SA diffraction due to Cs (see } Hirsch et al for instance). } } 5) You should focus Kikuchi-lines, not go for the smallest spot, } since these are real object in the diffraction pattern. What you } will see is then representative of you incident beam, sometimes } a Gaussian range of directions. } } 6) The simplest way to get more parallel illumination in general is } to reduce the condensor aperture size. } } 7) All bets are off if you have a FEG. Rather than having incoherent } illumination things like the Cs of the prefield (above the sample) } and postfield (below) add coherently making it much more complicated. } For instance, the illumination angle varies across the field of view. } While this is also there with LaB6, it is a weak effect (except for } certain classes of diffracted beams, e.g. those with a screw-axis). } With } a FEG you can get a "spot" pattern many ways, and most of these will } have } large aberrations (e.g. pincushion). } } ++++++++++++++++++++++++++++++++++++++++++++++++ } Laurence Marks } Department of Materials Science and Engineering } Northwestern University } fax: (847) 491-7820 } mailto:l-marks-at-nwu.edu } http://www.numis.nwu.edu } ++++++++++++++++++++++++++++++++++++++++++++++++ } } }
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University fax: (847) 491-7820 mailto:L-marks-at-nwu.edu http://www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
Greetings, Butyl methyl methacrylate is in our hands non fluorescent totally. You can use it for TEM, although it is not as good as epoxies. Hope this helps. Tobias } } } I am working on fluorescent imaging of thin sections cut from samples } embedded in TEM embedding resin, and am having a significant problem with } resin fluorescence. Does anyone know of an embedding resin that doesn't } autofluoresce? } } Matt Plantinga } Amway Corporation } matt_plantinga-at-amway.com
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Does anyone know of a current source for this chemical? It's an ingredient of an embedding medium made up by our group. We have a supply on hand but have to reserve it if it's no longer commercially available.
} } We are very excited about the possibilities of HACH as a solution for a } } problem } } that has troubled us for a couple of years with our PU grafts. We have, } } unfortunately, one serious problem in that we cannot find a source for HA. } } We have tried Sigma Aldrich and they informed us that they withdrew } } 2-Hydroxyhexane-dial from their range in 1990. We have tried a web search } } and are contacting our local suppliers, so far without success. Do you have } } any ideas where we can find it? We are very anxious to try HACH on our PU } } samples and will gratefully appreciate any additional help that you can give } } us.
Have I been cut off, or is nobody talking anymore?
-- Dr MP Roberts Phone: [+27](0)46 603 8313 Dept of Geology Fax: [+27](0)46 622 9715 Rhodes University e-mail: malc-at-rock.ru.ac.za 6140 Grahamstown SOUTH AFRICA
I have tried GoldEnhance for preembedding protocol to label intracellular structures. It is marvelous. Buy it and use it - you will not be unsatisfied. I have no any interest in Nanoprobes, I just like these dense gold particles. Practically all they claim is true: - light insensitive - no self-nucleation - do not react with buffer ions - is not dissolved by uranil and osmium (I have treat for 1 h)
Sincerely, Dr. Alexander A. Mironov Jr., MD, PhD Unit of Morphology Dept. of Cell Biology and Oncology Consorzio Mario Negri Sud Via Nazionale, S.Maria Imbaro (Ch) 66030 Italy
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Colleagues, } Has anyone used gold enhancement reagents from Nanoprobes, Inc. } (Stonybrook, NY - USA) as an alternative to silver enhancement for } enlarging 1 nm immunogold probes? } } Advertisements state that gold enhancement has lower background and is } compatible with osmium. Can anyone verify this from personal experience? } } In addition to these advantages, I am hoping that milder pH conditions will } be less damaging to ultrastructure in cultured neurons (using a } preembedment protocol). } } Thank you, } Michael } }
The comparison between Zeiss and Olympus '100x' objective lenses is impossible unless one knows the type of lens be compared.
Catagories used to determine the quality of such a lens include: aberration correction (spherical and chromatic), correction of flatness of field, numerical aperture, the presence of an iris diaphram, whether the system is based on a 160mm tube length or infinity correction, and if the lens is used for brightfield or phase contrast for instance.
Resolution is still basically related to half the wavelength utilized. Hey, let's look at the advantages of oblique illumination for contrast enhancement and a differerent formula for resolution!!
Is your client looking at a system or an indivdual lens? Do they have a Zeiss or an Olympus system? One does not mixed and match objectives from different systems merely for the convenience of pricing.
An additional consideration is the type and correction of the condenser lens. Check the numerical aperture of not only the objective, but the condenser lens as well.
More questions than answers...Let me know if I can further complicate the answers!
Cheers! Ken
------------ Ken Tiekotter Dept. of Biology The University of Portland 5000 N Willamette Blvd. Portland, OR 97303
On Thu, 3 Feb 2000, Steve Niemela wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } To: {Microscopy-at-MSA.Microscopy.Com} } Subject: A comparison of optics } Date: Thu, 3 Feb 2000 10:57:19 -0600 } MIME-Version: 1.0 } Content-Type: multipart/alternative; } boundary="----=_NextPart_000_001F_01BF6E35.70E8F2E0" } X-Priority: 3 } X-MSMail-Priority: Normal } X-Mailer: Microsoft Outlook Express 5.00.2615.200 } X-MimeOLE: Produced By Microsoft MimeOLE V5.00.2615.200 } } A client of ours wonders about a comparison between Zeiss and Olympus } optics. The Zeiss 100X lense cost $10,000, and the Olympus about $4,000: } does that mean the Olympus is inferior? That's what our client's } colleagues imply. What measure of lense function is relevant? Does Olympus } (or Zeiss) have a specification, like 'resolves particles to 0.1 micron' } or something like that? Is there a measure of distortion? Best } regards, Steve NiemelaVayTek, Inc.305 West Lowe AvenueFairfield, IA } 52556email: sniemela-at-vaytek.com Phone: 515.472.2227Fax: 515.472.8131web: } www.vaytek.comgeneral email: vaytek-at-vaytek.com } } } }
} We are making preparations for experiments on determination of the chemical } composition of semiconductor oxides, formed locally. The size of the } modified area should be about 1 micron, and we are not sure yet if it is } possible to make mentioned chemical analysis with Auger microprobe at all.
I can recommend you some former colleagues of mine performing auger spectroscopy and many other semiconductor surface analytics. Please contact Prof. Wolfgang Richter: richter-at-pinet.uni-jena.de. The URL is: http://www.physik.uni-jena.de/~layer/
W'ere all waiting to hear news of MSSA Conference 2000 { :-)
Tony
} } } Dr Malcolm Roberts {malc-at-rock.ru.ac.za} 02/04/00 09:04AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Have I been cut off, or is nobody talking anymore?
-- Dr MP Roberts Phone: [+27](0)46 603 8313 Dept of Geology Fax: [+27](0)46 622 9715 Rhodes University e-mail: malc-at-rock.ru.ac.za 6140 Grahamstown SOUTH AFRICA
} Does anyone out there have experience with the Technoorg-Linda IV3H ion beam } thinner. } We have their instruction manual but it has been a long time since we had a } training session and are having some problem with the set up. } Does anyone have a revised instruction manual or perhaps some tips on the } alignment procedure of the guns. (please please please please).
We have the IV3 here in Sheffield - I'll try and help you if I can. You need to be able to actually see the beams (cover your head and the viewing glass with a black sheet if you cannot darken the room). Get one gun to fire through the center of the sample holder (tilt it at right angles to the beam) and to hit the opposite gun where it's beam would exit. Do the same for the other gun. Now keep this gun orientation, load the specimen and tilt the holder to the desired milling angle.
Hope this helps,
Alan Walker
********************************************* Alan Walker Dept of Electronic and Electrical Engineering University of Sheffield Mappin Street Sheffield S1 3JD United Kingdom
A Kevex 8000 (with huge 8 or 10" disk drives), attached to an old Amray 1000, has been donated to us. All components were fully functional when they were crated up a year ago. (They are sitting in our basement until we move into our new building this June).
A colleague in Physics (I'm a biologist) wishes to use the system to quantify elemental composition (beryllium, gallium, etc) in semi-conductor ceramics. We already have (up and running) an Hitachi HS-510 (SEM) and an Hitachi H-7100 (STEM).
1) Is this (Kevex 8000) an appropriate instrument for her to do the analyses?
2) Which would be wiser: Trying to get the Amray installed and running or getting an adapter to use the Kevex in one of the Hitachi's? (Space is a consideration, so I'd prefer not to have to install (and keep in running condition) another scope.)
3) The Kevex is pretty old. Are we wasting money trying to get or keep running something that is this old? I realize that we may need to replace the detector, since it has been at room temp for about a year. Is there anything else we should automatically consider replacing or upgrading as part of the installation?
4) She also has been sending her samples out to do x-ray analysis of the material to determine it's crystalline structure. The place she sends it to has a special x-ray diffraction instrument. Could the x-ray diffraction on the H-7100 be used to get equal results, or does this other instrument have capabilities that the TEM doesn't.
Thanks for any advice.
Don ______________________________________________________________________ Donald L. Lovett e-mail: lovett-at-tcnj.edu Assoc. Professor, Dept. of Biology voice: (609) 771-2876 P.O. Box 7718 fax: (609) 637-5118 The College of New Jersey Ewing, NJ 08628-0718
The cost of an objective depends on a number of issues, not the least of which are the corrections for chromatic and spherical aberration as well as the size of the numerical aperture. I'd need more information to really compare. "Optimizing Light Microscopy" has a detailed discussion of all of this which might be helpful. Ordering information is available on our website.
The best test is to try each system with your applications. Since much of Vaytek's work centers on deconvolution of fluorescence images, fluorescent beads of varying sizes would be a good test object for you and high throughput and crisp imaging would probably be two key benchmarks. Since fluorescence detects objects beyond the resolution limits, the normal resolution tests are meaningless. By the way, the numerical aperture of the objective is the major deciding factor on a microscope's ability to resolve fine detail (as well as providing good edge information).
If you'd like to write or call me directly, I may be able to give you further guidance.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education}
Caveat: MME does have a commercial interest in the sale of "Optimizing Light MIcroscopy:"
At 04:04 PM 2/3/00 -0600, Steve Niemela wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
If your Kevex has a Quantum thin window detector you should be able to see about half of a Boron peak, but there is no commercially available detector (that I know of) that reliably sees all of the beryllium peak. And if you have a standard window, you will only be able to see sodium and above.
I suppose the detector may still be functional even after storage. I recall that it is not a trivial thing to switch detectors between scope models. The adapters can be quite different and can get pricey. But I have not priced one in over 10 years.
I suppose with rigorous use of standards, you might be able to analyze Be by difference, but it would be a trick.
I think the Kevex 8000 may be worth installing if you are primarily interested in x-ray analysis. We used a Kevex Delta for many years and found it quite adequate. I might be able to talk you through some of the technical issues.
However, if you are interested in digital imaging or x-ray mapping, the improvements in the new equipment is fantastic compared to the old systems. We run an IXRF Systems EDS on a JEOL 840A (having replaced our Kevex) and a Link ISIS on a Hitachi 2460N. I could hardly imagine giving up their capabilities and going back to the Delta or an 8000. Disk storage, the operating environment, digital imaging can hardly be compared between the old and the new.
FWIW, Warren
At 09:17 AM 2/4/2000 -0500, Donald L. Lovett wrote: } A Kevex 8000 (with huge 8 or 10" disk drives), attached to an old Amray } 1000, has been donated to us. All components were fully functional when } they were crated up a year ago. (They are sitting in our basement until we } move into our new building this June). } } A colleague in Physics (I'm a biologist) wishes to use the system to } quantify elemental composition (beryllium, gallium, etc) in semi-conductor } ceramics. We already have (up and running) an Hitachi HS-510 (SEM) and an } Hitachi H-7100 (STEM). } } 1) Is this (Kevex 8000) an appropriate instrument for her to do the } analyses? } } 2) Which would be wiser: Trying to get the Amray installed and running } or getting an adapter to use the Kevex in one of the Hitachi's? (Space is } a consideration, so I'd prefer not to have to install (and keep in running } condition) another scope.) } } 3) The Kevex is pretty old. Are we wasting money trying to get or keep } running something that is this old? I realize that we may need to replace } the detector, since it has been at room temp for about a year. Is there } anything else we should automatically consider replacing or upgrading as } part of the installation? } } 4) She also has been sending her samples out to do x-ray analysis of the } material to determine it's crystalline structure. The place she sends it } to has a special x-ray diffraction instrument. Could the x-ray } diffraction on the H-7100 be used to get equal results, or does this other } instrument have capabilities that the TEM doesn't. } } Thanks for any advice. } } Don
I responded to Eugenia's query on another listserver (MAS microprobe listserver), but since it is also discussed here, I will add a couple things.
I did some work on ammonium-bearing minerals (with ~ 3 - 5 wt%N), using BN and later synthetic buddingtonite (ammonium feldspar) as the N standard (see Microbeam Analysis 1994, New Orleans meeting). I avoided using any epoxy for the sample prep as a precaution against N artifacts from the mounting medium. The synthetic buddingtonite can be made ala Voncken et al., 1988 (Phys. Chem of Minerals, v. 15, p. 323).
We did "normal" carbon coating, using brass color as a guide to obtain 150-200A coating (standards and unknowns). I used a synthetic crystal, 2d 60A, and ran at 10kV, 25 nA probe current. I agree that a light C coat, done at same time/conditions as the standards is important. I don't recall a substantial problem with mobility at thnese operating conditions, although the N intensity yields were low and detection limits were on the order of 0.5wt% N. The minerals are feldpar and perhaps the ammonium ion stayed put better than it woud in zeolites. I agree though, it is a difficult analytical problem. But that makes it interesting.
Jim -- ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ James J. McGee (email: jmcgee-at-sc.edu) Department of Geological Sciences University of South Carolina Columbia, SC 29208
Tel: 803-777-6300 Fax: 803-777-6610
Carl Henderson wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Eugenia, } } You can avoid your concern with C coating by coating your standards } and unknowns at the same time. You should use the minimum coating } thickness--just enough to prevent beam charging. } } However, as stated below by Dr. Lachowski, ammonia volatility would } be a big problem, especially since you will need help from high beam } intensities and/or long counting times to get enough N counts to give } decent precision. The expected intensity measured with a 10kV beam } from a zeolite nominally of analcime composition (NaAlSi2O6.H2O) with } 5 wt% N would be about 2.5% that of the intensity from Si3N4. High N } intensity is difficult to achieve even with the best spectrometer and } crystal designs. And to add insult to injury, the addition of the C } coating absorbs even more of the N x-rays from the sample. The mass } absorption coefficient for N K-alpha emission with a C absorber is } huge (greater than 23,000). } } Plus, you should check for possible N peak shape differences between } Si3N4 (or other standards) and your ammonium-bearing zeolite. } } All in all, it is a very difficult analytical problem. } } Good luck! } } Carl } } At 4:24 PM +0000 2/3/00, Dr Eric Lachowski wrote: } } Eugenia, } } the microprobe is not the best tool for the analysis of } } ammonia in zeolites because ammonia is very volatile in the } } electron beam. If your zeolite is phase pure it would be } } far better to dissolve it and do a classical chemical } } analysis. } } } } regards, } } Eric } } On Thu, 3 Feb 2000 14:54:20 +0100 bengi {bengi-at-unimo.it} } } wrote: } } } } } } } } Dear listers, } } } I have started a research project on ammonium -bearing mineral (zeolites), } } } and now I'm trying to quantify N using electron microprobe analysis. My } } } problems of dealing with N are: } } } } } } a) The thickness of the coating (in the literateur = } } approximately 150 A°). } } } Does anyone have information on the model of coater to making these films? } } } Does anyone have experience with preparation of such } } } samples? } } } } } } b) Obtaining appropriate standards. (I have Si3N4, but it contains too much } } } N (=39.96%) towards my samples (5%). Does anyone known any good nitrogen } } } standards for such a sample ? } } } } } } Any other suggestions about quantitative nitrogen analysis would be greatly } } } appreciated !! } } } Thanks for your time. } } } } } } My best regards } } } } } } Eugenia } } } } } } } } } Eugenia Marchi } } } Università degli Studi di Modena } } } Dip. Scienze della Terra } } } P.le S.Eufemia n°.19 } } } 41100 Modena (Italy) } } } Tel. +39-059-417289 } } } Fax. +39-059-417399 } } } e-mail: bengi-at-unimo.it } } } } } } } } } } } } } ---------------------- } } Dr Eric Lachowski } } Department of Chemistry } } University of Aberdeen } } Meston Walk } } Old Aberdeen AB24 3UE } } Scotland } } +44 (0)1224 272934 fax 272921 } } e.lachowski-at-abdn.ac.uk } } ====================================== } Carl Henderson } Electron Microbeam Analysis Laboratory } University of Michigan } 2501 C.C. Little Bldg. } Ann Arbor, MI 48109-1063 USA } (734) 936-1550 FAX (734) 763-4690 } ======================================
Since the discussion on CM12 alignment shifted to several other problems, and eventually got some JEOL 2010 users involved (Larry Thomas), I thought of throwing in this alignment question regarding the OA in a 2010 with a LaB6 gun:
To keep the camera length at a consistent value when I am not using an internal standard, I record SA diffraction patterns with C2 fully defocused cw (this is by conventional jargon, although actually using the Brightness knob I am changing C3). This is supposed to make the illumination as parallel as possible and is easily reproducible. Then I focus the direct beam with the intermediate lens (diffraction focus). CBD and NBD are another problem.
The problem is to align the Objective Aperture under this condition. If the OA is centered around the direct beam in diffraction, when switching to image mode and increasing the illumination (Brightness) for imaging conditions, the aperture will be apparently out of center. This gives the impression that the voltage center changes between a fully defocused C2 and a moderately focused (still cw from crossover, converging the illumination on the sample) condenser. That shift is obvious in DIFF mode, the direct beam can be seen to shift as C2 is turned ccw from the fully cw position.
As was noted in the on-going discussion, the crossover planes move along the optic axis as the strength of C2 changes. The helicoidal path of the beam is straight down the optic axis of the lenses only piecewise, and at the level of the OA plane it stays centered only for a limited range of C2 settings--or convergence angles. In my case, the convergence changes considerably when I record images digitally in a 1kx1k CCD camera, which screams for brightness.
My two ways around this are to (1) recenter the OA in image mode, or (2) in DIFF mode, but keeping C2 at the setting I will use in MAG mode. Both methods are a pain, mostly when doing BF/DF work, and probably incorrect.
Is there an alignment procedure for the 2010 that will prevent this " OA shift " from DIFF to MAG modes?
Augusto Morrone Seagate Technology NRW-115 7801 Computer Ave S. (612) 844-5838 Fax: (612) 844-7301
Dear Paul, Why don't you use the Hitachi service from NSC? They are the best and the cheapest. At 12:12 PM 2/3/00 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
Can anyone recommend an aqueous mounting medium that polymerises (or whatever) to form a permanent hard mount, and that does not autofluoresce? Thanks.
We had been using silver enhancement kits from Amersham and Nanoprobe. Of these, we found the Nanoprobe kit to be very difficult to work with given it's high viscosity. We also use a pre-embed protocol, and found that both silver enhancement kits caused collagen fibrils to denature (unravel). However, during our protocol the tissue is incubated for fairly long duration in the enhancement medium (15 minutes on ice, then warmed to 25 C in a water bath for an additional 5 minutes, all in the dark).
Our more recent experience is with the Nanoprobe gold enhancement kit. It has HUGE advantages over the silver system.
First, you can Osmicate the tissue. Second, the viscosity of the components is comparatively very low, easing the use of the product. It is not so sensitive to light, and it does not cause denaturation of collagen fibrils. We do see some variability in particulate size, but this is probably a problem associated with diffusion of the media through the tissue. We have found no disadvantages of the Gold enhancement v.s. Silver and now use the Gold method exclusively.
I hope this helps,
Doug
On Thu, 03 Feb 2000 12:44:23 -0500 Michael Plociniak {plocinia-at-aecom.yu.edu} wrote:
} } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America To Subscribe/Unsubscribe -- Send Email } to ListServer-at-MSA.Microscopy.Com On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Colleagues, } Has anyone used gold enhancement reagents from Nanoprobes, } Inc. (Stonybrook, NY - USA) as an alternative to silver } enhancement for enlarging 1 nm immunogold probes? } } Advertisements state that gold enhancement has lower } background and is compatible with osmium. Can anyone } verify this from personal experience? } } In addition to these advantages, I am hoping that milder pH } conditions will be less damaging to ultrastructure in } cultured neurons (using a preembedment protocol). } } Thank you, } Michael
---------------------- Douglas R. Keene Associate Investigator Shriners Hospital Research Facilities 3101 S.W. Sam Jackson Park Road Portland, Oregon 97201 phone: 503-221-3434 FAX: 503-412-6894 (9-5 PST) e-mail: DRK-at-shcc.org
-----Original Message----- } From: Ken Tiekotter {tiekotte-at-up.edu} ..One does not mixed and match objectives from different systems merely for the convenience of pricing.... ---------------------------------------------------------------------------- ---------------------------------------------------------------------------
Of course, I've heard this sort of statement many times before. But has anyone ever done any comparisons of mixed systems to see if there are certain brands that are particularly incompatible or particularly compatible?
I ask this both as a general question and because I use a Wild M7 for which I need a pair of high eyepoint oculars. I don't need the adjusting collar built into current models and haven't been able to find an older pair used. But before I buy a Nikon/Olympus/ etc. pair I'd love to know how different the complete system will be from what I've got now.
I know many people who have used third party photo eyepieces, in just the situation where you'd think quality would be of the utmost concern. Dealers are often not the best source of info since they tend to represent one brand and, even when they are completely honest, rarely have experienc ewith a wide range of brands (especially mixed up.
Dan Freidus freidus-at-wwnet.com
P.S. Of course, if anyone has a pair of Wild 10x high eyepoint oculars for sale, that would also solve my problem (I'd also consider other magnifications). (I'm also looking for accessory objective lenses and various other accessories. Please contact me off-list if you've got anything for sale or trade.)
At 09:20 AM 2/4/00 , you wrote: } } A client of ours wonders about a comparison between Zeiss and Olympus } } optics. The Zeiss 100X lense cost $10,000, and the Olympus about $4,000: } } does that mean the Olympus is inferior? That's what our client's } } colleagues imply. What measure of lense function is relevant? Does Olympus } } (or Zeiss) have a specification, like 'resolves particles to 0.1 micron' } } or something like that? Is there a measure of distortion? Best } } regards, Steve NiemelaVayTek, Inc.305 West Lowe AvenueFairfield, IA } } 52556email: sniemela-at-vaytek.com Phone: 515.472.2227Fax: 515.472.8131web: } } www.vaytek.comgeneral email: vaytek-at-vaytek.com } }
The NA is very important in achieving high resolution. The other factor is the NA of the condenser. An Abbe condenser is easily out gunned by the aplanatic condenser (NA=1.4). Olympus makes two PlanAPO 100X oil objectives. One has an NA of 1.3 while the other is 1.4. The 1.4 lens costs today new about $6000. The lower NA lens is about $4500. So this is probably the one you are comparing. I had a Zeiss PlanAPO and recall that it had an NA of 1.3. It was terrible. I replaced it with the Olympus 1.4 and got major improvements. Of the Zeiss objectives, their 63X PlanAPO is probably the best one they have made. The rest are so so. Some are easily beaten by Olympus flourites. Zeiss is more expensive because it is Zeiss, not necessarily because it is better. The cost of manufacturing in Germany is very high compared to Japan and other places that Olympus uses. The emerging problem I see is that Olympus is moving much of its scope production to China. I have purchased all of the Olympus items I need and that were made in Japan before being stuck with Chinese Olympus. It may reach a point where Zeiss is a better product despite the higher cost.
BTW, the 1.4NA lens at $6000 reflects the recent 10% across the board price increase by Olympus.
There are important differences between older and more recent TEMs, which go to the heart of this discussion.
On older dedicated HREM's (LaB6) imaging is done with the beam at or close to crossover on the specimen, and beam convergence is determined by the user using the C2 aperture. On newer TEM's such as the JEOL 2010 (FEG or not), you need to set C2 so crossover is pretty far from the specimen plane in order to get decent coherence for imaging. Whenever this is the case, the C2 aperture isn't incoherently filled, and some of the older theory won't apply (regardless of whether the machine is or isn't a FEG).
For example, the standard method of determining convergence in an image is by switching to diffraction under the same conditions used to image, and measuring the disc diameter. (see e.g. O'Keefe and Sanders, Acta Cryst A31, p.307). This assumes the beam being near or at crossover on the specimen aperture. It won't work if the beam is far from crossover on the specimen, because now the incident angle depends on position on the specimen. The disc diameters in diffraction only indicate what the total range of incident angles is over the entire illuminated area. But the area visible in the HREM image is much smaller than this, and over this latter area the range of illuminating angles is smaller. This is just another way of saying that the C2 aperture isn't incoherently filled. If it were, would be impossible to form a shadow image of the C2 aperture at the specimen plane, as one does by spreading the beam.
Think of the illumination on the specimen as a convolution of the source image with the circular C2 aperture: When the beam is at crossover, the aperture part is approximating a delta function - we see the source in the image. When the beam is spread very far, the illumination is a nice shadow image of the C2 aperture, with a little blurriness at the edge because of convolution with a source which is not quite point-like.
In the latter case, the angular spread locally is given by the source size (demagnification) in the diffraction pattern. This can be imaged by switching to diffraction and focusing with the intermediate lens to obtain sharp source images.
This doesn't depend on whether the machine is FEG or not, though the source will be very much smaller if it is. I have posted an example in which a tungsten filament is imaged sharply by defocusing with the intermediate lens with the illumination just slightly defocused. This should be a square pattern, so note that the optics introduce significant distortions in this extreme case.
++++++++++++++++++++++++++++++++++++++++++++++++++ Wharton Sinkler Argonne National Laboratory West P. O. Box 2528 Idaho Falls, ID 83403
} } } On Thu, 3 Feb 2000, Thomas, Larry wrote: } } } Laurie, } } } } Now I'm really confused. I used to think I understood this somewhat. } } } } 1. In the JEOL 2010F FEG (the "all bets are off" case in your item#7), } } changing the illumination spread by changing C2 profoundly moves the } } "diffraction plane" relative to the objective aperture. [There are actually } } two } } objective apertures at different heights in this microscope, but that's } } beside } } the point]. By diffraction plane, I mean the plane below the objective lens } } where the diffraction spots are in sharp focus. In other words (assuming } } fixed } } OL and condenser minilens excitations), in this microscope there is only one } } C2 } } value that allows the diffraction spot pattern and OL aperture to be in focus } } simultaneously. If you focus on the OL aperture (with the intermediate } } lens), } } the spot pattern can be focused at only this one C2 setting. At any other C2 } } setting, you can focus the diffraction pattern with the intermediate lens } } ("diffraction focus") but the OL aperture will then be defocused. Changing } } the } } condenser minilens excitation changes the diffraction plane (relative to the } } OL } } aperture) to a different C2 setting. This is actually quite a useful feature } } of } } the microscope, e.g., for getting very intense focused diffraction patterns, } } but } } a different story. } } } OK, the bets off case. In a FEG you have a somewhat coherent } range of incident directions. The "diffraction pattern" is therefore } something which is effected by the coherent aberrations of the microscope, } unlike the case with incoherent illumination. Consider the case with } focussed illumination, when the diffraction pattern shows an image of } the condensor aperture. In a FEG As a you can "focus" this (coherent) } image to a small spot by going out of focus in a true sense for the } diffraction pattern. This gives you a spot-like pattern, but you will also } see severe distortions at higher angles. With incoherent illumination you } cannot do this and going out of focus (in the diffraction pattern) will } give in general a blurred image of the condensor aperture - life is } simple. } What you need to do is focus "real diffraction" features, e.g. } Kikuchi lines, then not play with the diffraction focus at all. As you } change C2 the size of the spots will grow or shrink, this is fine. } } } 2. The reason for choosing to focus the diffraction spots rather than } } the K-lines is --to put it simply-- that's where the intensity is. In } } amplitude } } contrast imaging (conventional brightfield and darkfield imaging), the } } intensity } } contribution to the image is limited by the OL aperture. It's really } } important } } to have the defining aperture and diffraction spot pattern in focus } } simultaneously. In addition, if I aim to measure diffraction spot patterns } } there is little incentive to focus the K-lines instead of the spots. } } } Yes, you can always get a "spot" pattern with a FEG, and it may } be prettier. However, beware the distortions which make doing } measurements from it very dangerous. } } } 3. Textbooks such as Hirsch et al. often discuss the positional error } } in SA diffraction due to Cs. What readers often miss is the consequence of } } the } } fact that the error depends strongly on the diffraction angle. If } } measurements } } are confined to the low-index (low-angle) reflections, the selecting area can } } be } } reduced (to 200 nm or so at 200 kV) without big positional errors. Maybe } } that's } } an example of differences in experimental and theoretical viewpoints. } } } Hirsch et al is just a good reference to read, about this as well } as many other things! } } } 4. Reducing the condensor aperture size will give less illumination, } } but it won't give parallel illumination. It will change the size of the } } diffraction spots, but not their focus. } } } It makes it closer to parallel. Of course, with really parallel } illumination there is no intensity! } } } 5. I'm not quite sure why the illumination spread has such a strong } } effect on the apparent position of the diffraction plane relative to the OL } } aperture, but suspect it arises from the action of the strong } } condensor-objective in this microscope. I would also question that it has } } much } } to do with coherent/incoherent beams in a FEG or with prefield/postfield Cs } } effects. } } } Sorry, I don't think that is correct. I would suggested that } people look at the distortions with a standard sample and see how bad they } can be. } } } Larry } } } } } } Larry Thomas } } Pacific Northwest National Laboratory } } MSIN P8-16 } } P.O. Box 999 } } Richland, WA 99352 } } Phone: (509)372-0793 Fax: (509)376-6308 } } Email: mailto: Larry.Thomas-at-pnl.gov } } } } } } } } ---------- } } From: L. D. Marks } } Reply To: L-marks-at-nwu.edu } } Sent: Thursday, February 3, 2000 8:06 AM } } To: Radostin Danev } } Cc: MSA listserver } } Subject: Re: TEM:finding the parallel beam } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Sorry, but I think we are getting a little confused here. } } } } 1) The diffraction plane is (by definition) the objective lens } } focal length below the sample - it is the focus of parallel } } directions coming from the sample. The focal length (and } } parameters such as Cs, Cc) change rapidly with the objective lens } } current so an eucentric position makes life simple but is not } } necessarily the best condition to use. (For HREM you generaly } } want to have the objective lens stronger and reduce Cs.) } } } } 2) In general C2 does not change the focal length at all. You } } can have some effect from coupling of the magnetic fields, but } } all C2 is supposed to do is change the convergence angle. } } } } 3) The objective aperature is (approximately) in the back-focal } } plane, given that it is finite and height adjustments are only } } done coarsely (for a specific value of the objective current } } only). } } } } 4) The selected area aperature is (approximately) in the same } } plane as the object for the same reasons as above. Remember that } } there are positional errors in SA diffraction due to Cs (see } } Hirsch et al for instance). } } } } 5) You should focus Kikuchi-lines, not go for the smallest spot, } } since these are real object in the diffraction pattern. What you } } will see is then representative of you incident beam, sometimes } } a Gaussian range of directions. } } } } 6) The simplest way to get more parallel illumination in general is } } to reduce the condensor aperture size. } } } } 7) All bets are off if you have a FEG. Rather than having incoherent } } illumination things like the Cs of the prefield (above the sample) } } and postfield (below) add coherently making it much more complicated. } } For instance, the illumination angle varies across the field of view. } } While this is also there with LaB6, it is a weak effect (except for } } certain classes of diffracted beams, e.g. those with a screw-axis). } } With } } a FEG you can get a "spot" pattern many ways, and most of these will } } have } } large aberrations (e.g. pincushion). } } } } ++++++++++++++++++++++++++++++++++++++++++++++++ } } Laurence Marks } } Department of Materials Science and Engineering } } Northwestern University } } fax: (847) 491-7820 } } mailto:l-marks-at-nwu.edu } } http://www.numis.nwu.edu } } ++++++++++++++++++++++++++++++++++++++++++++++++ } } } } } } } } ++++++++++++++++++++++++++++++++++++++++++++++++ } Laurence Marks } Department of Materials Science and Engineering } Northwestern University } fax: (847) 491-7820 } mailto:L-marks-at-nwu.edu } http://www.numis.nwu.edu } ++++++++++++++++++++++++++++++++++++++++++++++++ } }
We are looking to localize B-glucuronidase in the mouse cochlea. We'd like to stain for the enzyme and still be able to decalcify the cochleas and process them into Epon-Araldite for LM and TEM without losing the reaction product. Does anyone have any suggestions for staining or localization protocols for this enzyme? I'm not certain whether the tissue will be taken all the way to TEM, but I do know we'll be looking at LM (1 to 4 microns). I found a method by Hayashi and Hayashi ("Electron Microscopic Cytochemistry and Immunocytochemistry in Biomedicine", CRC Press, 1993) using hexazonium pararosanilin (red in LM and electron dense for EM) on 40 micron non-frozen sections (and then processed for EM) that sounds appropriate, but I'm concerned that it may not work en bloc (the cochlea is a twisty, windy beasty).
I posted this request a few months ago and received no assistance (just individuals also interested in similar protocols and a company promoting their antibody).
Thanks so much for any direction I can get,
Jaclynn M. Lett, Research Assistant jmlett-at-cid.wustl.edu
Fay and Carl Simons Center for Biology of Hearing and Deafness Central Institute for the Deaf 818 S. Euclid Ave. St. Louis, MO 63110
===} changing C3 (brightness) leads to a shift of the bright field (BF) beam in the back focal plane (BFP) causing the objective aperture to appear off centre after converging the beam?
If you have aligned the gun properly then I suspect that the (shift) wobble alignment has not been done properly. When this button is pressed the microscope goes over to diffraction and you should see a pair of discs or spots (which represent the two extremes in tilt when the beam is shifted to its own extremes). The beam tilts are adjusted to bring the two discs/spots into coincidence. This is done for both the X and Y shift coils separately. The X wobble button shifts the beam back and forth (X shift coils), but in the back focal plane the spot or disc should remain stationary. This is what you are trying to achieve with this alignment procedure. Afterwards this should give you a beam that expands about the same point in the back focal plane. You should find that the objective aperture remains centred in both diff and image modes after doing this alignment.
As for the crossovers, yes you are right, they do move up and down when changing the brightness. The back focal plane remains stationary (by definition as L.Marks said). Anything other than a parallel beam should give you a disc in the BFP (which represents the angular distribution of rays incident on the objective lens).
What did you mean by a voltage centre? My interpretation of this is that the source high tension is wobbled and any shift in the (previously focussed) image is minimised by tuning the beam tilts. This alignment would be used for energy filtered TEM (EFTEM) when doing elemental mapping on a Gatan Imaging Filter (GIF) for example.
******************************************************** Dr Jonathan Barnard
Analytical Materials Physics The Angstrom Laboratory, Uppsala University P O Box 534, SE-751 21 Uppsala, Sweden Phone: +46-(0)18-4716838 Fax: +46-(0)18-500131 http://www.angstrom.uu.se/analytical/home.html
The San Francisco Microscopical Society Announces its next regular meeting:
Wednesday, February 9, 2000
Randall Museum 199 Museum Way San Francisco, CA 94114-1429 (415) 554-9600
7:00 pm
Topic: Introduction to Phase Contrast Microscopy by Robert D. Griffin, President, SFMS And: Annual Business Meeting and Election of Officers
Further information, including an important Letter to the Members from President Griffin, is available at http://www.microdataware.com/sfms .
The San Francisco Microscopical Society (SFMS) is a small, informal group of light microscopists who meet on a monthly basis to discuss topics of common interest. All are invited and welcome, regardless of knowledge level or professional field.
Dear Steve, Because of the nomenclature and the way manufacturers determine their specifications, it is really difficult to compare optics on paper. When I find myself in these types of discussions, I find it more useful to compare optics of similar list price rather than specifications because generally speaking, in our industry, the more things cost, the more care is taken in their manufacture. Olympus has a new 100X objective with an NA of 1.65 that sells for $10k. That would be the optic I would compare to the $10k Zeiss.
Shane Collins Scientific Instruments
-----Original Message----- } From: Steve Niemela [mailto:sniemela-at-vaytek.com] Sent: Thursday, February 03, 2000 2:05 PM To: microscopy-at-sparc5.microscopy.com
To: {Microscopy-at-MSA.Microscopy.Com}
SECOND ANNOUNCEMENT: UBC LIVE-CELL COURSE: Please register by March!!
Hello all,
The faculty for the 2000 UBC 3D Live-cell Microscopy Course is now set and we welcome several new faces, including Stephan Hell, Andreas Kriete and Steve Potter. The whole list is below.
o Stephen Adams University of California-SD o Ping Chin Cheng SUNY, Buffalo o Elaine Humphrey Univ. of British Columbia o Stephan Hell Max Planck, Goettingen o Ted InouŽ Universal Imaging, PA o Iain Johnson Molecular Probes, OR o Larry Keenan Cell Robotics, NM o Ernst Keller Carl Zeiss, Thornwood, NY o Andreas Kriete University of Geissen o Felix Margadant University of Sydney o Tim Murphy University of British Columbia o Jim Pawley University of Wisconsin-Madison o Steve Potter California Institute of Tech o Jason Swedlow University of Aberdeen o Michael Weis Agriculture Canada
Basic info is below but you can get the entire brochure, including links to faculty home pages, at
Organized by Prof. James Pawley, (University of Wisconsin-Madison) (SEE SABBATICAL ADDRESS AT END OF MESSAGE)
in association with Dr. Elaine Humphrey UBC BioSciences Microscopy Facility: University of British Columbia Vancouver, BC, Canada
DATES
Applications must be received by March 1, 2000 Deposit due April 15, 2000 Registration 5:00 - 7:00 PM Sunday, June 18, 2000 First Lecture 7:30 PM Sunday, June 18, 2000 Live-cell Course ends, noon Thursday, June 29, 2000
APPLICATIONS DUE BY MARCH 1, 2000
More information at : http://www.cs.ubc.ca/spider/ladic/course/bulletin.html
or
REGARDING COURSE ORGANIZATION:
Prof. James B. Pawley, (on Sabbatical) Room LG 10, Madsen Building, F-09, University of Sydney, NSW, 2006 Australia
Dr. Elaine Humphrey, Biosciences EM Facility Biosciences Building Univ. of British Columbia 6270 University Blvd. Vancouver, BC, V6T-1Z4
THE PURPOSE OF THE COURSE
Modern methods of 3D light microscopy promise a revolutionary improvement in our ability to view living cells. To help convert this promise to reality for a wider selection of biological scientists, an intensive eleven-day residential course concentrating on all aspects of 3D Microscopy of Living Cells will be held at the University of British Columbia, in June of 2000. The course includes 4 days on 2D techniques, 6 days of 3D techniques and a summary presentation. It covers basic microscopy to the highest level confocal microscopy. (A half-day Pre-course is offered for any who may need to brush up on basic optics!).
Topics include: o Quantitative 2D light microscopy o How to keep your cells alive o 3D imaging in confocal microscopy o Widefield/deconvolution techniques o Two-photon excitation microscopy o Fluorescent and backscattered light signals o Dye design, characteristics and use o Pixelation: The Nyquist Criterion o Lasers and laser tweezers o Objectives and aberrations o Scanning-systems: AODs and mirrors o Optimal pinhole size/photon efficiency o Detectors: operation and performance o Video-rate confocal imaging o Measuring ion concentrations o Display and measurement of 3D data
Morning lecture/demonstrations lead to hands-on laboratory exercises each afternoon that will utilize most of the commercial instruments currently available for 3D microscopic imaging. Students will work in groups of 3 or 4 throughout the discussion and laboratory sessions, and may complete a live-cell 3D study on their own specimens. In the first four years, over 100 students from 22 countries have attended. Last year, 11 separate, 3D microscopical workstations were available for student use under the supervision of a 17-member international faculty. We expect to have even more workstations in 2000. Including manufacturers representatives, the teacher/ student ratio will be almost 2:1.
INTERNATIONAL FACULTY
o Stephen Adams University of California-SD o Ping Chin Cheng SUNY, Buffalo o Elaine Humphrey Univ. of British Columbia o Stephan Hell Max Planck, Goettingen o Ted InouŽ Universal Imaging, PA o Iain Johnson Molecular Probes, OR o Larry Keenan Cell Robotics, NM o Ernst Keller Carl Zeiss, Thornwood, NY o Andreas Kriete University of Geissen o Felix Margadant University of Sydney o Tim Murphy University of British Columbia o Jim Pawley University of Wisconsin-Madison o Steve Potter California Institute of Tech o Jason Swedlow University of Aberdeen o Michael Weis Agriculture Canada
TUITION
Course tuition is $2,150 US and includes lunches. On receipt of 50% deposit, students will receive preliminary group assignments and the textbook, Handbook of Biological Confocal Microscopy, (Plenum, 1995). The tuition fee includes the textbook, a binder of handouts, and tickets for the Opening Reception, the Manufacturer's Reception and the Beach Party. Accommodations and meals other than lunch are not included in the tuition fee. The Pre-course is $100 US.
APPLICATIONS
Applicants will complete a questionnaire to assess knowledge level and field of interest. Enrollment is limited to about 24 participants. Selection will be made on the basis of background and perceived need. Those without previous LM experience will be provided with basic texts on request to read before the course begins and are encouraged to take the Pre-course on the afternoon of June 18.
Application forms, and other course information from this and past years, can be downloaded from the WWW site at:
Dr. Elaine Humphrey, Biosciences EM Facility Biosciences Building Univ. of British Columbia 6270 University Blvd. Vancouver, BC, V6T-1Z4 Phone: 1-604.822-3354 FAX: 1-604.265-5315 Email: ech-at-unixg.ubc.ca.
Application deadlines:
Application forms are due by March 1, 2000! Deadlines are early to facilitate setting up groups. Successful applicants will be notified by April 1, and a deposit of 50% must be received by April 15, 2000 to reserve your position. In general, deposit refunds are only be possible if your position can be filled from the waiting list. The remainder of the fees is due at registration.
DATES
Applications must be received by March 1, 2000 Deposit due April 15 2000 Registration 5:00 - 7:00 PM Sunday, June 18, 2000 First Lecture 7:30 PM Sunday, June 18, 2000 Live-cell Course ends, noon Thursday, June 29, 2000
TEACHING PHILOSOPHY
As befits teaching in an area at the boundary of "what is now known," lecturers have been chosen based on their expertise as scientists working in the field rather than because they all agree. They are encouraged more to be provocative than to be prosaic. Students should expect discussion in areas where differences of opinion exist.
Prior to the course, students will be organized into groups and encouraged to communicate by email/phone, about the "Living-cell" group projects that they will pursue during the course and that will be presented to the class on the last day of the course. It has been found that group interactions make best use of students' prior experience and can be very effective in teaching the practical skills covered in a hands-on course of this type.
ARRANGEMENTS FOR LIVE SAMPLES
Students must contact Dr. Elaine Humphrey to make necessary arrangements for the transport and maintenance of cell lines etc. needed for their projects. Organisms linked in any way with human disease are not permitted because of safety considerations. Transport and customs arrangements for living specimens are entirely the responsibility of the student. Students also attending the 3D Image Processing Course, may be able to analyze, process and display some of the 3D data collected from their specimens
The workshop will cover 3D image processing for measurement and display. Enrollment is limited to those attending the 3D Microscopy course. Tuition : $850 US (lunches and snacks incl.)
WHO SHOULD ATTEND?
The course is designed for biologists working with multidimensional and possibly multicolor microscopical data sets. Getting the data is only half of the battle. Image data in 3, 4 or even 5 dimensions may be difficult to store let alone analyze or display. This course is to help students understand the hardware and software aspects of this problem and give them the techniques they need to make the best use of their data.
The course is designed for biologists who need to make measurements on 3D microscopical data sets and then display the results in an effective manner. The course will be taught on SGI, Macintosh and IBM-compatible computers. A wide variety of software designed for the 3D microscopy market will be described, demonstrated and available for use.
Workshop Organizers
o Andreas Kriete Giessen University, DE o Felix Margadant University of Sydney, Au
Faculty
o Ping Chin Cheng State U. of New York, Buffalo o Chris MacLean Vaytek Inc., IA
PLAN OF INSTRUCTION Classes will meet from 8:30-12:00 and 1:00-6:00 with lecture-demonstrations followed immediately by hands-on laboratory sessions using a variety of workstations. Students will "learn-by-doing" with two to a machine. Lab handouts will describe some specific exercises to be performed on "canned" data sets. Facilities and supervision will be available until 11:00 PM, for students to work on their own data. **************************************** Prof. James B. Pawley, (on Sabbatical) Ph. 61-2-9351-7548/2351 Room LG 10, Madsen Building, F-09, FAX 61-2-9351-7682 University of Sydney, NSW, 2006 Australia JBPAWLEY-at-FACSTAFF.WISC.EDU Come to the 5th 3D Live-cell Microscopy Course, June 19-29, 2000, UBC, Canada. Info and forms at: http://www.cs.ubc.ca/spider/ladic/course/bulletin.html "A scientist is not one who can answer questions but one who can question answers." Theodore Schick Jr., Skeptical Enquirer, 21-2:39
I recently inherited a used LKB Nova ultramicrotome from a medical facility in town. The previous owner could not locate the operating instructions and I am at a loss to figure out its proper operation. Does
anyone on the list have instructions for operation of this instrument?? I would be willing to reimburse anyone for a copy of the instruction manual.
Dr. Stanley A. Rice University of Tampa srice-at-alpha.utampa.edu (813) 253-3333 Ext. 3340
Content-Type: text/x-vcard; charset=us-ascii; name="vcard.vcf" Content-Transfer-Encoding: 7bit Content-Description: Card for Stan Rice Content-Disposition: attachment; filename="vcard.vcf"
I tried a Cy5 filtercube today on a upright flourescent microscope equiped with a Diagnostics Instrument Spot Camera. My tissue had been immunolabelled with Cy5 and I did not expect to see thru the microscope. So we took pictures with the Spot Camera and the immunolabeling seemed to work. The problem was that there was uneven illumination in the pictures, half moon of brightness and the rest of the field dark. The salesperson did check that the mercury bulb was aligned correctly. Also there was no ambient light from the room causing the shadow.
Anyone with an idea of what is causing this problem?
With emphasis on Tripod Polishing and Focused Ion Beam Specimen Preparation ....
..will be offered in conjunction with the Joint Annual Meeting of the Florida Society for Microscopy and the Florida American Vacuum Society in Orlando at the University of Central Florida March 15-17, 2000.
Instructors:
Ron Anderson, IBM Fred Stevie, Lucent Technologies Lucille Giannuzzi, UCF
Vendor Sponsors:
FEI Company Micro Optics South Bay Technology
For registration material please contact Lucille Giannuzzi at lag-at-mail.ucf.edu
******************************************************************* Lucille A. Giannuzzi, Ph.D.
Associate Professor, Dept. of Mechanical, Materials, and Aerospace Eng., University of Central Florida, PO Box 162450, 4000 Central Florida Blvd., Orlando, FL 32816-2450 USA phone (407) 823-5770 fax (407) 823-0208 email lag-at-mail.ucf.edu
Director, UCF/Cirent Materials Characterization Facility, 12443 Research Parkway, Suite 305 Orlando, FL 32826 phone (407) 275-4354,5,6 fax (407) 275-4321 -------------------------------------------------------------------- "Good judgement comes from experience.
Experience comes from making bad judgement."
Mark Twain ********************************************************************
The responses to this thread have been outstanding. Yet from my perspective the unasked and thus unanswerable question is: what is the purpose for the lens? A couple of the responses have brushed the issue, but it still hasn't been specifically stated. Assuming that resolution is the main issue, the concerns of flatness of field, NA, color correction, working distance, etc, all have to be factored into the evaluation of the different lenses. It should be possible to demo both (or any of a set of) lenses under the lab and experimental conditions of real life. Only then can one truly determine which lens is the "best" (or more correctly, the appropriate) lens. All of the theory and specifications in the world don't matter if the lens doesn't aid the microscopist in obtaining the data necessary to carry out the work. One thing that is unalterable is that even if one is considering a number of "infinity-corrected" lenses from different manufacturers, they may not be truly interchangeable. Make sure the lens works on the microscope body. Then make sure it provides the performance needed. That's the right lens.
Roger Moretz
On Thu, 3 Feb 2000 16:04:55 -0600, Steve Niemela wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } To: {Microscopy-at-MSA.Microscopy.Com} } Subject: A comparison of optics } Date: Thu, 3 Feb 2000 10:57:19 -0600 } MIME-Version: 1.0 } Content-Type: multipart/alternative; } boundary="----=_NextPart_000_001F_01BF6E35.70E8F2E0" } X-Priority: 3 } X-MSMail-Priority: Normal } X-Mailer: Microsoft Outlook Express 5.00.2615.200 } X-MimeOLE: Produced By Microsoft MimeOLE V5.00.2615.200 } } A client of ours wonders about a comparison between Zeiss and Olympus } optics. The Zeiss 100X lense cost $10,000, and the Olympus about $4,000: } does that mean the Olympus is inferior? That's what our client's } colleagues imply. What measure of lense function is relevant? Does Olympus } (or Zeiss) have a specification, like 'resolves particles to 0.1 micron' } or something like that? Is there a measure of distortion? Best } regards, Steve NiemelaVayTek, Inc.305 West Lowe AvenueFairfield, IA } 52556email: sniemela-at-vaytek.com Phone: 515.472.2227Fax: 515.472.8131web: } www.vaytek.comgeneral email: vaytek-at-vaytek.com } } }
Roger C Moretz, Ph.D.
_______________________________________________________ Get 100% FREE Internet Access powered by Excite Visit http://freeworld.excite.com
Lesley, How about CMC (Refractive Index=1.37). CMC10 (R.I.=1.39), CMCP-9 (R.I.=1.40) and CMCP-10 (R.I. 1.41)? This is a mixture of Polyvinyl alcohol (5%), phenol(25%), lactic acid (25%), and water (45%).
I believe you can get this from Polysciences Inc. This mountant causes problems with most stained specimens, but may work for you. Is glycerine jelly auto-fluorescening?
-Ken
-------------- Ken Tiekotter Dept. of Biology The University of Portland 5000N. Willamette Blvd. Portland, OR 97303
Dan, Mixing and matching objectives is one thing, while doing the same for oculars is probably not visually as big an issue. However, in highly corrected systems, compensating eyepieces are a critical consideration if one wishes to maintain image quality, i.e., plan apo objectives require compensating oculars to maintain the correction of the apochromatic objective. This is especially true for color reproduction in color photomicrograph. This is not as important in black and white photomicrography.
-Ken
---------------- Ken Tiekotter Dept. of Biology The University of Portland 5000 N Willamette Blvd. Portland, OR 97303
On Fri, 4 Feb 2000, Dan Freidus wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } -----Original Message----- } } From: Ken Tiekotter {tiekotte-at-up.edu} } ..One does not mixed and match objectives from different systems merely for } the convenience of pricing.... } ---------------------------------------------------------------------------- } --------------------------------------------------------------------------- } } Of course, I've heard this sort of statement many times before. But has } anyone ever done any comparisons of mixed systems to see if there are } certain brands that are particularly incompatible or particularly } compatible? } } I ask this both as a general question and because I use a Wild M7 for which } I need a pair of high eyepoint oculars. I don't need the adjusting collar } built into current models and haven't been able to find an older pair used. } But before I buy a Nikon/Olympus/ etc. pair I'd love to know how different } the complete system will be from what I've got now. } } I know many people who have used third party photo eyepieces, in just the } situation where you'd think quality would be of the utmost concern. Dealers } are often not the best source of info since they tend to represent one brand } and, even when they are completely honest, rarely have experienc ewith a } wide range of brands (especially mixed up. } } Dan Freidus } freidus-at-wwnet.com } } P.S. Of course, if anyone has a pair of Wild 10x high eyepoint oculars for } sale, that would also solve my problem (I'd also consider other } magnifications). (I'm also looking for accessory objective lenses and } various other accessories. Please contact me off-list if you've got anything } for sale or trade.) } } } } } }
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Reply to: RE: Aqueous mounting medium Dear Lesley,
You can make a very useful mounting medium from inexpensive ingredients. The main ingredient is poly(vinyl) alcohol which is soluble in water but will dry without shrinking too much. The water solubility means you can take previously mounted slides, rehydrate them and use them again.
Fading of fluorescent dyes can be retarded by including anti-fade compounds.
You can find the recipe at {http://www.hei.org/htm/moviol.htm} .
Details of anti-fade compounds can be found here (at the bottom of the page) {http://www.hei.org/htm/ifse.htm} . Ths site is getting a little outdated so if you feel there are details missing then please contact me with your questions.
Many thanks to Heinz Schwarz of Tuebingen for writing these protocols.
Regards,
Paul Webster
Paul Webster, Ph.D. Associate Scientist & Director Ahmanson Advanced Electron Microscopy & Imaging Center House Ear Institute 2100 West Third St. Los Angeles, CA 90057
by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id VAA13116 for dist-Microscopy; Sat, 5 Feb 2000 21:13:46 -0600 (CST) Received: from no_more_spam.com (sparc5.microscopy.com [206.69.208.10]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id VAA13113 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Sat, 5 Feb 2000 21:12:48 -0600 (CST) Received: from hme0.mailrouter01.sprint.ca (hme0.mailrouter01.sprint.ca [207.107.250.16]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id VAA13106 for {Microscopy-at-sparc5.Microscopy.Com} ; Sat, 5 Feb 2000 21:12:36 -0600 (CST) Received: from normandlaurier (spc-isp-mtl-58-8-589.sprint.ca [149.99.151.82]) by hme0.mailrouter01.sprint.ca (8.8.8/8.8.8) with SMTP id WAA18723; Sat, 5 Feb 2000 22:07:44 -0500 (EST) Message-ID: {000b01bf7046$84afd240$52976395-at-normandlaurier} "Dr. Gary Gaugler" {gary-at-gaugler.com} References: {v03007801b4bfabe09222-at-[129.78.149.225]} {4.2.2.20000204145256.00b2a820-at-pop.calweb.com}
I would call this a typical PARTIAL judgment. What you see is what you get. Thee only way, to the best of my experience, is to get both objectives, side by side if possible, and to do your own evaluation. I have honestly doubts concerning those values set on objectives. Things change and optic too and sometimes faster that we could expect. Norm ----- Original Message ----- } From: Dr. Gary Gaugler {gary-at-gaugler.com} To: MSA listserver {Microscopy-at-sparc5.Microscopy.Com} ; {sniemela-at-vaytek.com} Sent: Friday, February 04, 2000 5:02 PM
I am interested to know what the present state of the art is re: solid state electon emitters, total electron beam currents available and also beam current densities. If anyone could point out a recent review article, I would appreciate it. Thanks,
Don Marshall
Donald J. Marshall Relion Industries P.O. Box 12 Bedford, MA 01730 Ph: 781-275-4695 FAX: 781-271-0252 email dmrelion-at-world.std.com
Cathodoluminescence, mass spectroscopy, electron beam technology
"A weed is a flower out of place."
(Please note: Do not send email with attachments to this address. Instead, send it to donbarlen-at-aol.com. Thank you.)
In fact, if you're really interested in getting the BEST lens, try at least three examples of the ones you are interested in . All high NA lenses must meet some minimal standard but some are significantly better than that minimum and the only way to find one that is is to try THE lens you are going to buy - not just an example.
Bill Miller
At 10:04 PM 2/5/00 -0400, Norm wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Do not have difraction sofrware, but here are some URLs where you can download free simulation software I use for e-beam to solid interactions, hope it may be useful for you.
Casino is a freeware for monte carlo simulations: http://www.gme.usherb.ca/casino/
Monte carlo resources: http://dsa.dimes.tudelft.nl/pattern_definition/monte/monte.html
My FAVORITE monte carlo software: http://web.utk.edu/~srcutk/htm/simulati.htm
Valery Ray, MSEE, SEM engineer Applied Materials 001-208-8413744
Hi everybody While this has been an interesting discussion we seem to have got away from the original question.
This is the method I use - after of course ensuring normal alignments and adjustments have been carried out
Insert the objective aperture with a specimen in the beam. Focus the image of this aperture in Selected Area Diffraction using the first intermediate lens focus.
Focus the diffraction pattern (i.e. the smallest zero order spot) with the final condenser lens, this will give a parallel beam incident upon the specimen.
To test this, remove the objective aperture and insert the largest selected area aperture. If the beam is parallel moving this aperture will not move the diffraction spot.
Tilt - The purpose of this alignment is to ensure that when the beam is tilted it doesn't move on the specimen. This is double tilt correction and depends on using double deflector coils.
Shift - Double shift correction is meant to ensure that when the beam is shifted it doesn't tilt. The operation of this alignment is dependent upon the value of the first intermediate lens. The best value for this alignment is a parallel beam condition as described above. By the very nature of the beast it will be found to be impossible to perform this alignment correctly at some CBD conditions. This is meant to be a convergent beam mode after all not a parallel beam mode.
I understand that this is also referred to as tilt and shift purity.
Richard Hey
The views expressed herein are purely my own and do not reflect those of my employer
Richard Hey Technical Support Engineer Jeol (UK) Ltd Phone: (44) 1707 377117
Dear colleagues, I am now currently using TEM to observe a filamentous structure so-called hrp pilus produced by plant pathogen, Pseudomonas syringae which is grown on planta mimic medium agar plates. Even though I could easily find this structure through directly put the nickel-grid on the bacteria plate for a certain time,followed by the negative staining, I can't convince other people to believe it's the structure we are looking for as its size is around 7-10 nm, which is indistinguishable from that of bacterial flagellum. So we want to label this structure by using the specific antibody against its major structural protein followed by the 10 nm gold-conjugated second antibody. The difficulty I am encountering now is most pili are washed away after the whole immnuogoldlabeling process. Since I am a beginnner on TEM work, I don't know whether it's a very commonly encountered problem in immnogoldlabeling experiment or just a very special case happened to me, such as the pilus i'm interested in is too vulnerable to survive the exclusive wash, or I didn't master the whole immunogoldlabeling techinique. If it's a common problem, I hope someone could kindly provide me with your valuable experience on how to resolve this problem, how we can prevent them to be washed away. If it's due to my techinical problem, could you please send me a very detailed immunogoldlabeling protocol. Any kind of help will be highly appreciated have a nice week yours qiaoling jin jinq-at-msu.edu
Thank you for your responses. Please note that I do perform the TILT and SHIFT alignments (aka "pivot point alignment" in other instruments) on a regular basis, but this doesn't solve the problem.
Regarding: "...What did you mean by a voltage centre? My interpretation of this is that the source high tension is wobbled and any shift in the (previously focussed) image is minimised by tuning the beam tilts. This alignment would be used for energy filtered TEM (EFTEM) when doing elemental mapping on a Gatan Imaging Filter (GIF) for example".
This is a (standard) routine alignment for imaging as well. The voltage center alignment is done by first focusing the specimen at the optimum OL current (adjusting Z), then adjusting the Tilt (in image mode) and Shift (in diffraction mode), and finally adjusting the beam tilts to minimize the image shift at the center of the screen. The problem arises when the Brightness is changed, as is typically adjusted when optimizing the illumination on the sample to record an image (intense illumination) versus an SA diffraction pattern (fully overfocused condenser). Apparently this brightness change affects the voltage center as evidenced by a shift of the direct beam as that brightness change is effected in the SAD mode. This shift in the direct beam is equivalent to a beam tilt.
I regret to say that there are some important points which perhaps are being missed. Classic optics, i.e. most TEM textbooks and introductory optics texts are for one of two cases: a) Every point in the condensor aperture (CA) is incoherent (i.e is not related in phase in a statistical sense) with respect to any other point. b) Every point in the CA is coherent (i.e. has a fixed phase) with respect to any other point.
Approximation a) is used in most computer codes for HREM, approximation b) for STEM.
Alas, both a) and b) are only approximations! Current microscopes, particularly FEGs (but perhaps others as well) operate under conditions when neither of the two approximations are valid. While the analytic theory is well established (see the Chapter in Born and Wolff on partial coherence), it is not easy and there are no clean solutions that you can write down. (It can be solved numerically, but this would require big 4-D FFT's which are beyond most current computers.)
The simplest test is to fully focus the illumination. If the spot size is A nm, the coherence in the CA is about 1/A nm-1 (which you can convert to an angle as appropriate). If this is small relative to the CA radius you are working with case a) above. If it is about the same as the CA radius you have b). If it is 2-3 times less, neither approximation is correct!
The moral of the story: 1) If you have a), you can simulate an HREM image with confidence using any of the available programs. They are WRONG quantitatively otherwise. 2) If you have a), you can use simple "ray-diagram" optics, but not otherwise. 3) If you have b) the simple equations for STEM operation are valid, but not otherwise.
Sorry, but as microscopes improve the theory we need to understand them can change.
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University fax: (847) 491-7820 mailto:l-marks-at-nwu.edu http://www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
I am trying to get a handle on the scale of some old pictures. There are human RBC in theses pictures. Can anyone tell me the diameter of your average human RBC?
Thanks Mike
-- _________________________________________________ / Michael J. Herron / / U of MN,Medicine/Infectious Diseases / / herro001-at-maroon.tc.umn.edu / / http://128.101.243.213 / /_____________________________________________/
When a characteristic X-ray is given off from an atom, it carries 1 h-bar (Planck's constant divided by 2 pi) of angular momentum. The electronic transition that occurred for the X-ray to come off must conserve angular momentum of the system. Therefore, a transition from a 2s1/2 to a 1s1/2 state is forbidden by selection rules. You can't produce a Be X-ray. -Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Mike: According to Fawcett's CONCISE HISTOLOGY (page 32), "In the blood, erythrocytes are 8.5 micrometers in diameter, but in the dehydrated state, in dried blood smears, they measure 7.6 micrometers." --Kent
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ A. Kent Christensen, Professor Emeritus Department of Cell and Developmental Biology, Medical Science II Building University of Michigan Medical School, Ann Arbor, MI 48109-0616 Office: 2703A Medical Science II Building Tel (work) (734) 763-1287, Fax (work) (734) 763-1166 E-mail: akc-at-umich.edu http://www.umich.edu/~akc/ ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ --On Mon, Feb 7, 2000 9:20 AM -0600 Michael Herron {herro001-at-maroon.tc.umn.edu} wrote:
} All, } } I am trying to get a handle on the scale of some old pictures. There } are human RBC in theses pictures. Can anyone tell me the diameter of } your average human RBC? } } Thanks } Mike } } -- } _________________________________________________ } / Michael J. Herron / } / U of MN,Medicine/Infectious Diseases / } / herro001-at-maroon.tc.umn.edu / } / http://128.101.243.213 / } /_____________________________________________/
John, If they do, I hope that they will respond to the whole list because we have clients who have experienced similar problems, especially re: computers that are controlling VIS spectrometers collecting CL spectra. I know that there are in line RF filters available (e.g., Steward Co.) and there may be others but the final word is not in on how well these work yet.
Don Marshall
} From microprobe-at-microanalysis.org Mon Feb 7 11:43:28 2000 } } } From: John Hunt {hunt-at-ccmr.cornell.edu} } Subject: Computer lock-ups from static } To: microprobe-at-microanalysis.org } } } Dear Probe community, } It is winter in Central NY, and the room humidity is 33%. Static } electricity (I believe) has been causing frustrating computer lockups } requiring rebooting our microprobe workstation. } I am wondering if anyone has found the best solution for preventing these } problems. } } } } } John Hunt } Cornell Center for Materials Research } 607-255-0108 microprobe } -3789 SEM/TEM } -2365 fax }
Donald J. Marshall Relion Industries P.O. Box 12 Bedford, MA 01730 Ph: 781-275-4695 FAX: 781-271-0252 email dmrelion-at-world.std.com
Cathodoluminescence, mass spectroscopy, electron beam technology
"A weed is a flower out of place."
(Please note: Do not send email with attachments to this address. Instead, send it to donbarlen-at-aol.com. Thank you.)
I would like to tap the wealth of information out there.
We are experiencing a lot of problems with the tungsten filaments that we are currently using on our JEOL 5800 SEM. During long acquisitions (many hours) there is a lot of brightness drift and shifting, even when lengthy stabilization periods are given prior to the acquisitions. JEOL has checked out the SEM itself, and can't find the source for the drifting. I was thinking that it might be the filament. I was wondering if there are different types of tungsten filaments out there, and which is the most stable over long periods of time. I would like to try out all of them (considering trying to switch our entire system to a LaB6 filament is a bit much for now). Any recommendations or experiences you have would help us out a lot.
Thanks in advance!
Marisa Ahmad R&D Specialist Semiconductor Insights mahmad-at-semiconductor.com
weather report (for those interested): It's unusually sunny and warm for here in Ottawa.....only -10 degrees C (but it feels like -25 with the wind). It's such a nice day that I washed my car this morning since it's not really supposed to be brown - now all the doors and windows are frozen shut. {sigh}
In my experience, RBCs prepared for SEM by critical point drying or HMDS shrink even more and may be only 4 or 5 microns across.
Marie } } Mike: According to Fawcett's CONCISE HISTOLOGY (page 32), "In the blood, } erythrocytes are 8.5 micrometers in diameter, but in the dehydrated state, } in dried blood smears, they measure 7.6 micrometers." --Kent }
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology, U-2131 University of Connecticut Storrs, CT 06269-2131 Phone: 860-486-3588 Fax: 860-486-1936
It depends on the fixation/dehydration parameters. Somewhere in the nieghborhood of 4 microns, dependant on the angle of measurement and the angle of the sample with respect to the camera. } Date: Mon, 07 Feb 2000 09:20:44 -0600 } From: Michael Herron {herro001-at-maroon.tc.umn.edu} } X-Accept-Language: en } MIME-Version: 1.0 } To: Microscopy listserv {Microscopy-at-sparc5.Microscopy.Com} } Subject: Diameter of human RBC? } Content-Transfer-Encoding: 7bit } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Can anyone tell me what a Brandes dip is and how it is done? The only thing I know is that it can be used to detect virus particles from plant sap but I can't find any references on it.
Thank You!
Margaret
Margaret Dienelt
Plant Pathologist Electron Microscopy Lab
Floral and Nursery Plants Research Unit U.S. National Arboretum/Agricultural Research Service/USDA
B. 010A, Rm. 238, BARC-W 10300 Baltimore Avenue Beltsville MD. 20705 USA
Does anybody know any program capable of doing TEM simulation of dislocation network, e.g., misfit dislocations in plane view TEM? I know some programs can do one or two dislocations, but what I need is the simulation for a set of dislocations. Thanks a lot.
It has been very interesting reading about diffraction and electron beams but it is quite clear that we often give the instrument credit, "the instrument does it automatically", when credit is not due. Almost every "automatic" is a designers estimate or better still a calculation, it is a setting!
After 36 years with TEM I have to say that the only area that the instrument could set in the SA mode could be the approximate position of the first image plane. We normally move to the SA mode to free up the diffraction lens (1st intermediate) to enable it to be focussed on the diffraction (intermediate) aperture independent of the magnification system. We would then focus the image inside the aperture with the objective lens to ensure that the selected area IS the area in the diffraction pattern. Without this procedure electron rotation and misalignments could give you a diffraction pattern from an area not in the centre of the screen! Users are correct in that if they set the eucentric point or better still, adjust the specimen height to focus at the same objective lens current value, that done the first image plane will be at the same point within the diffraction (intermediate) lens.
So how could the Philips work? I would guess they set the diffraction lens at approximately the first image plane. Then as they do not use it within their lens scheme for the SA magnifications it is fixed at this focus, good enough I think?
A second point, parallel beams are very important if you are chasing the best image quality, biology or materials, most times we need high coherence. There are misunderstandings on how to obtain a parallel beam or high coherence, setting the final condenser underfocus is incorrect. The procedure for high coherence would be to use the smallest spot size you can tolerate (this probably means you must up the emission current). Once in this condition overfocus the final condenser (clockwise from crossover) the spot, what ever size it is now, becomes your virtual source. The further overfocus you go the greater the beam coherence and that is what we are after. You will deduce the smaller the condenser aperture the sharper the spot and the smaller the spot the greater the coherence for a given degree of overfocus.
Work with a design team and they expect everyone to overfocus, they expect everyone to use small condenser apertures and they expect everyone to use small spot sizes. They do not expect everyone to use too low an emission current (almost everyone does, we have talked before about filament life being the most important feature of many laboratories) because this makes the task too difficult! There must be people more knowledgeable than I who will cut out all the guesswork?
Steve Chapman Senior Consultant Protrain Electron microscopy courses world wide http://www.emcourses.com Tel & Fax 44 (0) 1280 814774
} I am trying to get a handle on the scale of some old pictures. There } are human RBC in theses pictures. Can anyone tell me the diameter of } your average human RBC?
When I was a lad...... in grad school, we were told it was a useful reference at 7um diameter.
Saturday, April 21, Sunday, April 22, 2000 Saturday, April 28, Sunday, April 29, 2000
An advanced course on polarized light microscopy which will cover the following topics: The nature of polarized light The origin and interpretation of interference colors Birefringence and crystal orientation The Indicatrix Compensation and variable compensators Interference figures and their interpretation
The workshop will consist of two consecutive weekends of lectures and hands on labs to cover the theoretical and practical aspects of polarized light microscopy.
The course instructors include;
Jan Hinsch of Leica, Inc.,
John Reffner of Trace Consulting,
N.Y.M.S. Instructor Donald O'Leary.
WHEN: April 22, 23, 28 & 29, 2000 from 10 A.M. to 4 P.M.
WHERE: Location To be Announced.
COST: $275 for N.Y.M.S. members, $295 for non-members (includes membership) Lunch and course materials are included. Checks made out to N.Y.M.S.
WHO: advanced course for those who have completed "The Use of the Microscope" or are experienced in microscopy and familiar with the theory of its use.
HOW: Register using the form below. Limited to the first 12 registrants.
FURTHER INFORMATION: Contact Donald O'Leary
eMail: mailto:donoleary-at-worldnet.att.net
(201) 797-8849 Voice Phone Number
PLEASE POST ------------------------------------------------------------------------------------------------------------
Return form to Donald O'Leary, 6 Chittenden Road, Fair Lawn, NJ 07410.
Registration Form
Polarized Light Microscopy, April 22, 23, 28 & 29, 2000
N.Y.M.S. Member _________________ ($275.00) Non-Member __________($295.00)
Name ___________________________________________________________________________
I'm just taking a guess. Is it possible, depending on the type of filters used in the cube, that infrared is passing through as flare if your first excitation filter is not blocking the IR? Or there may be a coating problem on your filter? We have an IR blocking filter right after the mercury lamp. The CCD cameras are extremely sensitive to IR and this has caused problems on our system. But in your situation I don't know why it would pass through. So it is probably something else.
Bob Morphology core U of Washington
On Fri, 4 Feb 2000, Patricia A. Glazebrook wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } I tried a Cy5 filtercube today on a upright flourescent microscope equiped } with a Diagnostics Instrument Spot Camera. My tissue had been } immunolabelled with Cy5 and I did not expect to see thru the microscope. So } we took pictures with the Spot Camera and the immunolabeling seemed to } work. The problem was that there was uneven illumination in the pictures, } half moon of brightness and the rest of the field dark. The salesperson did } check that the mercury bulb was aligned correctly. Also there was no } ambient light from the room causing the shadow. } } Anyone with an idea of what is causing this problem? } } Thank-you, Pat Glazebrook } } } } }
One more thought to support Gary's observation about the importance of the NA of the condenser: The full equation for the limit of resolution is R = (1.22)(wavelength)/(NAobjective + NAcondenser). However, if you don't oil it to the back of the slide, you might as well use a lower NA condenser. Reminds me of the hematology scope built by an otherwise sensible company, meant to be used in ultra high resolution applications. The manufacturer was very good about providing a quality, high NA, oil immersion objective as well as a high NA condenser. The only problem was that you could never raise the condenser high enough to oil it to the back of the slide and, therefore, use it at the NA inscribed... only at about .95! What a waste of time and money!
Also, a reminder that the NAs engraved are the maximum working limit. Closing down an aperture iris or an iris in the back focal plane of the objective reduces the NA accordingly.
One last fact: the NA affects both resolution AND edge information, so, for the crispest, highest resolution images, go for high NA in both objective and condenser.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education}
At 03:02 PM 2/4/00 -0600, Dr. Gary Gaugler wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Marisa Ahmad: } I would like to tap the wealth of information out there. } } We are experiencing a lot of problems with the tungsten filaments that we } are currently using on our JEOL 5800 SEM. During long acquisitions (many } hours) there is a lot of brightness drift and shifting, even when lengthy } stabilization periods are given prior to the acquisitions. JEOL has } checked out the SEM itself, and can't find the source for the drifting. I } was thinking that it might be the filament. I was wondering if there are } different types of tungsten filaments out there, and which is the most } stable over long periods of time. I would like to try out all of them } (considering trying to switch our entire system to a LaB6 filament is a bit } much for now). Any recommendations or experiences you have would help us } out a lot. } Dear Marisa, Do you have control over the filament voltage? If so, set the voltage just a hair over that necessary to saturate the filament. If the voltage is too low, parts of the filament will not emit electrons which get through the wehnelt and into the beam; if too high, there may be significant evaporation of the filament; either may cause drifting and/or shifting. I have had very good luck with the stabilities of both Agar and Ebtec (now Energy Beam Sciences) filaments; not that others may not be as good, I haven't tried them. Ebtec makes several shapes of filament, and one may be best for long-term stability. I have no interest in Agar or Ebtec except as a satisfied customer. Yours, Bill Tivol
Have you considered a "local" remedy, like putting up a humidifier? I know that static can be an issue, but I have never heard of it being this bad before. In more humid environments, we often put a small enclosure around a light microscope then put out a dish of Drierite. Perhaps some simple plastic sheeting, like they sell for exterior storm windows/wind breaks, can be hung from the ceiling around your microprobe plus the humidifier will keep the humidity immediately around your microprobe more balanced.
Best of luck on a tough problem.
Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education}
At 12:06 PM 2/7/00 -0500, donald j marshall wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Is this a Spot Jr. or a SpotII? The IR filter in the Jr. cuts off at 650 nm, the SpotII puts it on a slider. Is it all the way out or [partially occluding the image? Bob Fern caught a lot of the possible issues, but is your lamp properly focussed and aligned? Are you picking up light through the eyepieces? We have a Nikon that will show a semi-circle of light in from a recessed ceiling light that projects to the scope at an angle.
Regards, Glen
Glen MacDonald Research Scientist Hearing Research Laboratories of the Virginia Merrill Bloedel Hearing Research Center Box 35-7923 University of Washington Seattle, WA 98195-7923 (206) 616-4156 glenmac-at-u.washington.edu
On Fri, 4 Feb 2000, Patricia A. Glazebrook wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } I tried a Cy5 filtercube today on a upright flourescent microscope equiped } with a Diagnostics Instrument Spot Camera. My tissue had been } immunolabelled with Cy5 and I did not expect to see thru the microscope. So } we took pictures with the Spot Camera and the immunolabeling seemed to } work. The problem was that there was uneven illumination in the pictures, } half moon of brightness and the rest of the field dark. The salesperson did } check that the mercury bulb was aligned correctly. Also there was no } ambient light from the room causing the shadow. } } Anyone with an idea of what is causing this problem? } } Thank-you, Pat Glazebrook } } } } }
Out here in the dry West (here we have Summer humidity around the 33% value you mention, in Winter we go a lot further down) we have the same problem. There are a few methods to help:
1) increase humidity. Perhaps you can use a humidifier in your equipment room? 2) prevent charging. There are shoes available that prevent charging. 3) get rid of charging. There are several options here:
a) The easiest way and what I usually do if no other method is available, is to touch some metal part NOT of the PC but a chair or table before touching the PC. This requires a) that a suitable object is near the PC, b) that you're not afraid of "shocking" yourself, and c) that you don't move around a lot while working on the PC.
b) Purchase a grounding mat to place in front of the equipment. these mats are connected to the AC ground (use the same as the PC ground). If the mat is big enough so that everybody has to step on the mat first AND is wearing the conductive shoes, everything should be OK.
c) wear grounded wrist straps. These require of course that you put them on.
We use a combination of these techniques to protect our equipment both in the office and the production floor (believe me, it IS necessary here). As bad as it is for electronic equipment, though, it is just great for comfort levels outside. 90 degrees (F) feel just right for bicycling, climbing, rafting...
One company where you can buy this stuff is called "-at-once". Their web site is www.4atonce.com. I just happened to find their catalog first. There are many other companies out there that sell static control equipment. We have no financial interest in this company whatsoever.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
} ---------- } From: donald j marshall[SMTP:DMRELION-at-WORLD.STD.COM] } Sent: Monday, February 07, 2000 10:06:10 AM } To: hunt-at-ccmr.cornell.edu } Cc: Microscopy-at-sparc5.Microscopy.Com } Subject: Re: Computer lock-ups from static } Auto forwarded by a Rule } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
John, If they do, I hope that they will respond to the whole list because we have clients who have experienced similar problems, especially re: computers that are controlling VIS spectrometers collecting CL spectra. I know that there are in line RF filters available (e.g., Steward Co.) and there may be others but the final word is not in on how well these work yet.
Don Marshall
} From microprobe-at-microanalysis.org Mon Feb 7 11:43:28 2000 } } } From: John Hunt {hunt-at-ccmr.cornell.edu} } Subject: Computer lock-ups from static } To: microprobe-at-microanalysis.org } } } Dear Probe community, } It is winter in Central NY, and the room humidity is 33%. Static } electricity (I believe) has been causing frustrating computer lockups } requiring rebooting our microprobe workstation. } I am wondering if anyone has found the best solution for preventing these } problems. } } } } } John Hunt } Cornell Center for Materials Research } 607-255-0108 microprobe } -3789 SEM/TEM } -2365 fax }
Donald J. Marshall Relion Industries P.O. Box 12 Bedford, MA 01730 Ph: 781-275-4695 FAX: 781-271-0252 email dmrelion-at-world.std.com
Cathodoluminescence, mass spectroscopy, electron beam technology
"A weed is a flower out of place."
(Please note: Do not send email with attachments to this address. Instead, send it to donbarlen-at-aol.com. Thank you.)
I am planning to try EM immunolabelling with GFP antibodies. Does anyone out there know of the best source for GFP antibodies and had any luck with using them for TEM. I would like to try pre-embedding with a nanogold labelled secondary. Any suggestions would be appreciated. Thank you.
JoAnn Buchanan Molecular and Cellular Physiology Stanford University School of Medicine Stanford, CA 94305 650-723-5856
Just a quick comment for whatever it's worth. Your posting suggests that you are concerned about mechanical drifts of the filament -- the subject of a certain body of folklore in the microscopy community. It's been my experience that the principal drift in a tungsten filament is not the filament itself, but rather it is the mechanism supporting the filament which drifts.
The actual issue with drifting filaments is that mechanical stresses in the wire will gradually relieve under heat, causing the filament to distort slightly. This is a first-time-used issue. However, I believe most manufacturers include an annealing step in their process to remove these stresses before the filament is shipped to a customer (at least, that's what we used to do in another company when we manufactured our own filaments). In any case, the filament is operated at a very high temperature and it doesn't take long to anneal out these stresses under normal use, so long-term drift effects shouldn't be an issue.
My experience is that long-term mechanical drifts originate elsewhere in the gun mechanism. When you think about it, this makes sense. The filament itself has very little mass and quickly reaches its ultimate operating temperature distribution. However, the rest of the gun assembly is much more massive and, depending on various design considerations, may take hours to reach thermal equilibrium. Meanwhile, all of those intricate parts are expanding -- and many at different rates. Principal suspects, in my opinion, are lateral adjusting and clamping screws -- for example, a set of radial set screws is commonly used to center the ceramic base of the filament within a ring of some type. I used to work with a rather complex assembly which positioned the filament via lateral screws and sometimes had episodes of "filament drift". Subsequently, I have had nearly a decade of experience with a very simple design which supports the filament solely via axial pressure and find that "filament drift" isn't an issue. My conclusion is that a lot of what I used to call "filament drift" was actually mechanism drift. So if you are experiencing mechanical instabilities of the filament, I would suspect the way the filament is being supported more than the filament itself.
This opinion is based on my own experiences, however, and I will be very interested to hear if others have had different experience.
Fred Schamber RJ Lee Instruments Limited
Marisa Ahmad wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I would like to tap the wealth of information out there. } } We are experiencing a lot of problems with the tungsten filaments that we } are currently using on our JEOL 5800 SEM. During long acquisitions (many } hours) there is a lot of brightness drift and shifting, even when lengthy } stabilization periods are given prior to the acquisitions. JEOL has } checked out the SEM itself, and can't find the source for the drifting. I } was thinking that it might be the filament. I was wondering if there are } different types of tungsten filaments out there, and which is the most } stable over long periods of time. I would like to try out all of them } (considering trying to switch our entire system to a LaB6 filament is a bit } much for now). Any recommendations or experiences you have would help us } out a lot. } } Thanks in advance! } } Marisa Ahmad } R&D Specialist } Semiconductor Insights } mahmad-at-semiconductor.com } } weather report (for those interested): } It's unusually sunny and warm for here in Ottawa.....only -10 degrees C (but } it feels like -25 with the wind). It's such a nice day that I washed my } car this morning since it's not really supposed to be brown - now all the } doors and windows are frozen shut. {sigh}
We're interested in knowing a little bit more about the Hematology Slide Stainer RSG 61 commercialized by EMS. So if you have one can you please share your experience with us. Give the plus and also all the minus. If you have another equivalent brand in which you're fully satisfied of course say it. Naturally salesman can contact me (offline only please, so the whole community will not be annoided...)
Thank you for your help
Marc
PS hi Keith how is weather in Plymouth ? Here it's raining and we're singing...
------------------------------ SCHMUTZ Marc IGBMC 1 rue Laurent FRIES BP 163 F 67404 Illkirch Cedex FRANCE
Nasa has been working on micro-machined solid state electron emitters and Thomas George of that agency has published a design utilizing an array of them as the source for a "palm top" SEM in the August, 1998 issue of Nasa Tech Briefs. The proposed SEM relies upon the speculated use of an electon transparent membrane at the bottom of the lens column in this sealed, vacuum pumpless system. Resolution is anticipate in the micron range.
Nasa has also published designs for miniature x-ray tubes in an October 1995 issue of Nasa Tech Briefs.
As far as I know a solid state high energy electron gun exists only as a Star Wars project.
Nasa has been working on micro-machined solid state electron emitters and Thomas George of that agency has published a design utilizing an array of them as the source for a "palm top" SEM in the August, 1998 issue of Nasa Tech Briefs. The proposed SEM relies upon the speculated use of an electon transparent membrane at the bottom of the lens column in this sealed, vacuum pumpless system. Resolution is anticipate in the micron range.
Nasa has also published designs for miniature x-ray tubes in an October 1995 issue of Nasa Tech Briefs.
As far as I know a solid state high energy electron gun exists only as a Star Wars project.
Nasa has been working on micro-machined solid state electron emitters and Thomas George of that agency has published a design utilizing an array of them as the source for a "palm top" SEM in the August, 1998 issue of Nasa Tech Briefs. The proposed SEM relies upon the speculated use of an electon transparent membrane at the bottom of the lens column in this sealed, vacuum pumpless system. Resolution is anticipate in the micron range.
Nasa has also published designs for miniature x-ray tubes in an October 1995 issue of Nasa Tech Briefs.
As far as I know a solid state high energy electron gun exists only as a Star Wars project.
Just a friendly reminder about the upcoming American Chemical Society Course, "Applied Optical Microscopy". This course is not just for chemists!
If you need a refresher on a basic technique, need to know more about interpreting images gathered by a variety of contrast systems (including Hoffman Modulation Contrast and Polarized light), or are moving more into digital imaging, this is a great course for you!
Hands-on, 3 days of total immersion, basic principles through advanced techniques, quantitative polarized light.... and New Orleans, thrown in for good measure! $895 for ACS members; $995 for non-members (tuition and materials only).
Visit the MME website for details: {http://MME-Microscopy.com/education}
Best regards, Barbara Foster ACS Course Coordinator
Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ******************************************************
-----Original Message----- From: Michael Herron [SMTP:herro001-at-maroon.tc.umn.edu]
I am trying to get a handle on the scale of some old pictures. There are human RBC in theses pictures. Can anyone tell me the diameter of your average human RBC?
Mike-
I have recently subscribed to this list in the hopes of picking up some SEM pointers as I am beginning to dip my toes into the field. I certainly can't answer any SEM questions, but I CAN answer your question regarding the size of human RBCs. On a dried film, normal RBC's have a diameter of between 7.2-7.9 um. In certain disease states, they can deviate markedly from this. Microcytic RBC's can be well under 6 um, while macrocytes will have diameters greater than 9 um.
Good luck in your sizing!
{ {...} } Karen Hay, MS, MT(ASCP) Hematology Research, MC 2522 Loma Linda University Medical Center Loma Linda, CA 92354 Tel: (909) 558-4000 x45350 Fax: (909) 558-4189 Khay-at-ahs.llumc.edu {mailto:Khay-at-ahs.llumc.edu}
Michael, How about ~7.5 to 8.5um in diameter (depending on fixation/preparation) and ~2um in greatest thickness. -Ken
On Mon, 7 Feb 2000, Michael Herron wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } All, } } I am trying to get a handle on the scale of some old pictures. There } are human RBC in theses pictures. Can anyone tell me the diameter of } your average human RBC? } } Thanks } Mike } } -- } _________________________________________________ } / Michael J. Herron / } / U of MN,Medicine/Infectious Diseases / } / herro001-at-maroon.tc.umn.edu / } / http://128.101.243.213 / } /_____________________________________________/ } }
We have supplied a number of Cryostats & Microtomes to the automotive industry for sectioning paint finishes on steel body parts & plastic components e.g. bumper (fenders) door handles, door handle surrounds etc. Please view our Website to see our range of instruments.
If I can be of further assistance please come back to me.
Best Regards
Alan Bright
Bright Instrument Co.Ltd. St Margaret's Way Huntingdon PE18 6EB England
Tel No:+44 (0)1480 454528 Fax No:+44 (0)1480 456031 Email: AlanBright-at-brightinstruments.com Web Site: www.brightinstruments.com
-----Original Message----- } From: "csedax-at-alpha.arcride.edu.ar"-at-sparc5.Microscopy.Com [mailto:"csedax-at-alpha.arcride.edu.ar"-at-sparc5.Microscopy.Com] Sent: 08 February 2000 17:35 To: MICROSCOPY-at-sparc5.microscopy.com
Hi! My name is Jorgelina C. Altamirano. I'm an undegrated student of the licenciate in chemical sciences. I'm currently preparing my thesis, being its subjet: The morphological characterization and the chemical analysis of automotive paints with forensic purposes. I'm undergoing my work using SEM and EPMA techniques with a JEOL, JSM 35C and a microprobe EDAX PV9500. I've chosen some papers as references: - Beam et al, Analysis Protocol for discrimination for automotive paints by SEM-EDAX, Journal of Forensic Sciences, 35, 5, 1990 - Zieba-Palus et al, Mikrochim. Acta, 14, 1997.
I've taken same samples of automotive paints (such as Fiat, Peugeot, Renault, Ford, Chevrolet, Volkswagen) corresponding to the years 1994-99. I've found many difficulties while preparing them in: - separating the paint from the body work of the car; - cutting the specimen; - to stick specimen to the graphite specimen pedestal;
Though I've made the primary morphological and chemical analysis and I haven't observed great differences between the diferent automotive paints trades (while polyuretane paints and belayer metalizer paints already tested), but I'd found great differences between layers and layers of the paints specimen. As I'm not quite satisfied with the results obtained to be used for forensic purposes, I would greatly appreciate making contacts with specialist fo this subject, so as to receive any comments on this subject. Thank you in advance for your help. Jorgelina Altamirano CERIDE, Guemes 3450 (3000) Santa Fe, Argentina e-mail: csedax-at-arcride.edu.ar
Jorgelina SEM/EDS//EPMA is only one class of analytical methods that the Forensic community applies to paint analysis for comparison and association with original source. Although used routinely, probe methods are not used exclusively. The organic components in paints are generally more discriminating. Sample preparation usually entails some form of embedment and either microtomy or polishing in order to reveal internal structure. Removal of paint from an auto body is tricky, and requires practice.The manufacturers have designed their paint systems to prevent removal! I would be glad to provide you with additional resources.
Dennis
_________________________________________________________________________ Dennis C . Ward voice: 202-324-2982 FBI Laboratory fax: 202-324-4018 Microanalysis Laboratory email: DCWard-at-concentric.net
----- Original Message ----- } From: {"csedax-at-alpha.arcride.edu.ar"-at-sparc5.Microscopy.Com} To: {MICROSCOPY-at-sparc5.Microscopy.Com} Sent: Tuesday, February 08, 2000 9:35 AM
Jorgelina SEM/EDS//EPMA is only one class of analytical methods that the Forensic community applies to paint analysis for comparison and association with original source. Although used routinely, probe methods are not used exclusively. The organic components in paints are generally more discriminating. Sample preparation usually entails some form of embedment and either microtomy or polishing in order to reveal internal structure. Removal of paint from an auto body is tricky, and requires practice.The manufacturers have designed their paint systems to prevent removal! I would be glad to provide you with additional resources.
Dennis
_________________________________________________________________________ Dennis C . Ward voice: 202-324-2982 FBI Laboratory fax: 202-324-4018 Microanalysis Laboratory email: DCWard-at-concentric.net
----- Original Message ----- } From: "Bright, Alan" {ABright-at-brightinstruments.com} To: {"csedax-at-alpha.arcride.edu.ar"-at-sparc5.Microscopy.Com} ; {MICROSCOPY-at-sparc5.Microscopy.Com} Sent: Tuesday, February 08, 2000 12:59 AM
Jorgelina SEM/EDS//EPMA is only one class of analytical methods that the Forensic community applies to paint for characterization and association with original source. Although used routinely, probe methods are not used exclusively. The organic components in paints are generally more discriminating. Sample preparation usually entails some form of embedment and either microtomy or polishing in order to reveal layer structure. Removal of paint from an auto body is tricky, and requires practice.The manufacturers have designed their paint systems to prevent removal! I would be glad to provide you with additional resources.
Dennis
_________________________________________________________________________ Dennis C . Ward voice: 202-324-2982 FBI Laboratory fax: 202-324-4018 Microanalysis Laboratory email: DCWard-at-concentric.net
----- Original Message ----- } From: {"csedax-at-alpha.arcride.edu.ar"-at-sparc5.Microscopy.Com} To: {MICROSCOPY-at-sparc5.Microscopy.Com} Sent: Tuesday, February 08, 2000 9:35 AM
Dear all
I am planning to replace my Kr/Ar laser on a Zeiss 310 CSLM with two separate lasers: 488nm Ar ion and HeNe 543nm. Does anyone have experience of using a HeNe 543? I need this or something similar to excite Rhodamine for food structure applications. I'd appreciate any advice.
The reason I'm replacing the mixed gas laser is that has averaged 25% downtime over the past three years.
Mark
Mark Auty Dairy Products Research Centre Moorepark, Fermoy, Co. Cork, Ireland. mauty-at-moorepark.teagasc.ie tel 00353 2542447 fax 00353 2542340
Thank you for the many responses to my inquery about operation instructions for the LKB Nova ultramicrotome. For future reference, I have the instrument and the original instructions for the LKB Reichert ultramicrotome if anyone has acquired one of these without instructions. Thanks again for all of your responses.
On Mon, 7 Feb 2000 12:33:38 -0500 Marisa Ahmad {mahmad-at-semiconductor.com} wrote:
} We are experiencing a lot of problems with the tungsten filaments that } we are currently using on our JEOL 5800 SEM. During long acquisitions } (many hours) there is a lot of brightness drift and shifting, even when } lengthy stabilization periods are given prior to the acquisitions. } JEOL has checked out the SEM itself, and can't find the source for the } drifting
Marisa,
You can easily enough to see if it is filament drift. First, start the filament alignment program to ensure alignment. Then with the beam on, wait a few minutes for the filament to drift, then switch off the autobeam function, and manually check the alignment using the 2 filament alignment knobs. If you can make the image brighter, then there is filament drift. With my experiences with the 5800 though, I suspect this is not the case.
W. L. (Buddy) Steffens, Ph.D Dept. of Pathology University of Georgia
On Mon, 7 Feb 2000 16:35:56 -0500 (EST) William Tivol {tivol-at-wadsworth.org} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. }
} Dear Marisa, Do you have control over the filament voltage? If so, } set the voltage just a hair over that necessary to saturate the } filament. If the voltage is too low, parts of the filament will not } emit electrons which get through the wehnelt and into the beam; if too } high, there may be significant evaporation of the filament; either may } cause drifting and/or shifting. I have had very good luck with the } stabilities of both Agar and Ebtec (now Energy Beam Sciences) } filaments; not that others may not be as good, I haven't tried them. } Ebtec makes several shapes of filament, and one may be best for } long-term stability. I have no interest in Agar or Ebtec except as a } satisfied customer. Yours, } Bill Tivol
Marisa,
This is a good point that Bill makes...if you are slightly undersaturated, there will be fluctuations in the rate of electron emission from the filament. The 5800 automatically aligns and saturates the filament when the Autobeam mode is selected...and on our 5800, saturation in this mode is a bit low. If you turn off the autobeam switch and use the filament emission knob to slightly increase the emission current, this might solve the problem at a cost of only a small decrease in filament life.
W. L. (Buddy) Steffens, Ph.D Dept. of Pathology University of Georgia
John Hunt wrote: } } Dear Probe community, } It is winter in Central NY, and the room humidity is 33%. Static } electricity (I believe) has been causing frustrating computer lockups } requiring rebooting our microprobe workstation. } I am wondering if anyone has found the best solution for preventing these } problems. } John we had this same problem for many years while we were using the old RT-11 based Tracor Northern and Kevex x-ray analyzers. The problem was especially bad with the TN-5500. This was due to the keyboard with built in scroll knob. The motion of turning that knob on the keyboard was perfect for generating a little static. (This is pre- GUI and mouse days, except for the Mac users!) Because of the low humidity conditions and severe static accumulation, in winter months we were forced to set the entire keyboard on a grounded pad. We also attached an antistatic strip to the front of the keyboard. Because of the nature of the job, and the movement to and from the computer and the microscope controls, we could not wear the wrist straps. But with the conductive grounding strip across the entire front of the keyboard we found that we could very easily get into the habit of grounding our wrist or thumb on the strip before touching the rest of the computer or keyboard. Then as we used the keyboard we would make some effort to touch the grounding strip from time to time. This virtually eliminated the problem. By the way I believe it was such a problem that Tracor engineered a static discharge switch into the keyboard connection.
Brad Huggins BPAmoco, Naperville, Microscopy and Microanalysis
At 9:20 AM -0600 2/7/0, Michael Herron wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Confocal Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
Hi! My name is Jorgelina C. Altamirano. I'm an undegrated student of the licenciate in chemical sciences. I'm currently preparing my thesis, being its subjet: The morphological characterization and the chemical analysis of automotive paints with forensic purposes. I'm undergoing my work using SEM and EPMA techniques with a JEOL, JSM 35C and a microprobe EDAX PV9500. I've chosen some papers as references: - Beam et al, Analysis Protocol for discrimination for automotive paints by SEM-EDAX, Journal of Forensic Sciences, 35, 5, 1990 - Zieba-Palus et al, Mikrochim. Acta, 14, 1997.
I've taken same samples of automotive paints (such as Fiat, Peugeot, Renault, Ford, Chevrolet, Volkswagen) corresponding to the years 1994-99. I've found many difficulties while preparing them in: - separating the paint from the body work of the car; - cutting the specimen; - to stick specimen to the graphite specimen pedestal;
Though I've made the primary morphological and chemical analysis and I haven't observed great differences between the diferent automotive paints trades (while polyuretane paints and belayer metalizer paints already tested), but I'd found great differences between layers and layers of the paints specimen. As I'm not quite satisfied with the results obtained to be used for forensic purposes, I would greatly appreciate making contacts with specialist fo this subject, so as to receive any comments on this subject. Thank you in advance for your help. Jorgelina Altamirano CERIDE, Guemes 3450 (3000) Santa Fe, Argentina e-mail: csedax-at-arcride.edu.ar
Dear Scott, I've seen a Be Ka x-ray peak in a Link advertisement and we have generated one in the Quartz XOne applications lab with a really good light element deteector. You will find the Be Ka1 peak listed at 0.110 keV in most x-ray tables. At 10:52 AM 2/7/00 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
we are looking at desmosomes in mouse epidermis. My question: From looking at sections we got the impression that demosomes are rather uniform, round knob-like structures. As we did not do any serial sections, we are not sure if this is true. Does anybody know of a publication dealing with this?
Thanks for your help,
Christoph
Christoph Bauer Ph.D. University of Chicago Molecular Genentics and Cell Biology 5841 S. Maryland Ave/MC 1028 Chicago, Il 60637
I received the following fax from Ms. Irene Piscopo, EM Consultant, regarding electron diffraction procedures for the CM-12 or CM-120 TEM's from FEI/Philips. I'll copy the information for those interested.
ELECTRON DIFFRACTION PROCEDURES
There are three methods for doing electron diffraction in the CM-12 involving two different techniques: Method I is an aperture limiting method while Methods II and III are probe limiting. In all cases, the sample must be in the eucentric position and focused.
METHOD I: (area } 1 µm) SAD
1. Select the SA mode (the diffraction aperture and image are focused in the same plane). 2. Insert and center the SA aperture around the area of interest.* 3. Select D. 4. Remove the objective aperture. 5. Overfocus the second condenser to sharpen the pattern (i.e. parallel illumination conditions). 6. Focus the pattern with the focus control which is now changing the diffraction lens). 7. Change the size of the pattern (i.e. camera length) by changing the magnification.
Size of the SA aperture * Size of area selected = ---------------------------------------------------------------------------- ------------ Objective lens magnification in D aperture plane (~ 30x**)
** Actual magnification is focal length dependent.
METHOD II: (areas down to 40 nm with w filament; 20 nm with LaB6)
1. Locate an area of interest. 2. Remove the objective aperture. 3. Using Condensers 1 and 2, bring the beam down to a size slightly smaller than the area of the area from which the diffraction pattern is to be obtained. 4. Go to D and select the desired CL with the magnification control. 5. Focus the pattern with the focus control. Note: While it's sometimes difficult to determine exact focus, it's often easier to insert an objective aperture and focus the edge of the aperture to determine diffraction focus. 6. The size of the discs within the pattern can be decreased by decreasing the size of the condenser aperture. Be sure the aperture is well-centered.
METHOD III: NANOPROBE and/or STEM (to 2 nm)
For diffraction and/or chemical information in the TEM mode from regions of the sample less than 40 nm in diameter requires operating the instrument in either the NANOPROBE or STEM modes.
NANOPROBE:
1. Depress NANOPROBE. 2. Follow instructions in Method II.
STEM:
1. Obtain a focused STEM image using the smallest C2 aperture. 2. Stop the scan. 3. Lower the main screen (CBED pattern) 4. Slightly larger probes and more parallel beam conditions can be obtained using UUD and focusing with INTENSITY (C2 lens).
NOTES ON ELECTRON DIFFRACTION
1. The sample must always be eucentric and focused. 2. In METHOD I, the area from which the diffraction pattern is obtained is determined by the selected area - (diffraction) aperture, when in SA mode. To sharpen the ED pattern, one overfocuses the second condenser (C2) 3. In METHODS II and III, the area from which the diffraction pattern is selected depends on the size of the beam. The size of the beam can be altered by changing condenser lenses C1 and C2 (i.e. spot size and intensity. To sharpen the ED pattern , one reduces the size of the C2 aperture. 4. Spherical aberration limits METHOD I to ~1 µm. 5. In METHODS II and III, spherical aberration of the objective lens is not the limiting factor, the minimum probe size is. 6. For identifying an ED pattern, at least three rings are required. 7. Be sure the sample and the standard are in the eucentric position. Changes in the objective current will cause variations in CL.
Bruce F. Ingber, Biologist USDA-ARS, SRRC 1100 Robert E. Lee Blvd. New Orleans, LA 70124
I'm wrong. I guess I should have looked up at the X-ray periodic table above my head and saw the 0.108 keV value for Be. The reason that I was wrong was that I did not consider the solid state aspects. I am still right about not having a 2s1/2 to a 1s1/2 transition and that an X-ray carries away 1h-bar of angular momentum. What I didn't do was think about the band structure of a solid. If you have the solid, the 2p bands of Be most be spilling into or be the conduction band for the electrons. That must be the source for the electronic transition. I guess I should have engaged brain before typing. My most humble apologies. -Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies." --
} -----Original Message----- } From: microprobe [mailto:chris.salter-at-materials.oxford.ac.uk] } Sent: Tuesday, February 08, 2000 7:10 AM } To: Walck. Scott D. } Subject: Re: Be X-ray peaks -I don't think so } } } Dear Walck, } } You may not, in theory, be able to produce Be X-rays, but we } can detect them on our JEOL JXA 8800, they correspond to the position } in the tables for Be Ka!, and the target is pure Be. } } On Mon, 7 Feb 2000 10:52:29 -0500 "Walck. Scott D." {walck-at-ppg.com} } wrote: } } -----Original Message----- } From: Ekstrom, Harry [mailto:harry.ekstrom-at-honeywell.com] } Sent: Monday, February 07, 2000 7:40 PM } To: 'Walck. Scott D.' } Subject: RE: Be X-ray peaks -I don't think so } } } I don't agree Scott. An L or M line for any element would } not be located in } the area } of Be Ka. And the peak I normally get there with an ultra thin window } is a well defined peak with great resolution. } Please re evaluate your theory. } } Harry Ekstrom } } } } -------------------------------------------------------------- } ---------- } } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.} html } } } } -------------------------------------------------------------- } ---------. } } } } } } When a characteristic X-ray is given off from an atom, it } carries 1 h-bar } } (Planck's constant divided by 2 pi) of angular momentum. } The electronic } } transition that occurred for the X-ray to come off must } conserve angular } } momentum of the system. Therefore, a transition from a } 2s1/2 to a 1s1/2 } } state is forbidden by selection rules. You can't produce a } Be X-ray. } } -Scott } } } } Scott D. Walck, Ph.D. } } PPG Industries, Inc. } } Glass Technology Center } } Guys Run Rd. (packages) } } P. O. Box 11472 (letters) } } Pittsburgh, PA 15238-0472 } } } } Walck-at-PPG.com } } } } (412) 820-8651 (office) } } (412) 820-8161 (fax) } } } } } } } } ------------------- } microprobe } chris.salter-at-materials.ox.ac.uk } } * This e-mail message was sent with Execmail V5.0.x * }
It has been very interesting reading about diffraction and electron beams but it is quite clear that we often give the instrument credit, "the instrument does it automatically", when credit is not due. Almost every "automatic" is a designers estimate or better still a calculation, it is a setting!
After 36 years with TEM I have to say that the only area that the instrument could set in the SA mode could be the approximate position of the first image plane. We normally move to the SA mode to free up the diffraction lens (1st intermediate) to enable it to be focussed on thefirst image plane and the diffraction (intermediate) aperture, independent of the magnification system. We would then focus the image inside the aperture with the objective lens to ensure that the selected area IS the area in the diffraction pattern. Without this procedure electron rotation and misalignments could give you a diffraction pattern from an area not in the centre of the screen! Users are correct in that if they set the eucentric point or better still, adjust the specimen height to focus at the same objective lens current value, that done the first image plane will be at the same point within the diffraction (intermediate) lens.
So how could the Philips work? I would guess they set the diffraction lens at approximately the first image plane. Then as they do not use it within their lens scheme for the SA magnifications it is fixed at this focus, good enough I think?
A second point, parallel beams are very important if you are chasing the best image quality, biology or materials, most times we need high coherence. There are misunderstandings on how to obtain a parallel beam or high coherence, setting the final condenser underfocus is incorrect. The procedure for high coherence would be to use the smallest spot size you can tolerate (this probably means you must up the emission current). Once in this condition overfocus the final condenser (clockwise from crossover) the spot, what ever size it is now, becomes your virtual source. The further overfocus you go the greater the beam coherence . You will deduce the smaller the condenser aperture the sharper the spot and the smaller the spot the greater the coherence for a given degree of overfocus.
Work with a design team and they expect everyone to overfocus, they expect everyone to use small condenser apertures and they expect everyone to use small spot sizes. They do not expect everyone to use too low an emission current (almost everyone does, we have talked before about filament life being the most important feature of many laboratories) because this makes the task too difficult!
Steve Chapman Senior Consultant Protrain Electron microscopy courses world wide http://www.emcourses.com Tel & Fax 44 (0) 1280 814774
} I am planning to replace my Kr/Ar laser on a Zeiss 310 CSLM with two } separate lasers: 488nm Ar ion and HeNe 543nm. Does anyone have } experience of using a HeNe 543? I need this or something similar to } excite Rhodamine for food structure applications. I'd appreciate any } advice.
} The reason I'm replacing the mixed gas laser is that has } averaged 25% downtime over the past three years.
} Mark
} Mark Auty } Dairy Products Research Centre } Moorepark, Fermoy, Co. Cork, Ireland. } mauty-at-moorepark.teagasc.ie } tel 00353 2542447 } fax 00353 2542340
I have been using the Argon ion (488nm) and HeNe 543nm combination on different Zeiss LSM310's and 410's for the past 9 years. The HeNe 543nm is fairly low power, but it has been perfectly adequate for scanning the fluorochromes I have needed to see including TRITC, Texas Red, Cy3, propidium iodide, and Sirius Red. I have never had a problem with the laser going down either, but one would expect that with a HeNe laser.
Slàinte,
Matthew J. Schibler Ph.D. UCLA Brain Research Institute 1524A Gonda (Goldschmied) Center for Neuroscience and Genetics Los Angeles, CA 90095-1761
Hi All, Does anyone have experience with the Pelco UVC2 or UVC1 Cryo Chamber for UV curing of resins (especially LR White) or with something similar? We're contemplating buying something like that for UV polymerization of LR White at -20 C. Thanks for your help, Kristen Kristen A. Lennon Cell, Molecular & Developmental Biology Group Department of Botany & Plant Sciences University of California Riverside, CA 92521 kalen-at-citrus.ucr.edu
Scott, I'm not sure what brought this up, but my reaction is "thems fightin' words". I do not currently have a wavelength reflecting crystal to measure that low in the periodic table, but I did on another instrument. I distinctly
recall seeing a pretty hefty peak at the Be K-alpha position when I focused the beam down on a piece of pure Be metal. I guess rules are made to be broken. Are your calculations correct? Or am I misunderstanding something here?
Jim -- ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ James J. McGee (email: jmcgee-at-sc.edu) Department of Geological Sciences University of South Carolina Columbia, SC 29208
Tel: 803-777-6300 Fax: 803-777
"Walck. Scott D." wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } When a characteristic X-ray is given off from an atom, it carries 1 h-bar } (Planck's constant divided by 2 pi) of angular momentum. The electronic } transition that occurred for the X-ray to come off must conserve angular } momentum of the system. Therefore, a transition from a 2s1/2 to a 1s1/2 } state is forbidden by selection rules. You can't produce a Be X-ray. } -Scott } } Scott D. Walck, Ph.D. } PPG Industries, Inc. } Glass Technology Center } Guys Run Rd. (packages) } P. O. Box 11472 (letters) } Pittsburgh, PA 15238-0472 } } Walck-at-PPG.com } } (412) 820-8651 (office) } (412) 820-8161 (fax)
Dear Listers, I have responses to two of today's postings, but they are quite unrelated. 1. I did not see the original question about Be x-rays, only Scott Walk's reply, so I don't know what info was being sought. We used a Be-containing mineral to evaluate microprobes before buying. I won't embarrass those who failed, but only one manufacturer succeeded. The reason the others failed is probably that in compounds the x-ray energy is subject to largish chemical shifts relative to pure Be because there is no shielding by outer electrons and you need to search on either side of the tabulated value of about 0.1keV. So there IS a Be x-ray and a reasonable WDS spectrometer should detect it.
2. Filament drift. Fred Schamber's reply is right most of the time, but I did once have a batch of 6 JEOL-type filaments made by an EM supply house where 2 of them drifted until the tips were right at the edge of the hole and stayed there even when the filament was cooled down again. This was the only time in about 15 years of using JEOL microscopes.
Eric ---------------------- Dr Eric Lachowski Department of Chemistry University of Aberdeen Meston Walk Old Aberdeen AB24 3UE Scotland +44 (0)1224 272934 fax 272921 e.lachowski-at-abdn.ac.uk
Hi List I am using live blood staining light microscopy, is there anyone who has tried or is using this technique. I would like to swap notes. Dean Armytage PhD Hillstream Health Centre Australia
I'm getting more curious as I read on here. Speaking strictly energy now and leaving out wavelength, the characteristic energy of Be Ka is about .11 KeV like Mary stated. I've had an occasional peak show up there before and thought it may have been Be. Now I understand that it may have been "noise" all along. Would "noise" be a pretty defined and relatively well resolved peak at that energy level using an ultra thin window? Depending on the low energy cutoff setting, one might truly have Be and never detect it. Would lowering the KV reduce the noise artifact?
I guess I'll get out my Be planchet and try a few things.
Harry
-----Original Message----- } From: Jim McGee [mailto:mcgee-at-geol.sc.edu] Sent: Tuesday, February 08, 2000 3:09 PM To: 'Microscopy'
Scott, I'm not sure what brought this up, but my reaction is "thems fightin' words". I do not currently have a wavelength reflecting crystal to measure that low in the periodic table, but I did on another instrument. I distinctly
recall seeing a pretty hefty peak at the Be K-alpha position when I focused the beam down on a piece of pure Be metal. I guess rules are made to be broken. Are your calculations correct? Or am I misunderstanding something here?
Jim -- ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ James J. McGee (email: jmcgee-at-sc.edu) Department of Geological Sciences University of South Carolina Columbia, SC 29208
Tel: 803-777-6300 Fax: 803-777
"Walck. Scott D." wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } When a characteristic X-ray is given off from an atom, it carries 1 h-bar } (Planck's constant divided by 2 pi) of angular momentum. The electronic } transition that occurred for the X-ray to come off must conserve angular } momentum of the system. Therefore, a transition from a 2s1/2 to a 1s1/2 } state is forbidden by selection rules. You can't produce a Be X-ray. } -Scott } } Scott D. Walck, Ph.D. } PPG Industries, Inc. } Glass Technology Center } Guys Run Rd. (packages) } P. O. Box 11472 (letters) } Pittsburgh, PA 15238-0472 } } Walck-at-PPG.com } } (412) 820-8651 (office) } (412) 820-8161 (fax)
My personal experience with silver enhancement dates back to when Janssen Life Sciences first introduced the IntenSE M silver enhancement kit in late 80's (this product was later taken over by Amersham). At the time, I felt the IntenSE M was easy to use, but its fast reaction made it hard to control the particle growth. Just like you, the desire for a better results has kept me searching and testing whenever a new protocol or product became available.
Danscher's method gives wonderful intensification, but its low pH sometimes damages the ultrastructure when used for the pre-embedding immunogold labeling. Burry's method and the Nanoprobes HQ silver made progress with pH, however they inherited Danscher's high viscosity and light sensitivity, which limit their penetration in pre-embedding silver enhancement and render it more difficult to handle from a practical standpoint. I have also tested the Nanoprobes GoldEnhance kit for pre-embedding immunogold labeling recently. At the LM level the results were decent. I have not spent a lot of time evaluating its performance at the EM-level. So far, the particle size homogeneity is not as good as what I've found with Danscher's method. But I will refrain from further judgment until more work has been done.
In my opinion, the first breakthrough in intensifying gold particles since Danscher's method was realized when Aurion, a Dutch company, first introduced the R-gent SE-EM silver enhancement kit last June. As you wished for in your post, it has a near-neutral pH, low viscosity, and it is light insensitive. Moreover, it gives a great enhancement efficiency and even particle size and shape. Background remains minimal even after two hours enhancement on the bench. Morphology preservation after enhancement is superior to any other enhancement reagent I have used. I have been testing this kit profusely for both post- and pre-embedding immunogold labeling on various types of samples (including cell cultures and vibratome sections), with many different primary and secondary antibodies, and am very pleased with its performance. I use 0.5% OsO4 for 20 min and have never had any problem with silver disappearing. If you like, I would be very happy to e-mail some images to you so you can evaluate them yourself.
Hong ================= Hong Yi Emory University Department of Neurology 6215 Woodruff Memorial Research Building 1639 Pierce Dr. Atlanta, GA 30341 Phone: (404) 727-8692 Fax: (404) 727-3157 Email: hyi-at-emory.edu
On Thu, 3 Feb 2000, Michael Plociniak wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Colleagues, } Has anyone used gold enhancement reagents from Nanoprobes, Inc. } (Stonybrook, NY - USA) as an alternative to silver enhancement for } enlarging 1 nm immunogold probes? } } Advertisements state that gold enhancement has lower background and is } compatible with osmium. Can anyone verify this from personal experience? } } In addition to these advantages, I am hoping that milder pH conditions will } be less damaging to ultrastructure in cultured neurons (using a } preembedment protocol). } } Thank you, } Michael }
Here is the way I found yesterday for finding the diffraction plane. By diffraction plane I mean the plane at which diffraction pattern is formed (crossover, Fourier of the object wave) not the theoretical BFP of the objective.
First perform standard alignment for brightfield mode. Then go to 10 000X. There are several ways to perform:
1. Fixed illumination and specimen position (i.e. OL current)
a) Adjust the desired brightness with C2. Adjust the desired OL current and specimen position. b) Insert OL aperture and move it so that the edge of the aperture to cross through the middle of the screen. If the aperture is exactly at the diff. plane then you'll not be able to see its edge - just the whole beam will fade uniformly ... so you have the diff. plane. c) If you see aperture edge then it is not at diff. plane. Now two ways: - change the aperture z-position (not recommended ... but if it is very far ..) - If your microscope has condenser minilens you can try tweaking both C2 and CML in order to keep the intensity same and just make the aperture edge disappear (i.e. the whole screen uniformly darkened).
2. Fixed specimen (i.e. OL current).
a) Focus the specimen b) same as 1b c) Change C2 excitement in order to make the edge disappear
3. Fixed illumination
a) Set desired C2 excitement b) same as 1b c) Change the objective lens current until the aperture edge disappears. Then move specimen in z-direction until it is focused.
In all these methods after adjusting the aperture at diff. plane you can try tweaking C2 or OL ... you will see that the aperture is not at diff. plane anymore, The edge will appear on one or the other side (i.e. mirrored) depending on the direction of the lens current change. Another thing - after adjusting the aperture at diff. plane you can check if beam shift tilts the beam (by tilting here I mean - if the specimen is illuminated with plane wave then shifting the beam should not tilt the wave front). Shift the beam and if the brightness changes (when the aperture edge partially covers the zero diff. spot) then the beam is tilting. I wish there was a IL1 tweaking knob allowing for change of the IL1 current while in brightfield mode. This wold make the things much more easier.
About the methods for making the illumination at the specimen parallel. They will work only if: - The OL aperture is positioned by the manufacturer exactly at the theoretical BFP (I think that in most of the TEMs it is not) - The OL current should be set to zero deviation (i.e. at the value for which the BFP position was calculated) - The intermediate lens system should be properly calibrated by the manufacturer so that if both of the above conditions are satisfied and the specimen is at front focal plane of the OL then it to be in focus. In other words the theoretical BFP of the objective to coincide with the theoretical front focal plane of the IL1.
I don't think parallel illumination is so very important ... using spherical wave will give the same results but just the diff. planes will be shifted.
All of the above is just my opinion. I could be mistaking somewhere ...
Best wishes,
Rado
--------------------------------------------------------------------- Radostin Danev Laboratory of Ultrastructure Research National Institute for Physiological Sciences Myodaiji-cho, Okazaki 444-8585, JAPAN e-mail: rado-at-nips.ac.jp ---------------------------------------------------------------------
----- Original Message ----- } From: "Dennis Ward" {DCWard-at-concentric.net} } } Jorgelina } SEM/EDS//EPMA is only one class of analytical methods that the Forensic } community applies to paint analysis for comparison and association with } original source. Although used routinely, probe methods are not used } exclusively. The organic components in paints are generally more } discriminating. } Sample preparation usually entails some form of embedment and either } microtomy or polishing in order to reveal internal structure. } Removal of paint from an auto body is tricky, and requires practice.The } manufacturers have designed their paint systems to prevent removal! } I would be glad to provide you with additional resources.
I have never do this but this is what i would try first.
One thing I would try on would be removing the metal from the paint. A weak elecrolite solution and a low DC voltage connected to reverse electoplate the metal away sould get rid of almost all of it and the last bit could be removed with acid or evaporated as the cathode in a vacum chamber.
Once you get the metal down to a bunch of small islands you might be able to let it rust free of the paint.
I have no idea what this would do to the paint but most paint films I have delt with are pretty tough.
Gordon
Gordon Couger gcouger-at-couger.com
Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
we are massively working on analytical and structural TEM characterization of semiconductors, mainly InGaAs/GaAs, quantum well, wires and dots.
Specimen preparation is of course a crucial point of our work.
We have just switched to PIPS (we used to make our samples using the Gatan Duo Mill), and we are getting controversial results about the damage introduced by the milling procedure.
We are perfectly aware of all the differences between the two instruments, especially the absence of cooling stage and the higher beam current.
Has anyone performed a systematic and careful analysis to try to evaluate the milling damage on similar materials systems. We are willing to start a systematic work to assess this issue, but would like to know if anyone has already done anything on this topic. Also, any collaboration will be more than welcome.
in our lab we have prepared a number of GaAs, AlGaAs, InGaAs devices. Mechanical thinning is by the tripod wedge technique, but typically we end up with more damage than we find for Si samples so there is a need to leave samples thicker and use more extensive ion milling for final thinning.
We also have a PIPS and I have found its capabilities essential for achieving good thin samples in these materials. I found more success by using low angles and low KeV for long times. Typically I would be using 5-6 degrees and 4-4.5 KeV initially, with 3-4 degrees and 3-3.5 KeV for the final ion polishing. On samples 2-3 microns thick, this would amount to 2-3 hours total milling time. I did not observe any artefacts introduced with milling under these conditions, and the junction structures and inherent defects were clearly observed.
The conditions above should work with any ion miller capable of low angle milling, but our experience is only with the PIPS. If you use the wedge technique for mechanical thinning, I would recommend that you use Mo grids to mount your specimens as the extended milling times at low angles will really chew up a Cu grid. Also, Mo grids are considerably thinner, allowing angles as low as 3-4 degrees on the grid hole side for a sample positioned in the middle of a 1 mm grid hole.
If you would like, I can send you some typical images of devices we have prepared, including images showing the removal of the mechanical polishing damage.
Philip L. Flaitz IBM Analytical Services, Hopewell Junction, NY http://www.chips.ibm.com/services/asg Ph.......(914) 892-3094, FAX -2003 flaitz-at-us.ibm.com
Massimo Catalano {massimo.catalano-at-ime.le.cnr.it} on 02/09/2000 03:43:03 AM
To: Microscopy-at-sparc5.Microscopy.Com cc:
Dear Listservers,
we are massively working on analytical and structural TEM characterization of semiconductors, mainly InGaAs/GaAs, quantum well, wires and dots.
Specimen preparation is of course a crucial point of our work.
We have just switched to PIPS (we used to make our samples using the Gatan Duo Mill), and we are getting controversial results about the damage introduced by the milling procedure.
We are perfectly aware of all the differences between the two instruments, especially the absence of cooling stage and the higher beam current.
Has anyone performed a systematic and careful analysis to try to evaluate the milling damage on similar materials systems. We are willing to start a systematic work to assess this issue, but would like to know if anyone has already done anything on this topic. Also, any collaboration will be more than welcome.
Hello ..... we have an old ETEC Omniscan SEM, and we are searching for ETEC company or any people that can supply manuals ....
any help is welcome
best regards
=================================================== Fernando D. Balducci Laboratorio de Microscopia Electr—nica Facultad de Ingenier’a - Bioingenieria Universidad Nacional de Entre Rios Argentina e-mail: microsc-at-fi.uner.edu.ar tel: 54 43 975100 fax: 54 43 975077
I am working with archival specimens of formalin-fixed paraffin embedded tissues and I am getting tremendous autofluorescence. I looked at tissue sections after the sections have been de-waxed using either FITC or rhodamine filter sets and red cells and connective tissues are brightly fluorescent. Is there a way of suppressing the autofluorescence and still retain reactivity of tissue to antibodies? I would appreciate any comments or suggestions.
******************************************************* Corazon D. Bucana, Ph.D. Department of Cancer Biology U.T. M.D. Anderson Cancer Center 1515 Holcombe Blvd. Box 173 Houston, Texas 77030 Phone: (713) 792-8106 FAX: (713) 792-8747 Email:bucana-at-audumla.mdacc.tmc.edu FAX: (713) 792-8747
I don't have a UTW detector here, so I'll leave this to the EDS experts. But, on the JEOL 8800/8900 I used to run we had a thin window detector that could see B (not Be) if the low end noise peak (which is huge) was properly discriminated. Get out your C planchett while you are getting the Be one, and see if you get the noise peak at Be. Far better to use wavelength than EDS down at that end.
Jim McGee -- ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ James J. McGee (email: jmcgee-at-sc.edu) Department of Geological Sciences University of South Carolina Columbia, SC 29208
Tel: 803-777-6300 Fax: 803-777-6610
"Ekstrom, Harry" wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I'm getting more curious as I read on here. Speaking strictly energy now } and leaving out wavelength, the characteristic energy of Be Ka is about .11 } KeV like Mary stated. I've had an occasional peak show up there before and } thought it may have been Be. Now I understand that it may have been "noise" } all along. Would "noise" be a pretty defined and relatively well resolved } peak at that energy level using an ultra thin window? Depending on the low } energy cutoff setting, one might truly have Be and never detect it. Would } lowering the KV reduce the noise artifact? } } I guess I'll get out my Be planchet and try a few things. } } Harry } } -----Original Message----- } } From: Jim McGee [mailto:mcgee-at-geol.sc.edu] } Sent: Tuesday, February 08, 2000 3:09 PM } To: 'Microscopy' } Subject: Re: Be X-ray peaks -I don't think so } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Scott, } I'm not sure what brought this up, but my reaction is "thems fightin' } words". I do not currently have a wavelength reflecting crystal to measure } that low in the periodic table, but I did on another instrument. I } distinctly } } recall seeing a pretty hefty peak at the Be K-alpha position when I focused } the beam down on a piece of pure Be metal. I guess rules are made to be } broken. Are your calculations correct? Or am I misunderstanding something } here? } } Jim } -- } ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ } James J. McGee (email: jmcgee-at-sc.edu) } Department of Geological Sciences } University of South Carolina } Columbia, SC 29208 } } Tel: 803-777-6300 Fax: 803-777 } } "Walck. Scott D." wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } When a characteristic X-ray is given off from an atom, it carries 1 h-bar } } (Planck's constant divided by 2 pi) of angular momentum. The electronic } } transition that occurred for the X-ray to come off must conserve angular } } momentum of the system. Therefore, a transition from a 2s1/2 to a 1s1/2 } } state is forbidden by selection rules. You can't produce a Be X-ray. } } -Scott } } } } Scott D. Walck, Ph.D. } } PPG Industries, Inc. } } Glass Technology Center } } Guys Run Rd. (packages) } } P. O. Box 11472 (letters) } } Pittsburgh, PA 15238-0472 } } } } Walck-at-PPG.com } } } } (412) 820-8651 (office) } } (412) 820-8161 (fax)
-- ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ James J. McGee (email: jmcgee-at-sc.edu) Department of Geological Sciences University of South Carolina Columbia, SC 29208
I am looking for TEM images that show cross-sectional views of skeletal muscle sarcomeres during the *contracted state*. In particular, I am interested in seeing the spacing of the thin filaments when they **overlap each other** (i.e., when those from one side of the sarcomere overlap those from the other side of the sarcomere in the contracted state). In addition, I am interested in seeing the distribution of the thin filaments as they pass through the M line. It would be greatly appreciated if anyone could tell me if they know of any research papers or review articles that contain such TEM images.
Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 75 N. Eagleville Rd. U-125 Storrs, CT 06269 Knecht-at-uconnvm.uconn.edu 860-486-2200 860-486-4331 (fax) home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html ************************************************************
Christoph: I don't have any references at hand, but there is a whole literature on desmosomes. A search on Medline (or PubMed on the internet) should provide you with a good list of EM related publications on desmosomes--probably more than you ever wanted to know! As an aside, I think there may be some books that cover this as well, but since it is not an area of personal expertise or current research interest, I am afraid they have all gone from my memory banks.
Roger
On Tue, 08 Feb 2000 09:56:35 -0800, Christoph Bauer wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } we are looking at desmosomes in mouse epidermis. My question: From } looking at sections we got the impression that demosomes are rather } uniform, round knob-like structures. As we did not do any serial } sections, we are not sure if this is true. Does anybody know of a } publication dealing with this? } } Thanks for your help, } } Christoph } } } } Christoph Bauer Ph.D. } University of Chicago } Molecular Genentics and Cell Biology } 5841 S. Maryland Ave/MC 1028 } Chicago, Il 60637 }
Roger C Moretz, Ph.D.
_______________________________________________________ Get 100% FREE Internet Access powered by Excite Visit http://freeworld.excite.com
Christoph, I have examined alot of mouse and pig skin at the TEM level and the desmosomes and hemidesmosomes appear rather typical, that is, small, long bridge-like structures between the cells.
Mary Gail Engle Electron Microscopy & Imaging Facility University of Kentucky
At 09:56 AM 2/8/00 -0800, Christoph Bauer wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
The Department of Microscopy and Microanalysis at Abbott Laboratories has two summer internships available for 8-12 weeks this summer. One position is in Biological Microscopy, and one is in Materials Analysis and Microscopy. We're looking for students who are considering a career in microscopy, especially students interested in the pharmaceutical industry.
Abbott Laboratories is a diversified healthcare company that produces pharmaceuticals, diagnostic devices, and hospital products. The Microscopy department analyzes samples related to all these functions. We use polarized light, fluorescence, TEM, SEM, EDXS, confocal microscopy, and flow cytometry to solve problems related to products, as well as to provide support for pharmaceutical Discovery and Development basic research and drug safety.
Housing and a stipend are provided for the summer. We're located near Lake Michigan and the Wisconsin state border, about an hour's drive or train-ride north of Chicago. There's lots to see and do in the area, and there are planned activities with other interns, as well. The Abbott Summer Internship Program has been nationally recognized as one of the best in the country.
If you're interested, please send a resume to:
Jane A. Fagerland, Ph.D. Abbott Laboratories D-45M/AP31 200 Abbott Park Rd. Abbott Park IL 60064-6202
You may also fax a resume to me at (847) 938-5027 or e-mail it to me at jane.a.fagerland-at-abbott.com.
If you'd like further information, I can be reached by telephone at (847) 935-0104, or by e-mail. I prefer e-mail, but will be more than happy to talk to discuss our projects and laboratory by telephone.
There is an easy way to determine the milling damage between the two machines and the amount of ion milling damage (subjectively); compare them with samples prepared using the small angle cleavage technique. Look at John McCaffrey's paper on using the small angle cleavage technique in Ultramicrotomy 38, (149) 1991. He shows the difference between high angle milling, low angle milling and the small angle cleavage technique. Of course, the small angle cleavage technique showed no ion milling damage. It is perfect for these types of materials. If you don't need a site specific technique, it is the way to go for semiconductors.
You should also look at his paper in the MRS TEM Sample Prep series III (vol 254) that also shows a comparison for a SiGe/Si layer structure. We have a detailed pictorial outline with tips and tricks on how to do it in the MRS TEM Sample Prep series IV (vol 480).
I know that you invested a lot in your ion mill, but you can do these samples very cheaply. SouthBay Technology sells a Microcleave kit that gets you started with the technique.
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies." --
} -----Original Message----- } From: Massimo Catalano [mailto:massimo.catalano-at-ime.le.cnr.it] } Sent: Wednesday, February 09, 2000 3:43 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: PIPS and milling damage } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.} html } } } -------------------------------------------------------------- } ---------. } } } Dear Listservers, } } we are massively working on analytical and structural TEM } characterization } of semiconductors, mainly InGaAs/GaAs, quantum well, wires and dots. } } Specimen preparation is of course a crucial point of our work. } } We have just switched to PIPS (we used to make our samples } using the Gatan } Duo Mill), and we are getting controversial results about the damage } introduced by the milling procedure. } } We are perfectly aware of all the differences between the two } instruments, } especially the absence of cooling stage and the higher beam current. } } Has anyone performed a systematic and careful analysis to try } to evaluate } the milling damage on similar materials systems. We are } willing to start a } systematic work to assess this issue, but would like to know } if anyone has } already done anything on this topic. Also, any collaboration } will be more } than welcome. } } Thanks. } } Massimo } } } } } Dr. Massimo Catalano } CNR-IME } Campus Universitario } Via Arnesano } 73100 Lecce - ITALY } tel: + 39 0832 322362 } fax: + 39 0832 325299 } email: massimo.catalano-at-ime.le.cnr.it } http://www.ime.le.cnr.it } http://www.ime.le.cnr.it/sime/sime.htm } } }
It has been brought to my attention that no dates were attached to this course reminder:
March 10-12, 2000 Hyatt Regency New Orleans, LA.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education}
At 07:57 PM 2/7/00 -0500, Barbara Foster wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Suggest you contact Chuck Garber at Structure Probe: www.2spi.com I'm sure that he has some sort of derivative of his tacky dots which would be helpful
At 08:48 PM 8/27/70 +0000, Linda Chicoine wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I know this is off topic however a change like this could make this type of forum impossible.
} Date: Tue, 8 Feb 2000 08:13:53 -0700 } X-Sender: liperez-at-cnmailsvr.nmsu.edu } Mime-Version: 1.0 } To: all-at-biology.nmsu.edu } From: liperez-at-NMSU.Edu (LSPSaldana) } Subject: Congress to allow email charges } Sender: owner-all-at-biology.nmsu.edu } Precedence: bulk } X-Keywords: } X-UID: 43 } Status: O } } } } } Congress to allow email charges } } } } } } Please pass this on to all you know since many of us use e-mail for } } } business and to keep up with friends and family, I thought you'd like } } } to know the following. } } } } } } Please jump on it right away and forward this to others. } } } } } } CNN has reported that within the next two weeks Congress is going to } } } vote on allowing telephone companies to CHARGE A TOLL FEE for } } } internet access. } } } Translation: Every time we send long distance e-mail we will receive a } } } long distance charge. This will get costly. Please visit the following web } } } site and file a complaint. Complain to your Congressperson. We can't } } } allow this to pass. The following address will allow you to send an } } } e-mail on this subject DIRECTLY to your Congressperson. } } } } } } http://www.house.gov/writerep {http://www.house.gov/writerep} } } } } } } Pass this on to your friends. It is urgent! I hope all of you will pass } } } this on to all your friends and family. We should ALL have } } } an interest in this one. } } } } } } WAIT, THERE'S MORE. IN ADDITION, The last few months have revealed an } } } alarming trend in the Government of the United States attempting to } } } quietly push through legislation that will affect your use of the } } Internet. } } } Under } } } proposed legislation the U.S. Postal Service will be attempting to bill } } } email users out of "alternate postage fees". Bill 602P will permit the } } } Federal Govt. to charge a 5 cent surcharge on every email delivered, by } } } billing Internet Service Providers at source. The consumer would then be } } } billed in turn by the ISP. Washington D.C. lawyer Richard Stepp is } } } working without pay to prevent this legislation from becoming law. The } } U.S. } } } Postal Service is claiming that lost revenue due to the proliferation of } } } email } } } is costing nearly $230,000,000 in revenue per year. You may have noticed } } } their recent ad campaign "There is nothing like a letter". } } } Since the average citizen received about 10 pieces of email per day in } } } 1998, } } } the cost to the typical individual would be an additional 50 cents per } } } day, or over $180 dollars Per year, above and beyond there regular } } Internet } } } costs. } } } Note that this would be money paid directly to the U.S. Postal Service } } } for a service they do not even provide. The whole point of the Internet is } } } democracy and non-interference. If the federal government is permitted } } } to tamper with our liberties by adding a surcharge to email, who knows } } Where } } } it will end. You are already paying an exorbitant price for snail mail } } } because of bureaucratic inefficiency. It currently takes up to 6 days for } } a } } } letter to be delivered from New York to Buffalo. If The U.S. Postal } } Service } } } is } } } allowed to tinker with email; it will mark the end of the "free" } } } Internet in the United States. } } } One congressman, Tony Schnell has even suggested a "twenty to forty } } } dollar per month surcharge on all internet service" above and beyond the } } } government's proposed email Charges. Note that most of the major } } } newspapers have ignored the story, the only exception being the } } } Washingtonian } } } which called the idea of email surcharge "a useful concept who's time has } } } come" } } } (March 6, 1999) Editorial. } } } } } } Don't sit by and watch your freedoms erode away! Send this e-mail to } } } EVERYONE on your list, and tell all your friends and relatives to write } } } to their congressman and say "No!" to Bill 602P. } } } } } } } } } It will only take a few moments of your time, and could very well be } } } instrumental in killing a bill we don't want. } } } } } } PASS THIS ON TO EVERYONE YOU KNOW WHO USES EMAIL REMEMBER THESE ARE TWO } } } SEPARATE ISSUES THAT EFFECT ALL OF US ONLINE. LET YOU VOICE BE HEARD } } } NOW, NOT AFTER } } } } } } } } } } } } } } } } } } } } }
Dear Donald, the Institute of Applied Physics (http://www.sandy.ru/science/science/appl.new/frames.html) constructs, produces and uses solid state high energy electron gun for the High-power electronics and plasma physics researches. May be they will help you.
Best regards, Dr. S.A.Gusev
******************************************** * Institute for Physics of Microstrutures * * Russian Academy of Science * * ( IPM RAS ) * * * * Niznii Novgorod, GSP-105 * * 603600 * * RUSSIA * * * ********************************************
Hello, We have got a JEM3010 Transmission electron microscopy.I have got a one problem about water cooling system.I dont know,Which kind of water to used?I think so, We should use pure water for water cooling?Besides My city water is not clean.What do you think about this problem? Thanks for your interest
********************************** ********************************** ** Research.Asst.ERDEM YASAR ** ** University of Kirikkale ** ** Department of Physics ** ** TEM LABORATORY ** ** 71450 KIRIKKALE/TURKEY ** ** erdem.yasar-at-physics.org ** ** yasar-at-science.ankara.edu.tr ** ** yasar-at-turkuaz.kku.edu.tr ** ********************************** **********************************
I am looking for TEM images that show cross-sectional views of skeletal muscle sarcomeres during the *contracted state*. In particular, I am interested in seeing the spacing of the thin filaments when they **overlap each other** (i.e., when those from one side of the sarcomere overlap those from the other side of the sarcomere in the contracted state). In addition, I am interested in seeing the distribution of the thin filaments as they pass through the M line. It would be greatly appreciated if anyone could tell me if they know of any research papers or review articles that contain such TEM images.
Thank you!
Izak Paul Biological Sciences Mount Royal College
The following papers are classics and would make good starting point. Huxley, H.E. & Hanson, J. (1954). Nature, 173, 973-976. Huxley, A.F. & Niedergerke, R.M. (1954). Nature, 173, 971. Huxley, H.E. (1957). Biophys. & Biochem. Cytol. 3, 631-648.
Thin filaments 'pass through' the Z-line. Try John Squire and Pradeep Luther (3D reconstruction of fish Z-disc) and Mike Reedy (insect) or Bellinda Bullard.
Matt Walker School of Biomedical Sciences Leeds University, UK
I'm sorry to bring this topic up yet again, but I have just come across the specification for a flat-bed scanner which looks as if it would be suitable for scanning e.m. cut-film. It is the Epson Perfection 1200 Photo which includes a 5x4inch film adapter.
The UK web site address is: http://www.epson.co.uk/sohoprod/imaging/scanner/perf1200/perf1200photo/spec.htm
and the specification is: A4 Colour flatbed with 5"x4" film image scanner Single pass scanning Optical resolution: 1200 x 2400 (30600 pixels/line) Maximum output resolution of 9600x9600 dpi 36 bits per pixel in colour (24 bit output) 12 bits per pixel in black (8 bit output) Optical Density 3.2D USB (Type B)
Does anyone have any experience of this machine because it is about a fifth of the price of the Umax flatbed film scanners? My only reservation is that it is a SOHO product rather than professional.
Thanks in advance
Malcolm
Malcolm Haswell Electron Microscopy School of Sciences Fleming Building University of Sunderland SUNDERLAND SR1 3SD Tyne and Wear UK
Tel (0191) 515 2872 e-mail: malcolm.haswell-at-sunderland.ac.uk
This is a recurring post about nothing more than a hoax.
Search for internet hoaxes and so forth and you will find this one and many other interesting yet equally invalid assertions.
gary g.
At 10:58 PM 2/9/00 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
The statement that the smallest spot size gives the best coherence can be a misleading. The full statement should be the spot size with the smallest angular distribution should be used. Anyone who has tried to do off-axis holography in nanoprobe mode will testify to this. The spatial coherence envelope, roughly the distance over which the beam is considered 'coherent', is the Fourier transform of the angular distribution of intensity at the source (Van Cittert Vernike theorem).
Born & Wolf (section 10.4.2) "Hence if the linear dimensions of the source and the distance between P1 and P2 (points in the imaging plane) are small compared to the distance of these points from the source, the degree of coherence, |mu_12| is equal to the absolute value of the normalized Fourier transform of the intensity function of the source"
The reason holography and high resolution work is done with te most parallel beam possible becomes clear, the angular distribution of a converged (or diverged beam) is wide enough to reduce the spatial coherence envelope. In off-axis holography this is even more stringent since two interfering points (reference wave & object wave) can be hundreds of nanometers, microns even, apart. Elliptical illumination is used to preserve coherence in the one important direction (minimum angular width) and converged in the other to improve the intensity.
******************************************************** Dr Jonathan Barnard
Analytical Materials Physics The Angstrom Laboratory, Uppsala University P O Box 534, SE-751 21 Uppsala, Sweden Phone: +46-(0)18-4716838 Fax: +46-(0)18-500131 http://www.angstrom.uu.se/analytical/home.html
I read the following just the other day, from the US Postal Service
http://www.usps.gov/news/press/99/99045new.htm
FOR IMMEDIATE RELEASE May 21, 1999 Release No. 45
E-MAIL RUMOR COMPLETELY UNTRUE
WASHINGTON – A completely false rumor concerning the U.S. Postal Service is being circulated on Internet e-mail. As a matter of fact, the Postal Service has learned that a similar hoax occurred recently in Canada concerning Canada Post.
The e-mail message claims that a "Congressman Schnell" has introduced "Bill 602P" to allow the federal government to impose a 5-cent surcharge on each e-mail message delivered over the Internet. The money would be collected by Internet Service Providers and then turned over to the Postal Service.
No such proposed legislation exists. In fact, no "Congressman Schnell" exists.
The U.S. Postal Service has no authority to surcharge e-mail messages sent over the Internet, nor would it support such legislation.
-30-
Evex Analytical X-ray Analyzers and Digital Imaging Systems 857 StateRoad Princeton, NJ 08540 609-252-9192 T 609-252-9091 F www.evex.com info-at-evex.com
In a message dated 2/10/00 12:26:56 AM Eastern Standard Time, chbennet-at-nmsu.edu writes:
{ { Subj: FYI: Congress to allow email charges Date: 2/10/00 12:26:56 AM Eastern Standard Time From: chbennet-at-nmsu.edu (Christina bennett) To: microscopy-at-sparc5.microscopy.com
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html -----------------------------------------------------------------------.
I know this is off topic however a change like this could make this type of forum impossible.
} Date: Tue, 8 Feb 2000 08:13:53 -0700 } X-Sender: liperez-at-cnmailsvr.nmsu.edu } Mime-Version: 1.0 } To: all-at-biology.nmsu.edu } From: liperez-at-NMSU.Edu (LSPSaldana) } Subject: Congress to allow email charges } Sender: owner-all-at-biology.nmsu.edu } Precedence: bulk } X-Keywords: } X-UID: 43 } Status: O } } } } } Congress to allow email charges } } } } } } Please pass this on to all you know since many of us use e-mail for } } } business and to keep up with friends and family, I thought you'd like } } } to know the following. } } } } } } Please jump on it right away and forward this to others. } } } } } } CNN has reported that within the next two weeks Congress is going to } } } vote on allowing telephone companies to CHARGE A TOLL FEE for } } } internet access. } } } Translation: Every time we send long distance e-mail we will receive a } } } long distance charge. This will get costly. Please visit the following web } } } site and file a complaint. Complain to your Congressperson. We can't } } } allow this to pass. The following address will allow you to send an } } } e-mail on this subject DIRECTLY to your Congressperson. } } } } } } http://www.house.gov/writerep {http://www.house.gov/writerep} } } } } } } Pass this on to your friends. It is urgent! I hope all of you will pass } } } this on to all your friends and family. We should ALL have } } } an interest in this one. } } } } } } WAIT, THERE'S MORE. IN ADDITION, The last few months have revealed an } } } alarming trend in the Government of the United States attempting to } } } quietly push through legislation that will affect your use of the } } Internet. } } } Under } } } proposed legislation the U.S. Postal Service will be attempting to bill } } } email users out of "alternate postage fees". Bill 602P will permit the } } } Federal Govt. to charge a 5 cent surcharge on every email delivered, by } } } billing Internet Service Providers at source. The consumer would then be } } } billed in turn by the ISP. Washington D.C. lawyer Richard Stepp is } } } working without pay to prevent this legislation from becoming law. The } } U.S. } } } Postal Service is claiming that lost revenue due to the proliferation of } } } email } } } is costing nearly $230,000,000 in revenue per year. You may have noticed } } } their recent ad campaign "There is nothing like a letter". } } } Since the average citizen received about 10 pieces of email per day in } } } 1998, } } } the cost to the typical individual would be an additional 50 cents per } } } day, or over $180 dollars Per year, above and beyond there regular } } Internet } } } costs. } } } Note that this would be money paid directly to the U.S. Postal Service } } } for a service they do not even provide. The whole point of the Internet is } } } democracy and non-interference. If the federal government is permitted } } } to tamper with our liberties by adding a surcharge to email, who knows } } Where } } } it will end. You are already paying an exorbitant price for snail mail } } } because of bureaucratic inefficiency. It currently takes up to 6 days for } } a } } } letter to be delivered from New York to Buffalo. If The U.S. Postal } } Service } } } is } } } allowed to tinker with email; it will mark the end of the "free" } } } Internet in the United States. } } } One congressman, Tony Schnell has even suggested a "twenty to forty } } } dollar per month surcharge on all internet service" above and beyond the } } } government's proposed email Charges. Note that most of the major } } } newspapers have ignored the story, the only exception being the } } } Washingtonian } } } which called the idea of email surcharge "a useful concept who's time has } } } come" } } } (March 6, 1999) Editorial. } } } } } } Don't sit by and watch your freedoms erode away! Send this e-mail to } } } EVERYONE on your list, and tell all your friends and relatives to write } } } to their congressman and say "No!" to Bill 602P. } } } } } } } } } It will only take a few moments of your time, and could very well be } } } instrumental in killing a bill we don't want. } } } } } } PASS THIS ON TO EVERYONE YOU KNOW WHO USES EMAIL REMEMBER THESE ARE TWO } } } SEPARATE ISSUES THAT EFFECT ALL OF US ONLINE. LET YOU VOICE BE HEARD } } } NOW, NOT AFTER } } } } } } } } } } } } } } } } } } } } }
----------------------- Headers -------------------------------- Return-Path: {Microscopy-request-at-sparc5.Microscopy.Com} Received: from rly-yd01.mx.aol.com (rly-yd01.mail.aol.com [172.18.150.1]) by air-yd01.mail.aol.com (v67_b1.24) with ESMTP; Thu, 10 Feb 2000 00:26:56 -0500 Received: from ultra5.microscopy.com ([206.69.208.10]) by rly-yd01.mx.aol.com (v67_b1.24) with ESMTP; Thu, 10 Feb 2000 00:26:44 -0500 Received: (from daemon-at-localhost) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id XAA23514 for dist-Microscopy; Wed, 9 Feb 2000 23:10:53 -0600 (CST) Received: from no_more_spam.com (sparc5.microscopy.com [206.69.208.10]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id XAA23511 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Wed, 9 Feb 2000 23:09:56 -0600 (CST) Received: from bubba.NMSU.Edu (bubba.NMSU.Edu [128.123.3.39]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id XAA23504 for {microscopy-at-sparc5.Microscopy.Com} ; Wed, 9 Feb 2000 23:09:44 -0600 (CST) Received: from dns1.NMSU.Edu (dns1.NMSU.Edu [128.123.3.5]) by bubba.NMSU.Edu (8.9.3/8.9.0) with ESMTP id WAA15989 for {microscopy-at-sparc5.Microscopy.Com} ; Wed, 9 Feb 2000 22:04:53 -0700 (MST) Received: from nestor.NMSU.Edu (nestor.NMSU.Edu [128.123.34.146]) by dns1.NMSU.Edu (8.9.3/8.9.0) with ESMTP id WAA29784 for {microscopy-at-sparc5.Microscopy.Com} ; Wed, 9 Feb 2000 22:04:54 -0700 (MST) Received: from [128.123.62.47] (analog-ts3-11.NMSU.Edu [128.123.62.47]) by nestor.NMSU.Edu (8.9.3/8.9.0) with ESMTP id WAA266830 for {microscopy-at-sparc5.Microscopy.Com} ; Wed, 9 Feb 2000 22:04:53 -0700 X-Sender: chbennet-at-cnmailsvr.nmsu.edu Message-Id: {v0310280db4c7f4c978e0-at-[128.123.62.47]} Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii" Date: Wed, 9 Feb 2000 21:58:18 -0700 To: microscopy-at-sparc5.microscopy.com From: Christina bennett {chbennet-at-nmsu.edu} Subject: FYI: Congress to allow email charges Errors-to: Microscopy-request-at-sparc5.Microscopy.Com
Is this yet another hoax eating time & bandwidth by being posted without confirmation (like the last 3-4 times I saw this message in the pas year or two), or is it real?
I could find no reference to it at the FCC site.... Those people who are REALLY in charge of telecommunication regulations....
Woody
New format, more pix: http://www.geocities.com/capecanaveral/3722
Weelll... For most systems, if the low end noise is not inhibited (most are), the noise generated, apparent x-ray intensity, tends to increase with lower ev. This would not produce a gaussian-like peak, but a monotonic rise in intensity with lower ev. OTOH, pulse processor circuitry *could* be designed/setup to produce such an effect. Seeing Be x-rays from 100% element is difficult to impossible with most EDS systems. If compounded, the odds get worse.
In the early 1980's Philips believed that their research labs had successfully developed a new electron source suitable for electron microscopes. Indeed there was a time when they were planning to sell microscopes with the new sources. I never heard what went wrong. There were rumors that the lifetime was not good enough.
Anyone who wants details can find them in their library: "An Efficient Silicon Cold Cathode for High Current Densities" Van Gorkom and Hoeberechts Philips Journal of Research 39 (1984) 51-60
Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvannia 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
This hoax is designed, like many of the others, to eat up bandwidth and get people involved in a general uproar.
Hopefully, no one panicked here! It may be worth the time to bookmark this hoax site, or a similar one, so that when we get messages such as this in our email, we can check their validity, before contributing to its spread. No insult intended towards anyone who falls victim to these emails, as I'm sure most of us have at one time or another.
Sincerely,
David Bell Scientist Electron Microscopy Lab Millipore Corporation 80 Ashby Road Bedford, MA 01730 (781) 533-2108
This is one of those internet hoaxes. It is not true, do not send it to your friends, do not write your congressperson. Mining Co has a great web page that debunks these hoaxes and myths {http://urbanlegends.miningco.com/culture/urbanlegends/library/blhoax.htm?pid=27 33&cob=home} and is a good place to check before sending on any email that urges you to send it to everyone you know. This message is a combination of two old and popular hoaxes, see {http://urbanlegends.miningco.com/culture/urbanlegends/library/blemtax2.htm} and {http://urbanlegends.about.com/culture/urbanlegends/library/weekly/aa012099.htm} Hopefully this will die here. Scott
} } } } Congress to allow email charges } } } } } } } } Please pass this on to all you know since many of us use e-mail for } } } } business and to keep up with friends and family, I thought you'd like } } } } to know the following. } } } } } } } } Please jump on it right away and forward this to others. } } } } } } } } CNN has reported that within the next two weeks Congress is going to } } } } vote on allowing telephone companies to CHARGE A TOLL FEE for } } } } internet access. } } } } Translation: Every time we send long distance e-mail we will receive a } } } } long distance charge. This will get costly. Please visit the following web } } } } site and file a complaint. Complain to your Congressperson. We can't } } } } allow this to pass. The following address will allow you to send an } } } } e-mail on this subject DIRECTLY to your Congressperson. } } } } } } } } http://www.house.gov/writerep {http://www.house.gov/writerep}
..sniped... } } } } } } } } PASS THIS ON TO EVERYONE YOU KNOW WHO USES EMAIL REMEMBER THESE ARE TWO } } } } SEPARATE ISSUES THAT EFFECT ALL OF US ONLINE. LET YOU VOICE BE HEARD } } } } NOW, NOT AFTER
-------------------note: new mailing address---------------------- Scott Wight e-mail: scott.wight-at-nist.gov NIST 222/A113 W voice: 301-975-3949 100 Bureau Dr STOP 8371 | fax: 301-417-1321 Gaithersburg, MD 20899-8371 \|/ disclaimer: Any opinion expressed is my own and does not represent those of my employer.
A technical position is available in the Neurobiology of Aging program at the Mount Sinai School of Medicine in New York. We are seeking an experienced Electron Microscopist with a BS/BA or MS in Biology/Life Sciences. Applicants should have excellent communication and organizational skills, an understanding of basic laboratory procedures, and the ability to manage a large and varied workload. The successful candidate will participate in ultrastructural studies focusing on the effects of: estrogen and aging on hippocampal circuitry which is implicated in learning and memory, quantitative excitatory amino acid receptor (NMDA and AMPA) distribution within the central nervous system of transgenic models, as well as manipulated primate and rodent models. Qualifications include at least 2 years of experience in routine transmission electron microscopy procedures, ultramicrotomy, immunogold labelling, specimen preparation, digital photography, and routine maintenance of equipment. Experience with immunofluorescence and confocal microscopy is an asset.
We offer a salary commensurate with experience and excellent benefits. For consideration, please mail/email your resume to:
Bill Janssen Neurobiology of Aging Box 867, EB9-02 Mount Sinai School of Medicine, One Gustave L. Levy Place, New York, NY 10029-6574
We are an equal opportunity employer fostering diversity in the workplace. Bill Janssen Neurobiology of Aging Laboratories Mount Sinai School of Medicine, Box 1639 One Gustave L. Levy Place New York, NY 10029
I think the best solution would be to use a water recirculatory system. Fill it with clean water and it should stay clean for a long time. We use a 'Neslab'
Hope this helps
Alan Walker.
********************************************* Alan Walker Dept of Electronic and Electrical Engineering University of Sheffield Mappin Street Sheffield S1 3JD United Kingdom
this is of course a recurring theme in image analysis. There really is no easy answer to this. Counting particles is easier as you can perhaps separate the particles and arrive at the correct count, but if you don't know what is occluded, you cannot measure it. However, if you can use some information that is available to make some guesses or extrapolations what the shape is, it can be done. A simple example: If you look at a heap of coins, they may overlap. It is nevertheless possible to measure the individual coins because I know that they are all round. So I can take the "protruding" part of a coin and simply fit a circle and that should give me the size pretty accurately.
Are ice crystals like snowflakes? Snowflakes, if I remember correctly, have a six-fold symmetry. Can't you just measure one "branch" of a snowflake (the one that you can see), and multiply that by 6? I am not an expert on ice crystals (although I love to ski), so this may be oversimplified. You may have to think about all the information you have about ice crystals and can then perhaps make some guesses about the occluded part.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
} ---------- } From: Rosemary Walsh[SMTP:RW9-at-PSU.EDU] } Sent: Tuesday, February 08, 2000 3:24:35 PM } To: microscopy-at-sparc5.Microscopy.Com } Subject: Size determination of overlapping particles } Auto forwarded by a Rule } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Listers, I have a student interested in sizing ice crystals which overlap. Any suggestions? Rosemary
We have a tedious task of liquid nitrogen transfer and fill-up on our instruments such as SEMs, TEM and for cryo-ultramicrotomes. Is there any compact unit on market which would condense nitrogen from air into liquid form? I know that Liquid oxygen would be a problem. TIA;
I just read the two excellent replies from Scott and Phillip and fully agree with them. Small angle cleavage technique (SACT) and the tripod polisher are two very useful TEM prep techniques. In my experience, SACT is easier to learn than tripod polishing but tripodding is applicable to more materials and yields larger thin areas than does SACT.
As Phillip mentions, low energy milling in the final step of the process is extremely important for reducing artifacts. I would finish using the lowest possible energy that still yields effective milling.
Another approach is to use reactive ion beam etching (RIBE) or chemically assisted ion beam etching (CAIBE). In both these techniques a reactive gas (usually Iodine) is used. In RIBE, iodine is actually ionized and is used as the milling gas. In CAIBE, iodine gas is introduced into the milling chamber directly adjacent to the sample during argon ion milling. Both these techniques have been shown to eliminate the ion beam damage for binary type III-V compound semiconductors containing InP. Chew and Cullis (Ultramicroscopy 23/1987/175-198) also report improvements of ion milled surfaces for ternary type III-V semiconductors containing InGaAs (materials you mentioned) using RIBE.
I believe that Gatan offers an optional CAIBE attachment for the PIPS. This may be the easiest thing to try first. Remember that iodine is very corrosive and can damage ion milling parts if not used properly. Check with Gatan for their recommendations.
Hope this helps,
Eric W.
Eric S. Windsor Physical Scientist NIST 100 Bureau Dr. Stop 8371 Gaithersburg, MD 20899-8371 (301) 975-3930 Fax: (301) 417-1321 Eric.Windsor-at-NIST.Gov
On Feb 9 -at- 09:43 Massimo Catalano wrote:
Dear Listservers, we are massively working on analytical and structural TEM characterization of semiconductors, mainly InGaAs/GaAs, quantum well, wires and dots. Specimen preparation is of course a crucial point of our work. We have just switched to PIPS (we used to make our samples using the Gatan Duo Mill), and we are getting controversial results about the damage introduced by the milling procedure. We are perfectly aware of all the differences between the two instruments, especially the absence of cooling stage and the higher beam current. Has anyone performed a systematic and careful analysis to try to evaluate the milling damage on similar materials systems. We are willing to start a systematic work to assess this issue, but would like to know if anyone has already done anything on this topic. Also, any collaboration will be more than welcome. Thanks. Massimo
Eric S. Windsor Physical Scientist NIST 100 Bureau Dr. Stop 8371 Gaithersburg, MD 20899-8371 (301) 975-3930 Fax: (301) 417-1321 Eric.Windsor-at-NIST.Gov
I have found the basic description on the US/Canada website so I assume its available there too - see address below: http://www.epson.com/cam_scan/scanners/perfection1200/index.html
The price inclusive of transparency adapter appears to be about 200 UK pounds over here. There seems to be three models: 1200s (standard print scanner - SCSI interface) 1200u (standard print scanner - USB interface) 1200photo (standard print scanner + 5x4inch transparency adapter - USB interface) It's the last of these that is of interest.
Malcolm Haswell ------------------------------------------ Maureen Petersen wrote: } } Malcolm: } } Will you divulge what this gem costs? Do you know if it is available in the } US? } } Thank you, Maureen Petersen } Dept Plant Pathology } University of Florida } Gainensville, FL 32611 } } I'm sorry to bring this topic up yet again, but I have just come across } the specification for a flat-bed scanner which looks as if it would be } suitable for scanning e.m. cut-film. It is the Epson Perfection 1200 } Photo which includes a 5x4inch film adapter. } } The UK web site address is: } http://www.epson.co.uk/sohoprod/imaging/scanner/perf1200/perf1200photo/spec. } htm } } and the specification is: } A4 Colour flatbed with 5"x4" film image scanner Single pass scanning } Optical resolution: 1200 x 2400 (30600 pixels/line) } Maximum output resolution of 9600x9600 dpi } 36 bits per pixel in colour (24 bit output) } 12 bits per pixel in black (8 bit output) } Optical Density 3.2D } USB (Type B) } } Does anyone have any experience of this machine because it is about a } fifth of the price of the Umax flatbed film scanners? My only } reservation is that it is a SOHO product rather than professional. } } Thanks in advance } } Malcolm } } Malcolm Haswell } Electron Microscopy } School of Sciences } Fleming Building } University of Sunderland } SUNDERLAND SR1 3SD } Tyne and Wear } UK } } Tel (0191) 515 2872 } e-mail: malcolm.haswell-at-sunderland.ac.uk
I am considering the purchase of a new digital camera. Some of the new cameras like the Spot-RT and Magnafire have the option of running as three shot color or one shot gray-scale acquisition. If someone wants to do dual channel fluorescence like FITC/Rhod or annexin/PI, we can do that as either acquiring each image with a separate filter cube and combining in software, or designing a dual filter cube and acquiring in color (or I guess acquiring separate color images iwth the single cubes and combining??). What is people real world experience with the relative merits of these two approaches. It seems that most of what I read is done by separate filter cubes combined in software, but there also seems to be a push toward fast cooled color cameras. The advantage I can see to the color approach would be using a color chip camera you get multiple channels simultaneously. However, do you have to expose significantly longer to acheive this compared to multiple gray scale acquisitions? I can't see an obvious advantage to the three shot color approach because I suspect the light throughput is lower and the filters sets I suspect are expensive to add on if you already have the single fluor filters. The only real advantage I see to three shot acquisition is the ability to easily image stained/histology transmitted light slides and not for fluorescence. Any thoughts appreciated. Thanks- Dave
Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 75 N. Eagleville Rd. U-125 Storrs, CT 06269 Knecht-at-uconnvm.uconn.edu 860-486-2200 860-486-4331 (fax) home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html ************************************************************
This nonsense resurfaces every few months. There has never been any truth to it. (Of course, there's no assurance that Congress will not act so foolishly in the future.)
Kal
On Wed, 9 Feb 2000, Christina bennett wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I know this is off topic however a change like this could make this type of } forum impossible. } } } } } } } Date: Tue, 8 Feb 2000 08:13:53 -0700 } } X-Sender: liperez-at-cnmailsvr.nmsu.edu } } Mime-Version: 1.0 } } To: all-at-biology.nmsu.edu } } From: liperez-at-NMSU.Edu (LSPSaldana) } } Subject: Congress to allow email charges } } Sender: owner-all-at-biology.nmsu.edu } } Precedence: bulk } } X-Keywords: } } X-UID: 43 } } Status: O } } } } } } } } Congress to allow email charges } } } } } } } } Please pass this on to all you know since many of us use e-mail for } } } } business and to keep up with friends and family, I thought you'd like } } } } to know the following. } } } } } } } } Please jump on it right away and forward this to others. } } } } } } } } CNN has reported that within the next two weeks Congress is going to } } } } vote on allowing telephone companies to CHARGE A TOLL FEE for } } } } internet access. } } } } Translation: Every time we send long distance e-mail we will receive a } } } } long distance charge. This will get costly. Please visit the following web } } } } site and file a complaint. Complain to your Congressperson. We can't } } } } allow this to pass. The following address will allow you to send an } } } } e-mail on this subject DIRECTLY to your Congressperson. } } } } } } } } http://www.house.gov/writerep {http://www.house.gov/writerep} } } } } } } } } Pass this on to your friends. It is urgent! I hope all of you will pass } } } } this on to all your friends and family. We should ALL have } } } } an interest in this one. } } } } } } } } WAIT, THERE'S MORE. IN ADDITION, The last few months have revealed an } } } } alarming trend in the Government of the United States attempting to } } } } quietly push through legislation that will affect your use of the } } } Internet. } } } } Under } } } } proposed legislation the U.S. Postal Service will be attempting to bill } } } } email users out of "alternate postage fees". Bill 602P will permit the } } } } Federal Govt. to charge a 5 cent surcharge on every email delivered, by } } } } billing Internet Service Providers at source. The consumer would then be } } } } billed in turn by the ISP. Washington D.C. lawyer Richard Stepp is } } } } working without pay to prevent this legislation from becoming law. The } } } U.S. } } } } Postal Service is claiming that lost revenue due to the proliferation of } } } } email } } } } is costing nearly $230,000,000 in revenue per year. You may have noticed } } } } their recent ad campaign "There is nothing like a letter". } } } } Since the average citizen received about 10 pieces of email per day in } } } } 1998, } } } } the cost to the typical individual would be an additional 50 cents per } } } } day, or over $180 dollars Per year, above and beyond there regular } } } Internet } } } } costs. } } } } Note that this would be money paid directly to the U.S. Postal Service } } } } for a service they do not even provide. The whole point of the Internet is } } } } democracy and non-interference. If the federal government is permitted } } } } to tamper with our liberties by adding a surcharge to email, who knows } } } Where } } } } it will end. You are already paying an exorbitant price for snail mail } } } } because of bureaucratic inefficiency. It currently takes up to 6 days for } } } a } } } } letter to be delivered from New York to Buffalo. If The U.S. Postal } } } Service } } } } is } } } } allowed to tinker with email; it will mark the end of the "free" } } } } Internet in the United States. } } } } One congressman, Tony Schnell has even suggested a "twenty to forty } } } } dollar per month surcharge on all internet service" above and beyond the } } } } government's proposed email Charges. Note that most of the major } } } } newspapers have ignored the story, the only exception being the } } } } Washingtonian } } } } which called the idea of email surcharge "a useful concept who's time has } } } } come" } } } } (March 6, 1999) Editorial. } } } } } } } } Don't sit by and watch your freedoms erode away! Send this e-mail to } } } } EVERYONE on your list, and tell all your friends and relatives to write } } } } to their congressman and say "No!" to Bill 602P. } } } } } } } } } } } } It will only take a few moments of your time, and could very well be } } } } instrumental in killing a bill we don't want. } } } } } } } } PASS THIS ON TO EVERYONE YOU KNOW WHO USES EMAIL REMEMBER THESE ARE TWO } } } } SEPARATE ISSUES THAT EFFECT ALL OF US ONLINE. LET YOU VOICE BE HEARD } } } } NOW, NOT AFTER } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } }
Someone at a remote location within our company has decided to purchase an RG Lee SEM. I am looking for information and opinions from people who have one of these microscopes. I'm interested in any service issues, reliability issues and usability concerns that anyone may have. The model they are interested in is the PSM 75LS. Also, if anyone has the URL for their website, that would be appreciated, as well.
Thanks in advance,
David Bell Scientist Electron Microscopy Lab Millipore Corporation 80 Ashby Road Bedford, MA 01730 (781) 533-2108
A poster does not mean anything. I have seen them with the Lithium-Ka energy listed, although quantum physics does forbid this X-ray line. But I can assure you the Be-K line does exist.
On modern EDX systems Be can be detected, provided you have either a windowless detector or a detector with a modern polymer type window (SUTW, SATW, Norvar, or whatever the EDX supplier calls it. My apologies if I break any trade-marks here...). On most demonstration setups your EDS specialist will be pleased to show you a Be peak. But probably from a pure Be sample only. The absorption coefficient (MAC) of Be in basically any matrix is so huge, and the X-ray fluorescence yield so small, that in any compound with less than 50 atomic percent Be you will not detect a visible Be peak. For this reason most EDX installations "in the field" are not setup to detect Be-K radiation.
WDS has a better P/B ratio, so if you have the proper multilayer crystal you can get results. But peak shifts and peak shape alterations will make quantification extremely difficult, not to mention the fact that for most MACs of Be-K in any matrix we only have a ball-park figure.
Other techniques, like PEELS, are much more suitable for Li and Be analysis. Best regards,
Hans Dijkstra
Disclaimer: This is my opinion, and not necessarily the one from EDAX Inc. ------------------------------------------------------------- EDAX Europe www.edax.com Ringbaan Noord 103 Tel.: +31-13-5364000 P.O. Box 4144 Fax.: +31-13-5356279 5004 JC Tilburg E-mail: hans.dijkstra-at-edax.nl the Netherlands -------------------------------------------------------------
} -----Original Message----- } From: Walck. Scott D. [mailto:walck-at-ppg.com] } Sent: Tuesday, February 08, 2000 6:57 PM } To: 'microprobe'; 'harry.ekstrom-at-honeywell.com' } Cc: 'Microscopy' } Subject: RE: Be X-ray peaks -I don't think so } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I'm wrong. I guess I should have looked up at the X-ray periodic table } above my head and saw the 0.108 keV value for Be. The reason that I was } wrong was that I did not consider the solid state aspects. I am still right } about not having a 2s1/2 to a 1s1/2 transition and that an X-ray carries } away 1h-bar of angular momentum. What I didn't do was think about the band } structure of a solid. If you have the solid, the 2p bands of Be most be } spilling into or be the conduction band for the electrons. That must be the } source for the electronic transition. I guess I should have engaged brain } before typing. } My most humble apologies. } -Scott } } Scott D. Walck, Ph.D. } PPG Industries, Inc. } Glass Technology Center } Guys Run Rd. (packages) } P. O. Box 11472 (letters) } Pittsburgh, PA 15238-0472 } } Walck-at-PPG.com } } (412) 820-8651 (office) } (412) 820-8161 (fax) } } } "The opinions expressed are those of Scott D. Walck and not of PPG } Industries, Inc. nor of any PPG-associated companies." } -- }
The first meeting of the millenium for NESM (New England Society for Microscopy) will be held on March 8, 2000 at Polaroid Corporation in Wayland, MA. Registration will begin at 5:00pm with tours of the (new) facility following at 5:30pm. Attendees will also be asked to fill outa questionnaire (optional) on "Professional Imaging" at this time.
A buffet supper will be served from 6-7pm followed by three scientific presentations. The speakers are : Gabriel Rojano, Staff Reliability Engineer-MIA-Com (Tyco Electronics Company), Paul Bain from the Harvard Medical Library and Dr. Hjalmar Brismer from the Karolinska Institute in Stockhold, Sweden.
Advance registration is encouraged (by March 3rd). The registration fee for members is $5.00 and $20.00 for non-members (this includes a current one-year membership in the society). Interested parties should contact: Peggy Sherwood, Corresponding Secretary (MESnesm-at-aol.com).
Different subject title in case other posting got filtered out.
} Date: Thu, 10 Feb 2000 08:08:10 -0800 } To: Microscopy-at-MSA.Microscopy.Com } From: "Dr. Gary Gaugler" {gary-at-gaugler.com} } Subject: urban myths and legends - Congress to allow e-mail charges } } Here are two URLs showing that this is just another hoax; } } } http://www.urbanmyths.com/email_internettax602p.html } } } } http://www.ulrc.com.au/html/report.asp?CaseFile=ULRR0027&Page=1&View=Request } } gary g.
The insidious nature of this hoax/scam/... is that not only does the original hoax get circulated, using up bandwidth, but then the replies of the people who identified the hoax, many with the full hoax message attached, also get circulated. And then the triple play (baseball and spring training is near) is when people like me comment on the aforesaid. Please, please don't reply to this note (which should have no attachments!) and make it a home run!
Don Marshall
Donald J. Marshall Relion Industries P.O. Box 12 Bedford, MA 01730 Ph: 781-275-4695 FAX: 781-271-0252 email dmrelion-at-world.std.com
Cathodoluminescence, mass spectroscopy, electron beam technology
"A weed is a flower out of place."
(Please note: Do not send email with attachments to this address. Instead, send it to donbarlen-at-aol.com. Thank you.)
This is not true. It is an internet hoax. Here's the info from an urban legends website: --------------------------- Claim: Congress will soon be voting on whether or not your phone company will be allowed to charge you long-distance rates for accessing the Internet.
Status: False.
Example: [Collected on the Internet, 1999]
Please forward this to everyone you can...
There is a new bill in the US Congress that will affect ALL INTERNET USERS. CNN stated that the Government would in two weeks time decide whether to allow or not allow a Charge to YOUR phone bill equal to a long distance call each time you access the Internet.
This affects us all! We cannot allow this to happen! Please visit the following URL and fill out the necessary form to let your Congressman know how you feel! The address is http://www.house.gov/writerep/. Write your representative!
Synopsis: Neither the FCC nor Congress is considering -- much less voting on -- legislation that would impose (or allow phone companies to impose) per-minute access fees on Internet users. Recent decisions by the FCC in this area have dealt only with the issue of how phone companies reimburse each other for handling calls placed to Internet service providers, not with the prospect of allowing phone companies to charge their customers long-distance rates for Internet use.
Origins: As soon as we get hold of something we really like at a reasonable price, we start worrying that it will be banned, taxed, or made too expensive for us to afford. The Internet is no exception, as our old friend -- the capitalized, exclamation pointed, "send this to everyone you know" anonymous e-mail message -- is here to tell you.
Way back when in 1987, the Federal Communications Commission did consider imposing a surcharge for transmitting data over the public telephone network, but they ultimately rejected the idea (thanks in part to the more than 10,000 letters of complaint they received). Unable to believe our good fortune (the government wasn't going to make us pay through the nose for dialing up all those neat computer bulletin boards we'd discovered), we couldn't leave well enough alone, and in 1991 a flood of urgent messages warning us that the FCC was again considering a proposal they'd rejected three years earlier hit e-mail systems all over the country (and the nascently popular Internet). Like the ubiquitous Craig Shergold message, the "modem tax" warning would long outlive the validity of the information it conveyed.
Fast forward to 1998. On-line services, the Internet, the World Wide Web, e-mail, and chat rooms are more popular than ever, a daily part of many people's lives. Somebody -- the government, the phone companies, Bill Gates, the Grinch -- must be on his way to spoil the party. Sure enough, we're now being told the phone companies and the government are in cahoots to ruin our good time.
First of all, a little background. Most of us still have to dial up over a modem and connect to an Internet Service Provider (ISP) to access the Internet. If your ISP is in your local dialing area, you probably don't pay anything at all for the call, no matter how long you stay connected. This means you get to tie up a phone line with your modem for hours and hours on end, every day, at no charge beyond the price of basic phone service. And the party at the other end of the line -- your ISP -- isn't paying anything extra, either. It's easy to see that somebody has a chance to reap some windfall profits here. If the phone companies were allowed to charge you a per-minute fee for accessing the Internet (or the government were allowed to tax your use of the Internet), their coffers would soon overflow with cash.
Scary thought, isn't it? All the phone companies need, we hear, is to get the FCC to reclassify and/or regulate ISPs, and then the phone companies can charge gobs of extra money for handling Internet traffic. And Congress is just about to vote on that very issue, we're told.
In fact, there is no such proposal before Congress, and there never has been.
The only real issue before the FCC concerning Internet usage (and the genesis of this latest round of scaremongering) is the subject of "reciprocal compensation." In short, reciprocal compensation means that when you place a local call to someone who is serviced by a different phone company, your phone company has to compensate his phone company for completing the call. (On the other hand, when you place a long-distance call, the long distance carrier who handles the traffic has to pay access charges to your phone company for originating the call.) But if the "person" you're calling is an ISP, should your phone company have to compensate the ISP's phone company?
The subject of recpirocal compensation has been a hot issue of late because new local phone companies have been springing up all over the place. The bigger phone companies, figuring that they would have many more customers than their newer (and smaller) competitors, negotiated reciprocal compensation agreements with the new phone companies. Every time one of these little phone companies' customers placed a call to a destination outside his local service that ended up on the bigger phone company's network, the big phone company would get to collect money from the little phone company. Not a bad scheme, the big phone companies thought.
Ah, but some of the little guys had a neat trick up their sleeves. They started offering their services to Internet service providers -- ISPs with banks and banks of modems that received thousands and thousands of calls every day, but never made any outgoing calls. All the reciprocal compensation started flowing one direction, from the big phone companies to the little phone companies, which wasn't what the big guys had in mind at all. "Foul," they cried. "Internet traffic flows all over the world," they noted. "Internet traffic is therefore interstate in nature and should be classified as long-distance, so the little guys should be paying us for originating the calls," they insisted. "We're not paying," they sputtered.
Enter the FCC to resolve the dispute, which they did (for now) on 25 February 1999 by ruling that phone companies are bound by whatever reciprocal compensation agreements they've negotiated with each other, whether they think they're fair or not. That was the only issue before the FCC. But most of us are already struggling with a glut of information, and we don't have the time to familiarize ourselves with details like interconnection agreements and reciprocal compensation, so when we hear reports with buzzwords like "Internet," "FCC," "access fees," and "long distance," we assume the worst, even though the real story is something quite different. And even if we make the effort to understand the whole story, we find all too often that we're reading information that has been misreported by others -- often the mainstream media -- who didn't make enough of an effort to understand the whole story themselves. If we can't even depend upon the people whose business it is to supply us with accurate information to get the facts straight, what more can we do? (See, for example, the misleading headline on the CNN article referenced in the "Additional information" section below.)
A few important points related to the recent FCC decision:
Didn't the FCC rule that Internet connections are long-distance calls?
Sort of. The FCC declared that "Internet traffic is jurisdictionally mixed and appears to be largely interstate in nature," which is the technical definition of "long distance." But that doesn't mean -- as is often misreported -- that Internet users will be paying long distance rates for dial-up connections, since the Internet has been, and still is, exempt from interstate access charges. The FCC did nothing to abolish that exemption.
Won't the phone companies just pass the cost of carrying Internet traffic to customers by raising their phone rates?
There is no guarantee that phone rates won't go up in the future, of course. However, since most states require phone companies to charge a flat rate for unlimited local usage, you won't have to pay per-minute charges for accessing the Internet (as long as your ISP has a dial-up number within your local service area).
What if the FCC changes their mind?
The possibility exists that the FCC might someday decide that additional fees can be imposed for Internet access. But as Bill Kennard, the chairman of the FCC, has stated on more than one occasion: "I want to say this as clearly as I can . . . as long as I'm chairman of the Federal Communications Commission this agency will not regulate the Internet. It's not going to happen. The FCC has no intention of making computer users pay long distance fees for dial-up access to the Internet, as people now pay when they make long-distance telephone calls. These rumors get on the Internet that the big bad FCC is going to impose all this regulation on the Internet. Now I know this painfully because every so often when one of these rumors flares up I get, literally, about 600 e-mail messages a day by people who are telling me to keep my hands off the Internet." [Note: this is not a direct quote from Kennard; it is pieced together from multiple statements of his.]
Additional information:
No Consumer Per-Minute Charges to Access ISPs (FCC)
Users, Advertisers Await FCC Decision on Internet Charges (CNN)
Update: In April 1999, a Canadian version of this message was unleashed on the Internet. Unlike its American counterpart, this version is not mere misinformation based on a flawed understanding of actual events or legislation -- it is an outright hoax:
Please read the following carefully if you intend to stay online and continue using email:
The last few months have revealed an alarming trend in the Government of Canada attempting to quietly push through legislation that will affect your use of the Internet. Under proposed legislation Canada Post will be attempting to bilk email users out of "alternate postage fees".
Bill 602P will permit the Federal Govt to charge a 5 cent surcharge on every email delivered, by billing Internet Service Providers at source. The consumer would then be billed in turn by the ISP. Toronto lawyer Richard Stepp QC is working without pay to prevent this legislation from becoming law.
The Canada Post Corporation is claiming that lost revenue due to the proliferation of email is costing nearly $23,000,000 in revenue per year. You may have noticed Canada Post's recent ad campaign "There is nothing like a letter". Since the average citizen received about 10 pieces of email per day in 1998, the cost to the typical individual would be an additional 50 cents per day, or over $180 dollars per year, above and beyond their regular Internet costs. Note that this would be money paid directly to Canada Post for a service they do not even provide. The whole point of the Internet is democracy and non-interference. If the Canadian Government is permitted to tamper with our liberties by adding a surcharge to email, who knows where it will end. You are already paying an exhorbitant price for snail mail because of beaurocratic inefficiency. It currently takes up to 6 days for a letter to be delivered from Mississauga to Scarborough. If Canada Post Corporation is allowed to tinker with email, it will mark the end of the "free" Internet in Canada. One back-bencher, Liberal Tony Schnell (NB) has even suggested a "twenty to forty dollar per month surcharge on all Internet service" above and beyond the government's proposed email charges. Note that most of the major newspapers have ignored the story, the only exception being the Toronto Star that called the idea of email surcharge "a useful concept who's time has come" (March 6th 1999 Editorial) Don't sit by and watch your freedoms erode away! Send this email to all Canadians on your list and tell your friends and relatives to write to their MP and say "No!" to Bill 602P.
Kate Turner Assistant to Richard Stepp QC Berger, Stepp and Gorman Barristers at Law 216 Bay Street Toronto, ON MlL 3C6
Yes, Americans are not alone in their belief that when the need for the primary service provided by a governmental agency diminishes or disappears, that agency will come up with draconian schemes to perpetuate its existence at taxpayer expense rather than modernizing or closing up shop. In this case, however, the message is simply too riddled with errors to be anything but a hoax:
There is no "Bill 602P" currently before the Canadian parliament. The designations of Canadian parliamentary bills take the form of the letter 'C' or 'S' followed by a number (depending upon whether they originated in the House of Commons or the Senate). Besides having the wrong prefix, this purported bill is assigned a number far too high to be one currently being considered in parliament, as you can see on the list of Canadian government bills.
Despite the claim in the message, nothing about this alleged bill is to be found. Also, there was no editorial about this on the 6 March 1999 OpEd page of Toronto Star. (Maybe "major papers have ignored the story" because it's a work of fiction?)
There is no Richard Stepp QC; law firm by the name of Berger, Stepp and Gorman; or 216 Bay Street in Toronto.
A list of Canadian Members of Parliament contains no MP by the name of Tony Schnell.
Of course, it didn't take long before the same thing started circulating in an Americanized version:
Dear Internet Subscriber:
Please read the following carefully if you intend to stay online and continue using email: The last few months have revealed an alarming trend in the Government of the United States attempting to quietly push through legislation that will affect your use of the Internet. Under proposed legislation the U.S. Postal Service will be attempting to bilk email users out of "alternate postage fees".
Bill 602P will permit the Federal Govt to charge a 5 cent surcharge on every email delivered, by billing Internet Service Providers at source. The consumer would then be billed in turn by the ISP. Washington D.C. lawyer Richard Stepp is working without pay to prevent this legislation from becoming law.
The U.S. Postal Service is claiming that lost revenue due to the proliferation of email is costing nearly $230,000,000 in revenue per year. You may have noticed their recent ad campaign "There is nothing like a letter". Since the average citizen received about 10 pieces of email per day in 1998, the cost to the typical individual would be an additional 50 cents per day, or over $180 dollars per year, above and beyond their regular Internet costs. Note that this would be money paid directly to the U.S. Postal Service for a service they do not even provide. The whole point of the Internet is democracy and non-interference. If the federal government is permitted to tamper with our liberties by adding a surcharge to email, who knows where it will end. You are already paying an exorbitant price for snail mail because of bureacratic efficiency. It currently takes up to 6 days for a letter to be delivered from New York to Buffalo. If the U.S. Postal Service is allowed to tinker with email, it will mark the end of the "free" Internet in the United States. One congressman, Tony Schnell (r) has even suggested a "twenty to forty dollar per month surcharge on all Internet service" above and beyond the government's proposed email charges. Note that most of the major newspapers have ignored the story, the only exception being the Washingtonian which called the idea of email surcharge "a useful concept who's time has come" (March 6th 1999 Editorial) Don't sit by and watch your freedoms erode away!
Send this email to all Americans on your list and tell your friends and relatives to write to their congressman and say "No!" to Bill 602P.
Kate Turner Assistant to Richard Stepp Berger, Stepp and Gorman Attorneys at Law 216 Concorde Street, Vienna, Va.
Does anyone know where I can find a troubleshooting manual/guide for paraffin sectioning? I am having trouble with the tissue "smearing" and with it looking chopped-up. Is this a vibration issue? I welcome any comments from those of you with experience in this area!
} Date: Thu, 10 Feb 2000 09:03:00 -0800 } To: microscopy-at-sparc5.microscopy.com } From: Laurie Wallin {lwallin-at-ucsd.edu} } Subject: paraffin sectioning troubleshooting guide } Cc: } Bcc: } X-Attachments: } } Does anyone know where I can find a troubleshooting manual/guide for } paraffin sectioning? I am having trouble with the tissue "smearing" and } with it looking chopped-up. Is this a vibration issue? I welcome any } comments from those of you with experience in this area! } } Thanks in advance. } } Sincerely.
Laurie Wallin UCSD Department of Anesthesiology 9500 Gilman Drive, 0629, La Jolla, CA 92093 (858) 822-3271
There has been some very significant work done by Dr. Arpad Barna et al in Hungary. Some of the papers he has presented include:
Ion Energy Effect on Surface Amorphisation of Semiconductor Crystals/A. Barna, et al
Amorphisation and Surface Morphology Development at Low Energy Ion Milling/A. Barna, B. Pecz.
Analysis of the Development of Large Surface Topography During Ion Etching/A. Barna; P. Barna; et al
Model Considerations of Ion Beam Thinning for Preparing TEM Samples/A.Barna; et al
Possibility of Surface Polishing by Ion Beam Thinning/ A. Barna
Ion Beam Thinning on the Basis of Topographic Kinetics/A. Barna
Low Angle and Low Energy Ion Beam Etching for TEM Sample Preparation/A. Barna
TEM Sample Preparation by Ion Milling / Amorphization/A. Barna, B. Pecz, M. Menyhard
I also have the following paper that may be of interest:
Preparation of InGaAs/GaAs Multilayered Materials for TEM by One Side Non-Rotation Ion Beam Thinning/J.Y. Yao; G.L. Dunlop
I do not have the complete references available in front of me, but we do have copies of all of these papers in our technical library. I would be pleased to mail copies to you if that would be of interest. We also have a list of over 250 papers dealing with various aspects of sample preparation (much of it TEM sample preparation) which may be of interest. If you would like to review that list, I can send it over to you in MS Excel format and you can select any other papers that would be of interest.
I hope this helps.
DISCLAIMER: South Bay Technology, Inc. markets the IV3 Ion Mill in both high and low energy (down to 200ev) versions which is based on the work of Dr. Barna. We also produce the XLA 2000 computer controlled Low Angle Ion Mill so we have a vested interest in promoting their use.
David Henriks Vice President TEL: 800-728-2233 (toll free in the USA) South Bay Technology, Inc. +1-949-492-2600 1120 Via Callejon FAX: +1-949-492-1499 San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by Massimo Catalano } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Listservers,
we are massively working on analytical and structural TEM characterization of semiconductors, mainly InGaAs/GaAs, quantum well, wires and dots.
Specimen preparation is of course a crucial point of our work.
We have just switched to PIPS (we used to make our samples using the Gatan Duo Mill), and we are getting controversial results about the damage introduced by the milling procedure.
We are perfectly aware of all the differences between the two instruments, especially the absence of cooling stage and the higher beam current.
Has anyone performed a systematic and careful analysis to try to evaluate the milling damage on similar materials systems. We are willing to start a systematic work to assess this issue, but would like to know if anyone has already done anything on this topic. Also, any collaboration will be more than welcome.
Well, this urban legend has finally made it to the microscopy listserver! And that is just what it is--a legend, a myth, a fable. As a stamp collector and one who keeps abreast of all of this kind of stuff, this urban legend has been debunked in both Linn's Stamp News and Stamp Collector.
All of us have important issues to consider. Please, put this one to rest.
Roger Moretz
On Wed, 9 Feb 2000 21:58:18 -0700, Christina bennett wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I know this is off topic however a change like this could make this type of } forum impossible. } } } } } } } Date: Tue, 8 Feb 2000 08:13:53 -0700 } } X-Sender: liperez-at-cnmailsvr.nmsu.edu } } Mime-Version: 1.0 } } To: all-at-biology.nmsu.edu } } From: liperez-at-NMSU.Edu (LSPSaldana) } } Subject: Congress to allow email charges } } Sender: owner-all-at-biology.nmsu.edu } } Precedence: bulk } } X-Keywords: } } X-UID: 43 } } Status: O } } } } } } } } Congress to allow email charges } } } } } } } } Please pass this on to all you know since many of us use e-mail for } } } } business and to keep up with friends and family, I thought you'd like } } } } to know the following. } } } } } } } } Please jump on it right away and forward this to others. } } } } } } } } CNN has reported that within the next two weeks Congress is going to } } } } vote on allowing telephone companies to CHARGE A TOLL FEE for } } } } internet access. } } } } Translation: Every time we send long distance e-mail we will receive a } } } } long distance charge. This will get costly. Please visit the following web } } } } site and file a complaint. Complain to your Congressperson. We can't } } } } allow this to pass. The following address will allow you to send an } } } } e-mail on this subject DIRECTLY to your Congressperson. } } } } } } } } http://www.house.gov/writerep {http://www.house.gov/writerep} } } } } } } } } Pass this on to your friends. It is urgent! I hope all of you will pass } } } } this on to all your friends and family. We should ALL have } } } } an interest in this one. } } } } } } } } WAIT, THERE'S MORE. IN ADDITION, The last few months have revealed an } } } } alarming trend in the Government of the United States attempting to } } } } quietly push through legislation that will affect your use of the } } } Internet. } } } } Under } } } } proposed legislation the U.S. Postal Service will be attempting to bill } } } } email users out of "alternate postage fees". Bill 602P will permit the } } } } Federal Govt. to charge a 5 cent surcharge on every email delivered, by } } } } billing Internet Service Providers at source. The consumer would then be } } } } billed in turn by the ISP. Washington D.C. lawyer Richard Stepp is } } } } working without pay to prevent this legislation from becoming law. The } } } U.S. } } } } Postal Service is claiming that lost revenue due to the proliferation of } } } } email } } } } is costing nearly $230,000,000 in revenue per year. You may have noticed } } } } their recent ad campaign "There is nothing like a letter". } } } } Since the average citizen received about 10 pieces of email per day in } } } } 1998, } } } } the cost to the typical individual would be an additional 50 cents per } } } } day, or over $180 dollars Per year, above and beyond there regular } } } Internet } } } } costs. } } } } Note that this would be money paid directly to the U.S. Postal Service } } } } for a service they do not even provide. The whole point of the Internet is } } } } democracy and non-interference. If the federal government is permitted } } } } to tamper with our liberties by adding a surcharge to email, who knows } } } Where } } } } it will end. You are already paying an exorbitant price for snail mail } } } } because of bureaucratic inefficiency. It currently takes up to 6 days for } } } a } } } } letter to be delivered from New York to Buffalo. If The U.S. Postal } } } Service } } } } is } } } } allowed to tinker with email; it will mark the end of the "free" } } } } Internet in the United States. } } } } One congressman, Tony Schnell has even suggested a "twenty to forty } } } } dollar per month surcharge on all internet service" above and beyond the } } } } government's proposed email Charges. Note that most of the major } } } } newspapers have ignored the story, the only exception being the } } } } Washingtonian } } } } which called the idea of email surcharge "a useful concept who's time has } } } } come" } } } } (March 6, 1999) Editorial. } } } } } } } } Don't sit by and watch your freedoms erode away! Send this e-mail to } } } } EVERYONE on your list, and tell all your friends and relatives to write } } } } to their congressman and say "No!" to Bill 602P. } } } } } } } } } } } } It will only take a few moments of your time, and could very well be } } } } instrumental in killing a bill we don't want. } } } } } } } } PASS THIS ON TO EVERYONE YOU KNOW WHO USES EMAIL REMEMBER THESE ARE TWO } } } } SEPARATE ISSUES THAT EFFECT ALL OF US ONLINE. LET YOU VOICE BE HEARD } } } } NOW, NOT AFTER } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } }
Roger C Moretz, Ph.D.
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Does anyone know if GFP is quenched after OsO4 postfixation? Hank Adams Integrated Microscopy Core Molecular and Cell Biology Baylor College of Medicine Houston, TX 77030
I'm working with a InSb sample prepared with tripod polishing. Since the layer is quite thick, more than 3 micron plus a 2 micron buffer layer, I am experimenting some difficult to prepare a good sample for cross section observation. The final polishing is done in a Gatan ion milling set using a low angle. The problem with my samples is how to get a uniform thickness from the top of the sample to the substrate. Usually the top of the sample is milled much faster than near to the substrate. I can't mechanically polish the sample too much, since the sample is somewhat fragile (brittle). Even in old papers people report this kind of problem when preparing InSb samples for TEM using ion milling. Does someone else suffered from this kind of problem: the top part of a sample being milled away? Oh yes, when I polish the sample I try to make wedge perpendicular to the sample surface, so the thickness of the sample is the same from the InSb layer to the substrate after the mechanical polishing. There is some correlation between the ion milling angle/energy with this effect? The Gatan epoxy I use to glue a Si piece on top of the sample should have some influence (charging)? Thanks.
Kazuo
o-------------------------------------------------------o | Carlos Kazuo Inoki | | Department of Physics - University at Albany | | 1400 Washington Ave.- Albany - NY - 12222 | o-------------------------------------------------------o
I think that I know understand the reason why you get Be X-ray and why there are energy shifts when it is in different compounds. My next question then is do the elements between B and F have measurable energy shifts depending on what material they are in? The key word here is measurable. The Ka1 and Ka2 peaks are due to transitions form the 2p1/2 and 2p3/2 (LII and LIII) to the 1s1/2 state. But these would be valence/conductance bands for the pure elements. (Assuming you collect your X-rays from solid N2, O2, or F2) The band structure would be different for different materials. For example, do you see any shifts between B2O3? and BN or C(diamond) and C(graphite) or TiC? Any microprobers following this thread?
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
} -----Original Message----- } From: Hans Dijkstra [mailto:hans.dijkstra-at-edax.nl] } Sent: Thursday, February 10, 2000 11:01 AM } To: Walck. Scott D.; 'microprobe' } Cc: 'Microscopy' } Subject: RE: Be X-ray peaks -I don't think so } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Dear Scott, } } A poster does not mean anything. I have seen them with the } Lithium-Ka energy } listed, although quantum physics does forbid this X-ray line. } But I can } assure you the Be-K line does exist. } } On modern EDX systems Be can be detected, provided you have either a } windowless detector or a detector with a modern polymer type } window (SUTW, } SATW, Norvar, or whatever the EDX supplier calls it. My } apologies if I break } any trade-marks here...). } On most demonstration setups your EDS specialist will be } pleased to show you } a Be peak. But probably from a pure Be sample only. The absorption } coefficient (MAC) of Be in basically any matrix is so huge, } and the X-ray } fluorescence yield so small, that in any compound with less } than 50 atomic } percent Be you will not detect a visible Be peak. For this } reason most EDX } installations "in the field" are not setup to detect Be-K radiation. } } WDS has a better P/B ratio, so if you have the proper } multilayer crystal you } can get results. But peak shifts and peak shape alterations will make } quantification extremely difficult, not to mention the fact } that for most } MACs of Be-K in any matrix we only have a ball-park figure. } } Other techniques, like PEELS, are much more suitable for Li } and Be analysis. } Best regards, } } Hans Dijkstra } } Disclaimer: This is my opinion, and not necessarily the one } from EDAX Inc. } ------------------------------------------------------------- } EDAX Europe www.edax.com } Ringbaan Noord 103 Tel.: +31-13-5364000 } P.O. Box 4144 Fax.: +31-13-5356279 } 5004 JC Tilburg E-mail: hans.dijkstra-at-edax.nl } the Netherlands } ------------------------------------------------------------- } } } } -----Original Message----- } } From: Walck. Scott D. [mailto:walck-at-ppg.com] } } Sent: Tuesday, February 08, 2000 6:57 PM } } To: 'microprobe'; 'harry.ekstrom-at-honeywell.com' } } Cc: 'Microscopy' } } Subject: RE: Be X-ray peaks -I don't think so } } } } } } } -------------------------------------------------------------- } ---------- } } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.} html } } } } -------------------------------------------------------------- } ---------. } } } } } } I'm wrong. I guess I should have looked up at the X-ray } periodic table } } above my head and saw the 0.108 keV value for Be. The } reason that I was } } wrong was that I did not consider the solid state aspects. } I am still } right } } about not having a 2s1/2 to a 1s1/2 transition and that an } X-ray carries } } away 1h-bar of angular momentum. What I didn't do was } think about the } band } } structure of a solid. If you have the solid, the 2p bands } of Be most be } } spilling into or be the conduction band for the electrons. } That must be } the } } source for the electronic transition. I guess I should } have engaged brain } } before typing. } } My most humble apologies. } } -Scott } } } } Scott D. Walck, Ph.D. } } PPG Industries, Inc. } } Glass Technology Center } } Guys Run Rd. (packages) } } P. O. Box 11472 (letters) } } Pittsburgh, PA 15238-0472 } } } } Walck-at-PPG.com } } } } (412) 820-8651 (office) } } (412) 820-8161 (fax) } } } } } } "The opinions expressed are those of Scott D. Walck and not of PPG } } Industries, Inc. nor of any PPG-associated companies." } } -- } } } }
I did InSb and AlInSb by the small angle cleavage technique when I was at Wright Patterson AFB. I assume since you are using the Tripod Technique that you require site-specific results. If not, try it. It works really well on this material.
I am getting to sound like a broken record on this topic, aren't I? See my post yesterday.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies." --
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
} -----Original Message----- } From: Carlos Kazuo Inoki [mailto:kazuo-at-csc.albany.edu] } Sent: Thursday, February 10, 2000 1:05 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: TEM: sample preparation for InSb } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.} html } } } -------------------------------------------------------------- } ---------. } } } Hi, } } I'm working with a InSb sample prepared with tripod polishing. } Since the layer is quite thick, more than 3 micron plus a 2 } micron buffer } layer, I am experimenting some difficult to prepare a good sample for } cross section observation. The final polishing is done in a Gatan ion } milling set using a low angle. The problem with my samples is } how to get a } uniform thickness from the top of the sample to the } substrate. Usually the } top of the sample is milled much faster than near to the substrate. I } can't mechanically polish the sample too much, since the sample is } somewhat fragile (brittle). Even in old papers people report } this kind of } problem when preparing InSb samples for TEM using ion milling. Does } someone else suffered from this kind of problem: the top part of a } sample being milled away? Oh yes, when I polish the sample I } try to make } wedge perpendicular to the sample surface, so the thickness } of the sample } is the same from the InSb layer to the substrate after the mechanical } polishing. There is some correlation between the ion milling } angle/energy } with this effect? The Gatan epoxy I use to glue a Si piece on } top of the } sample should have some influence (charging)? Thanks. } } Kazuo } } o-------------------------------------------------------o } | Carlos Kazuo Inoki | } | Department of Physics - University at Albany | } | 1400 Washington Ave.- Albany - NY - 12222 | } o-------------------------------------------------------o } }
I hate to add to the bandwidth clutter but these are very old hoaxes.
Damian
} Is this yet another hoax eating time & bandwidth by being posted without } confirmation (like the last 3-4 times I saw this message in the pas year or } two), or is it real?
Yes, after OsO4 GFP is quenched, moreover it is quenched significantly (three times or more) even with 0.05% of glutaraldehyde (1Ž0 min.
Dr. Alexander A. Mironov Jr., MD, PhD Unit of Morphology Dept. of Cell Biology and Oncology Consorzio Mario Negri Sud Via Nazionale, S.Maria Imbaro (Ch) 66030 Italy
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Does anyone know if GFP is quenched after OsO4 postfixation? } Hank Adams } Integrated Microscopy Core } Molecular and Cell Biology } Baylor College of Medicine } Houston, TX 77030 } } }
Thanks to all of you who have shown an interest in our internships. I'd like to provide a few more details and answer some questions I've received so far about the summer internships at Abbott.
First: Are the internships open to international students? Technically, yes. However, interns are employees of Abbott Laboratories during their time here. Thus, the logistics of processing the necessary paperwork for non-US citizens for a 12-week employment period would not be practical. So, in truth, the answer has to be that, except in highly unusual circumstances, we will limit applications to students in the United States.
Second: Are the internships intended for college or high school students? The internships are for college students working towards a Bachelor's degree or higher. Students in the Associate degree programs at Madison Area Technical College and San Joaquin Delta College are eligible, but priority will be given to students who already have a Bachelor's degree.
Third: How to apply? Go to the Abbott Laboratories website at abbott.com and follow the links: Careers, Entry Level Positions, Summer Internship Program. There you will find an electronic resume form that can be submitted from your computer. THE DEADLINE FOR SUBMISSION IS MARCH 1.
Along with submitting the electronic resume, please send me either an e-mailed or faxed copy of your resume, or a hardcopy via snail mail. The electronic resume ends up in Corporate Staffing, where your skills will be matched with the internship openings at Abbott. If I have a copy of your resume, I can make sure that you are considered for positions in the microscopy department.
If there are any other questions, please feel free to contact me.
Thanks,
Jane A. Fagerland, Ph.D. Dept. Microscopy and Microanalysis D-45M/AP31 200 Abbott Park Rd. Abbott Park IL 60064-6202 voice: (847) 935-0104 fax: (847) 938-5027 e-mail: jane.a.fagerland-at-abbott.com
There was a paper by Cowen, Haven and Burnstock 1985, using Pontamine Sky Blue couterstain to reduce autofluorescence.
Also a paper by Kittleburger, Davis, and Stephans in Acta Histochem 89, using Eriochrome Black T.
Bob Morphology Core U of Washington
On Wed, 9 Feb 2000, Corazon Bucana wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I am working with archival specimens of formalin-fixed paraffin embedded } tissues and I am getting tremendous autofluorescence. I looked at tissue } sections after the sections have been de-waxed using either FITC or } rhodamine filter sets and red cells and connective tissues are brightly } fluorescent. Is there a way of suppressing the autofluorescence and still } retain reactivity of tissue to antibodies? I would appreciate any comments } or suggestions. } } ******************************************************* } Corazon D. Bucana, Ph.D. } Department of Cancer Biology } U.T. M.D. Anderson Cancer Center } 1515 Holcombe Blvd. Box 173 } Houston, Texas 77030 } Phone: (713) 792-8106 } FAX: (713) 792-8747 } Email:bucana-at-audumla.mdacc.tmc.edu } FAX: (713) 792-8747 } } }
At 8:51 AM +0100 4/2/00, Alexander Mironov Jr. wrote: } I have tried GoldEnhance for preembedding protocol to label intracellular } structures. It is marvelous. Buy it and use it - you will not be } unsatisfied. I have no any interest in Nanoprobes, I just like these dense } gold particles. } Practically all they claim is true: } - light insensitive } - no self-nucleation } - do not react with buffer ions } - is not dissolved by uranil and osmium (I have treat for 1 h)
I have been using BioCell's silver enhancement kit for years and particularly like it for the light insensitivity - one can monitor the reaction under the light microscope. Is it possible to do this with the GoldEnhance as well?
GoldEnhance sounds perfect - has anyone had any problems with it?
Diana
Diana van Driel Department of Clinical Ophthalmology Sydney University GPO Box 4337 Sydney NSW AUSTRALIA 2001
Reply to: Re:FYI: Congress to allow email charges -Hoax? I too had my suspicions. It looks very much like the "Internet Access Tax" letter from a few years ago. See: {http://urbanlegends.miningco.com/culture/urbanlegends/library/weekly/aa010798.htm} for details.
There are many sites documenting this sort of stuff. {http://www.hoaxkill.com/} is an example .
Disclaimer: I have no affiliation to this site.
Regards, Paul Webster
Paul Webster, Ph.D. Associate Scientist & Director Ahmanson Advanced Electron Microscopy & Imaging Center House Ear Institute 2100 West Third St. Los Angeles, CA 90057
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Woody, } } I hate to add to the bandwidth clutter but these are very old hoaxes. } } Damian } } } } Is this yet another hoax eating time & bandwidth by being posted without } } confirmation (like the last 3-4 times I saw this message in the pas year or } } two), or is it real? } } Damian's response is indeed correct -- see posted responses by "David_Bell-at-millipore.com", "EvexAnalyt-at-aol.com", Scott Wight, Kalman Rubin, Dr. Gary Gaugler, donald j. marsh, Roger Moretz, Laurie Wallin and Damian. Being new to the MSA listserver, I'm very relieved that such notices (err, BANDWIDTH-OCCUPYING, ANNOYING HOAXES) are determined to be false by very astute readers. I fell for this, and, upon reading other people's notices, I am upset that someone would suggest spreading such hoaxes around like the classic junk chain mail activities done at school (or even college). Such notices clog up the space needed for important issues regarding microscopy. Anyway .. I'm responding merely to alert other GULLIBLE people (like myself) not to believe such notices, as well as be thankful that this has been a recurring event in the MSA listserver. Nelson Conti
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IMHO, the main problem with GoldEnhance is some inhomogenity in particle size but for my purposes it is irrelevant. Also the kit works good in my hands at pH 7.4 but does not at pH 6.5 (as claimed Nanoprobes).
Sincerely, Dr. Alexander A. Mironov Jr., MD, PhD Unit of Morphology Dept. of Cell Biology and Oncology Consorzio Mario Negri Sud Via Nazionale, S.Maria Imbaro (Ch) 66030 Italy
Dear readers, could anyone give me some hints or tips how I can separate a formvarfilm (0.5% in chloroform) from the glassslide ? Breathing or freezing doesn't help at all. It works better with non-washed slides, but then I get an uneven and dirty surface of the film. I appreciate every kind of support. Best regards
Guido Luond Institute of Oral Microbiology and General Immunology Univ. of Zurich Plattenstr. 11 CH-8028 Zurich, Switzerland E-mail: lueoend-at-zzmk.unizh.ch
Carlos, sounds like you may be having a problem with differential milling rates. Try milling over a restricted angle; you can use oscillation (if your ion mill is set up to do this) or make C-shaped Ta plate shields which you fix on the sample holding plates of your ion mill, with the sample at the centre of the 'C'. If you put your sample in the plates such that the interface is parallel to the opening of the shield, the average direction of the ion beam will be perpendicular to the interface. This will prevent the ion beam milling parallel to the glue line and stop the layers disappearing before the substrate. If you stick to angles below 10 degrees you will get better results, and of course use liquid nitrogen cooling and/or iodine. My apologies if this isn't very clear - it's hard to describe everything with just words! I can send you a picture of the sample holder I use if you like.
Best regards,
Richard Beanland
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi, } } I'm working with a InSb sample prepared with tripod polishing. } Since the layer is quite thick, more than 3 micron plus a 2 micron buffer } layer, I am experimenting some difficult to prepare a good sample for } cross section observation. The final polishing is done in a Gatan ion } milling set using a low angle. The problem with my samples is how to get a } uniform thickness from the top of the sample to the substrate. Usually the } top of the sample is milled much faster than near to the substrate. I } can't mechanically polish the sample too much, since the sample is } somewhat fragile (brittle). Even in old papers people report this kind of } problem when preparing InSb samples for TEM using ion milling. Does } someone else suffered from this kind of problem: the top part of a } sample being milled away? Oh yes, when I polish the sample I try to make } wedge perpendicular to the sample surface, so the thickness of the sample } is the same from the InSb layer to the substrate after the mechanical } polishing. There is some correlation between the ion milling angle/energy } with this effect? The Gatan epoxy I use to glue a Si piece on top of the } sample should have some influence (charging)? Thanks. } } Kazuo } } o-------------------------------------------------------o } | Carlos Kazuo Inoki | } | Department of Physics - University at Albany | } | 1400 Washington Ave.- Albany - NY - 12222 | } o-------------------------------------------------------o }
============================================================== Richard Beanland Caswell Technology, Caswell, Towcester, Northants NN12 8EQ
e-mail richard.beanland-at-gecm.com Tel. +44 1327 356363 Fax. +44 1327 356398 ============================================================== "The information contained in this message is legally privileged and confidential information intended for the eyes of the individual or entity named above. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution or copying of this message is strictly prohibited. If you have received this message in error, please notify us immediately by telephone. Caswell Technology is the trading name of Marconi Caswell Limited. Registered in London No. 3694360 Registered Office: One Bruton Street London W1X 8AQ. Holding Company: Marconi plc."
Is your end goal hi res imaging or ratioing? If the latter, I saw an interesting technology at Cell Bio which might be of help: the PARISS Spectral imaging system from LightForm. I saw very early versions of this technology some years ago at PITTCON but it has really matured and offers an interesting alternative. PARISS can acquire 240 spectra simultaneously, full field or in a region of interest. It really crashes through the old barriers imposed by filter cubes. Also, despite the radical differences in intensity, it can acquire the transmitted light image simultaneously with the fluorescence image and present the combined results with perfect registration. The system retrofits to existing microscopes and is moderately priced, considering all the accessories (2 cameras, spectrometer, motorized stage, software). If you are interested, visit their website at lightforminc.com or contact Jeremy Lerner (908-281-9098).
Caveat: MME has no financial interest in this product.
Hope this is helpful.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education}
At 10:23 AM 2/10/00 -0500, David Knecht wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Sorry, I think a small correction/clarification is requires. This is ONLY for an incoherently filled condensor aperture, i.e. a large beam with C2 fully focussed. This approximation is not true with the newer microscopes!
On Thu, 10 Feb 2000, Jonathan Barnard wrote:
} Born & Wolf (section 10.4.2) } "Hence if the linear dimensions of the source and the distance between P1 } and P2 (points in the imaging plane) are small compared to the distance of } these points from the source, the degree of coherence, |mu_12| is equal to } the absolute value of the normalized Fourier transform of the intensity } function of the source" }
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University fax: (847) 491-7820 mailto:l-marks-at-nwu.edu http://www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
WDS spectra of Boron, Carbon, Nitrogen and Oxygen in many compounds have been extensively recorded by Bastin and Heijligers in the 1980's and early 1990's. They report that peak shifts and peak shape alterations are clearly present in Boron compounds. In the case of TiB crystals it was even found that the crystal orientation influences the peak shape and position, i.e. if you rotate the sample 90 degrees you get a different peak shape. For Carbon the peak shapes are practically unaffected by the bonding with other elements.
Please check "Quantitative Electron Probe Microanalysis of Boron in Binary Borides" by Bastin and Heijligers, University of Technology Eindhoven, the Netherlands, 1997, ISBN 90-3860-898-5.
In EDX the peak shifts are undetectable. Modern EDX systems give a FWHM of around 60-65 eV for these very light elements, so a peak shift of a few eV is basically undetectable. For many users this makes EDX the prefered technique to quantify Boron compounds, since with WDS you need to record either the full peak (tedious) or use fixed area-peak-fraction correction methods. In EDX we can simple ignore peak shape and peak position changes.
Best regards,
Hans Dijkstra
Disclaimer: This is my opinion, and not necessarily the one from EDAX Inc. ------------------------------------------------------------- EDAX Europe www.edax.com Ringbaan Noord 103 Tel.: +31-13-5364000 P.O. Box 4144 Fax.: +31-13-5356279 5004 JC Tilburg E-mail: hans.dijkstra-at-edax.nl the Netherlands -------------------------------------------------------------
} -----Original Message----- } From: Walck. Scott D. [mailto:walck-at-ppg.com] } Sent: Thursday, February 10, 2000 7:51 PM } To: 'Microscopy' } Subject: RE: Be X-ray peaks, solid state physics, how about 2nd row } elements } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I think that I know understand the reason why you get Be X-ray and why } there are energy shifts when it is in different compounds. My next question } then is do the elements between B and F have measurable energy shifts } depending on what material they are in? The key word here is measurable. } The Ka1 and Ka2 peaks are due to transitions form the 2p1/2 and 2p3/2 (LII } and LIII) to the 1s1/2 state. But these would be valence/conductance bands } for the pure elements. (Assuming you collect your X-rays from solid N2, O2, } or F2) The band structure would be different for different materials. For } example, do you see any shifts between B2O3? and BN or C(diamond) and } C(graphite) or TiC? Any microprobers following this thread? } } -Scott } } Scott D. Walck, Ph.D. } PPG Industries, Inc. } Glass Technology Center } Guys Run Rd. (packages) } P. O. Box 11472 (letters) } Pittsburgh, PA 15238-0472 } } Walck-at-PPG.com } } (412) 820-8651 (office) } (412) 820-8161 (fax) }
first of all I would like to thank of the people who spent part of their time trying to share their knowledge with me, and also to the manufacturers who of course replied my messages.
All messages contained very important suggestions, that I need to study and evaluate, and I will contact personally the people that offered to share their previous experience and/or to collaborate.
Our work on InGaAs/GaAs quantum wells, single and stacked quantum wires, single and stacked quantum dots, consists of conventional TEM, high resolution transmission electron microscopy and high spatial resolution analytical electron microscopy, that we perform in close collaboration with Arizona State University, using a FEG/STEM machine equipped with EELS and EDX. One of the goals of the research is to evaluate compositional fluctuations in the well and in the dots at nanoscale.
As everybody knows, the requirements from a TEM sample for imaging are completely different from the requirements necessary to perform high spatial resolution analytical work, were effects such as amorphisazion, surface contamination are critical.
We have a pretty well estabilished specimen preparation technique, but before publishing results into the scientific community, we want to be sure that what we observe is what is in the sample (isn't this the main concern of every microscopist?).
That's why all suggestions, opinions and critics are strongly appreciated.
I don't wash the slides, just wipe the dry slide with a Kim-Wipe a few times. Works like a charm, whatever the weather! (Very grey and nasty here on "the island" today, by the way).
Tamara Howard CSHL
On Fri, 11 Feb 2000, GUIDO L[ISO-8859-1] üöND wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear readers, } could anyone give me some hints or tips how I can separate a formvarfilm } (0.5% in chloroform) from the glassslide ? Breathing or freezing doesn't } help at all. It works better with non-washed slides, but then I get an } uneven and dirty surface of the film. } I appreciate every kind of support. } Best regards } } Guido Luond } Institute of Oral Microbiology and } General Immunology } Univ. of Zurich } Plattenstr. 11 } CH-8028 Zurich, Switzerland } E-mail: lueoend-at-zzmk.unizh.ch } } }
There is a product called "Victawet", which is available here in the U.S. from various EM suppliers. It comes in a small vial that lasts forever and is an off-white, waxy or soapy substance. We used to take our slides and clean them very well with lint-free papers, then place them in a rack inside a vacuum evaporator, with the side intended for the Formvar film facing a tungsten basket connected to the electrodes. Then we place a piece of Victawet about the size of a grain of rice in the basket, pump down the system to high vacuum, and slowly heat the tungsten basket until it glows dull red. (If you increase the current too quickly the little piece will jump out of the basket.) The slides will begin to look fogged, as if someone had breathed moisture onto them. You can stop at that point---you only need a little.
Store these coated slides until needed, then take them as required and polish them very carefully with lint-free paper. They should feel slippery. Use these cleaned slides to dip into the Formvar solution and the film should separate easily.
Please note that the type of slide and the humidity levels in the workplace also have a large effect. I have worked in places where films would separate from any slide, every day, with no Victawet coating. I have also worked in places where we could only get Formvar films on occasion (who knows why?), and then we made a bunch of them at once.
Hope this helps.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 (573) 882-8304 (573) 884-5414 (fax) email: tindallr-at-missouri.edu {mailto:tindallr-at-missouri.edu} http://biotech.missouri.edu/emc {mailto:tindallr-at-missouri.edu}
Dear readers,
could anyone give me some hints or tips how I can separate a formvarfilm
(0.5% in chloroform) from the glassslide ? Breathing or freezing doesn't
help at all. It works better with non-washed slides, but then I get an
uneven and dirty surface of the film.
I appreciate every kind of support.
Best regards
Guido Luond
Institute of Oral Microbiology and
General Immunology
Univ. of Zurich
Plattenstr. 11
CH-8028 Zurich, Switzerland
E-mail: lueoend-at-zzmk.unizh.ch
Randy Tindall EM Specialist Electron Microscopy Core Facility W122 Veterinary Medicine University of Missouri Columbia, MO 65211 (573) 882-8304 (573) 884-5414 (fax) email: tindallr-at-missouri.edu {mailto:tindallr-at-missouri.edu} http://biotech.missouri.edu/emc {mailto:tindallr-at-missouri.edu}
Hasn't anyone realized that the "hoax" email is a success even as a failure. Everyone is responding so much about the email being a hoax that we're using up band width and cluttering email accounts. We all know it's a hoax..., let's leave it at that.....
Regards,
Mike Santana, Jr. Fab25 Product Engineering Advanced Micro Devices Desk - (512) 602-4172 Pager - (512) 622-2494 email - mike.santanajr-at-amd.com
On Thursday, February 10, 2000 10:27 PM, Paul Webster [SMTP:pwebster-at-hei.org] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Reply to: Re:FYI: Congress to allow email charges -Hoax? } I too had my suspicions. It looks very much like the "Internet Access Tax" letter from a few years ago. See: {http://urbanlegends.miningco.com/culture/urbanlegends/library/weekly/aa 010798.htm} for details. } } There are many sites documenting this sort of stuff. {http://www.hoaxkill.com/} is an example . } } Disclaimer: I have no affiliation to this site. } } Regards, } Paul Webster } } Paul Webster, Ph.D. } Associate Scientist & Director } Ahmanson Advanced Electron Microscopy & Imaging Center } House Ear Institute } 2100 West Third St. } Los Angeles, CA 90057 } } Phone: (213) 273-8026 } Fax: (213) 413-6739 } e-mail: pwebster-at-hei.org } http://www.hei.org/htm/aemi.htm }
I would like to contact users of Bal-Tec RES 010 ion thinner to comment on performance of this equipment. We have found over the years frequent stability problems of the guns, which are not solved only with cleaning. We are at the moment checking any possible source of unstability. We would like advise from anyone who has had similar problems with this apparatus, to know if we are missing something and what would be the best course of action.
Thanks in advance
Jesœs Ricote ------------------------------------------------------------- Dr. Jesœs Ricote Departamento de Materiales FerroelŽctricos Instituto de Ciencia de Materiales de Madrid Consejo Superior de Investigaciones Cientificas (CSIC) Cantoblanco 28049 Madrid SPAIN
first of all I would like to thank of the people who spent part of their time trying to share their knowledge with me, and also to the manufacturers who of course replied my messages.
All messages contained very important suggestions, that I need to study and evaluate, and I will contact personally the people that offered to share their previous experience and/or to collaborate.
Our work on InGaAs/GaAs quantum wells, single and stacked quantum wires, single and stacked quantum dots, consists of conventional TEM, high resolution transmission electron microscopy and high spatial resolution analytical electron microscopy, that we perform in close collaboration with Arizona State University, using a FEG/STEM machine equipped with EELS and EDX. One of the goals of the research is to evaluate compositional fluctuations in the well and in the dots at nanoscale.
As everybody knows, the requirements from a TEM sample for imaging are completely different from the requirements necessary to perform high spatial resolution analytical work, were effects such as amorphisazion, surface contamination are critical.
We have a pretty well estabilished specimen preparation technique, but before publishing results into the scientific community, we want to be sure that what we observe is what is in the sample (isn't this the main concern of every microscopist?).
That's why all suggestions, opinions and critics are strongly appreciated.
Ah. Bit of a problem, that. I didn't mention how far away the shield is from the sample - about 3 or 4 mm - and that for low angles I use a wedge-shaped shield to deflect ions away from the specimen, rather than back on to it. Did you really try it that quickly? I only sent the e-mail a few hours ago! I did expect anyone who was going to have a go to ask me for a picture of my holder. As to cleaning your sample, I think you can only ion mill it some more, with amended shields...
Richard
} Dear Richard, } } I did as you suggested in my sample preparation. However, the Ta was } sputtered on the surface of the } specimen. Could you suggest me how to avoid or how to clean the Ta on the } specimen surface. } } Chengge Jiao } H.H.Wills Physics Lab } University of Bristol, U.K } c.g.jiao-at-bristol.ac.uk
============================================================== Richard Beanland Caswell Technology, Caswell, Towcester, Northants NN12 8EQ
e-mail richard.beanland-at-gecm.com Tel. +44 1327 356363 Fax. +44 1327 356398 ============================================================== "The information contained in this message is legally privileged and confidential information intended for the eyes of the individual or entity named above. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution or copying of this message is strictly prohibited. If you have received this message in error, please notify us immediately by telephone. Caswell Technology is the trading name of Marconi Caswell Limited. Registered in London No. 3694360 Registered Office: One Bruton Street London W1X 8AQ. Holding Company: Marconi plc."
One of the benefits from the small angle cleavage technique on the III-V compounds is that often a "step-like" sample is produced. When this happens, you have a perfect sample for both High resolution and analytical work. In the very thin steps, you can do HREM and EELS. In the thicker steps, you can do EDS analysis. -Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
} -----Original Message----- } From: Massimo Catalano [mailto:massimo.catalano-at-ime.le.cnr.it] } Sent: Friday, February 11, 2000 10:23 AM } To: Microscopy-at-sparc5.Microscopy.Com } Subject: PIPS and milling damage: Second message } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Dear Listservers: } } first of all I would like to thank of the people who spent } part of their } time trying to share their knowledge with me, and also to the } manufacturers } who of course replied my messages. } } All messages contained very important suggestions, that I } need to study and } evaluate, and I will contact personally the people that } offered to share } their previous experience and/or to collaborate. } } Our work on InGaAs/GaAs quantum wells, single and stacked } quantum wires, } single and stacked quantum dots, consists of conventional TEM, high } resolution transmission electron microscopy and high spatial } resolution } analytical electron microscopy, that we perform in close } collaboration with } Arizona State University, using a FEG/STEM machine equipped } with EELS and } EDX. One of the goals of the research is to evaluate compositional } fluctuations in the well and in the dots at nanoscale. } } As everybody knows, the requirements from a TEM sample for } imaging are } completely different from the requirements necessary to perform high } spatial resolution analytical work, were effects such as } amorphisazion, } surface contamination are critical. } } We have a pretty well estabilished specimen preparation } technique, but } before publishing results into the scientific community, we } want to be sure } that what we observe is what is in the sample (isn't this the } main concern } of every microscopist?). } } That's why all suggestions, opinions and critics are strongly } appreciated. } } Best regards } } Massimo } } } Dr. Massimo Catalano } CNR-IME } Campus Universitario } Via Arnesano } 73100 Lecce - ITALY } tel: + 39 0832 322362 } fax: + 39 0832 325299 } email: massimo.catalano-at-ime.le.cnr.it } http://www.ime.le.cnr.it } http://www.ime.le.cnr.it/sime/sime.htm } } }
I've always had good luck getting the film off by polishing the slide first with Ross Optical Lens Tissue. My major professor taught me this, and his explaination was that it has silicone in it. Once you have cast the film onto the slide and allowed it to dry, cut through the film at the edges and bottom of the slide by running a different slide along them. Then breathe on the film and lower it into the water dish.
Good luck, Heather Owen
p.s. I have no affiliation with the company that makes, or any of the EM supply houses that sell Ross lens paper.
Heather A. Owen, Director Electron Microscope Laboratory Department of Biological Sciences University of Wisconsin - Milwaukee (414)229-6816
Dear Scott, In the Microbeam Analysis 1985, G.F. Bastin and H.J.M. Heijligers have the first article, entitled: "Quantitative Electtron-Probe Microanalysis of Very Light Elements" (page 1). They have several articles in that and succeeding years in which they try to analyse every element and compound in the B to F range material that they can get to sit still under the EPMA beam. They did a very valuable set of studies that is worth looking into if you want to do any very light element analysis. They cover peak shifts, peak shape changes, even crystallographic direction sensitivities. At 01:51 PM 2/10/00 -0500, you wrote:
} } I think that I know understand the reason why you get Be X-ray and why } there are energy shifts when it is in different compounds. My next question } then is do the elements between B and F have measurable energy shifts } depending on what material they are in? The key word here is measurable. } The Ka1 and Ka2 peaks are due to transitions form the 2p1/2 and 2p3/2 (LII } and LIII) to the 1s1/2 state. But these would be valence/conductance bands } for the pure elements. (Assuming you collect your X-rays from solid N2, O2, } or F2) The band structure would be different for different materials. For } example, do you see any shifts between B2O3? and BN or C(diamond) and } C(graphite) or TiC? Any microprobers following this thread? } } -Scott Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
Hello Casey/Marty I cannot answer for the first question as we use Diotome knives and I have no experience with the other.
But the answer to the second question is "Yes." At least for us it is. We are a facility used by the Faculty of Medicine and Science so many and varied samples come through here for electron, light, and confocal microscopy. We have been working up some protocols for difficult specimens such as nematodes, drosophila larvae and plant material. The power supply allows control so that you don't have to do any other processing to the specimens than toss them in the fixative. Since the fix and post fix enter the specimens at low power ( {200 watts) without damaging the specimens (full power is usually about 800 watts) we can fix in about 45 minutes for something which would normally be in 24 hours. And we don't have to punch holes in the specimen.
We have just been playing with the introduction of fluorescent dyes for light microscopy and confocal in C. elegans, without the use of agents to distrupt the membranes to get the dye in. So far we have got the timings for beautiful staining of the tail region and partial staining of the thicker sections. We still have to get it right but the initial experiments are very positive.
No-one seems to know why the microwaves work, but the hypothesis is that at low power, the cell membranes are "jiggled about" to let the fix/dye in without distrupting them.
We are a very busy lab, and our experiments are constantly getting interupted so it is taking longer than we'd like to pin down a finished protocol.
I would be interested in receiving protocols from labs who have already got it to work for their specimens. We are finding that every specimen is different and the protcols have to be tweaked for each one. Elaine
} } A colleague would like your responses to these questions. I believe that I } say something on the first question about a year ago and apologize for } asking again.
} } Here's what I/we need to know. } } } } 1) Are Microstar diamond knives equal to Diatome. We have an offer that } } will save us about $1200 on three knives ( one 3mm ultra with 45 degree } } angle, one 3mm ultra with 35 degree angle, one 10 mm histo knife). } } } } } } 2) Is the power controller feature on the Pelco Microwave Tissue processor } } (model #3451) worth it? (costs about $1300 additional compared to the } } Pelco 3450 without the power controller). I plan to do immunolocalization } } work, but am not sure this power controller feature is necessary or worth } } the cost. } } } } Casey
Marty Reed
Dr. Elaine Humphrey Biosciences Electron Microscopy Facility University of British Columbia 6270 University Blvd, mail-stop Botany Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-unixg.ubc.ca
Dear Guido, I have learned long ago not to use films cast on slides. It is easier and much cleaner to spread your films on the surface of water in a trough. You can vary the thickness by the amount of drops you use and chose the area you want to place your grids. This method gave me beautiful and sturdy films. Regards, Maria
Maria Fazio-Zanakis Bioimaging and Molecular Histology Aventis Pharmaceuticals 1-908-231-3357 Fax: 1-908-231-3962 Email: Maria.Fazio-Zanakis-at-Aventis.com
-----Original Message----- } From: GUIDO LŸšND [mailto:lueoend-at-zzmk.unizh.ch] Sent: Friday, February 11, 2000 4:31 AM To: EM-Tips
Dear readers, could anyone give me some hints or tips how I can separate a formvarfilm (0.5% in chloroform) from the glassslide ? Breathing or freezing doesn't help at all. It works better with non-washed slides, but then I get an uneven and dirty surface of the film. I appreciate every kind of support. Best regards
Guido Luond Institute of Oral Microbiology and General Immunology Univ. of Zurich Plattenstr. 11 CH-8028 Zurich, Switzerland E-mail: lueoend-at-zzmk.unizh.ch
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Reply to: RE: TEM Formvar Film Cast on Glass Hi Guido,
Here is my recipe which has worked on four continents at altitudes up to 2,000m and in all weathers (humid to dry, cold to hot). Plus, it does not use those very unscientific body fluids I often see being used on this list!
Dissolve the formvar in chloroform as a concentration higher than will make the correct film thickness (film thickness will vary with the speed the glass is drawn out of the solution as well as with formvar concentration).
Take a glass slide (it is important to use precleaned 25x75 mm slides, VWR Scientific, Cat number 48312-002) and dip it in absolute ethanol. Immediately dry it with Ross lens tissue. If you wash it any other way or dry it with anything other than the Ross lens tissue, the film will not come off the glass.
Coat the slide immediately and let the film dry. Score the edges of the film so that it will easily detach from the glass and float it onto the water surface. Evaluate the film thickness and adjust the formvar concentration by adding chloroform to the solution.
I know there are many versions of this method that work. However, this is the only method I have found that is completely oblivious to the weather (still sunny in LA) and has been reproduced by many people. in my experience, the formvar dissolved in choroform does not seem to have the short shelf life I read about recently. I have used the same solution for years, topping it up with chloroform to keep it at a concentration that will produce thin films.
Regards,
Paul Webster
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm
GUIDO LŸšND wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear readers, } could anyone give me some hints or tips how I can separate a formvarfilm } (0.5% in chloroform) from the glassslide ? Breathing or freezing doesn't } help at all. It works better with non-washed slides, but then I get an } uneven and dirty surface of the film. } I appreciate every kind of support. } Best regards } } Guido Luond } Institute of Oral Microbiology and } General Immunology } Univ. of Zurich } Plattenstr. 11 } CH-8028 Zurich, Switzerland } E-mail: lueoend-at-zzmk.unizh.ch } } } } } RFC822 header } ----------------------------------- } } Received: from ultra5.microscopy.com [206.69.208.10] by mailhouse.hei.org } with ESMTP } (SMTPD32-5.05) id A182210B0080; Fri, 11 Feb 2000 10:14:26 -0800 } Received: (from daemon-at-localhost) } by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id DAA27990 } for dist-Microscopy; Fri, 11 Feb 2000 03:36:30 -0600 (CST) } Received: from no_more_spam.com (sparc5.microscopy.com [206.69.208.10]) } by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id DAA27987 } for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Fri, 11 Feb 2000 } 03:35:32 -0600 (CST) } Received: from alpha.zzmk.unizh.ch (zzmkmail.unizh.ch [130.60.67.4]) } by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id DAA27980 } for {Microscopy-at-sparc5.microscopy.com} ; Fri, 11 Feb 2000 03:35:21 -0600 (CST) } Received: from [192.168.7.192] by alpha.zzmk.unizh.ch } (8.8.8/1.1.22.3/03Jan00-0358PM) } id KAA0000026325; Fri, 11 Feb 2000 10:30:33 +0100 (MET) } User-Agent: Microsoft Outlook Express Macintosh Edition - 5.0 (1513) } Date: Fri, 11 Feb 2000 10:31:14 +0100 } Subject: TEM Formvar Film Cast on Glass } From: GUIDO L=?ISO-8859-1?B?/PY=?=ND {lueoend-at-zzmk.unizh.ch} } To: EM-Tips {Microscopy-at-sparc5.microscopy.com} } Message-ID: {B4C99572.725%lueoend-at-zzmk.unizh.ch} } Mime-version: 1.0 } Content-type: text/plain; charset="US-ASCII" } Content-transfer-encoding: 7bit } Errors-to: Microscopy-request-at-sparc5.Microscopy.Com } X-RCPT-TO: {pwebster-at-hei.org} } X-UIDL: 242867993 } Status: U } Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm
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hi all-
just got a request to view surface structure of the inside of a 12" metal pipe. its too big for my chamber and they'd prefer a non-destructive technique. will replication of the surface with acetate/acetone and SEM work. i've never tried this before so any pointers would be welcome...
thx! brian ---------------------------------------------- Brian McIntyre Electron Microscopy Lab- River Campus Univ of Rochester Rochester, NY 14627 716-275-3058/4875
Laurie, Frieda Carson's book about histologic technique is the "gold standard for histotechs right now. It is available at Amazon.com and Barnes and Noble's website, and can be special ordered at the major bookstores, as it is currently in print. (Unfortunately I am at home and do not have the actual title in front of me) Wanda Shotsberger ----- Original Message ----- } From: Laurie Wallin {lwallin-at-ucsd.edu} To: {microscopy-at-sparc5.Microscopy.Com} Sent: Thursday, February 10, 2000 11:03 AM
I was asked a couple of questions about fluorescent microscopy and I didn't have a clue. Are all the dyes used with UV or are there other wavelengths that can fluoresce them? Are the dyes used dangerous/hazardous? Are they transparent to visible? Are they expensive? Can you get them in quantity?
-Clueless in Pittsburgh
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
I knew a very good, reputable company in the Southern California region that serviced ETEC SEMs. The name was Scan Service in California, Earl Weltmer 714 area code.
- Jeff
-----Original Message----- } From: LAB. DE MICROSCOPIA ELECTRONICA - FI - UNER [mailto:microsc-at-fi.uner.edu.ar] Sent: Wednesday, February 09, 2000 5:10 AM To: microscopy-at-sparc5.microscopy.com
Hello ..... we have an old ETEC Omniscan SEM, and we are searching for ETEC company or any people that can supply manuals ....
any help is welcome
best regards
=================================================== Fernando D. Balducci Laboratorio de Microscopia Electr—nica Facultad de Ingenier’a - Bioingenieria Universidad Nacional de Entre Rios Argentina e-mail: microsc-at-fi.uner.edu.ar tel: 54 43 975100 fax: 54 43 975077
In regards to the diamond knives, both companies make an excellent product and both provide good service. Because of price, the last 6 knives I've bought have been from Microstar (two well-used but dull Diatome knives have been traded during these transactions) . All six knives have been more than suitable for the intended purpose. I have no commercial interest in either company.
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A colleague would like your responses to these questions. I believe that I say something on the first question about a year ago and apologize for asking again.
} Here's what I/we need to know. } } 1) Are Microstar diamond knives equal to Diatome. We have an offer that } will save us about $1200 on three knives ( one 3mm ultra with 45 degree } angle, one 3mm ultra with 35 degree angle, one 10 mm histo knife). } } } 2) Is the power controller feature on the Pelco Microwave Tissue processor } (model #3451) worth it? (costs about $1300 additional compared to the } Pelco 3450 without the power controller). I plan to do immunolocalization } work, but am not sure this power controller feature is necessary or worth } the cost. } } Casey } } } Thanks for your time } Marty Reed Equipment Technician Biology Department Humboldt State University Arcata CA 95521 707-826-3234 707-826-3201 FAX mmr7001-at-axe.humboldt.edu
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Date sent: Fri, 11 Feb 2000 14:01:04 -0500 To: Microscopy-at-sparc5.Microscopy.Com } From: Brian McIntyre {mcintyre-at-optics.rochester.edu}
Depends on the detail needed, but acetate replication should work for almost all situations. Of course, everything you see will be inside out... Often, selecting inverse video helps one perceive thing properly. If the pipe is dirty or corroded, you will likely pick up some deposit with the replica. This can be distracting, but can also be helpful if x-ray analysis is required... Woody White McDermott Technology My place: http://www.geocities.com/capecanaveral/3722
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hi all-
just got a request to view surface structure of the inside of a 12" metal pipe. its too big for my chamber and they'd prefer a non-destructive technique. will replication of the surface with acetate/acetone and SEM work. i've never tried this before so any pointers would be welcome...
thx! brian ---------------------------------------------- Brian McIntyre Electron Microscopy Lab- River Campus Univ of Rochester Rochester, NY 14627 716-275-3058/4875
by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id RAA00487 for dist-Microscopy; Fri, 11 Feb 2000 17:29:13 -0600 (CST) Received: from no_more_spam.com (sparc5.microscopy.com [206.69.208.10]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id RAA00484 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Fri, 11 Feb 2000 17:28:14 -0600 (CST) Received: from postoffice.mail.cornell.edu (POSTOFFICE.MAIL.CORNELL.EDU [132.236.56.7]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id RAA00477 for {Microscopy-at-sparc5.microscopy.com} ; Fri, 11 Feb 2000 17:28:00 -0600 (CST) Received: from [132.236.129.190] ([132.236.129.190]) by postoffice.mail.cornell.edu (8.9.3/8.9.3) with ESMTP id SAA19931 for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 11 Feb 2000 18:23:10 -0500 (EST) X-Sender: mvp2-at-postoffice.mail.cornell.edu Message-Id: {v04003000b4ca4820943f-at-[132.236.129.190]} Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
If any one has a PGT IMIX X-ray Analysis/Imaging system 4-40 (SUN Sparc) that is in storage, or not in use, I would be interested in acquiring its Image Signal Processor ISP module (Part# MO328D-00).
Stop destroying your pipe. With our large chamber SEM we can analyse your pipe up to a length of 35 inches and a diameter of 28 inches. Maximum weight of the sample should not exceed 500 pounds.
Because of the positionable electron gun it is easy to observe the inside surface.
Best regards Martin Klein
Disclaimer: VisiTec is the manufacturer of large chamber SEMs. And we like large samples.
----- Original Message ----- } From: Brian McIntyre {mcintyre-at-optics.rochester.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Friday, February 11, 2000 8:01 PM
I am in desperate need of a control unit for a LKB ultratome III, since mine is beyond repair. I would appreciate if anyone coud sell me an unused unit from old apparatus, even if it does not work, I might be able to repair mine with the pieces. Please reply to the e-mail below
Thanks
Dr. A.P. Alves de Matos Dental Medical School Lisbon University apmatos-at-ip.pt
ELECTRON MICROSCOPY MATERIALS SPECIALIST UNL Center for Materials Research and Analysis
Analyze/characterize materials using electron microscopy, materials preparation and computer instrumentation. Supervise/train students, faculty and visiting researchers utilizing these methods. Bachelor's with major in physical science, engineering or related field plus three years experience in the operation, repair or design of electron microscopes or other scientific instrumentation. Master's preferred. Must have excellent computer and interpersonal/communication skills. Knowledge of x-ray diffraction, TEM/SEM principles and sample preparation experience preferred. Excellent benefits. Submit cover letter, resume and names, addresses and telephone numbers of three professional references to Professor David J. Sellmyer, 112 Brace Lab, Lincoln, NE 68588-0113. Review of resumes will begin March 1. Position will remain open until a suitable candidate is found. UNL is committed to AA/EEO and ADA/504. If you require accommodation, please call (402) 472-8762.
Try a different brand of slide. Corning is the only brand we use in our EM Class. There must be different kinds of surfactants used on them. Some other brands of slides just don't release the formvar. We usually wipe them off thoroughly with a Kimwipe only. Dip the slide into the formvar and let dry. But don't dry for too long. Try using a diamond tipped (or carbide) pen to score a "rectangle" on the slide. Slowly insert the slide at about a 30 degree angle into the water. Wait until you see the formvar start to release before you continue inserting into the water. This method usually works in our lab and for students.
Dear Linda, I have a DuPont Sorvall MT2B or a Porter Blum MT1 I would be interested in selling if the price is right. If you are interested, let me know via my email address: tiekotte-at-up.edu. Cheers! Ken ----------- Ken Tiekotter Dept. of Biology The University of Portland 5000 N. Willamette Blvd. Portland, OR 97303
On Mon, 27 Aug 1956, Linda Chicoine wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Does anyone in the Connecticut area have an old microtome they would } like to sell? I would be interested in anything from an LKB to RMC to } Reichert/ and Leica. Please contact me by email. Thanks. Linda } Chicoine } } }
} To: MICROSCOPY BB {Microscopy-at-sparc5.microscopy.com} } Subject: SEM pipe replication } Date: Sat, 12 Feb 2000 10:44:32 -0500 } From: "Garber, Charles A." {cgarber-at-2spi.com} } X-Mailer: E-Mail Connection v3.1a } Content-Length: 3106
} } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } Brian McIntyre wrote: } ========================================= } just got a request to view surface structure of the inside of a 12" metal } pipe. its too big for my chamber and they'd prefer a non-destructive } technique. will replication of the surface with acetate/acetone and SEM } work. i've never tried this before so any pointers would be welcome... } ========================================== } You did not indicate how far into the pipe you want to do your looking, but } cellulose acetate is certainly one approach. The biggest problem you are } going to have is to balance the need to do this all "nondestructively" vs. } the need to see what might be underneath material covering the top surface
} (after all, it is the metal substrate that you really want to see). In } other words, if the acetone should be a solvent for the contaminating } material, (or if it lifts off metal oxide or other aspects of a scale) you } will not be doing this very non-destructively. One technique is to make } repetitive replicas on the same area, each additional one taking off more of } the contaminating material, in order to get to the bare metal surface. But } this might not be considered, by a court of law, to be nondestructive. And } if you are under an order to look at it nondestructively, this could result } in a problem with the court. } } When we have been faced with this dilemma, in order to first document what } was there originally, we like to use our SPI "Wet Replica" kit, it is a } silicone based system, and is described on our website. Certain dental } replication systems such as "Sil-Flow" might work as well, but in general, } for this kind of work, we believe our own kit is better. The silicone resin } in general won't remove surface contamination or scale. The drawback of this } technique is that after about 800X in the SEM you start to see "structure" } from the replication system itself. A "plus" however is that you can make a } "replica of the replica", doing your SEM viewing on a positive, which does } look just like the original inside surface. } } Once you have documented the state of the inside surface that way, you } should be able to get permission to try the cellulose acetate replication } method. There is additional information on the SPI Supplies website relative } to cellulose acetate replication methods. } } Disclaimer: SPI Supplies offers both cellulose acetate replicating tapes } and sheets as well as the silicone based SPI "Wet Replica Kit" for use in } electron microscopy applications. } } Chuck } } ============================================ } } Charles A. Garber, Ph. D. Ph: 1-610-436-5400 } President 1-800-2424-SPI } SPI SUPPLIES FAX: 1-610-436-5755 } PO BOX 656 e-mail:cgarber-at-2spi.com } West Chester, PA 19381-0656 USA } Cust.Service: spi2spi-at-2spi.com } } Look for us! } ######################## } WWW: http://www.spi.cc } ######################## } ============================================ } } } }
Hi, I made an embedding of the endodermis of an iris root with LR-Wihte and would like to make ultrathin sections and stain them. My problem is how to stain the sections and how to receive permantly stained sections. Is there anyone who has experiences with the staining of suberin in the 3rd endodermis? Please inform me about the handling. Thank you so much Dagmar Preusse
I run a 1830 Amray S.E.M. and work with failed aircraft components. From time to time I am asked to replicate a surface using the acetate tape & acetone method. The coating I use is a combination of Au and Pd. My problem is that when I have to stay in one particular area of the replica, the beam leaves a nice little square embedded into the surface of the replica. I try to rapidly focus in full field instead of partial field, and use between 6 - 10Kv. Does anyone have any advise for me on how to avoid the embedded square?
It is the total current that you put into the specimen that is causing the material to erode.
If you use a smaller spot size, smaller final aperture or smaller emission current you should be better off? I do not know how good your SEM is at low kV but if it was me I would use 2kV on this type of specimen; it will be very sensitive even though it is coated.
To run happily at 2kV you need a filament to cathode distance that will give you at least 30uA with your normal bias settings. If you do a good deal of work on this type of specimen I would be inclined to lift the anode by 5mm also. This will make the gun more efficient and make the job easier.
Good luck
Steve Chapman Protrain for Electron Microscopy Courses World Wide E-mail - protrain-at-emcourses.com Web Site - www.emcourses.com
I have been trying to develop a styrene replica fluid so I can use polorized light.So far the styrene clumps ups in little islands insted of making a nice film. I am using acetone as a solvent
Has anyone had any luck with this?
Gordon
Gordon Couger gcouger-at-couger.com
Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
} A colleague would like your responses to these questions. I believe that I
} say something on the first question about a year ago and apologize for
} asking again.
}
}
} } Here's what I/we need to know.
} }
} } 1) Are Microstar diamond knives equal to Diatome. We have an offer that
} } will save us about $1200 on three knives ( one 3mm ultra with 45 degree
} } angle, one 3mm ultra with 35 degree angle, one 10 mm histo knife).
} }
} }
} } 2) Is the power controller feature on the Pelco Microwave Tissue processor
} } (model #3451) worth it? (costs about $1300 additional compared to the
} } Pelco 3450 without the power controller). I plan to do immunolocalization
} } work, but am not sure this power controller feature is necessary or worth
} } the cost.
} }
} } Casey
} }
} }
} }
} Thanks for your time
} }
} Marty Reed
} Equipment Technician
} Biology Department
} Humboldt State University
} Arcata CA 95521
} 707-826-3234
} 707-826-3201 FAX
} mmr7001-at-axe.humboldt.edu
}
}
{/fontfamily} Cheers, Mark **************************************************** Mark A. Sanders University of Minnesota Program Director Twin Cities Campus Imaging Center St. Paul, MN 55108 23-35 Snyder Hall ph: 612-624-3454 1475 Gortner Ave. fax: 612-624-1799 http://biosci.umn.edu/imagingcenter President: Minnesota Microscopy Society http://resolution.umn.edu/MMS/
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------- Start of forwarded message -------
Rules, FAQ Docs - Microscopy ListServer To: NewSub-at-sparc5.Microscopy.Com } From: "Nestor J. Zaluzec" {zaluzec-at-sparc5.Microscopy.Com}
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To: NewSub-at-MSA.Microscopy.Com } From: "Nestor J. Zaluzec" {zaluzec-at-microscopy.com}
Here is a final reminder and IMPORTANT CORRECTION for the Call for Papers for Microscopy and Microanalysis 2000.
The title for symposium #05 in the Call for Papers was listed incorrectly. The correct title should be "Phase Transitions" (If you check the symposium titles on the electronic data submission forms it is listed correctly). X-ray microanalysis papers should be directed to symposia 25 and 26.
The deadline for papers is Tuesday February 15, which is almost upon us.
Stuart McKernan (Program Chair)
__________________ Stuart McKernan stuartm-at-tc.umn.edu Director Office:(612) 626-7594 IT Characterization Facility, University of Minnesota Desk:(612) 624-6009 100 Union Street S. E., Minneapolis, MN 55455 NEW-} Fax:(612) 625-5368
} X-Sender: mcintyre-at-optics.rochester.edu } Date: Fri, 11 Feb 2000 14:01:04 -0500 } To: Microscopy-at-sparc5.Microscopy.Com } From: Brian McIntyre {mcintyre-at-optics.rochester.edu} } Subject: metal pipe- inside surface } Errors-to: Microscopy-request-at-sparc5.Microscopy.Com } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
The best way to make replicas is to use dental casting compound.
It has the ability to replicate very fine detail and accepts even damp surfaces.
We used "Kerr Extrude Wash" (two part polyvinylsiloxane impression material) to replicate (damp) tuna fish skin.
Once cured, we made epoxy positive casts from the negative casts, then mounted, gold coated etc. for regular SEM.
Your local dental supply house will have this.
Kerr USA is at 28200 Wick Road, Romulus MI 48174-2600
Dr. Mel Dickson, Deputy Director, The Electron Microscope Unit, Adjunct Associate Professor, School of Microbiology & Immunology The University of New South Wales Sydney, Australia. Phone +612 9385 6383 Fax +612 9385 6400
True optical resolution 1600x3200. Dynamic range 3.3 Although it is not from a scientific point of view the authors claim that this is the best scanner for the price they've ever seen.
Good luck,
Rado
Disclaimer: I'm not related to Epson company in any way.
--------------------------------------------------------------------- Radostin Danev Laboratory of Ultrastructure Research National Institute for Physiological Sciences Myodaiji-cho, Okazaki 444-8585, JAPAN e-mail: rado-at-nips.ac.jp --------------------------------------------------------------------- ----- Original Message ----- } From: Malcolm Haswell {malcolm.haswell-at-sunderland.ac.uk} To: Microscopy (MSA) {microscopy-at-sparc5.microscopy.com} Sent: Thursday, February 10, 2000 8:26 PM
On Sat, 12 Feb 2000, Gordon Couger wrote:
} I have been trying to develop a styrene replica } fluid so I can use polarized light.So far the } styrene clumps up in little islands instead of } making a nice film. I am using acetone as } a solvent
Polystyrene is only partially soluble in acetone, so the low MW goes into solution while the high MW forms clumps or whatever. BUTANONE (MEK) should work fine, but you'll have to give it perhaps 4 times the drying time you'd need with acetone. For samples that are sensitive to polar solvents such as these, you can also use toluene, but that takes an even longer drying time.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
} I run a 1830 Amray S.E.M. and work with failed aircraft components. From } time to time I am asked to replicate a surface using the acetate tape & } acetone method. The coating I use is a combination of Au and Pd. My } problem is that when I have to stay in one particular area of the replica, } the beam leaves a nice little square embedded into the surface of the } replica. I try to rapidly focus in full field instead of partial field, and } use between 6 - 10Kv. Does anyone have any advise for me on how to avoid } the embedded square?
Polystyrene MIGHT be a better material to use than cellulose acetate, which tends to chemically self-destruct under electron beams: in fact, when there is acetate remaining on replicas for TEM I sometimes burn it off SLOWLY with the electron beam (too fast and you leave carbonaceous stuff). The aromatic nature of PS also allows it to take up more energy.
You could replicate with PS, as follows:
Make a solution of PS in butanone (MEK) in a glass Petri dish. Allow to evaporate slowly (if you can fume it off from a closed dish in an oven at about 50^C, that's better). This film can then either be cast onto the component with butanone, or you can melt-form it above the glass transition of PS, say about 150^C. Don't use PS windows directly from envelopes, because the films contain a lot of built-in-strain.
I'm not sure of it's performance in SEM, because I've used it for TEM replication of acetone-sensitive specimens.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
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So your companyÕs SEM operator just retired. You have used light microscopes for years and they have asked you to run the SEM also. So now what do you do? The SEM manuals are long gone and do you really want to crack open that 800 page SEM/x-ray microanalysis textbook?
DonÕt worry, consider taking Electron Microscopy and Microanalysis a one-day short course to be held on Sunday, March 12, at PITTCON 2000 in New Orleans. The course is an introduction to SEM, TEM and x-ray microanalysis and is designed for analytical chemists and others who need to use these techniques for industrial problem-solving and product research and development.
For further information and online registration visit www.pittcon.org (Course #2029) or contact me offline.
Mark S. Germani, Ph.D. MicroMaterials Research, Inc. 136 Shore Drive Suite 200 Burr Ridge, IL 60521
by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id IAA06667 for dist-Microscopy; Mon, 14 Feb 2000 08:52:52 -0600 (CST) Received: from no_more_spam.com (sparc5.microscopy.com [206.69.208.10]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id IAA06661 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Mon, 14 Feb 2000 08:51:54 -0600 (CST) Received: from mercury.uwe.ac.uk (mercury.uwe.ac.uk [164.11.132.23]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id IAA06651 for {microscopy-at-sparc5.Microscopy.Com} ; Mon, 14 Feb 2000 08:51:37 -0600 (CST) Received: from fas655.uwe.ac.uk ([164.11.205.145]) by mercury.uwe.ac.uk (2.0.4/SMS 2.0.4-devel) with SMTP id OAA04217; Mon, 14 Feb 2000 14:46:35 GMT
Electron Optical Services Ltd. make a replacement control box. Their address is given below.
Dave
52 Higher Road, Urmston, Manchester, M41 9AP England UK
Dave On Fri, 11 Feb 2000 19:12:34 -0600 "A.P. Alves de Matos" {apmatos-at-ip.pt} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I am in desperate need of a control unit for a LKB ultratome III, since } mine is beyond repair. } I would appreciate if anyone coud sell me an unused unit from old } apparatus, even if it does not work, I might } be able to repair mine with the pieces. } Please reply to the e-mail below } } Thanks } } Dr. A.P. Alves de Matos } Dental Medical School } Lisbon University } apmatos-at-ip.pt } } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
Dear Microscopy Listserver members, can someone provide me: - a detailed protocol for embedding Chlamydomonas into the Lowicryl Resin K4M (fixation media, desiccation solutions, times and concentrations that are suitable for Chlamydomonas) - a fixation protocol for cryosectioning of Chlamydomonas
Thank you very much, Yvonne Nemcova
-- ***************************************** Mgr. Yvonne Nemcova Department of Botany, Charles University, Prague Benatska 2. 128 01 Praha 2, Czech Republic tel: 00-420-2-21953127, fax: 00-420-2-21953125 e-mail address: ynemcova-at-natur.cuni.cz *****************************************
Thank you for the information on the MSA/MAS format that you sent me some weeks ago.
The MSA/MAS spectra file format is now implemented in the EELS and XAS database, therefore the spectra can be downloaded in this format. Please let me know if you have any other problems. Any more suggestions ?
We are having an Edwards Vacuum Evaporator model E12E2 with a control unit EVM052 in excess. This instrument has not been used for several year and it may need some maintanance work. If you are interested please contact Tea Meulia (meulia.1-at-osu.edu) or Pat Ashbaugh (ashbaugh.11-at-osu.edu).
Tea Meulia
Tea Meulia Research Scientist and Head Molecular and Cellular Imaging Center OSU/OARDC 1680 Madison Ave. Wooster OH 44691
Thanks to all that responded. I have a lot better handle on how to do it. I will report back my results.
Thanks to all for providing a great source of infomation.
Gordon
Gordon Couger gcouger-at-couger.com
Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00 ----- Original Message ----- } From: "Robert H. Olley" {r.h.olley-at-reading.ac.uk} } On Sat, 12 Feb 2000, Gordon Couger wrote: } } } I have been trying to develop a styrene replica } } fluid so I can use polarized light.So far the } } styrene clumps up in little islands instead of } } making a nice film. I am using acetone as } } a solvent } } Polystyrene is only partially soluble in acetone, so the low MW goes into } solution while the high MW forms clumps or whatever. BUTANONE (MEK) } should work fine, but you'll have to give it perhaps 4 times the drying } time you'd need with acetone. For samples that are sensitive to polar } solvents such as these, you can also use toluene, but that takes an even } longer drying time. }
Epson has a new ink jet printer out-Stylus pro 5000-that is expensive but they claim it is much better than the much cheaper models many of us know and love. Would anyone know whether this is true of just hype-hard to tell from the published specs...? Thanks...Tom Reese
Dear Gordon, Try xylene. It works quite nicely as a mounting medium of a differenting refractive index from that of say, canada balsam or Permount. Email me if you have questions. Good luck! Cheers, Ken
------------- Ken Tiekotter Dept. of Biology The Univerisity of Portland 5000 N. Willamette Blvd. Portland, OR 97303
On Sat, 12 Feb 2000, Gordon Couger wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I have been trying to develop a styrene replica } fluid so I can use polorized light.So far the } styrene clumps ups in little islands insted of } making a nice film. I am using acetone as } a solvent } } Has anyone had any luck with this? } } Gordon } } Gordon Couger gcouger-at-couger.com } } Stillwater, OK www.couger.com/gcouger } 405 624-2855 GMT -6:00 } } } }
You wrote "I am working with archival specimens of formalin-fixed paraffin embedded tissues and I am getting tremendous autofluorescence. I looked at tissue sections after the sections have been de-waxed using either FITC or rhodamine filter sets and red cells and connective tissues are brightly fluorescent. Is there a way of suppressing the autofluorescence and still retain reactivity of tissue to antibodies? I would appreciate any comments or suggestions."
It is an unfortunate fact that most fixatives cause high autofluorescence making the studies with conventional fluorescence microscope hard. The disturbance of autofluorescence can be avoided by using time-resolved fluorescence microscope and special labels (See the following articles: VŠisŠlŠ M, SevŽus L, Kuusisto A, Harju R, and Soini E: Time- resolved fluorescence microscopy: Elimination of autofluorescence in tissue specimens for Image cytometry with fluorescent labelled probes. Micron and Microscopica Acta 21; 3, (1990).
SevŽus L, Kuusisto A, VŠisŠlŠ M, HemmilŠ I., and Soini E: Lanthanide chelates as labels in in situ hybridization and immunohistochemistry, Micron and Microscopica Acta 21; 3, (1990).
SevŽus L, VŠisŠlŠ M, SyrjŠnen S, Sandberg M, Kuusisto A, Harju R, Salo J, HemmilŠ I, Kojola H, and Soini E: Time-Resolved Fluorescence Imaging of Europium Chelate Label in Immunohistochemistry and In Situ Hybridization, Cytometry 13; 329, (1992).
SevŽus L, Kuusisto A, VŠisŠlŠ M, HemmilŠ I., Kojola H., Roomans G. M. and Soini E.: Use of Fluorescent Europium Chelates as Labels in Microscopy Allows Glutaraldefyde Fixation and Permanent Mounting and Leads to Reduced Autofluorescence and Good Long-Time Stability, Microscopy Research and Technique 27; 00, (1994).
I saw the Epson web spec and it looks quite impressive - especially as they seem to be quoting speeds for real picture printing, not 5% coverage or text.
My only concern is that Epson now market two printers with "10 years light fastness" (the Epson Stylus photo 870 - A4 and 1270 A3) so you have to wonder which is better for e.m. prints.
Malcolm Haswell Electron Microscopy School of Sciences Fleming Building University of Sunderland SUNDERLAND SR1 3SD Tyne and Wear UK
Tel (0191) 515 2872 e-mail: malcolm.haswell-at-sunderland.ac.uk
Tom Reese wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Epson has a new ink jet printer out-Stylus pro 5000-that is expensive } but they claim it is much better than the much cheaper models many of } us know and love. Would anyone know whether this is true of just } hype-hard to tell from the published specs...? Thanks...Tom Reese
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Dear Carol,
We were looking up to 70 mm deep inside cylinder bores and engine blocks several times during the last months with our large-chamber SEM. Roughness was typically very small (less than 10 µm), but for small diameter tubes the positioning system needs large travels, tilts, and rotary axes to inspect every interesting area.
Disclaimer: Visitec manufactures the large-chamber SEM.
Best regards Peter ___________________________ Dr. Peter Marienhoff VisiTec Microtechnik GmbH Karl-Marx-Str. 14 D-23936 Grevesmuehlen Germany
"Orth99-at-aol.com"-at-sparc5.Microscopy.Com wrote: } How often does this community need to look at the inside of metal pipes or } pipes in general? } } And, what is the typical surface roughness you are dealing with, i.e. would } an instrument with 100 um to 200 um of Z range be able to handle it? } } And, what is the typical x,y range, i.e. would an instrument with 2 mm image } field be acceptable? } } Just wondering. Thanks for any input you are willing to share. } } } Carol Rabke, Ph.D. } Surface Metrology Marketing & Applications Consultant } 716-425-0976 }
The Epson 5000 is a printer/proofer using 6 color inkjet technology. The 6 color technology expands the available color gamut significantly. It is available both with or without a Fiery RIP. The Fiery RIP is used for Postscript processing of complex files from Quark, Pagemaker, Illustrator,etc. Without the Fiery box, an Epson software RIP is included. Print quality is excellent although without the Fiery RIP, printing times can be long,especially for 11x17 or larger prints. For more information see: http://prographics.epson.com/products/stypro5000/index.html
Canon also has an excellent printer, the BJC-8500 that also uses 6 color technology. The BJC-8500 also handles up to 13x19 paper with excellent print quality. Canon utilizes a technology that optimizes ink drying and water resistance. For more information see: http://www.bjc8500.com/index2.html
Neither of these printers are speed demons in terms of page throughput, but quality is excellent. I will be happy to answer any questions anyone may have regarding output devices.
George Laing National Graphic Supply
-----Original Message----- } From: Tom Reese [mailto:treese-at-mbl.edu] Sent: Monday, February 14, 2000 6:57 PM To: Microscopy-at-sparc5.microscopy.com
Epson has a new ink jet printer out-Stylus pro 5000-that is expensive but they claim it is much better than the much cheaper models many of us know and love. Would anyone know whether this is true of just hype-hard to tell from the published specs...? Thanks...Tom Reese
} -----Original Message----- } From: Wright, Cheryl W [mailto:CWright-at-Sikorsky.com] } Sent: Saturday, February 12, 2000 7:25 AM } To: 'Microscopy-at-MSA.Microscopy.Com' } Subject: Photography of Coated Replicas } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListser} ver/FAQ.html } } } -------------------------------------------------------------- } ---------. } } } I run a 1830 Amray S.E.M. and work with failed aircraft } components. From } time to time I am asked to replicate a surface using the } acetate tape & } acetone method. The coating I use is a combination of Au and Pd. My } problem is that when I have to stay in one particular area of } the replica, } the beam leaves a nice little square embedded into the surface of the } replica. I try to rapidly focus in full field instead of } partial field, and } use between 6 - 10Kv. Does anyone have any advise for me on } how to avoid } the embedded square? } } Thanks, Cheryl }
I am a technical recruiter. Currently, I am working on a search for one of my Northeast clients for a Senior Metallographic Technician:
Will perform metallographic evaluation of coatings including work with SEM (Scanning Electron Microscopy) and X-ray. Will be responsible for the metallurgy laboratory, including organization of work, and operation and calibration of equipment. Will write technical and engineering reports. Will prepare projects for the metallurgy laboratory. Will maintain communication with customers.
AAS/BS plus fours years experience in a metallography laboratory.
Resumes should be sent by email, mail or fax. For best results, please send emails as attached Word.docs - I have Word 97.
Pearl Martin Image Associates Inc. 5254 Merrick Road Massapequa, NY 11758 Phone (516)798-3993 Fax (516)797-8703 Email: pearl-at-jobspot.com
With regard to diamond knives, I think it depends on what you're cutting and what quality of section you require.
We cut steel and cross sections of coatings (some very hard and brittle) on steel, and the only knife that approaches satisfactory result is Diatome 35 degree, sharpened to extra fine edge. We traded our big old Microstar knife for a 2nd Diatome, because it couldn't make good, electron transparent sections.
On the other hand, if your material cuts as well with a glass knife as with a diamond but you don't want to bother making your own glass knives, then go ahead and save some dollars.
(In my view, the same reasoning applies to the brand of ultramicrotome to use. My company is very frugal, and there is always strong pressure to save a dollar here and there, but we found that only Reichert/Leica can consistently produce good samples in our demanding application; and the higher initial cost has been more than offset by savings in time and effort, and by the quality of results. (I.e., often the ability to just get any kind of a result).)
Valdemar Furdanowicz, Ph.D. Research Labs - G165 Bethlehem Steel Bethlehem PA 18016
No interest in Microstar, Diatome, Leica; and this not reflect an opinion or position of my employer....etc.
-----Original Message----- From: Chuck Butterick [mailto:cbutte-at-ameripol.com] Sent: Friday, February 11, 2000 5:11 PM To: Microscopy-at-sparc5.Microscopy.Com; mmr7001-at-humboldt.edu Subject: Re: questions for MSA members
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In regards to the diamond knives, both companies make an excellent product and both provide good service. Because of price, the last 6 knives I've bought have been from Microstar (two well-used but dull Diatome knives have been traded during these transactions) . All six knives have been more than suitable for the intended purpose. I have no commercial interest in either company.
Chuck Butterick Engineered Carbons, Inc.
______________________________ Reply Separator _________________________________ Subject: questions for MSA members Author: Marty Reed {mmr7001-at-humboldt.edu} at INTERNET-MAIL Date: 1/4/80 8:23 AM
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A colleague would like your responses to these questions. I believe that I say something on the first question about a year ago and apologize for asking again.
} Here's what I/we need to know. } } 1) Are Microstar diamond knives equal to Diatome. We have an offer that } will save us about $1200 on three knives ( one 3mm ultra with 45 degree } angle, one 3mm ultra with 35 degree angle, one 10 mm histo knife). } } } 2) Is the power controller feature on the Pelco Microwave Tissue processor } (model #3451) worth it? (costs about $1300 additional compared to the } Pelco 3450 without the power controller). I plan to do immunolocalization } work, but am not sure this power controller feature is necessary or worth } the cost. } } Casey } } } Thanks for your time } Marty Reed Equipment Technician Biology Department Humboldt State University Arcata CA 95521 707-826-3234 707-826-3201 FAX mmr7001-at-axe.humboldt.edu
We are considering the purchase of a separate deconvolution system to augment our imaging lab's confocal system Any users or vendors with information about such systems, please contact me offline.
thanks in advance
steve
Dr. Steven Barlow, Associate Director EM Facility/Biology Department San Diego State University 5500 Campanile Drive San Diego CA 92182-4614 phone: (619)594-4523 fax: (619) 594-5676 email: sbarlow-at-sunstroke.sdsu.edu website: http://www.sci.sdsu.edu/emfacility/
Chairman, Educational Outreach subcommittee promoting access to microscopes Microscopy Society of America http://www.msa.microscopy.com/
We are in the market for a new oven for curing blocks.... Any suggestions? This lab uses as a standard Eponate 12 as the epoxy resin of choice.... The oven we currently have after being fixed several times does not see to keep a constant temp.....
One of the members of this listserver group just asked me privately about the fact that he had noticed that the reading of the thermocouple gauge on his vacuum evaporator changed markedly when he jiggled the wires running from the gauge tube to the gauge control unit. This is a matter that is discussed in some detail on page 87 of my book 'Vacuum Methods in Electron Microscopy' (see http://www.2spi.com/catalog/books/book48.html for a description)
Actually, this is a rather common problem with thermocouple gauges which all people using them should be aware of. It arises because the sensing element in a thermocouple gauge is an arrangement of very fine wires that form one or more thermocouple junctions. The output of these gauges is thus the electrical potential produced by these junctions that is of the order of only 10 or 15 millivolts DC. As a consequence, the signal detected by the gauge controller can be strongly influenced by minor changes the resistance of the electrical lines connecting the gauge to the controller. Typically, unplugging one of the connectors in this line and reconnecting it can change the resistance of the line sufficiently to cause a significant change in the reading produced by the controller.
For this reason, considerable care must be exercised in installing and maintaining thermocouple gauges if acceptable readings are to be obtained. As a minimum, the same cables used in calibrating a gauge must subsequently be used when it is put into operation, and the pins and sockets of the plugs involved in the circuit must be kept clean and firmly connected.
Because of the low DC voltages that must be detected, the meters used in the gauge circuit must be rather delicate and sensitive. We have encountered situations when a static electric charge on the plastic window of a meter affected the movement of the needle inside the meter enough to prevent an accurate response. Coating the meter window with an anti-static solution cured this situation.
As discussed in my book, Thermocouple gauges are rugged, inexpensive, and very convenient for use in many applications, but they must be handled with care if they are to give acceptable service.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
A graduate student is getting ready for a rather large scale preparation of rat brains for TEM using vascular perfusion. Luckily (since I'm a botanist) her major professor has had experience with the procedure. He doesn't know for sure whether he used EM or Biological grade glutaraldehyde previously, but considering the large volume that will be required, and the cost difference, I'm hoping that the Biological grade will suffice.
I would appreciate advice from those of you that have experience with this method of fixation.
Thank you in advance for any information.
Heather Owen
Heather A. Owen, Director Electron Microscope Laboratory Department of Biological Sciences University of Wisconsin - Milwaukee (414)229-6816
I have attempted to purchase an epifluorescence nose from 'Carl Zeiss' to fit the old "Universal" model scopes, but was dismayed to find out that it is no longer available. If anyone has one that is not being used and wishes to sell it, I would be very interested.
Thanks
JOSE ULLAN SERRANO, Ph.D., M.D. ÑÑÑÑÑÑÑÑÑÑÑÑÑÑÑÑ E-mail: jullan-at-mixcoac.upmx.mx
Departamento de ANATOMIA Escuela de MEDICINA Universidad Panamericana c/ Donatello, 59 Col. Insurgentes-Mixcoac 03920 MEXICO, D.F. ÑÑÑÑÑÑÑÑÑÑÑÑÑÑÑ http://www.mixcoac.upmx.mx Telef. 55 98 33 02 / 55 63 13 43 FAX: 56 11 35 85
Thermocouple gauges are more like gross indicators of vacuum. As a resistive bridge, they are driven by constant current sources and the bridge or sensor output is fed to a differential amplifier. This amplifier has a gain and zero adjust potentiometer. If either of these are dirty or defective, readings will be off. But if a thermocouple sensor is exhibiting erratic readings, and it uses the standard vacuum tube socket connector, I'd replace the detector right off the bat. They do eventually fail. A $25 throw away isn't worth the hassle.
Most instruments, like SEMs, use the TC gauge to indicate that the mechanical pump is working. Once the turbo pump spins up to } 80% RPM, the TC gauge is rather useless. Better readings are from cold cathode gauges....which tend not to fire very regularly. Hence, the optimum readings are from the ion pump current.
At 01:11 PM 2/15/00 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We perfuse 4% formaldehyde in 1x PBS only, than prepared the brain's slices .3-.4 mm thick and fix them in a mixture 3-4% formaldehyde 2% GA (all - EM grade) in 1x PBS. If you would like to add GA into perfusion solution - it is only 0.1% - not a big deal. From another hand, I don't think Biological grade GA will affects fixation quality. But, who knows, every time it is a "game": if fixation is good - no problem, if - no, we will start to think what's going on and "biological" grade may be a "case"... To avoid such troubles I always used fresh GA and formaldehyde from non-opened ampoules.
Good luck, Sergey
} Date: Tue, 15 Feb 2000 18:02:57 -0600 (CST) } From: Heather A Owen {owenha-at-csd.uwm.edu} } Subject: TEM - glutaraldehyde for perfusion } To: MSA Listserver {Microscopy-at-sparc5.Microscopy.Com} } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
As for my knowledge (mostly from physic's course at 7th grade Russian school) the thermocouple gauge has two "thermocouples": the probe itself and reference thermocouple inside of gauge (separate "references" for each type of probe, I believe). All together they generate enough response to be easily measured. I agree that the major problem in your case is a bad electrical contact in the connectors or even broken wire (it is well possible, because some material uses for thermocouples are very fragile). In my point of view, thermocouple gauges is very reliably and most accurate devices and they do not need any special care.
I am using BARNANT-100 Thermocouple Gauge and REX-P96 Thermocouple Controller with type "T" thermocouple sensors (copper/constantan, I believe, but not sure) for many years without any problem. The sensor goes into the vacuum chamber through regular 8-pin feed-thought. I was using same as thermocouple material for screws to connect thermocouple's wires to the feed-though wiring. For some reason, soldering does not work. I took that screws from disassembled thermocouple connectors (looks like small plugs) most suppliers sells for thermocouple probes (a dollar each). That connectors should correspond to the thermocouple probe you are using. Usually those connectors are color-coded. For instance connectors for my type "T" probe was coded in blue. BARNANT-100 Thermocouple Gauge and REX-P96 Controller performs self-calibration test every time you shut it ON or disconnect/connect probe. During that calibration they "compensate" some electrical abnormalities in the circuit like difference in the probe length or as in my case the current deviation on the enter and end of the feed-through. In theory, the "break" in thermocouple wiring on feed-through should affect the gauge accuracy. In practice, I did not find a difference between "solid" and with-feed-though probe up to 0.1oC accuracy (which is sufficient for my needs). REX-P96 Thermocouple Controller is just amazing. You may program complicated profiles for cooling/heating cycles and it holds temperature pretty well. It has special auto-calibration function to adjust heat rate depending from parameters of your system. In practice it mean that your sample will not overheated because of heater inertia. I am using that Controller in my high-resolution shadowing device to control temperature from -150 to +50oC.
I have no interest in described products (only happy user) mostly because I even don't know the names of real manufacturers. I remember it was distributed by Cole-Parmer.
Sergey
} Date: Tue, 15 Feb 2000 15:11:33 -0400 } From: Wil Bigelow {bigelow-at-engin.umich.edu} } Subject: Thermocouple gauges } X-Sender: bigelow-at-srvr5.engin.umich.edu } To: Microscopy Listserver {microscopy-at-sparc5.Microscopy.Com} } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
MEK worked great. I broke up the clear plastic from a CDROM case and made a thick solution by adding MEK. cleaned a slide and spread a drop of the solution on the slide and put a celery leaf on it I covered the whole thing with saran warp and gently pressed another slide on it to get good contact. Then removed the slide and Saran wrap.
After drying about 30 minutes I pealed the leaf off and took a look under a Nomarski phase scope. It was the best image I have seen on the scope. The Nomarski colors were extremely strong and I could get three bands of color in some features/artifacts. The effect of the styrene on the polarized light were not as strong as I had hoped for but they do increase the contrast and Nomarski colors. It will make lovely photos. Now if my color CCD camera would just get here. Monochrome just won't do it justice.
I has been a long time since I looked at a leaf but the stoma showed up just like the books show and I remember. Individual cells were very clearly visible.
The film is extremely fragile and probably needs a little plastizer added and probably go to a go to a multi coat system if the film needs to be pealed from a surface and mounted. with the first coats being thin with little or no plastizier and the following coats being thicker and carrying more plastisizer.
My thanks to all the helped on this. I will probably try other solvents but MEK does not set off my asthma like xylene and toluene do. So I will probably put up with the slower drying times.
I was very impressed with the detail that was replicated.
Thanks Gordon
Gordon Couger gcouger-at-couger.com
Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
} From: Robert H. Olley [mailto:r.h.olley-at-reading.ac.uk] } On Sat, 12 Feb 2000, Gordon Couger wrote: } } } I have been trying to develop a styrene replica } } fluid so I can use polarized light.So far the } } styrene clumps up in little islands instead of } } making a nice film. I am using acetone as } } a solvent } } Polystyrene is only partially soluble in acetone, so the low MW goes into } solution while the high MW forms clumps or whatever. BUTANONE (MEK) } should work fine, but you'll have to give it perhaps 4 times the drying } time you'd need with acetone. For samples that are sensitive to polar } solvents such as these, you can also use toluene, but that takes an even } longer drying time.
Dear Josi, Try contacting John Oren at Vermont Optechs. I cannot tell you immediately what his email or web site address is but his mail address is: P.O. Box 69, Charlotte, VT 05445: Tel. (802) 425-2040; FAX. (802) 425-2074. Good luck! -Ken
On Tue, 15 Feb 2000, Jos[ISO-8859-1] é Ullán Serrano wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Dear Colleagues: } } I have attempted to purchase an epifluorescence nose from 'Carl Zeiss' } to fit the old "Universal" model scopes, but was dismayed to find out that } it is no longer available. } If anyone has one that is not being used and wishes to sell it, I would be } very interested. } } Thanks } } JOSE ULLAN SERRANO, Ph.D., M.D. } ‹‹‹‹‹‹‹‹‹‹‹‹‹‹‹‹ } E-mail: jullan-at-mixcoac.upmx.mx } } Departamento de ANATOMIA } Escuela de MEDICINA } Universidad Panamericana } c/ Donatello, 59 } Col. Insurgentes-Mixcoac } 03920 MEXICO, D.F. } ‹‹‹‹‹‹‹‹‹‹‹‹‹‹‹ } http://www.mixcoac.upmx.mx } Telef. 55 98 33 02 / 55 63 13 43 } FAX: 56 11 35 85 } }
I have heard recently that Diatome invent some coating of their diamond knives that significantly improve the quality of cryosections (they appear more smooth, less compressed, practically without folds). Is it true or just usual story to increase sales?
Sincerely, Dr. Alexander A. Mironov Jr., MD, PhD Unit of Morphology Dept. of Cell Biology and Oncology Consorzio Mario Negri Sud Via Nazionale, S.Maria Imbaro (Ch) 66030 Italy
Speaking of thermocouples, I need to replace (probably) the T1 and T2 thermocouples in our OLD Denton evaporator. Where, other than Denton, can I look for these? I also need service work done on our old Ultracut (the model before the "E" series). I refuse to work with Leica. Is there anyone in the "real" midwest area?
I've also seen this type of behavior caused by a "cold" solder joint on one or more of the wires in the guage connector socket. Re-flowing the joints did wonders for stability.
============================================================ Bede Willenbring Phone: (651)236-5470 H.B. Fuller Company FAX: (651)236-5430 Research Chemist E-mail: Bede.Willenbring-at-HBFuller.com P.O. Box 64683 St. Paul, MN 55164-0683
} } } Wil Bigelow {bigelow-at-engin.umich.edu} 02/15 1:11 PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
One of the members of this listserver group just asked me privately about the fact that he had noticed that the reading of the thermocouple gauge on his vacuum evaporator changed markedly when he jiggled the wires running from the gauge tube to the gauge control unit. This is a matter that is discussed in some detail on page 87 of my book 'Vacuum Methods in Electron Microscopy' (see http://www.2spi.com/catalog/books/book48.html for a description)
Actually, this is a rather common problem with thermocouple gauges which all people using them should be aware of. It arises because the sensing element in a thermocouple gauge is an arrangement of very fine wires that form one or more thermocouple junctions. The output of these gauges is thus the electrical potential produced by these junctions that is of the order of only 10 or 15 millivolts DC. As a consequence, the signal detected by the gauge controller can be strongly influenced by minor changes the resistance of the electrical lines connecting the gauge to the controller. Typically, unplugging one of the connectors in this line and reconnecting it can change the resistance of the line sufficiently to cause a significant change in the reading produced by the controller.
For this reason, considerable care must be exercised in installing and maintaining thermocouple gauges if acceptable readings are to be obtained. As a minimum, the same cables used in calibrating a gauge must subsequently be used when it is put into operation, and the pins and sockets of the plugs involved in the circuit must be kept clean and firmly connected.
Because of the low DC voltages that must be detected, the meters used in the gauge circuit must be rather delicate and sensitive. We have encountered situations when a static electric charge on the plastic window of a meter affected the movement of the needle inside the meter enough to prevent an accurate response. Coating the meter window with an anti-static solution cured this situation.
As discussed in my book, Thermocouple gauges are rugged, inexpensive, and very convenient for use in many applications, but they must be handled with care if they are to give acceptable service.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
} Heather A Owen wrote: } } } Hello everyone, } } } } A graduate student is getting ready for a rather large scale preparation } } of rat brains for TEM using vascular perfusion. Luckily (since I'm a } } botanist) her major professor has had experience with the procedure. He } } doesn't know for sure whether he used EM or Biological grade } } glutaraldehyde previously, but considering the large volume that will be } } required, and the cost difference, I'm hoping that the Biological grade } } will suffice. } } } } Heather, } } It's been a while but, back when I did such things, I had bouts of } non-uniform fixation when using biological grade glutaraldehyde solutions } even in plant tissues. However, the solutions had been kept much longer than } I'd now use and there are other preparation technique changes that confound } the determination of causality. In the cases where I've perfused small } animals, I've used EM grade GA since, as expensive as it is, it's cheaper } than repeating the experiment. } } However, I did a full systemic perfusion. One random thought ( I haven't } tried it): What's the vascular volume of a rat brain? If you just need just } the brain, I'd think a little creative clamping (really small hemostats?) in } the thorax should let you reduce the volume needed to make even a large } experiment affordable with "the good stuff". } } cheers and good luck, } John }
Heather:
I agree with John on both counts. We clamp the aorta (and anything eles that gets in the way) high in the abdomen to avoid perfusing organs we are not interested in. Saves time and money. Also we tilt the animal once perfusion is underway so that the head is lower.
Geoff -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
I have had a lot of years' experience doing vascular perfusion of the CNS of rodents (rat and mouse). We have used the EM Sciences Biological Grade glutaraldehyde, 50%, as our primary source for perfusion without problems related to fixation. However, we have not used straight glutaraldehyde, but found that 2% paraformaldehyde + 1% glutaraldehyde in 0.1M cacodylate buffer at pH 7.0 to 7.2 gave consistent results with excellent preservation of structures, morphology and cytoplasmic and nuclear components. We have used cacodylate primarily because we use uranyl acetate stain either in the dehydration/processing protocol and/or on the thin sections. Use of phosphate buffers invariably results in precipitate formation, even when used only on sections. Another reason for the paraform/glut/cacodylate combination is that we have studied the leakage of tracers (e.g. HRP, microperoxidase) across the blood brain barrier, and had to be able to move into Tris buffer for DAB reactions, and phosphate buffer gave consistently poorer results.
Hope this helps.
Roger Moretz Dept. of Toxicology BI Pharmaceuticals, Inc.
I have no personal or financial interest in EM Sciences--just a satisfied user.
On Tue, 15 Feb 2000 18:02:57 -0600 (CST), Heather A Owen wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello everyone, } } A graduate student is getting ready for a rather large scale preparation } of rat brains for TEM using vascular perfusion. Luckily (since I'm a } botanist) her major professor has had experience with the procedure. He } doesn't know for sure whether he used EM or Biological grade } glutaraldehyde previously, but considering the large volume that will be } required, and the cost difference, I'm hoping that the Biological grade } will suffice. } } I would appreciate advice from those of you that have experience with this } method of fixation. } } Thank you in advance for any information. } } Heather Owen } } Heather A. Owen, Director } Electron Microscope Laboratory } Department of Biological Sciences } University of Wisconsin - Milwaukee } (414)229-6816 } } }
Roger C Moretz, Ph.D.
_______________________________________________________ Get 100% FREE Internet Access powered by Excite Visit http://freeworld.excite.com
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello everyone, } } A graduate student is getting ready for a rather large scale preparation } of rat brains for TEM using vascular perfusion. Luckily (since I'm a } botanist) her major professor has had experience with the procedure. He } doesn't know for sure whether he used EM or Biological grade } glutaraldehyde previously, but considering the large volume that will be } required, and the cost difference, I'm hoping that the Biological grade } will suffice. } } I would appreciate advice from those of you that have experience with this } method of fixation. } } Thank you in advance for any information. } } Heather Owen } } Heather A. Owen, Director } Electron Microscope Laboratory } Department of Biological Sciences } University of Wisconsin - Milwaukee } (414)229-6816
Heather,
It's been a while but, back when I did such things, I had bouts of non-uniform fixation when using biological grade glutaraldehyde solutions even in plant tissues. However, the solutions had been kept much longer than I'd now use and there are other preparation technique changes that confound the determination of causality. In the cases where I've perfused small animals, I've used EM grade GA since, as expensive as it is, it's cheaper than repeating the experiment.
However, I did a full systemic perfusion. One random thought ( I haven't tried it): What's the vascular volume of a rat brain? If you just need just the brain, I'd think a little creative clamping (really small hemostats?) in the thorax should let you reduce the volume needed to make even a large experiment affordable with "the good stuff".
cheers and good luck, John
John Heckman MSM Department Michigan State University
We use Biological grade to perfuse rats (whole body, we haven't tried clamping bits off, though that might be a good idea since we're interested in just the head). After the perfusion is finished, we remove the interesting bit, which is huge by EM standards, and fix some more in Karnowski's. Then we go on to osmium, and continue processing. We get acceptable EM pictures at high magnification, even though our blocks are so large and processing times are therefore extended. So I can recommend Biological grade glutaraldehyde. Hope this helps.
Lesley Weston.
On Tue, 15 Feb 2000, Heather A Owen wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello everyone, } } A graduate student is getting ready for a rather large scale preparation } of rat brains for TEM using vascular perfusion. Luckily (since I'm a } botanist) her major professor has had experience with the procedure. He } doesn't know for sure whether he used EM or Biological grade } glutaraldehyde previously, but considering the large volume that will be } required, and the cost difference, I'm hoping that the Biological grade } will suffice. } } I would appreciate advice from those of you that have experience with this } method of fixation. } } Thank you in advance for any information. } } Heather Owen } } Heather A. Owen, Director } Electron Microscope Laboratory } Department of Biological Sciences } University of Wisconsin - Milwaukee } (414)229-6816 } } } }
} From: RLevenson-at-cri-inc.com Subject: RE: autofluorescence quenching: spectral imaging can help
You asked about strategies for reducing autofluorescence.
In addition to various strategies which can include using near-IR dyes like Cy5.5 and exciting far in the red, you can also treat tissues with sodium borohydride or toluidine blue (these latter observations I pass on from others--have not tried them myself).
I have recently had very promising results using spectral imaging to discriminate autofluorescence from desired fluorescence. [Caution: I am describing a product sold by my company]. CRI makes a liquid crystal tunable filter (LCTF) that can be used to collect a stack of images at different wavelengths. This yields a spectrum at every pixel of an image. (A product with similar capabilities is sold by Applied Spectral Imaging, but uses a different technology.) CRI's new LCTF, the FluoroSpec, uses special optics to combine polarization states; along with other improvements, it has more than twice the light throughput compared to previous models.
The sample was formalin-fixed brain probed with a Cy2-labeled antibody to a low-abundance protein. By eye it was very difficult to distinguish the Cy2 signal from the bright green to green-orange autofluorescence. Using a spectral data set (15 images from 510 nm to 650 nm at 10 nm intervals) and linear combination algorithms for pixel-unmixing, it was possible to completely separate the Cy2 and autofluorescence images.
I would be glad to share these images with any interested parties.
Richard
Richard Levenson, MD Technical Director, Biomedical Systems Cambridge Research & Instrumentation (CRI), Inc. 80 Ashford St. Boston, MA 02134
I just purchased one for printing images, x-ray maps, etc. obtained from my new JEOL 8900 microprobe. Previous to this I was utilizing an Epson Photo 700 printer with my older Cameca MBX probe. I was very pleased with the 700 and for a few hundred dollars sure was worth it! Not a fast printer but gave high quality photo-type prints. After reading the 5000 specs I decided to get one for the new probe. In a "nutshell" here are my observations:
I purchased just the 5000 printer (not the Fiery RIP server that interfaces to it for network printing). Cost: around $3000. for the 5000 only. The 5000 is a much more robust printer than the 700 type Epson models. It can print up to 13"X19" formats. It is faster than the 700 series and the ink cartridges and paper capacity are much greater.
To realistically compare the printers I ran a speed and print quality comparison between my two models. I printed a full 8 1/2" X 11" full color bleed consisting of 3 x-ray maps, a BSE image and an EDS spectrum, along with some text. Printing on the same PC, under the same conditions the print quality is about the same. (A little disappointing here). However the print speed was a full 4 times faster on the 5000 as compared to the 700. Actual print time with the 5000 was about 2.5 min. at 1440 DPI. I found an excellent web site which compares various Epson Photo printers, including the 5000 and 700 series, and demonstrates print quality issues quite vividly:
http://www.tssphoto.com/sp/dg/news/dot_comp.html
All in all I believe the 5000 is a much higher caliber printer than the 700 series, but unfortunately the print quality is about the same.
One final note: the paper makes ALL the difference in the world with this class of printers. Email me if you need info and suggestions on paper issues...
Chuck Burilla United Technologies Research Center 400 Silver Lane East Hartford, CT 06108
(860) 610-7388 burilact-at-utrc.utc.com
-----Original Message----- } From: Malcolm Haswell [mailto:malcolm.haswell-at-sunderland.ac.uk] Sent: Tuesday, February 15, 2000 9:03 AM To: Microscopy (MSA)
I saw the Epson web spec and it looks quite impressive - especially as they seem to be quoting speeds for real picture printing, not 5% coverage or text.
My only concern is that Epson now market two printers with "10 years light fastness" (the Epson Stylus photo 870 - A4 and 1270 A3) so you have to wonder which is better for e.m. prints.
Malcolm Haswell Electron Microscopy School of Sciences Fleming Building University of Sunderland SUNDERLAND SR1 3SD Tyne and Wear UK
Tel (0191) 515 2872 e-mail: malcolm.haswell-at-sunderland.ac.uk
Tom Reese wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Epson has a new ink jet printer out-Stylus pro 5000-that is expensive } but they claim it is much better than the much cheaper models many of } us know and love. Would anyone know whether this is true of just } hype-hard to tell from the published specs...? Thanks...Tom Reese
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Hello,
Does anyone know the name of a supplier of wall posters suitable for an EM lab. I am thinking of cut-out diagrams of TEM and SEM, history of EM (I think JEOL once had one like that), specimen preparation etc. Regards,
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm
by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) id UAA14463 for dist-Microscopy; Wed, 16 Feb 2000 20:00:35 -0600 (CST) Received: from no_more_spam.com (sparc5.microscopy.com [206.69.208.10]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with SMTP id TAA14458 for "MicroscopyFilteredEmail-at-msa.microscopy.com"; Wed, 16 Feb 2000 19:59:38 -0600 (CST) Received: from mendota.terracom.net (mendota.terracom.net [208.170.71.129]) by ultra5.microscopy.com (8.9.1b+Sun/8.9.1) with ESMTP id TAA14451 for {microscopy-at-sparc5.microscopy.com} ; Wed, 16 Feb 2000 19:59:26 -0600 (CST) Received: from [208.170.95.54] (3C2-24.terracom.net [208.170.95.24]) by mendota.terracom.net (8.9.1a/8.8.7) with SMTP id UAA06155 for {microscopy-at-sparc5.microscopy.com} ; Wed, 16 Feb 2000 20:19:02 -0600 (CST) X-Sender: oshel-at-pop.terracom.net Message-Id: {v02120d16b4d105b345a8-at-[208.170.95.54]} Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
My apologies to the list, but: could Gordon Cougar email me? I get a fatal error when trying to email you: The following addresses had permanent fatal errors ----- } {gcouger-at-rfdata.net} [...] } 550 {gcouger-at-rfdata.net} ... User unknown
Thanks!
Phil
*** Be famous! Send a Tech Tip or article to Microscopy Today! *** Philip Oshel Technical Editor, Microscopy Today P.O. Box 620068 Middleton, WI 53562-0068 Voice: days (608) 263-4162, evening (608) 833-2885 Fax (608) 836-1969 (please make sure my name is on any fax)
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Paul, My favorite has always been the DIATOME calendars. Next are the Gatan calanders. Dennis Kunkel has great original SEMs of diatomes etc, but I don't know if he has any posters. David Sharpe has the old standby insect SEMs. The students at SanJoaquin Delta College have put together some good calendars also.
} Date: 16 Feb 00 15:54:53 -0800 } From: Paul Webster {pwebster-at-mailhouse.hei.org} } Subject: General:Wall Posters } To: MSA listserver submission {Microscopy-at-sparc5.microscopy.com} } X-Priority: 3 } MIME-Version: 1.0 } Reply-To: Paul Webster {pwebster-at-mailhouse.hei.org} } X-MIME-Autoconverted: from quoted-printable to 8bit by } ultra5.microscopy.com id RAA14282 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Very best regards, Steve D'Angelo Equipment Resurrection 650-738-0351
Does anyone out there have direct experience of any benefit(s) gained by the use of Santovac 5 in the diffusion pumps of a JEOL 840, instead of the recommended Lion S?
And did they have to fiddle with either the heating elements or heater supply voltages in order to acheive the benefits?
I know that Santovac is well thought of, but it's expensive, and my experience of Lion S in an older JEOL (JXA-5A) has been very positive. I think there's something in there about optimising the pump design to a particular fluid.
As an economical alternative, mentioned in Wil Bigelow's book, has anyone tried NEOVAC SY, marketed by Varian, in an 840?
Does anyone know what chemical family Lion S belongs to?
Perhaps someone from JEOL, or even from the Lion company might care to reply.
cheers
Ritchie
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Hello, I'm cleaning out some things in the laboratory and we have a Mikros Vacuum Evaporator Manual C-20, 1964 here. We no longer have this evaporator, so we no longer need the manual. If anyone would have need of it, please email me personally and we'll send it out to you.
I'm amazed at the quality of this older manual - actual blue prints included!
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
There was a fairly extensive discussion about Dp oils on the listserver a while ago. That should be in the archives somewhere.
} } Does anyone out there have direct experience of any benefit(s) gained } by the use of Santovac 5 in the diffusion pumps of a JEOL 840, } instead of the recommended Lion S? }
"Yes". We have been using Santovac 5 in our early 6400 machine for about 5 months now. The diffusion pumps appear to be the same as on an 840 and the 6400 had Lion S in it beforehand.
} And did they have to fiddle with either the heating elements or } heater supply voltages in order to acheive the benefits? }
No. The vacuum seems to be as good as ever without doing any of this. It all looks very clean.
} I know that Santovac is well thought of, but it's expensive, and my } experience of Lion S in an older JEOL (JXA-5A) has been very } positive. I think there's something in there about optimising the } pump design to a particular fluid. } } As an economical alternative, mentioned in Wil Bigelow's book, has } anyone tried NEOVAC SY, marketed by Varian, in an 840? } } Does anyone know what chemical family Lion S belongs to?
That should all be in the archives somewhere, or perhaps on certain web sites. Jim Darley and/or Will Bigelow had the answers.
} } Perhaps someone from JEOL, or even from the Lion company might care } to reply. }
We were told that the Santovac 5 is more tolerant of the occasional gulp of air.....so presumably it should last longer.
Cheers
Arthur Day, Electron Microscope Unit Phone: 61-2-9717-3457 Ansto Materials Division Fax: 61-2-9543-7179 PMB 1, Menai (Sydney), NSW, 2234 Email: ard-at-ansto.gov.au Australia www: http://www.ansto.gov.au/
I am using Photoshop 5.5 on a G3 Macintosh and can no longer get a preview of the image when the file is selected in FILE } OPEN. What gives?
In previous versions (say 4.0 and earlier) when the file was selected, I got a thumbnail preview of the image prior to opening it. This is very convenient and I miss this capability.
Thanks,
John
#################################################################### John J. Bozzola, Ph.D., Director Micro-Imaging and Analysis Center 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
Santovac is a great oil to use in Jeol instruments. Backstreaming is less and it will withstand a lot more abuse than Lion oil if the vac system has a glitch. Just put a little less charge in than the normal Lion oil. Regards Steve Parkins
WOW, thanks soooo much for all the responses to my question. I have alot of techniques to try so that should keep me busy for awhile. This is a very informative site and I appreciate the help!!
The Department of Biological Sciences at the University of Iowa must find a home for a Philips 300 Transmission Electron Microscope by June 2000. It is in excellent condition and has been under service contract for nearly 30 years. We are remodeling our building and will no longer maintain support for this instrument. Free to anyone willing to come and take it away (plus Haskris chiller, carbon evaporator, film dessicator, etc.).
Please contact: Dean Abel Biological Sciences 138BB University of Iowa Iowa City IA 52242 USA 319-335-1070 (fax 319-335-1069) dean-abel-at-uiowa.edu
We are currently using Cool Prep in our Coolwell cooler system. This is a liquid cleaner & slime preventer that helps keep mineral deposits to a minimum in the cooling system. It also helps prevent corrosion in the water passages without harming hoses, gaskets or seals. Since Coolwell is no longer in business does anyone know of a replacement product for Cool Prep.
Can we simply switch to ethylene glycol? Robert Champaign Raytheon Electronic Systems Labs - Texas Region r-champaign-at-raytheon.com 2501 W.University, MS 8011 McKinney Texas, 75070 972-952-3165
Hi Y'all: We have an JEOL 845 SEM (TMP pumped) and we have been having problems with the TMP REV light staying on. Because of the microscopes' timing circuit, the scope will shut off after 20 minutes. The shutting off/timing issue can be overridden by keeping the key turned clockwise: the microscope will eventually go to the correct vacuum, but the darn TMP light never goes out. JEOL told us of a trick of depressing the standby button on the TMP controller, sometimes remedies this situation, but it didn't work in our case. We have been in contact with JEOL and so far we/they haven't figured out the problem. The TMP pump and the rough pump both seem OK: the TMP 80 % relay kicks in at 80% RPM as it's supposed to and we changed the rough pump oil, just to be sure). I have a few questions to ask of you 845 owners: 1) Have you seen this type of problem, and if so, how did you remedy it? 2) Does anyone out there have wiring diagrams, manuals for the TMP 845 (not the DP, we already have that), if so can we get a copy? JEOL doesn't seem to have anything.
Your help will be greatly appreciated. Mike Coviello Lab Manager UT Arlington Arlington, TX 817 272-5496
We have an old Photomicroscope from Zeiss. It's grey in color and about 20 years old, but works pretty good. Until now we have an old 1 inch video camera on top. We want to change this video camera to a state of art CCD-camera with 1/2 inch CCD-chip. To enlarge the field of view, we want to use a video adapter with a magnification 0.5x. Unfortunately several oprtical corrections are made within this adapter. Such an adapter is not offered by Zeiss. Does anybody out there knows a company that sells such a special adapter or can build one?
We have an old Photomicroscope from Zeiss. It's grey in color and about 20 years old, but works pretty good. Until now we have an old 1 inch video camera on top. We want to change this video camera to a state of art CCD-camera with 1/2 inch CCD-chip. To enlarge the field of view, we want to use a video adapter with a magnification 0.5x. Unfortunately several oprtical corrections are made within this adapter. Such an adapter is not offered by Zeiss. Does anybody out there knows a company that sells such a special adapter or can build one?
FORWARDED TO LISTSERVER, PLEASE RESPOND TO THE ADDRESS SHOWN BELOW:
{randy_anderson-at-ameritech.net}
I am a member of MSA and I work for the University of Chicago in the section of Nephrology. I was wondering if you would know of any universities or other types of institutions that would offer summer courses for the beginner and advance areas of immunofluorescence microscopy. Any help that you can give me in matter would greatly be appreciated.
Thank you
Randy Anderson
#################################################################### John J. Bozzola, Ph.D., Director Micro-Imaging and Analysis Center 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
The JEOL Schematics are somewhat ambiguous as the logic from the turbo pump activates several relays. The relays are NOT connected in the manner that one would think, i.e. when the turbo pump speed is 80% the relay is activated and signals the vacuum logic that all is well.
Earl Weltmer
Michael Coviello wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi Y'all: } We have an JEOL 845 SEM (TMP pumped) and we have been having problems with the } TMP REV light staying on. Because of the microscopes' timing circuit, the } scope will shut off after 20 minutes. The shutting off/timing issue can be } overridden by keeping the key turned clockwise: the microscope will } eventually go to the correct vacuum, but the darn TMP light never goes out. } JEOL told us of a trick of depressing the standby button on the TMP } controller, sometimes remedies this situation, but it didn't work in our } case. We have been in contact with JEOL and so far we/they haven't figured } out the problem. The TMP pump and the rough pump both seem OK: the TMP 80 % } relay kicks in at 80% RPM as it's supposed to and we changed the rough pump } oil, just to be sure). I have a few questions to ask of you 845 owners: } 1) Have you seen this type of problem, and if so, how did you remedy it? } 2) Does anyone out there have wiring diagrams, manuals for the TMP 845 (not } the DP, we already have that), if so can we get a copy? JEOL doesn't seem to } have anything. } } Your help will be greatly appreciated. } Mike Coviello } Lab Manager } UT Arlington } Arlington, TX } 817 272-5496
ELECTRON MICROSCOPY TECHNICIAN A full time position for a senior research technician is available at the University of Chicago in the Department of Molecular Genetics and Cell Biology. The lab of Elaine Fuchs Ph.D. is interested in the cytoskeleton, in cell-cell adhesion and in the cell biology of skin development. The successful candidate will perform routine transmission electron microcopy, including specimen acquisition, tissue processing, ultramicrotomy, operation of a Philips CM120 electron microscope and darkroom procedures/digital imaging. In addition he/she will be trained in on-section immunolabeling using Lowicryl K4M. Applicant Qualifications: Minimum requirements are a Bachelors degree and 2 years of relevant experience. If you are motivated to work in an excellent scientific environment with state of the art instrumentation please contact:
Christoph Bauer Ph.D. University of Chicago, Department of Molecular Genetics and Cell Biology 5841 S. Maryland Ave/MC 1028 Chicago, Il 60637
I'm reinitializing the server. You may see some old and/or duplicate messages. I will try to restore those that I think did not get through. If you tried to post a message in the last day or so and haven't seen it please wait ~ 12 hours after that time if you do not see it appearby then , you should presume it was lost and repost your original question/comment.
} } Ms. Lalini Pillayi Would like to take both a TEM and SEM short } } course. She is located at the at the US ARmy Dental Research Great } } Lakes Lab. } } Her applications will be both biological and biological } } combined with material. If you offer such a course please contact her } } directly at } } lalini70-at-hotmail.com } } } } Thanks
Gregory W. Erdos, Ph.D. Ph. 352-392-1295 Assistant Director, Biotechnology Program PO Box 110580 Fax: 352-846-0251 University of Florida Gainesville, FL 32611
The JEOL Schematics are somewhat ambiguous as the logic from the turbo pump activates several relays. The relays are NOT connected in the manner that one would think, i.e. when the turbo pump speed is 80% the relay is activated and signals the vacuum logic that all is well.
Earl Weltmer
Michael Coviello wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi Y'all: } We have an JEOL 845 SEM (TMP pumped) and we have been having problems } with the } TMP REV light staying on. Because of the microscopes' timing circuit, the } scope will shut off after 20 minutes. The shutting off/timing issue can be } overridden by keeping the key turned clockwise: the microscope will } eventually go to the correct vacuum, but the darn TMP light never goes out. } JEOL told us of a trick of depressing the standby button on the TMP } controller, sometimes remedies this situation, but it didn't work in our } case. We have been in contact with JEOL and so far we/they haven't figured } out the problem. The TMP pump and the rough pump both seem OK: the TMP 80 % } relay kicks in at 80% RPM as it's supposed to and we changed the rough pump } oil, just to be sure). I have a few questions to ask of you 845 owners: } 1) Have you seen this type of problem, and if so, how did you remedy it? } 2) Does anyone out there have wiring diagrams, manuals for the TMP 845 (not } the DP, we already have that), if so can we get a copy? JEOL doesn't seem to } have anything. } } Your help will be greatly appreciated. } Mike Coviello } Lab Manager } UT Arlington } Arlington, TX } 817 272-5496
We would like to resurrect 2 older Kevex SiLi detectors for scintillation counting, and have lost or misplaced the manuals for the preamp output connectors, ie. voltages and pinouts. The model numbers of the preamps are 2002 and 2003.
If anyone has this info, could you please fax or email?
Thanks in advance
Fred Pearson
******************************************************** Fred Pearson Brockhouse Institute for Materials Research McMaster University 1280 Main St. West Hamilton, Ontario Canada L8S 4M1
ELECTRON MICROSCOPY TECHNICIAN A full time position for a senior research technician is available at the University of Chicago in the Department of Molecular Genetics and Cell Biology. The lab of Elaine Fuchs Ph.D. is interested in the cytoskeleton, in cell-cell adhesion and in the cell biology of skin development. The successful candidate will perform routine transmission electron microcopy, including specimen acquisition, tissue processing, ultramicrotomy, operation of a Philips CM120 electron microscope and darkroom procedures/digital imaging. In addition he/she will be trained in on-section immunolabeling using Lowicryl K4M. Applicant Qualifications: Minimum requirements are a Bachelors degree and 2 years of relevant experience. If you are motivated to work in an excellent scientific environment with state of the art instrumentation please contact:
Christoph Bauer Ph.D. University of Chicago, Department of Molecular Genetics and Cell Biology 5841 S. Maryland Ave/MC 1028 Chicago, Il 60637
Hi Y'all: We have an JEOL 845 SEM (TMP pumped) and we have been having problems with the TMP REV light staying on. Because of the microscopes' timing circuit, the scope will shut off after 20 minutes. The shutting off/timing issue can be overridden by keeping the key turned clockwise: the microscope will eventually go to the correct vacuum, but the darn TMP light never goes out. JEOL told us of a trick of depressing the standby button on the TMP controller, sometimes remedies this situation, but it didn't work in our case. We have been in contact with JEOL and so far we/they haven't figured out the problem. The TMP pump and the rough pump both seem OK: the TMP 80 % relay kicks in at 80% RPM as it's supposed to and we changed the rough pump oil, just to be sure). I have a few questions to ask of you 845 owners: 1) Have you seen this type of problem, and if so, how did you remedy it? 2) Does anyone out there have wiring diagrams, manuals for the TMP 845 (not the DP, we already have that), if so can we get a copy? JEOL doesn't seem to have anything.
Your help will be greatly appreciated. Mike Coviello Lab Manager UT Arlington Arlington, TX 817 272-5496
Heather A Owen wrote: } } Hello everyone, } } A graduate student is getting ready for a rather large scale preparation } of rat brains for TEM using vascular perfusion. Luckily (since I'm a } botanist) her major professor has had experience with the procedure. He } doesn't know for sure whether he used EM or Biological grade } glutaraldehyde previously, but considering the large volume that will be } required, and the cost difference, I'm hoping that the Biological grade } will suffice. } } I would appreciate advice from those of you that have experience with this } method of fixation. } } Thank you in advance for any information. } } Heather Owen } } Heather A. Owen, Director } Electron Microscope Laboratory } Department of Biological Sciences } University of Wisconsin - Milwaukee } (414)229-6816
Heather,
It's been a while but, back when I did such things, I had bouts of non-uniform fixation when using biological grade glutaraldehyde solutions even in plant tissues. However, the solutions had been kept much longer than I'd now use and there are other preparation technique changes that confound the determination of causality. In the cases where I've perfused small animals, I've used EM grade GA since, as expensive as it is, it's cheaper than repeating the experiment.
However, I did a full systemic perfusion. One random thought ( I haven't tried it): What's the vascular volume of a rat brain? If you just need just the brain, I'd think a little creative clamping (really small hemostats?) in the thorax should let you reduce the volume needed to make even a large experiment affordable with "the good stuff".
cheers and good luck, John
John Heckman MSM Department Michigan State University
Santovac is a great oil to use in Jeol instruments. Backstreaming is less and it will withstand a lot more abuse than Lion oil if the vac system has a glitch. Just put a little less charge in than the normal Lion oil. Regards Steve Parkins
We use Biological grade to perfuse rats (whole body, we haven't tried clamping bits off, though that might be a good idea since we're interested in just the head). After the perfusion is finished, we remove the interesting bit, which is huge by EM standards, and fix some more in Karnowski's. Then we go on to osmium, and continue processing. We get acceptable EM pictures at high magnification, even though our blocks are so large and processing times are therefore extended. So I can recommend Biological grade glutaraldehyde. Hope this helps.
We have an old Photomicroscope from Zeiss. It's grey in color and about 20 years old, but works pretty good. Until now we have an old 1 inch video camera on top. We want to change this video camera to a state of art CCD-camera with 1/2 inch CCD-chip. To enlarge the field of view, we want to use a video adapter with a magnification 0.5x. Unfortunately several oprtical corrections are made within this adapter. Such an adapter is not offered by Zeiss. Does anybody out there knows a company that sells such a special adapter or can build one?
Does anyone have the geometry settings for a Kevex Quantum detector on a JEOL 2000FX? I am using DTSA and need the values. I would like to have the sample to detector distance and the azimuthal angle. I am using +45 for the azimuthal angle and 90 for the detector angle. Are these correct? I know that the takeoff angle is 70. -Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
} From: RLevenson-at-cri-inc.com Subject: RE: autofluorescence quenching: spectral imaging can help
You asked about strategies for reducing autofluorescence.
In addition to various strategies which can include using near-IR dyes like Cy5.5 and exciting far in the red, you can also treat tissues with sodium borohydride or toluidine blue (these latter observations I pass on from others--have not tried them myself).
I have recently had very promising results using spectral imaging to discriminate autofluorescence from desired fluorescence. [Caution: I am describing a product sold by my company]. CRI makes a liquid crystal tunable filter (LCTF) that can be used to collect a stack of images at different wavelengths. This yields a spectrum at every pixel of an image. (A product with similar capabilities is sold by Applied Spectral Imaging, but uses a different technology.) CRI's new LCTF, the FluoroSpec, uses special optics to combine polarization states; along with other improvements, it has more than twice the light throughput compared to previous models.
The sample was formalin-fixed brain probed with a Cy2-labeled antibody to a low-abundance protein. By eye it was very difficult to distinguish the Cy2 signal from the bright green to green-orange autofluorescence. Using a spectral data set (15 images from 510 nm to 650 nm at 10 nm intervals) and linear combination algorithms for pixel-unmixing, it was possible to completely separate the Cy2 and autofluorescence images.
I would be glad to share these images with any interested parties.
Richard
Richard Levenson, MD Technical Director, Biomedical Systems Cambridge Research & Instrumentation (CRI), Inc. 80 Ashford St. Boston, MA 02134
Hi all. I have just joined this list and am posting without the usual "lurking time" due to time constraints - please forgive if recently covered. I am also a novice in all microscopy techniques, especially fluorescence.
I am trying to detect GFP-containing cells in liver tissue and having some problems.
1. I can't determine the spectra for the filter blocks I am using (on a Nikon Eclipse TE300 with TE-FM Epifluorescence attachment). The blocks are marked "UV", "B-2" (blue from source, green in field) and "G" (green from source, red in field). I have e-mailed Nikon and to be fair they've only had a few days but I'm under some pressure. No help from the manual. I have been using B-2 for GFP.
2. I get quite a lot of background fluorescence even with frozen sections (fixed in neutral buffered formalin). Can anyone suggest a way to reduce this? (eg any extra filters?)
3. I have read conflicting opinions on fixation. Most previous work has used thick sections (50 microns cut eg with Vibratome, ? to avoid having to embed tissue blocks). GFP is interfered with by acetone (and probably other organic solvents) but I would have thought that after fixation with formalin it should be stabilized and resistant to the xylene/alcohol used in paraffin embedding. I have seen fluorescence retained after this treatment but perhaps it can be improved.
4. For those with liver fluoro experience - on "UV" setting I see bright fluorescence which photobleaches. I believe I am looking at retinoids in stellate cells, although the bleaching is incomplete and a bit slower than I have seen before. Can anyone confirm this?
Apologies for long post. Any help much appreciated.
David Lockwood University of Qld Dept of Surgery PA Hospital Brisbane Australia
Hello Robert, Preparation of the water in water chillers is a strange world of its own and you may get many responses to your question. The majority of my experience has been with the Haskris systems but the chemistry may be the same. In about 30 years of looking after many chillers, my formula for success goes like this: 1 - Use deionized or distilled water. The water will leach enough products from the hoses, microscope lenses, pumps and so forth to reach a equilibrium and then be "ideally suited" for your system. 2 - Don't use any fungicide or other bug killer because it will slime up the works. 3 - Put 4 or 5 drops of oil on top of the water reservoir. This will form a barrier by creating a monolayer on top of the water and keep things from growing in the water. I have used this proceedure in tropical climates and in the north and in both dry and humid environments and it has always worked very well. I can almost gurantee you that ethyline glycol will gum up the works.
Best wishes and good luck.
Alex Greene SCIENTIFIC INSTRUMENTATION SERVICES, INC. PMB-499, 1807 West Slaughter Lane, Number 200 Austin, Texas 78748-6200 Phone:512-282-5507 FAX 512/280-0702
QUALITY ELECTRON MICROSCOPE REPAIRS
-----Original Message----- } From: Robert Champaign {r-champaign-at-raytheon.com} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
I'm using a CM30 FEG with GIF PEELS atached to it. I'm getting confused about focussing an energy filtered image.
GATAN proposes to focus the image on the TV camere at a loss of 100eV.Apparently it will be focused more or less OK at a higher loss say 741eV. DM uses the voltage offset which changes the HT of the microscope. So as far as I understand the focus should be OK for every energy loss(no refocussing should be needed at all) . Although in practice it is not. The trick with focussing at 100eV works better than not focussing at all but it is not satisfactory for high spatial resolution.
In the Phd of M.Schenner (TU Wien "The quantification problem in EELS Microanalysis") I found a callibration procedure to find a defocus to correct for chromatic abberation as a function of the energy loss you are interested in. But he seems to use the drift tube in the GIF.
The question now: How should I focus my ESI images and what is the explanation behind it?
Thanks in advance,
Jo
************************************************************* * Jo Verbeeck * * University of Antwerp * * Dept. EMAT (Electron Microscopy for Materials Research) * * e-mail: joverbee-at-ruca.ua.ac.be * * tel: +32(0)3 218 02 49 * * fax: +32(0)3 218 02 57 * *************************************************************
Dear All, We are considering trying to label aspects of innervation using a quinacrine fluorescence technique. So far the preliminary trials have not gone well. Could anyone give advice on this, especially on a source for a reliable fluorescent quinacrine dye?
Thanks in advance
Alex Black Department of Anatomy National University of Ireland, Galway
We have specialists in this area who can offer customized, on-site courses, using your equipment and speaking specifically to your application. If you are interested in further details, or in how U. Chi can sponsor such a course, please contact me directly.
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At 04:59 PM 2/21/00 -0600, John J. Bozzola wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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I perfused dozens of rat brains over the years. I would recomend using the purified glutaraldehyde. However, you can use a slightly less concentrated solution since the fixation is so much more efficient tham immersion fixation. We primarily fixed for ICC but only the choice of fix would vary not the method. For normal ultrastructure, I would try 2-3% Glut or half-strength Karnovsky's in a phosphate buffer system with added NaCl and Mg at physiological concentrations and see which fixation I prefered by examining the desired tissues prior to fixing critical animals.
We would anesthetize the animal, open the chest cavity and clamp off the descending aorta, thus limiting the perfusion to the upper half of the animal. The cannula would then be inserted into a small slit in the left ventricle, positioned up into the aorta, and held with a hemostat. Perfusion lines should have been filled with normal saline or ringers so that as soon as the cannula is in place, the flow can be started to wash out the blood. A small slit is made in the right atrium to permit release of venous blood, etc. As soon as the return is relatively pale, switch to your fixative and run the desired amount through. The condition of the liver is a reasonable criteria for the efficiency of the perfusion. It should be very pale yellow as the blood is removed and should stiffen up within a few minutes. In fact the entire upper part of the animal should become quite stiff as the tissues are fixed. Figure on a couple of hundred mls. of fix per animal.
A couple of suggestions are to use the initial ringer wash and fixatives at room temperature or slightly above so as not to cause vessels to contract. Do any further washes and post fix the portions of interest at 4oC. Also make sure you have a consistant but gental perfusion rate so as not to break delicate capillaries. Usually this can be accomplished just by raising the vessels containing the buffer and fix above the animal and let gravity do the rest. Good luck, Debby
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
--------------------------------------
Heather A Owen wrote: } } Hello everyone, } } A graduate student is getting ready for a rather large scale preparation } of rat brains for TEM using vascular perfusion. Luckily (since I'm a } botanist) her major professor has had experience with the procedure. He } doesn't know for sure whether he used EM or Biological grade } glutaraldehyde previously, but considering the large volume that will be } required, and the cost difference, I'm hoping that the Biological grade } will suffice. } } I would appreciate advice from those of you that have experience with this } method of fixation. } } Thank you in advance for any information. } } Heather Owen } } Heather A. Owen, Director } Electron Microscope Laboratory } Department of Biological Sciences } University of Wisconsin - Milwaukee } (414)229-6816
Heather,
It's been a while but, back when I did such things, I had bouts of non-uniform fixation when using biological grade glutaraldehyde solutions even in plant tissues. However, the solutions had been kept much longer than I'd now use and there are other preparation technique changes that confound the determination of causality. In the cases where I've perfused small animals, I've used EM grade GA since, as expensive as it is, it's cheaper than repeating the experiment.
However, I did a full systemic perfusion. One random thought ( I haven't tried it): What's the vascular volume of a rat brain? If you just need just the brain, I'd think a little creative clamping (really small hemostats?) in the thorax should let you reduce the volume needed to make even a large experiment affordable with "the good stuff".
cheers and good luck, John
John Heckman MSM Department Michigan State University
I have a Beseler 45M Enlarger and Kodak Royal Print Processor for sale. Both are in working condition. We are going digital! Equipment is located in Easton, PA.
Well the DNS (Domain Name Server) failed and was probably pretty sick over most of last week. Alot of mail probably just ended up in a black hole somewhere on the Net. Some of you may have gotten messages, but I see that most of the mail I sent out during my trip DownUnder to the ACEM-16 meeting in Canberra never made it to it's destination.
The DNS "looks" fixed now but I'm not confident enough to proclaim all as back to normal yet. Let's see if this message actually gets through to everyone this time.
Sorry gang....
Nestor Your Friendly (Frustrated) Neighborhood SysOp
I have 2 queries pertaining to the use of the lipid colourant Sudan Black B, I hope someone can be of some help:
1. I have been able to stain oil traces in the glands of citrus fruit BUT the epoxy resin Procure/Araldite also stains quite strongly. Is this typical of epoxy resins and would Spurrs show the same problem?
- Should I reduce the staining time from 30 mins? - Adjust the temperature from 60 degrees C for staining? - Rinse my sections for longer with 70% ethanol after staining?
2. I have used osmium tetroxide to fix lipids in the citrus rind tissues, with mixed results. I have also seen some success in oil fixation by applying Sudan Black B to the tissue prior to chemical fixation. I assume it binds and reduces the oil solubility. I am going to test this by treating tissue with SBB prior and post chemical fix, and comparing.
- SBB is made up in 70% ethanol. As a postfix, would it be most logical to introduce it at the 70% step in my ethanol dehydration series? (10 to 100%)
Department of Horticulture, Viticulture and Oenology The University of Adelaide Plant Research Centre Waite Campus, PMB 1 Glen Osmond, SA 5064 AUSTRALIA
has anyone out there any experiences with immuno-labeling of osmificated and Spurr embedded samples. We have tried out etching, oxidizing sections with hydrogen peroxide (10% for 15 min) or sodium methylat (1% for 15 min) without any significant increase of label intensity (Ab used is polyclonal from rabbit, protein A purified)
It made it her ok. I thought I was getting pretty good service all week. But I know what evils hide in name servers. Not nearly to the extent you do:)
As always you efferots are welcome. You provide the pipe for one of the best resorces on the net.
Gordon
G. C,. Couger 624 Cheyenne Stillwater, OK 74075-1411 405 624 2855 between 3:00 pm to 1:00 am CST 405 742 2758 cell ----- Original Message ----- } From: "Nestor J. Zaluzec" {zaluzec-at-sparc5.microscopy.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, February 22, 2000 7:20 PM
} } Dear All: } } I was wondering if anyone out there is aware of a company } that sells pinhole grids. I am looking for grid(2D) of } pinholes that are approximately 15-20 microns in diameter } with a center to center separation of about 150-200 microns. } } I would greatly appreciate any information that you could } provide regarding the commercial availability of similar } pinhole grids. } } TIA, } Fatima Merchant } } } {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} } } Fatima Merchant, Ph.D. } Senior Research Engineer } Perceptive Scientific Instruments, Inc. } 2525 South Shore Blvd., Suite 100 } League City, Texas 77573 } } Telephone: (281) 334-3027 Ext: 230 } Toll Free: (800) 288-3027 Ext: 230 } Facsimile: (281) 538-2222 } Email: merchant-at-persci.com } } {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} } }
-----Original Message----- } From: Valdes Erica R Dr. SBCCOM Sent: Tuesday, February 22, 2000 1:26 PM To: 'microscopy-at-sparc5.Microscopy.Com'
We got a sputter-coater system (VG Microtech SC500) that we would like to convert into a glow discharge. Does anybody have done it already? We got electronic schemes to do the High Voltage unit control, but we miss the mecanical design to fit into our vacuum chamber (of the SC500). Does anybody know how to do? We know that an acessory (for the Microtech) have been developped some years ago (GD350A) but it is not anymore available. What are the modification made to use this chamber? Jean-Jacques LACAPERE INSERM U 410 FacultŽ de MŽdecine X. Bichat Paris FRANCE
I'm using a CM30 FEG with GIF PEELS atached to it. I'm getting confused about focussing an energy filtered image.
GATAN proposes to focus the image on the TV camere at a loss of 100eV.Apparently it will be focused more or less OK at a higher loss say 741eV. DM uses the voltage offset which changes the HT of the microscope. So as far as I understand the focus should be OK for every energy loss(no refocussing should be needed at all) . Although in practice it is not. The trick with focussing at 100eV works better than not focussing at all but it is not satisfactory for high spatial resolution.
In the Phd of M.Schenner (TU Wien "The quantification problem in EELS Microanalysis") I found a callibration procedure to find a defocus to correct for chromatic abberation as a function of the energy loss you are interested in. But he seems to use the drift tube in the GIF.
The question now: How should I focus my ESI images and what is the explanation behind it?
Thanks in advance,
Jo
************************************************************* * Jo Verbeeck * * University of Antwerp * * Dept. EMAT (Electron Microscopy for Materials Research) * * e-mail: joverbee-at-ruca.ua.ac.be * * tel: +32(0)3 218 02 49 * * fax: +32(0)3 218 02 57 * *************************************************************
Material fixed like yours works only rarely. You might try etching with saturated sodium metaperiodate for one hour followed by water washes before incubating with the primary. That is assuming that your antigen is not periodate sensitive.
At 07:26 AM 02/23/2000 +0100, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Gregory W. Erdos, Ph.D. Ph. 352-392-1295 Assistant Director, Biotechnology Program PO Box 110580 Fax: 352-846-0251 University of Florida Gainesville, FL 32611
} } I was wondering if anyone out there is aware of a company } } that sells pinhole grids. I am looking for grid(2D) of } } pinholes that are approximately 15-20 microns in diameter } } with a center to center separation of about 150-200 microns. } } } } I would greatly appreciate any information that you could } } provide regarding the commercial availability of similar } } pinhole grids. } }
Dear Fatima, I asked about the same question and got a reply from Janko Otto (otto-at-quantifoil.com). He wrote me:
"I read a discussion about making holey films . We had solving the problem using semiconductor lithographic techniques.
We have been producing QUANTIFOIL, a perforated foil with pre-definable hole size shape and arrangement. The variation among the hole sizes is small: the standard deviation for their distribution is well below 0.1 µm within a batch; the reproducibility for the average hole size between batches is 0.2 µm. The shapes of the holes in the foils produced so far are circular and square. Until now, their arrangements have been orthogonal.
Foils with any desired hole size in the micrometer range, and with any shape and arrangement can be fabricated. Any of these three parameter can also be varied in a defined manner within one foil.
Until now, we produce three different types of QUANTIFOIL:
Type Hole Repetition distancee um um R1.2/1.3 ~1.2 2.5 (circular holes) R2/2 ~2 4 (circular holes) S7/2 ~7 9 (square holes)
QUANTIFOIL is generally delivered as a carbon foil; it can also be obtained strengthened with plastic film.
If you interested in these perforated foil, I can send you some samples."
He may be able to produce what you want. I have not yet tried his grids out, but I intend to when I get my specimens ready. I am not affiliated with Quantifoil, just a potential user. Good luck. Yours, Bill Tivol
************************************************************* * Jo Verbeeck * * University of Antwerp * * Dept. EMAT (Electron Microscopy for Materials Research) * * e-mail: joverbee-at-ruca.ua.ac.be * * tel: +32(0)3 218 02 49 * * fax: +32(0)3 218 02 57 * *************************************************************
---------- Forwarded message ----------
} We are currently using Cool Prep in our Coolwell cooler system. This is a } liquid cleaner & slime preventer that helps keep mineral deposits to a } minimum in the cooling system. It also helps prevent corrosion in the } water passages without harming hoses, gaskets or seals. Since Coolwell is } no longer in business does anyone know of a replacement product for Cool } Prep. } } Can we simply switch to ethylene glycol?
Dear Robert, The suggestion of using distilled water and letting the system come to equilibrium might work in a truly closed system composed of only one material, but if you need to add water, or if you have several metals in the system--as we do--this will be unsatisfactory in the long run. We have found that Aqua-Treet 42 works very well in our Haskris system. This product is made by Aqua Laboratories, Inc., PO Box 645, 8 Indus- trial Way, Amesbury MA, (978) 388-3989. I have no relationship to this company except as a satisfied user of their product. Yours, Bill Tivol
I am running two copies of DTSA, one is an older version and one is the newer one that will run on a power Mac. I am also fairly new to DTSA. The old version does not have the MSA format capability for reading and writing files, but the newer one does. However, the new one will read two versions of MSA format, 1.0 and 1.1 but will only save in 1.1. Now I'm confused. I asked Nestor where I could get the latest MSA description of the MSA format and he sent me what I already have, EMMFF.total from of the EMMPDL/MAS-Lib files. I know that Nestor was on that committee. So what gives with MSA version 1.1 in the latest version of DTSA? Why don't they have the 1.0 option? The keywords are not even the same.
-Confused in Pittsburgh.
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Why not use a liquid that has NO water in it. That solved my problem in vibratory polishing of dissimilar metals in an alumina slurry. Ethylene glycol or propylene glycol, (less toxic), are possibilities.
Dear Bernie,
That will work unless there is some microorganism which will
grow in ethylene glycol. One could also just buy commercial anti-freeze
and get some additional corrosion protection. I don't think that there
would be any problem with the circulation pump, since the viscosity
of glycol is similar to that of water. Keeping the fluid in the system for
some decades has different requirements from using it over a period
Look for a "Write MSA 1.0" plug-in in the "extras Folder" of your DTSA download and move it to the "Plug-ins" folder at the same level as DTSA.
At 11:42 AM -0500 2/23/00, Walck. Scott D. wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
====================================== Carl Henderson Electron Microbeam Analysis Laboratory University of Michigan 2501 C.C. Little Bldg. Ann Arbor, MI 48109-1063 USA (734) 936-1550 FAX (734) 763-4690 ======================================
Greetings, I wonder if anyone could explain the following observation to me. I have been sputter coating with platinum and viewing the sample with a FESEM (Hitatchi 4700, below lens type). In a certain type of sample, the image appears darker than the background where there is sample. Now these samples are perhaps a bit unusual. They are very thin, starting off life as 1.75 um methacrylate sections of a plant root adhered to a glass coverslip, followed by extraction in hot acidic peroxide to remove all organic material except crystalline cellulose. The cellulosic cell walls left behind on the coverslip are nominally 100 nm thick and perhaps flattened further by the processing. The cell walls are present in the sample as either cross sections or as small regions laying in the plane of the coverslip. These regions can be anywhere from the size of a cell (10 x 40 or so microns) down to small portions thereof. All of this remaining cell wall is darker than the surrounding background. I am putting on a thin coat of Pt, somewhere around 2 nm, and imaging at 2 kV (I have seen the same thing at 5 kV). It really puzzles me that my sample is darker than background. Any explanations???
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Hi, Well I guess we never run out of new challenges when it comes to specimen prep. This one concerns a product that is made of a cereal bound together with a sugar syrup. The investigators want to see how the syrup behaves as to structural integrety over a period of time with and without the addition of various stabilizing components. Information would be collected at the light microscopic level. We first thought of cryosectioning followed by examination using LM. However, the product is filled with air pockets and literally disintegrates when microtomed. It is easily cut into pieces at temperatures above freezing as long as the pieces are no thinner than3-4 mmAccempts to infiltrate it with OCT compoound prior to freezing were not satisfactory. New thought is to use a low temperature resin to infiltrate the sample followed by UV or chemical polymerization. butThe resulting blocks could then be microtomed at room temperature and imaged. It would be desirable to cut sections of 4-8 µm in thickness which is too thick for an ultramicrotome with glass or diamond knives. Also UV polymerization would probably limit us to very small sample size which may be a problem. Any suggestions would be appreciated as to resins to use which have low viscosity, could be used to infiltrate pieces of ~0.5-1cm sq, polymerize at low temperatures (+4 to 0oC range) using UV or chemical polymerization, and are preferably soft enough to section with a metal microtome blade. Any other suggestions of preparations techniques to try would be very appreciated.
Thanks in advance for your input. Debby
Debby Sherman, Manager Phone: 765-494-6666 Microscopy Center in Agriculture FAX: 765-494-5896 Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu Purdue University 1057 Whistler Building West Lafayette, IN 47907-1057
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Dear listservers, I need to determine the value of a Philips 410 that has been on service contract until it was decommissioned last year. I need this information for an insurance claim. Is there a company or dealer of used em equipment that can help me? Thanks Hank Adams Integrated Microscopy Core Molecular and Cellular Biology Baylor College of Medicine Houston, Tx
Sorry if this is a second post, but we just got an e-mail saying the first was rejected as spam, so here goes:
Hi:
I want to be able to "Glow Discharge" carbon coated copper grids for the purpose of making them hydrophilic and negatively charged, so that I can spread aqueous suspensions on them. I have a Denton Vacuum Desk ll sputter coater with a "etch" mode. The question I have for my fellow list servers is: will "etching" the grids do the same thing as "Glow Discharging" them? Many Thanks, Tim
Timothy Schneider, Director of Electron Microscopy Department of Pathology Thomas Jefferson University Room 229 Jeff Hall 1020 Locust St. Philadelphia, Pa. 19107 215-503-4798 Timothy.Schneider-at-Mail.TJU.EDU
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We are interested in localizing arsenic in biological tissues (toxicology research), but have been unable to come up with a technique that will give us the cellular localization information we're looking for (sub-cellular would be even better).
We have tried TEM with energy dispersive x-ray microanalysis and our concentrations are apparently too low, so we are unable to detect arsenic.
A few years ago we looked into using the autometallographic techniques that have been published by Dr. Gorm Dansher & his colleagues. We were told that the photographic emulsions required were no longer available.
In a brainstorming session yesterday several techniques were bantered about, but in truth we are not familiar enough with the techniques to know if they would suit our needs. Does EELS have anything to offer, or perhaps wavelength dispersive spectoscopy (WDS)? How about some of the new FT-IR instruments? Are there any other techniques that might be able to localize arsenic and/or indicate its state of oxidation?
Vendors are welcome to reply.
Yours, Doug Cromey .................................................................... : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : Research Specialist, Principal University of Arizona : : (office: AHSC 4212A) P.O. Box 245044 : : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: doug-cromey-at-ns.arizona.edu): :...................................................................: http://www.pharmacy.arizona.edu/exp_path.html Home of: "Microscopy and Imaging Resources on the WWW"
Go to http://www.cstl.nist.gov/div837/837.02/dtsa.html and click on "supporting files" page then download "preferences and plugins" you will find 2 versions of the MSA write 1.0 (one is Kevex specific). It would make life much easier on us microscopists if all manufacturers adopted the MSA format. Good luck, Scott
} I am running two copies of DTSA, one is an older version and one is the } newer one that will run on a power Mac. I am also fairly new to DTSA. The } old version does not have the MSA format capability for reading and writing } files, but the newer one does. However, the new one will read two versions } of MSA format, 1.0 and 1.1 but will only save in 1.1. Now I'm confused. I } asked Nestor where I could get the latest MSA description of the MSA format } and he sent me what I already have, EMMFF.total from of the EMMPDL/MAS-Lib } files. I know that Nestor was on that committee. So what gives with MSA } version 1.1 in the latest version of DTSA? Why don't they have the 1.0 } option? The keywords are not even the same. } } -Confused in Pittsburgh. } } Scott D. Walck, Ph.D. } PPG Industries, Inc. } Glass Technology Center } Guys Run Rd. (packages) } P. O. Box 11472 (letters) } Pittsburgh, PA 15238-0472 } Walck-at-PPG.com } (412) 820-8651 (office) } (412) 820-8161 (fax)
For the weather watchers: Mostly sunny, 60 F, snow is melted away.
-------------------note: new mailing address---------------------- Scott Wight e-mail: scott.wight-at-nist.gov NIST 222/A113 W voice: 301-975-3949 100 Bureau Dr STOP 8371 | fax: 301-417-1321 Gaithersburg, MD 20899-8371 \|/ disclaimer: Any opinion expressed is my own and does not represent those of my employer.
Dear all, I am doing the image analysis of SEM micrographs. The micrographs contain both dense (light) and air cell (dark) areas. But two areas are not discrete. I'm having a difficulty of quantifying those separated fractions. I wonder if anyone has any ideas of how to solve the problem. Looking forward for your suggestions. Regards, Supanee
Hello all, I need to know how to draw an ellipse to measure at Image Tool software. Thanks a lot. Rejane Pimentel Rejane Magalh‹es Pimentel Galindo Functional Plant Morphology Universidade Federal Rural de Pernambuco ggalindo-at-elogica.com.br Rua Maria Carolina, 417/804 51020-220 Boa Viagem
I thought I'd try this again in case it got lost in space:
Hi-
We are going to be processing rat eyes (from perfused animals) for embedding in epoxy and I was wondering if anyone has any suggestions for processing. We are interested in the retina, optic nerve, cornea and uveal tract.
Ahh! That is always the rub, trying to get the areas of interest to stand out as a distinct range of gray scales.
If it is just a question of noise pixels in what is otherwise a distinct region, then you might try filtering (smoothing) your image before thresholding. Or you might try changing your image acquisition protocol to get less noisy images.
You might try a different signal to find something that is less subject to overlaps. Usually I resort to backscattered electron imaging since the dependence on atomic number of that signal typically results in distinct gray levels.
If that won't do it, you might be able to perform some sort of image processing to separate the regions. There are techniques available, but there seems to be as much art as science to them. I would refer you to John Russ' "Image Processing Handbook" or a similar text, because there is not a short answer to that question.
Warren Straszheim
At 03:40 PM 2/23/2000 -0500, you wrote: } Dear all, } I am doing the image analysis of SEM micrographs. The } micrographs contain both dense (light) and air cell (dark) areas. But two } areas are not discrete. I'm having a difficulty of quantifying those } separated fractions. I wonder if anyone has any ideas of how to } solve the problem. } Looking forward for your suggestions. } Regards, } Supanee
} From: {"wft03-at-health.state.ny.us"-at-sparc5.microscopy.com} To} Why not use a liquid that has NO water in it. That solved my problem in } vibratory polishing of dissimilar metals in an alumina slurry. Ethylene } glycol or propylene glycol, (less toxic), are possibilities. } } } Dear Bernie, } } That will work unless there is some microorganism which will } } grow in ethylene glycol. One could also just buy commercial anti-freeze } } and get some additional corrosion protection. I don't think that there } } would be any problem with the circulation pump, since the viscosity } } of glycol is similar to that of water. Keeping the fluid in the system for } } some decades has different requirements from using it over a period } } of hours, though.
Glycols won't have near the heat capacity of water. I doubt you will find a bug that will live in ethylene glycol but you can probably find many that can live in propylene glycol.
Any cooling system needs to have the coolant changed periodically. Following the manufactures recommendation is the safe way to go.
Gordon
Gordon Couger gcouger-at-couger.com
Stillwater, OK www.couger.com/gcouger 405 624-2855 GMT -6:00
This is something that we can probably help you with. We do similar jobs for several other companies. If you could fax a drawing with the material, OD, Thickness, number of holes, etc. to 1-802-878-8074 we can get you a quote.
Thanks,
John Arnott Chairman --
LADD RESEARCH 131 Dorset Lane Williston, VT 05495
TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net web site http://www.ladd.cc
--
LADD RESEARCH 131 Dorset Lane Williston, VT 05495
TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net web site http://www.ladd.cc
We are considering to purchase a cryotransfer system for our 2010F TEM. There are two major suppliers: Gatan and Oxford. We are not sure which one works better since this is the first time dealing with this technique.
Could you please send me your comments and experiences on these two vendors? Our samples are mostly soft tissues: Polymers, surfactants, cell membranes, and proteins. Your help is highly appreciated.
Maoxu Qian **************************** * Maoxu Qian, Ph.D. * * Dept of MSE, box 352120 * * University of Washington * * Seattle, WA 98195 * * mxq-at-u.washington.edu * * (206)616-3973(phone) * * (206)543-3100(fax) * ****************************
This course will cover all aspects of pre-thinning and focus on final thinning via Tripod Polishing. Due to the limited class size and the extensive hands-on opportunities, this course is well suited to novices as well as advanced Tripodders. Attendees will also learn the latest techniques available in ion milling, plasma cleaning and ion beam sputter deposition and etching for TEM and SEM samples.
The workshop will be held in San Clemente, CA on May 26-27, 2000. Please call for more information or visit our web site ( www.southbaytech.com ) for registration information.
David Henriks Vice President TEL: 800-728-2233 (toll free in the USA) South Bay Technology, Inc. +1-949-492-2600 1120 Via Callejon FAX: +1-949-492-1499 San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
*************************************************************************** } } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
} } } Hi, There, } } We are considering to purchase a cryotransfer system for our 2010F TEM. } There are two major suppliers: Gatan and Oxford. We are not sure which one } works better since this is the first time dealing with this technique. } Could you please send me your comments and experiences on these two } vendors? Our samples are mostly soft tissues: Polymers, surfactants, cell } membranes, and proteins. Your help is highly appreciated. } } Maoxu Qian } **************************** } * Maoxu Qian, Ph.D. * } * Dept of MSE, box 352120 * } * University of Washington * } * Seattle, WA 98195 * } * mxq-at-u.washington.edu * } * (206)616-3973(phone) * } * (206)543-3100(fax) * } **************************** } } } } }
A related question from India. We have been using DM (demineralised) water in our JEOL 2000 EX II cooled by a Neslab chiller for the last ten years. This requires change every 6 months approximately. What additives can be used to prevent algae (? it is green and slimy and I am not a biologist) growth? Something obtainable locally (chemical name rather than trade names) will be most useful to me. Thanks very much.
-----Original Message----- } From: William Tivol [SMTP:tivol-at-wadsworth.org] Sent: Thursday, February 24, 2000 10:25 AM To: Robert Champaign Cc: microscopy-at-sparc5.microscopy.com
} We are currently using Cool Prep in our Coolwell cooler system. This is a } liquid cleaner & slime preventer that helps keep mineral deposits to a } minimum in the cooling system. It also helps prevent corrosion in the } water passages without harming hoses, gaskets or seals. Since Coolwell is } no longer in business does anyone know of a replacement product for Cool } Prep. } } Can we simply switch to ethylene glycol?
Dear Robert, The suggestion of using distilled water and letting the system come to equilibrium might work in a truly closed system composed of only one material, but if you need to add water, or if you have several metals in the system--as we do--this will be unsatisfactory in the long run. We have found that Aqua-Treet 42 works very well in our Haskris system. This product is made by Aqua Laboratories, Inc., PO Box 645, 8 Indus- trial Way, Amesbury MA, (978) 388-3989. I have no relationship to this company except as a satisfied user of their product. Yours, Bill Tivol
Dear Colleagues Does anybody have experience on the Allied MultiPrep Polishing System, especially in terms of getting a good TEM sample? How well do the Allied's Precision Cross-Section Tool and TEM Wedge Polisher work on this semi automatic multiprep polisher? Also, how does their TEM Wedge Polisher compare with the South Bay Tripod Polisher? TIA Anita ******************************************* Dr. Anita Garg NASA Glenn Research Center at Lewis Field Advanced Metallics Branch Mail Stop 49-3 21000 Brookpark Road Cleveland, OH 44135 Phone : (216) 433-8908 Fax : (216) 977-7132 E-mail : Anita.Garg-at-grc.nasa.gov *******************************************
Fatima Merchant, I used to work for a company called Buckbee Mears. They have been making tight tolerance electroformed Ni pinholes for many, many years. They are located in St. Paul, MN and their phone number is 651-228-6400. Good luck. Dan May
} Hi- } } We are going to be processing rat eyes (from perfused animals) for } embedding in epoxy and I was wondering if anyone has any suggestions for } processing. We are interested in the retina, optic nerve, cornea and uveal } tract. } } Thank you very much, } Sandy Perkins
**************************** Hi Sandra,
Eyes are a real pain. There are so many layers of different cell types, and each seems to infiltrate differently. Having worked on a fir number of eyes, I woauld make the following suggestions: Process the cornea separately. It is almost pure collagen. It will not osmicate all that noticeably, so don't be surprised when it doesn't turn dark brown or black. Give it long infiltration times, in vaccuum if possible.
If you don't need the optic nerve attached to the eye, dissect it free and process it. Give it slighlty longer than usual dehydration and infiltration steps to be sure you'e got the myelin taken care of.
As for the rest: remove the lens and the vitreous so that you have good acess to the retina. Be careful not to disrupt the retina when you remove the vitreous. Once the lens is out, you can usually get the vitreous out by gently blotting against a Kimwipe. You may want to split the orb into hemispheres. Just be sure not to take any sections from the cut edge, since you will disturb the structure there when you cut. Again, you'll nees longer dehyration and infiltration times, since the sclera is really tough. I use 30 minutes in each step of graded acetone, propylene oc=xide then a graded infiltraion with PO:resin (Epon analog) 2:1 for 30 min, 1:1 overnight, 1:2 2 hours, pure resin overnight, on a rotator.
I'm sure there ore others out there, with their own "pet protocol" for eyes. Its all a bit of magic and luck, isn't it?
Good luck, Lee
Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Confocal Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
Not sure this went out earlier...I've inherited an Olympus BH-2 RFCA upright that I'd like to equip with WI lenses for live cell fluorescence work. This is a 160mm tube length scope, and Olympus is now only manufacturing the 60X WI fluor objective (I think/hope). Any leads to sources of compatible WI objectives in the 10-60X range much appreciated. Please reply offline. Thanks.
Mike
Michael Ferrari Department of Biological Sciences 629 Science Engineering (SCEN) University of Arkansas Fayetteville, Arkansas 72701 Ph: 501-575-6372 Fax: 501-575-4010
We are interested in selling our Perkin-Elmer Model 1010M microdensitometer at a very reasonable price. It was purchased as a reconditioned model from Perkin-Elmer eight years ago and is in very good condition. Technical manual is included.
If interested, please contact:
Amy Kendall Department of Molecular Biology Vanderbilt University Box 1820, Station B Nashville TN 37232 (615) 322-2012 amy-at-helix.molbio.vanderbilt.edu
R. Divakar wrote, "A related question from India. We have been using DM (demineralised) water in our JEOL 2000 EX II cooled by a Neslab chiller for the last ten years. This requires change every 6 months approximately. What additives can be used to prevent algae (? it is green and slimy and I am not a biologist) growth? Something obtainable locally (chemical name rather than trade names) will be most useful to me."
I quote here recommendations from the Haskris Co. (we have several Haskris chillers):
1. If algae is present, flush the system with one of two alternatives: add 1 cup of chlorine bleach or one pint of hydrogen peroxide per 15 gallons of water. Circulate 20-30 minutes. Drain the system and refill with clean water.
2. To prevent regrowth of algae, one of the following additives are recommended: a. 10% solution of lab grade (no additives) ethylene glycol. b. Dichlorophene. This is an insoluble white powder which is a fungicide/bacteriacide. It should be sprinkled evenly across the entire surface of the water in the chiller's tank. c. 8 parts per million chlorine.
---------------------------------------------------------------------- Russell E. Cook, Ph.D. Electron Microscopy Center for Materials Research Argonne National Laboratory Materials Science Division, Building 212 9700 South Cass Avenue Argonne, IL 60439-4838 (630)252-7194 FAX: (630)252-4289 recook-at-anl.gov
Dear Tobias, The most logical explanation for this phenomenon is that the carbon-based cell material is of a lower average atomic number, even with the thin Pt coating, than the glass coverslip (SiO2) it is located on, also with the Pt coat. The secondary electron yield is directly proportional to the average atomic number, if topographic effects are not in evidence, which they won't be on such a thin sample. At 11:39 AM 2/23/00 -0600, you wrote: } Greetings, } I wonder if anyone could explain the following observation } to me. I have been sputter coating with platinum and viewing the } sample with a FESEM (Hitatchi 4700, below lens type). In a certain } type of sample, the image appears darker than the background where } there is sample. Now these samples are perhaps a bit unusual. They } are very thin, starting off life as 1.75 um methacrylate sections of } a plant root adhered to a glass coverslip, followed by extraction in } hot acidic peroxide to remove all organic material except crystalline } cellulose. The cellulosic cell walls left behind on the coverslip are } nominally 100 nm thick and perhaps flattened further by the } processing. The cell walls are present in the sample as either cross } sections or as small regions laying in the plane of the coverslip. } These regions can be anywhere from the size of a cell (10 x 40 or so } microns) down to small portions thereof. All of this remaining cell } wall is darker than the surrounding background. I am putting on a } thin coat of Pt, somewhere around 2 nm, and imaging at 2 kV (I have } seen the same thing at 5 kV). It really puzzles me that my sample is } darker than background. Any explanations??? } } THanks, } Tobias
Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
You have a basic problem - free lipids interfere with the polymerization of epoxies. Since you are having mixed results with osmium not all lipids may be fixing adequately. If possible, try keeping the specimens in osmium overnight, on a rotator. (After the first hour of exposure to osmium, change to fresh osmium). I have done this very successfully with lipid rich structures. Hope it works.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Microscopist, } } has anyone out there any experiences with immuno-labeling of osmificated and } Spurr embedded samples. We have tried out etching, oxidizing sections with } hydrogen peroxide (10% for 15 min) or sodium methylat (1% for 15 min) } without any significant increase of label intensity (Ab used is polyclonal } from rabbit, protein A purified) } } Any suggestions are welcome. Manfred } } } Hello,
You may have a problem which you do not recognize. Spurr's really has no place in immunocytochemistry. It is highly crosslinked. As you might know, epoxies not only crosslink, they also chemically combine with tissue proteins making them unavailable. Also, a highly stable, highly crosslinked embeddding medium will cut a shiny, very smooth surface on sections. The surface relief of such a section is extremely low, and, of course, the more surface relief, the more chance for the antigen to be exposed to markers. You might also have a paucity of antigen which compounds all problems.
If you want to stay with epoxies, I would suggest switching to a soft Epon-Araldite for starters. If that does not work, then my suggestion is to go to LR Gold. A block of LR Gold does not "cut", but cleaves, causes something like a triple increase in surface relief thus giving more antigens the chance to be exposed.
If I am not making myself clear, please e-mail me. I have just gone through 2yrs of frustration with stuff like this, but this week the paper is going out!!!!
Bye, Hildy Crowley University of Denver Denver, CO
P.S. I am not making all this up. I can reference everything, but do not have the time to do that in this format
Before adding glycol to your Haskris water chiller check with the manufacture of the microscope. If you use glycol to cool a Hitachi microscope you will damage the plastic water lines and they will all need to be replaced after a few years. I have seen this happen a few times now after people have used this advice from the list server. Please check with your service engineer first.
To those users of Hitachi S2500/S570, glycol is used to cool the objective lense but only on these 2 instruments.
I am a service engineer for Hitachi in Canada and the above opinion is my opinion and not that of Hitachi.
} I quote here recommendations from the Haskris Co. (we have several Haskris } chillers): } } 1. If algae is present, flush the system with one of two alternatives: add } 1 cup of chlorine bleach or one pint of hydrogen peroxide per 15 gallons of } water. Circulate 20-30 minutes. Drain the system and refill with clean } water. } } 2. To prevent regrowth of algae, one of the following additives are } recommended: } a. 10% solution of lab grade (no additives) ethylene glycol. } b. Dichlorophene. This is an insoluble white powder which is a } fungicide/bacteriacide. It should be sprinkled evenly across the entire } surface of the water in the chiller's tank. } c. 8 parts per million chlorine. } } ---------------------------------------------------------------------- } Russell E. Cook, Ph.D. } Electron Microscopy Center for Materials Research } Argonne National Laboratory } Materials Science Division, Building 212 } 9700 South Cass Avenue } Argonne, IL 60439-4838 } (630)252-7194 } FAX: (630)252-4289 } recook-at-anl.gov } }
I've been using dichlorophene in our Haskris chillers following the incident related below. If you want to buy some, dichlorophene is listed in several chemical company catalogs as:
2,2'-Methylenebis(4-chlorophenol)
Considering how I was planning to use it, I bought technical grade (90%), since it was substantially less expensive than higher grades and have not experienced any problems.
A complete replacement of our chiller water (along with the water in the scope and lines) was required about a year ago when the water pump in our TEM Haskris had to be replaced. Our sevice engineer flushed the system using the concentrated hydrogen peroxide as mentioned above, and a truly amazing amount of material spewed forth, so much so that small connectors to the power supply clogged up a few weeks down the line, causing our HV to shut down. Cleaning out the connectors and flushing the system again with distilled water took care of the problem.
At the time, we were cautioned (by Haskris, I think) to use distilled water and not deionized water in the chillers. Does anyone know why?
Heather Owen
Heather A. Owen, Director Electron Microscope Laboratory Department of Biological Sciences University of Wisconsin - Milwaukee (414)229-6816
What is the real answer? When we had our Hitachi S-3200N installed two years ago, the service engineer used an ethylene glycol/water solution in the Haskris chiller. And it was pretty strong, too. Something like 1:3 glycol:water.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
====================================== Carl Henderson Electron Microbeam Analysis Laboratory University of Michigan 2501 C.C. Little Bldg. Ann Arbor, MI 48109-1063 USA (734) 936-1550 FAX (734) 763-4690 ======================================
TEM Technician A position is available at White Oak Semiconductor, Richmond VA, USA. The successful candidate will perform TEM operation and sample preparation in support of DRAM fabrication and TEM engineering. The candidate should have at least an associates degree and a minimum of 3 years experience in the semiconductor industry; of which at least 1 year should pertain to semiconductor TEM sample preparation. Additionally, they should understand the fundamental tools and techniques of TEM sample preparation and the semiconductor process from a top down and cross sectional perspective. Good eye-hand coordination, communication skills, and attention to detail are required. Candidates should be highly motivated, self directed, good at problem solving, interested in learning new skills and effective working alone or in a team environment. Interested candidates please contact
Kim Christensen Failure Analysis Laboratory White Oak Semiconductor 6000 Technology Boulevard Sandston, Virginia 23150 Ph: 804 952 7307 Fax: 804 952 7902 Pager: 1 800 759 8888, pin# 130 3382
If you can puncture the eye, or preferably take off the anterior chamber, so that the chemicals can get in, there is absolutely no problem. Standard embedding, using longish times, works well. For instance, 3 hrs in osmium, 1 hr in UAc, 20 mins in each of the alcohols/acetone, 1 hr in resin/acetone (dilute), overnight infiltration in resin/acetone (more concentrated), 1 hr in warm resin, embed. If you need to have the eye unpunctured, it may be difficult. I've never embedded a discrete eye, but I do know that it is really difficult to get even isolated sclera well embedded, as it seems to act as a very effective barrier to resin. The eye is a well designed ball that doesn't want to deflate, but that also means nothing much will get in.
Contact me if you want further info. I've embedded more eyes than I care to remember, including some rats eyes.
Diana
Diana van Driel Department of Clinical Ophthalmology Sydney University GPO Box 4337 Sydney NSW AUSTRALIA 2001
You are correct Carl you can use glycol to cool your sem(all Hitachi sems) as only the diffusion pump is being cooled and Hitachi uses a vinyl hose that isn't effected by the Glycol. For those that have Hitachi TEM's the water lines for the lenses and diffusion pumps are the smaller black appearing lines, these will be adversely effected by the glycol.
Sorry for the confusion I will be more specific in the future. So in the sem's that use the clear reinforced vinyl hoses only, glycol is okay. In Hitachi TEM which use a small black plastic hose for the lenses and diffusion pumps, do not use glycol.
} OK, now I am concerned.
} What is the real answer? When we had our Hitachi S-3200N installed } two years ago, the service engineer used an ethylene glycol/water } solution in the Haskris chiller. And it was pretty strong, too. } Something like 1:3 glycol:water.
I am pleased to bring to your attention the following TEM technician position at Motorola's Process and Materials Characterization Laboratory (PMCL) I encourage all persons interested to respond ASAP.
Position: TEM Technician/S9001P Employer: Motorola-Semiconductor Products Sector's Process and Materials Characterization Laboratory (PMCL) Location: Mesa, AZ Employment type: Full-time Employment status: Full-employee (non-contractor) Number of Positions: 2 Shift: all (compressed) Relocation: Available
Duties/Responsibilities: Experience in Transmission Electron Microscopy techniques including basic operation, specimen preparation, maintenance, and troubleshooting of TEM's. Work effectively as a team player in multiple projects providing routine and non-routine TEM analysis in support of semiconductor product manufacturing. Exposure to many different types of processes and technologies, and working knowledge of FIB and EDS are a plus.
Specific knowledge: AA degree preferred. A higher or lower classification will be established depending upon qualification and experience.
Contact info: Send resume or questions via email to Brian_Wajdyk-at-email.mot.com or fax to 480-655-4316 C/O Brian Wajdyk. -- ******************************************************************** Brian Wajdyk Team Leader / Electron Microscopist (FESEM, EDS, SAM) Motorola - Process and Materials Characterization Laboratory (PMCL) 2200 W. Broadway Rd., Mesa AZ 85202 Mail Drop: M360 Tel: 480-655-4337 Fax: 480-655-4316 Email: brian_wajdyk-at-email.mot.com Pager: 1-800-313-5960 ********************************************************************
I am interested in doing some PC based image analysis of fibers and fiber properties on crimping. Is there anyone doing IA of crimp angle/amplitude or crimps per inch? Is there commercial IA software that will do these types of measurements?
Any help would be appreciated. Thanks.
Duane Krueger 7635 Harold Avenue Golden Valley, Minnesota 55427
} Glycols won't have near the heat capacity of water. I doubt } you will find a bug that will live in ethylene glycol but you can } probably find many that can live in propylene glycol. }
Dear Gordon, I've been trying to look up the heat capacity of glycols in my Handbook of Chemistry and Physics. It doesn't have the data I want, but it does have the Cp for water (~4 j/gK), butane (~0.006 j/molK = ~10^-4 j/gK, if I'm reading my cal's and Cal's right), and 1-propanol (~0.01 j/molK with the same caviat). } From these, I'd guess Cp for ethylene glycol is ~0.014 j/molK, which is, indeed, much less than that of water. Propylene glycol's Cp should be similar. Yours, Bill Tivol
} A related question from India. We have been using DM (demineralised) water in our JEOL 2000 EX II cooled by a Neslab chiller for the last ten years. This requires change every 6 months approximately. What additives can be used to prevent algae (? it is green and slimy and I am not a biologist) growth? Something obtainable locally (chemical name rather than trade names) will be most useful to me. Thanks very much.
Dear Divakar, Russell's suggestion of dichlorophene is good--it's also what we use. To prevent growth in the tubing, we added some CuSO4--being careful to keep the pH slightly above 7.5. We have copper tubing in the system, so the Cu++ should not be a problem, and it will inhibit algal growth. At that pH, CuSO4 is not too soluble, so go slowly (if at all). Yours, Bill Tivol
} A complete replacement of our chiller water (along with the water in the } scope and lines) was required about a year ago when the water pump in our } TEM Haskris had to be replaced. Our sevice engineer flushed the system } using the concentrated hydrogen peroxide as mentioned above, and a truly } amazing amount of material spewed forth, so much so that small connectors } to the power supply clogged up a few weeks down the line, causing our HV } to shut down. Cleaning out the connectors and flushing the system again } with distilled water took care of the problem. } }
We added inline filters--the ones with the fiber cartridges--to trap all the particulate material in the lines, and over a period of several months, we probably got as much material as you got all at once. I highly recommend adding filters.
At the time, we were cautioned (by Haskris, I think) to use distilled
} water and not deionized water in the chillers. Does anyone know why? }
Hard to say. Deionized water can contain material leached out of the resin used, and maybe that is bad for the system, while volatiles (mostly organics, I'd guess) from distillation may not be. Yours, Bill Tivol
A student wishes to take a number of serial sections through a bee brain mounted in glycerine. The total distance is around 500µm and she would like to include three reference points which enable the sections to be correctly orientated throughout the brain. Embedding three small rods seems to be the most likely option but the material would need to be stable and able to be cut by a vibratome. Keeping the rods parallel would also be beneficial.
If anyone has tried this sort of thing or has any suggestions as to what we could use for the markers I'd appreciate hearing your bright ideas.
Regards
Andrew McNaughton
______________________________________________________________________________ Andrew McNaughton South Campus Electron Microscope Unit C/-Department of Anatomy and Structural Biology School of Medical Sciences University of Otago PO Box 913, Dunedin NEW ZEALAND
We are using sputter coater in "etch" mode for this purpose. It is doing the job flawlessly. I don't know about your coater but I'll give you the settings we are using here. The coater has maximum operating voltage of 1400V. The grids are put on glass plate (so they do not make contact with the lower electrode) and then the plate is put on the lower electrode. The current we use is 5 mA for 20 seconds (the current depends strongly on the pressure and the voltage). The pressure is about 20 Pa.
Good luck !
Rado
--------------------------------------------------------------------- Radostin Danev Laboratory of Ultrastructure Research National Institute for Physiological Sciences Myodaiji-cho, Okazaki 444-8585, JAPAN e-mail: rado-at-nips.ac.jp --------------------------------------------------------------------- ----- Original Message ----- } From: Timothy Schneider {Timothy.Schneider-at-mail.tju.edu} To: {microscopy-at-sparc5.microscopy.com} Sent: Thursday, February 24, 2000 4:29 AM
Dear Microscopy users,
I just want to inform you about a new web site (http://www.nanofactory.com) with some useful information on scanning probe microscopy (SPM)
There are, for example: - Weekly update of SPM research articles - Suggestions of some SPM literature - Links to many SPM research groups - Links to almost all SPM companies
There are also some interesting mpeg-movies with TEM-STM measurements of two tunneling tips jumping into each other.
Best regards, Hakan Olin -- Dr Hakan Olin Physics and Engineering Physics Chalmers University of Technology SE-412 96 Goteborg Sweden tel. +46-31-772 3338 fax +46-31-772 3367 mobile +46-70-30 88 99 0 e-mail hakan.olin-at-fy.chalmers.se home page http://fy.chalmers.se/~f4aolin
An effective way to prevent your cooling system from growing green or blue-green algae is to ensure that it is totally light-tight. Algae require light for photosynthesis. The Philips engineers who installed our CM120 Biotwin recommended we connect up the chiller with the black-lined vinyl hose used in drinks vending machines, where microorganism growth in the water lines is obviously undesirable. The product we used is trademarked "Aquavend", is opaque white outside and black inside. It appears to have worked so far, though the ethylene glycol we use would also discourage algae. Yours Chris Jeffree
Date sent: Thu, 24 Feb 2000 18:08:15 -0500 } From: William Tivol {tivol-at-wadsworth.org} To: microscopy-at-sparc5.microscopy.com
Dear Debby, Some ten yaers ago we were doing research on sugar syrup in cooperation with a sugar company. They were interested in the stucture of mixtures of sugar syrups. W e had some succes using freeze-fracture and looking at the replicas in the TEM. Of course these were not mixtures of cereals with sugar syrup and the specimens were allowed to be a lot smaller but this could be a suggestion. Succes ! Wim Jacob Ctr.of Electron Microscopy University of Antwerp Belgium
Debby Sherman wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi, } Well I guess we never run out of new challenges when it comes to specimen prep. This one concerns a product that is made of a cereal bound together with a sugar syrup. The investigators want to see how the syrup behaves as to structural integrety over a period of time with and without the addition of various stabilizing components. Information would be collected at the light microscopic level. } We first thought of cryosectioning followed by examination using LM. However, the product is filled with air pockets and literally disintegrates when microtomed. It is easily cut into pieces at temperatures above freezing as long as the pieces are no thinner than3-4 mmAccempts to infiltrate it with OCT compoound prior to freezing were not satisfactory. } New thought is to use a low temperature resin to infiltrate the sample followed by UV or chemical polymerization. butThe resulting blocks could then be microtomed at room temperature and imaged. It would be desirable to cut sections of 4-8 µm in thickness which is too thick for an ultramicrotome with glass or diamond knives. Also UV polymerization would probably limit us to very small sample size which may be a problem. } Any suggestions would be appreciated as to resins to use which have low viscosity, could be used to infiltrate pieces of ~0.5-1cm sq, polymerize at low temperatures (+4 to 0oC range) using UV or chemical polymerization, and are preferably soft enough to section with a metal microtome blade. Any other suggestions of preparations techniques to try would be very appreciated. } } Thanks in advance for your input. } Debby } } Debby Sherman, Manager Phone: 765-494-6666 } Microscopy Center in Agriculture FAX: 765-494-5896 } Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu } Purdue University } 1057 Whistler Building } West Lafayette, IN 47907-1057
Dear Doug, Why don't you try imaging SIMS? I would not surprised if such instrument(s) are present at the University of Arizona. Of course this would restrict the resolution, I gess, to 0.1 - 0.5 micrometer, with the newer instruments. Regards Wim Jacob Ctr. of Electron Microscopy University of Antwerp Belgium
Doug Cromey wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Colleagues, } } We are interested in localizing arsenic in biological tissues (toxicology } research), but have been unable to come up with a technique that will give } us the cellular localization information we're looking for (sub-cellular } would be even better). } } We have tried TEM with energy dispersive x-ray microanalysis and our } concentrations are apparently too low, so we are unable to detect arsenic. } } A few years ago we looked into using the autometallographic techniques that } have been published by Dr. Gorm Dansher & his colleagues. We were told } that the photographic emulsions required were no longer available. } } In a brainstorming session yesterday several techniques were bantered } about, but in truth we are not familiar enough with the techniques to know } if they would suit our needs. Does EELS have anything to offer, or perhaps } wavelength dispersive spectoscopy (WDS)? How about some of the new FT-IR } instruments? Are there any other techniques that might be able to localize } arsenic and/or indicate its state of oxidation? } } Vendors are welcome to reply. } } Yours, } Doug Cromey } .................................................................... } : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : } : Research Specialist, Principal University of Arizona : } : (office: AHSC 4212A) P.O. Box 245044 : } : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : } : (FAX: 520-626-2097) (email: doug-cromey-at-ns.arizona.edu): } :...................................................................: } http://www.pharmacy.arizona.edu/exp_path.html } Home of: "Microscopy and Imaging Resources on the WWW"
Please visit my new homepage at http://syli.homepage.com at your convinience! It contains not only my current research work, but also LINKS to useful journal informations, instruction for authors, on-line EELS database, periodic table, physical constants, online TEM course and etc.
I am looking forward to your invaluable suggestions! Yours sincerely,
Shu-You Li
************************************************** Shu-You Li, Dr. Institut fuer Physikalische Chemie Johannes Guttenberg Universitaet Jakob-Welder-Weg 11 D-55099 Mainz, Germany
Becky, I do not know if it is okay in the S4700 to use glycol. If it is working as in why change?
X-From_: r-holdford-at-ti.com Thu Feb 24 19:53:24 2000
X-From_: Patrick.Kilcran-at-hii-hitachi.com Thu Feb 24 23:55:28 2000 } From: "Kilcran, Patrick-HIIEX" {Patrick.Kilcran-at-hii-hitachi.com} To: dhoyle-at-istar.ca Cc: "Lima, Rick-HIIEX" {Rick.Lima-at-hii-hitachi.com}
Dear Researcher:
The University of Florida College of Medicine Electron Microscopy Facility is hosting a two-day workshop on immunogold technique. The workshop will include two identical sessions on April 10/11 and 13/14. Dr. Jan Leunissen from the Aurion Immunogold Reagent & Accessories, an internationally known expert in the field, will be the instructor for the workshop. Attached is the program information. If you are interested in attending this workshop, please contact Dr. Verlander or Ms. Hong Yi at the addresses provided below. Due to practical considerations, the workshop will be limited to a maximum of 20 participants for each session. Therefore, we encourage you to register as soon as possible. If you are not able to attend, we would appreciate if you could pass the message onto someone else who might be interested in learning about immunogold techniques. Thank you very much.
Dr. Jill W. Verlander Reed (workshop faculty) Director, University of Florida College of Medicine Electron Microscopy Facility Phone: (352) 846-0820 Fax: (352) 846-3299 verlaj-at-medicine.ufl.edu
Hong Yi (technical coordinator) Supervisor, Emory Neurology Microscopy Core Laboratory Phone: (404) 727-8692 Fax: (404) 727-3157 Email: hyi-at-emory.edu
University of Florida, College of Medicine, Electron Microscopy Facility Aurion Immunogold Reagents & Accessories / Electron Microscopy Sciences
Present Immunogold Workshop
April 10-14, 2000 Gainesville, FL
WORKSHOP HOST Dr. Jill Verlander Reed Director, Electron Microscopy Facility University of Florida College of Medicine Phone: (352) 846-0820 Fax: (352) 846-3299 verlaj-at-medicine.ufl.edu
WORKSHOP INSTRUCTORS
Dr. Jan L.M. Leunissen is an internationally known scientist in the field of ultrastructural localization. He received his Ph.D in molecular cell biology at the University of Utrecht in the Netherlands. Dr. Leunissen is the inventor of ultra small colloidal gold probes and also the founder and president of the company Aurion, which specializes in immunogold reagents and custom immunogold labeling. He has been invited to many international microscopy conferences and workshops and is especially experienced in providing hands-on training. His previous workshops in Europe, Asia, and the US have been fully attended and very well received.
Dr. Jill W. Verlander is the Director of the Electron Microscopy Facility for the University of Florida College of Medicine. Dr. Verlander has 15 years of experience in ultrastructural research in kidney and is known internationally for her work examining structure-function relationships in renal transport epithelia. Her work has been published in such journals as the Journal of Clinical Investigation, the American Journal of Physiology, Journal of the American Society of Nephrology, and Kidney International. These studies have been accomplished using light microscopic immunohistochemistry and in situ hybridization, scanning and transmission electron microscopy, morphometric analyses, and immunogold and immunoperoxidase cytochemistry at the ultrastructural level.
GENERAL INFORMATION Objectives To provide researchers the opportunity to learn the theory and practice of immunogold labeling To permit participants to process their own samples using these techniques under expert guidance To promote technology exchange and research collaboration
Rooms will be reserved in the above hotel under the name "UF EM workshop". Shuttle buses are available between the hotel and UF campus
Enrollment Note Two sessions (A; April 10-11, B: April 13-14) will be hosted at UF. Each session will be limited to a maximum of 20 participants. The workshops will provide samples to those who prefer not to bring their own. Each participant will be provided with a sample of gold conjugate of their choice at the end of the workshop.
MAIN CURRICULUM The properties of gold particles and their protein conjugates. Theories underlying immunogold labeling protocols. Silver enhancement of gold particles Imunogold labeling on a variety of sample preparations for LM. Immunogold labeling for EM a. Pre-embedding immunogold labeling using ultra_small gold conjugates and silver enhancement. b. Post-embedding immunogold labeling using ultra small and conventional colloidal gold conjugates.
CONTACT PERSON AND TECHNICAL COORDINATOR Hong Yi Phone: (404) 727-8692 Fax: (404) 727-3157 Email: hyi-at-emory.edu
Jill Verlander Reed, D.V.M. Associate Scientist Office of the Dean University of Florida College of Medicine P.O. Box 100215 Gainesville, FL 32610 verlaj-at-medicine.ufl.edu Phone: (352) 846-0820 Fax: (352) 846-3299
Molecular Imaging is updating our contact information.
If you are interested in scanning probe microscopy, would you please take a moment and update your information at http://www.molec.com/info/form.html
Thank you
George
_______________________ George Sibbald, President Molecular Imaging: Technology Leader In Advanced Bio-AFM Systems 9830 South 51 Street, Ste 124A Phoenix AZ 85044 (480) 753-4311 www.molec.com
In fact, we have an S-4700 and the service technician told us that there are tons of problems with using glycol in the chillers. We have a Haskris chiller. Apparently, the glycol volatilizes some of the hose components.
} } Looking to find and to determine what is the best printer (photographic and } archive quality) } for TEM digital images. } } Currently considering Kodak 8670PS, Codonics, or Fuji Printers } Would be interested in comments from people using photoraphic qualtiy } printers } for TEM applications } thoughts, "next time's" etc. } } Thank You } } Gregory Argentieri } Novartis Pharmacuticals Corp } gregory.argentieri-at-pharma.novartis.com } } } }
Debby Sherman Talk to Mark Cavaleri at 3M. He made a polished cross sectgion of M%M candies and preserved tyhe sugr layer
Sam Purdy
} ---------- } From: Debby Sherman } Sent: February 2000 1:17 PM } To: message to: MSA list } Subject: Food Science Prep. } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi, } Well I guess we never run out of new challenges when it comes to } specimen prep. This one concerns a product that is made of a cereal bound } together with a sugar syrup. The investigators want to see how the syrup } behaves as to structural integrety over a period of time with and without } the addition of various stabilizing components. Information would be } collected at the light microscopic level. } We first thought of cryosectioning followed by examination using LM. } However, the product is filled with air pockets and literally } disintegrates when microtomed. It is easily cut into pieces at } temperatures above freezing as long as the pieces are no thinner than3-4 } mmAccempts to infiltrate it with OCT compoound prior to freezing were not } satisfactory. } New thought is to use a low temperature resin to infiltrate the sample } followed by UV or chemical polymerization. butThe resulting blocks could } then be microtomed at room temperature and imaged. It would be desirable } to cut sections of 4-8 µm in thickness which is too thick for an } ultramicrotome with glass or diamond knives. Also UV polymerization would } probably limit us to very small sample size which may be a problem. } Any suggestions would be appreciated as to resins to use which have low } viscosity, could be used to infiltrate pieces of ~0.5-1cm sq, polymerize } at low temperatures (+4 to 0oC range) using UV or chemical polymerization, } and are preferably soft enough to section with a metal microtome blade. } Any other suggestions of preparations techniques to try would be very } appreciated. } } Thanks in advance for your input. } Debby } } Debby Sherman, Manager Phone: 765-494-6666 } Microscopy Center in Agriculture FAX: 765-494-5896 } Dept. of Botany & Plant Pathology E-mail: sherman-at-btny.purdue.edu } Purdue University } 1057 Whistler Building } West Lafayette, IN 47907-1057 } }
I received this today. If anyone could help, please reply to the sender.
Thanks.
ML
} Date: Fri, 25 Feb 2000 17:14:04 -0600 } From: Howard Hsu {hhsu-at-kumc.edu} } Organization: KU Medical Center } X-Accept-Language: en } MIME-Version: 1.0 } To: wong-at-msg.ucsf.edu } Subject: sem } } I am a researcher at the University of Kansas Med. Center and would like } study a biological sample for the presence of mineral using SEM } approach. If you can provide external service on cost or collaborative } basis, please let me know. Otherwise, I would greatly appreciate if you } would let me know any academic or industrial sources that provide this } type of service. Thank you. Howard Hsu, Ph.D. } Mei Lie Wong Electron Microscope Laboratory Department of Biochemistry HHMI-UCSF Ph. 415-476-4441 Fax 415-476-1902 http://util.ucsf.edu/agard/wong/index.html email wong-at-msg.ucsf.edu
We are a hospital based research lab. starting to use the electron microscope to identify or confirm pathogens that cause gastrointestinal disturbances. We would like to know if someone out there could recommend appropriate specific EM protocols (negative staining, processing of the gastrointestinal contents, etc....) to visualize these pathogens (ranging from viruses to bacteria and protozoans). We have a Jeol 1010 EM and perform the routine EM preparations designed to examine tissues.
In the interest of bring quality healthcare to our people, we will be very much obliged,
Mei Lie Wong wrote: ================================================== I received this today. If anyone could help, please reply to the sender.
Thanks.
ML
} Date: Fri, 25 Feb 2000 17:14:04 -0600 } From: Howard Hsu {hhsu-at-kumc.edu} } Organization: KU Medical Center } X-Accept-Language: en } MIME-Version: 1.0 } To: wong-at-msg.ucsf.edu } Subject: sem } } I am a researcher at the University of Kansas Med. Center and would like } study a biological sample for the presence of mineral using SEM } approach. If you can provide external service on cost or collaborative } basis, please let me know. Otherwise, I would greatly appreciate if you } would let me know any academic or industrial sources that provide this } type of service. Thank you. Howard Hsu, Ph.D. } Mei Lie Wong Electron Microscope Laboratory Department of Biochemistry HHMI-UCSF Ph. 415-476-4441 Fax 415-476-1902 http://util.ucsf.edu/agard/wong/index.html email wong-at-msg.ucsf.edu ============================================================ Although you have specifically asked about SEM, I am aware only of a TEM approach for this kind of analysis. We at Structure Probe, Inc. do perform this kind of work for clients on a commercial basis. You can see an example of the final sample that is analyzed by TEM on our website at page URL http://www.2spi.com/catalog/instruments/etchers4.html
The method is pretty much described so anyone really could do with, provided they did have access by a plasma etcher such as the SPI Plasma Prep II. If you wanted to do it yourself and were having difficulties, let us know, perhaps we could give you assistance. We have found that the removal of the organics is usually quite important in order to remove the source of the unwanted Bremsstrahlung radiation.
We have heard from persons trying to do this by SEM/EDS, where by the section is deposited on a polished beryllium mount, and then the organics etched away but they seem to have not been successful.
Disclaimer: SPI Supplies manufactures the plasma etching equipment needed to practice this procedure and our Structure Probe® analytical services part of our business performs this kind of sample preparation and analysis for clients. We realize there might be other methods for doing this, but we have favored the approach that works for us.
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.spi.cc ######################## ============================================
We have some experience in using micro-PIXE (proton induced X-ray emission) and the so-called nuclear microprobe in toxicological research. It is more sensitive than EDS and WDS, is very easy to quantify (standardless technique). In principle you can have detection limits of the order of 1 ppm in point analysis mode. Specimens can be of any thickness and their actual thickness is assessed by the simultaneous use of the BS technique (proton backscatering spectrometry). What in my opinion makes it really attractive for this purpose is its very good scanning capability. Scan sizes are from ca. 2.5 mm x 2.5 mm down to the beam resolution, which is of the order of few micrometer (can be down to 1 micrometer or even less, but in most of our uses few micrometers is good enough). The sample preparation is similar as for EDS, using cryotechniques to avoid elemental losses and redistribution. We have so far done studies on Ni, Mo, Cd. The detection limits for As are better than for Mo and Cd. Of course, this technique cannot indicate the state of oxidation, you can really treat it as a "more sensitive EDS".
If you want more information, I can refer you to our earlier work and perhaps you might be interested in contacting me.
Best regards
xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx Dr Wojciech J. Przybylowicz Materials Research Group National Accelerator Centre PO Box 72 Faure 7131 South Africa E-mail: przybylowicz-at-nac.ac.za Fax: +27-21-8433543 Phone: +27-21-8433820 http://www.nac.ac.za/IBA/ http://www.nac.ac.za/materials/ xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx
We have some experience in using micro-PIXE (proton induced X-ray emission) and the so-called nuclear microprobe in toxicological research. It is more sensitive than EDS and WDS, is very easy to quantify. In principle you can have detection limits of the order of 1 ppm in point analysis mode. Specimens can be of any thickness and their actual thickness is assessed by the simultaneous use of the BS technique (proton backscatering spectrometry). What in my opinion makes it really attractive for this purpose is its very good scanning capability. Scan sizes are from ca. 2.5 mm x 2.5 mm down to the beam resolution, which is of the order of few micrometer (can be down to 1 micrometer or even less, but in most of our uses few micrometers is good enough). The sample preparation is similar as for EDS, using cryotechniques to avoid elemental losses and redistribution. We have so far done studies on Ni, Mo, Cd. The detection limits for As are better than for Mo and Cd. Of course, this technique cannot indicate the state of oxidation, you can really treat it as a "more sensitive EDS".
If you want more information, I can refer you to our earlier work and perhaps you might be interested in contacting me.
Best regards
xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx Dr Wojciech J. Przybylowicz Materials Research Group National Accelerator Centre PO Box 72 Faure 7131 South Africa E-mail: przybylowicz-at-nac.ac.za Fax: +27-21-8433543 Phone: +27-21-8433820 http://www.nac.ac.za/IBA/ http://www.nac.ac.za/materials/ xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx
An information about a new principle in vibration damping to all microscopy users, who have problems with particularly low frequency vibrations from natural environment.
I invite you kindly to visit our home page: http://www.hwlbioanalytic.com
We show there active vibration isolation systems for various applications with damping frequencies from 0,6 Hz to 100/200 Hz and load capacities (depending from system) from max. 90 kg up to 330/660 kg and max. 1.500 kg. Thank you for your time.
We use many of the techniques and the atlas from the following excellent book. "EM in Diagnostic Virology" by Doan and Anderson,1987, Cambridge University Press, ISBN 0 521 24311 4.
-- Tracy Gales Fox Chase Cancer Center Electron Microscopy Facility 7701 Burholme Ave. Philadelphia PA 19111 Tel. 215-728-2484 Fax 215-728-2412
A friend recently retired and gave me a TEM diffraction plate reader. This was made by Ernest F. Fullam, Inc. in the early 1970's. It is still in the wooden shipping crate, is in unused condition and looks complete except there is no manual. It is a basic no-frills manual version. It has a high quality Starrett vernier scale and could be used for measurement of any electron micrograph, or other type of transparency, or it may be useful for teaching the history and theory of diffraction pattern measurement.
I would be willing to send it to someone willing to pay the shipping costs.
Thanks, Louie
Louie Kerr Research and Education Support Coordinator Marine Biological Laboratory 7 MBL Street Woods Hole, MA 02543 508-289-7273 508-540-6902 (FAX) 508-292-0289 (Cell phone)
The University of Kentucky invites applications for a Research Associate in the area of electron microscopy. The Electron Microscopy Facility houses two SEMs, two TEMs one AFM and extensive sample preparation equipment that are used by undergraduate and graduate students in support of research and education. The primary responsibility of the Research Associate will be to oversee the JEOL 2010F Field Emission TEM and auxiliary equipment, including an Oxford energy dispersive x-ray spectrometer (EDS), Gatan electron energy loss imaging filter and acquisition hardware and software. S/he will be responsible for coordinating and administering maintenance, access, training and use of the equipment by internal and external users. S/he will also be encouraged to develop sponsored research programs and collaborate with university faculty members. The qualified candidate will have a Ph.D. and practical experience in analytical electron microscopy and demonstrate good communication skills. The salary will be commensurate with qualifications and experience. The University of Kentucky offers comprehensive insurance and benefits packages and is an equal opportunity employer. Please send applications to:
We perform about 1000 exams/yr for viruses using EM. Below are some refs (check out especially 2, 5, 9, 11, 14). Also, at the bottom is a synopsis I give to our medical students; maybe it will be useful.
As for enteric viruses, EM is the best way to look for them. Many of them do not grow in tissue culture, and those that can be coaxed, do so under special conditions that are not employed in the routine culture lab. Biochemical identification (e.g., immunological kits, PCR, Western Blots) require a specific reagent to recognize the virus and if you don't use the right reagent, you will miss viruses (e.g., if you run a test for rotavirus, you will miss adenovirus, etc.) Furthermore, there aren't biochemical reagents for all viruses. With EM, you can see a wide variety of viruses by doing a negative stain of an aqueous extract of stool.
Good luck, Sara
REFERENCES ON VIRUS IDENTIFICATION BY ELECTRON MICROSCOPY
1. Cegielski JP, Msengi AE, Miller SE. 1994. Enteric viruses associated with HIV infection in Tanzanian children with chronic diarrhea. Ped AIDS HIV Inf 5:296-299.
2. Doane FW, Anderson N. 1986. Electron Microscopy in Diagnostic Virology: A Practical Guide and Atlas, Cambridge University Press, New York.
3. Lew JF, Glass, RI, Petric M, LeBaron CW, Hammond GW, Miller SE, Robinson C, Boutilier J, Riepenhoff-Talty M, Payne CM, Franklin R, Oshiro LS, and Jaqua M-J. 1990. Six year retrospective surveillance of gastroenteritis viruses identified at ten electron microscopy centers in the United States and Canada Pediatr Infect Dis J 9:709-714.
4. Mclean DM, Wong KK. 1984. Same Day Diagnosis of Human Virus Infections, CRC Press, Boca Raton, FL.
5. Miller SE. 1986. Detection and identification of viruses by electron microscopy. J Electron Microsc Tech 4:265-301.
5. Miller SE, Howell DN. 1988. Viral infections in the acquired immunodeficiency syndrome. J Electron Microsc Tech 8:41-78.
7. Miller SE. 1988. Diagnostic virology by electron microscopy. ASM News 54:475-481.
8. Miller SE. 1989. Electron microscopy in rapid viral diagnosis. EMSA Bull 19:53-59.
9. Hayat MA, Miller SE. 1990. Negative Staining: Applications and Methods. McGraw-Hill, New York.
10. Miller SE. 1991. Evaluation of electron microscopic information available from clinical samples. In de la Maza LM, Peterson EM (eds.), Medical Virology, Vol 10. Irvine, CA. pp 21-54.
11. Miller SE. 1995. Diagnosis of viral infection by electron microscopy. In Lennette EH, Lennette DA, Lennette ET (eds), Diagnostic Procedures for Viral, Rickettsial and Chlamydial Infections, American Public Health Assoc, Washington, DC. pp 35-76.
12. Oshiro LS, Miller SE. 1992. Application of electron microscopy to the diagnosis of viral infections. In Lennette EH (ed), Laboratory Diagnosis of Viral Infections. Marcel Dekker, New York. pp 45-68.
13. Palmer E, Martin ML. 1982. An Atlas of Mammalian Viruses, CRC Press, Boca Raton, FL.
14. Palmer E, Martin ML. 1988. Electron Microscopy in Viral Diagnosis, CRC Press, Boca Raton, FL.
15. Murphy FA, Fauquet CM, Bishop DHL, Ghabrial SA, Jarvis AW, GP Martelli, Mayo MA, Summers MD. 1995. Virus Taxonomy. Sixth Report of the International Committee on Taxonomy of Viruses. Springer-Verlag, New York.
2, 5, 9, 11-14 = Methods and micrographs 2, 14 = Excellent atlases 7, 8 = Brief synopses of technique and basis of virus ID 15 = Good general reference, but don't memorize!! Lists all viruses, not just human.
TWO WAYS TO LOOK AT VIRUSES BY EM: Negative staining Used with liquid samples Shows whole virus (size, surface structure important) Thin sectioning Used with cells and tissues Shows a slice of an infected cell Shows relationship of viruses to cell organelles (internal structure and cell organelle association important) Location within the cell is a clue to nucleic acid type DNA viruses are usually seen in the nucleus (some exceptions). RNA viruses are usually seen in the cytoplasm (some exceptions). Exceptions: Naked viruses get out of the cell by lysis; if the cell is very sick, there may be virions everywhere, e.g., adenovirus (DNA). Poxviruses are DNA viruses, but are constructed in the cytoplasm. Hepatitis B core particles (DNA) may be in the nucleus and cytoplasm. Dane particles (complete viruses) are only cytoplasmic. Some paramyxovirus (RNA) nucleocapsids, not the complete virion, can be seen in the nucleus (e.g., measlesvirus). Enveloped viruses are associated with cell membranes; the type of cell membrane (plasma membrane, nuclear membrane, endoplasmic reticulum, vacuoles) can be a clue to virus identification.
SUMMARY OF VIRUS TYPES (based on morphology): Naked viruses All human naked viruses are icosahedral (spherical). There are 3 size ranges from 20-35 nm, 45-55 nm, 70-90 nm. Examples: Small: parvovirus, enterovirus, rhinovirus Medium: polyomavirus, papillomavirus Large: rotavirus/reovirus, adenovirus Enveloped viruses All have a lipoprotein membrane on the outside of the nucleocapsid. Most derive this layer by budding through cell membranes; this pliable layer makes the virion pleomorphic. Poxviruses synthesize their shell de novo; sometimes they pick up an extra covering by budding. They are not pleomorphic, but different varieties may be more ovoid, others, brick-shaped. Most range in size from 40-400 nm. (Filoviruses are 80 nm by up to 1400 nm.) The envelope has proteins on the outside that may look like spikes or fuzz (e.g., influenza virus, coronavirus). Sometimes the spikes are too short to be distinguished by EM, making the virion appear smooth (e.g., herpesviruses, rubella virus). The nucleocapsid shape can be a. icosahedral, like the naked viruses; however, some are morphologically nondescript as seen by EM; b. filamentous, like a "slinky." c. complex (poxvirus cores look like dumbbells with lateral bodies) d. morphologically nondescript (just dense without any particular shape). Enveloped DNA viruses (except pox) construct their nucleocapsids in the nucleus. Enveloped RNA viruses construct their nucleocapsids in the cytoplasm; (measles nucleocapsids can sometimes be seen in the nucleus).
On Sat, 26 Feb 2000, Ronald R. Matias wrote:
} Date: Sat, 26 Feb 2000 08:19:49 +0800 } From: Ronald R. Matias {rrm-at-skyinet.net} } To: Microscopy-at-sparc5.microscopy.com } Subject: Need help on EM negative staining } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Hello and Help! } } We are a hospital based research lab. starting to use the electron } microscope to identify or confirm pathogens that cause gastrointestinal } disturbances. We would like to know if someone out there could recommend } appropriate specific EM protocols (negative staining, processing of the } gastrointestinal contents, etc....) to visualize these pathogens (ranging } from viruses to bacteria and protozoans). We have a Jeol 1010 EM and perform } the routine EM preparations designed to examine tissues. } } In the interest of bring quality healthcare to our people, we will be very } much obliged, } } ronnie matias } }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
We perform about 1000 exams/yr for viruses using EM. Below are some refs (check out especially 2, 5, 9, 11, 14). Also, at the bottom is a synopsis I give to our medical students; maybe it will be useful.
As for enteric viruses, EM is the best way to look for them. Many of them do not grow in tissue culture, and those that can be coaxed, do so under special conditions that are not employed in the routine culture lab. Biochemical identification (e.g., immunological kits, PCR, Western Blots) require a specific reagent to recognize the virus and if you don't use the right reagent, you will miss viruses (e.g., if you run a test for rotavirus, you will miss adenovirus, etc.) Furthermore, there aren't biochemical reagents for all viruses. With EM, you can see a wide variety of viruses by doing a negative stain of an aqueous extract of stool.
Good luck, Sara
REFERENCES ON VIRUS IDENTIFICATION BY ELECTRON MICROSCOPY
1. Cegielski JP, Msengi AE, Miller SE. 1994. Enteric viruses associated with HIV infection in Tanzanian children with chronic diarrhea. Ped AIDS HIV Inf 5:296-299.
2. Doane FW, Anderson N. 1986. Electron Microscopy in Diagnostic Virology: A Practical Guide and Atlas, Cambridge University Press, New York.
3. Lew JF, Glass, RI, Petric M, LeBaron CW, Hammond GW, Miller SE, Robinson C, Boutilier J, Riepenhoff-Talty M, Payne CM, Franklin R, Oshiro LS, and Jaqua M-J. 1990. Six year retrospective surveillance of gastroenteritis viruses identified at ten electron microscopy centers in the United States and Canada Pediatr Infect Dis J 9:709-714.
4. Mclean DM, Wong KK. 1984. Same Day Diagnosis of Human Virus Infections, CRC Press, Boca Raton, FL.
5. Miller SE. 1986. Detection and identification of viruses by electron microscopy. J Electron Microsc Tech 4:265-301.
5. Miller SE, Howell DN. 1988. Viral infections in the acquired immunodeficiency syndrome. J Electron Microsc Tech 8:41-78.
7. Miller SE. 1988. Diagnostic virology by electron microscopy. ASM News 54:475-481.
8. Miller SE. 1989. Electron microscopy in rapid viral diagnosis. EMSA Bull 19:53-59.
9. Hayat MA, Miller SE. 1990. Negative Staining: Applications and Methods. McGraw-Hill, New York.
10. Miller SE. 1991. Evaluation of electron microscopic information available from clinical samples. In de la Maza LM, Peterson EM (eds.), Medical Virology, Vol 10. Irvine, CA. pp 21-54.
11. Miller SE. 1995. Diagnosis of viral infection by electron microscopy. In Lennette EH, Lennette DA, Lennette ET (eds), Diagnostic Procedures for Viral, Rickettsial and Chlamydial Infections, American Public Health Assoc, Washington, DC. pp 35-76.
12. Oshiro LS, Miller SE. 1992. Application of electron microscopy to the diagnosis of viral infections. In Lennette EH (ed), Laboratory Diagnosis of Viral Infections. Marcel Dekker, New York. pp 45-68.
13. Palmer E, Martin ML. 1982. An Atlas of Mammalian Viruses, CRC Press, Boca Raton, FL.
14. Palmer E, Martin ML. 1988. Electron Microscopy in Viral Diagnosis, CRC Press, Boca Raton, FL.
15. Murphy FA, Fauquet CM, Bishop DHL, Ghabrial SA, Jarvis AW, GP Martelli, Mayo MA, Summers MD. 1995. Virus Taxonomy. Sixth Report of the International Committee on Taxonomy of Viruses. Springer-Verlag, New York.
2, 5, 9, 11-14 = Methods and micrographs 2, 14 = Excellent atlases 7, 8 = Brief synopses of technique and basis of virus ID 15 = Good general reference, but don't memorize!! Lists all viruses, not just human.
TWO WAYS TO LOOK AT VIRUSES BY EM: Negative staining Used with liquid samples Shows whole virus (size, surface structure important) Thin sectioning Used with cells and tissues Shows a slice of an infected cell Shows relationship of viruses to cell organelles (internal structure and cell organelle association important) Location within the cell is a clue to nucleic acid type DNA viruses are usually seen in the nucleus (some exceptions). RNA viruses are usually seen in the cytoplasm (some exceptions). Exceptions: Naked viruses get out of the cell by lysis; if the cell is very sick, there may be virions everywhere, e.g., adenovirus (DNA). Poxviruses are DNA viruses, but are constructed in the cytoplasm. Hepatitis B core particles (DNA) may be in the nucleus and cytoplasm. Dane particles (complete viruses) are only cytoplasmic. Some paramyxovirus (RNA) nucleocapsids, not the complete virion, can be seen in the nucleus (e.g., measlesvirus). Enveloped viruses are associated with cell membranes; the type of cell membrane (plasma membrane, nuclear membrane, endoplasmic reticulum, vacuoles) can be a clue to virus identification.
SUMMARY OF VIRUS TYPES (based on morphology): Naked viruses All human naked viruses are icosahedral (spherical). There are 3 size ranges from 20-35 nm, 45-55 nm, 70-90 nm. Examples: Small: parvovirus, enterovirus, rhinovirus Medium: polyomavirus, papillomavirus Large: rotavirus/reovirus, adenovirus Enveloped viruses All have a lipoprotein membrane on the outside of the nucleocapsid. Most derive this layer by budding through cell membranes; this pliable layer makes the virion pleomorphic. Poxviruses synthesize their shell de novo; sometimes they pick up an extra covering by budding. They are not pleomorphic, but different varieties may be more ovoid, others, brick-shaped. Most range in size from 40-400 nm. (Filoviruses are 80 nm by up to 1400 nm.) The envelope has proteins on the outside that may look like spikes or fuzz (e.g., influenza virus, coronavirus). Sometimes the spikes are too short to be distinguished by EM, making the virion appear smooth (e.g., herpesviruses, rubella virus). The nucleocapsid shape can be a. icosahedral, like the naked viruses; however, some are morphologically nondescript as seen by EM; b. filamentous, like a "slinky." c. complex (poxvirus cores look like dumbbells with lateral bodies) d. morphologically nondescript (just dense without any particular shape). Enveloped DNA viruses (except pox) construct their nucleocapsids in the nucleus. Enveloped RNA viruses construct their nucleocapsids in the cytoplasm; (measles nucleocapsids can sometimes be seen in the nucleus).
On Sat, 26 Feb 2000, Ronald R. Matias wrote:
} Date: Sat, 26 Feb 2000 08:19:49 +0800 } From: Ronald R. Matias {rrm-at-skyinet.net} } To: Microscopy-at-sparc5.microscopy.com } Subject: Need help on EM negative staining } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Hello and Help! } } We are a hospital based research lab. starting to use the electron } microscope to identify or confirm pathogens that cause gastrointestinal } disturbances. We would like to know if someone out there could recommend } appropriate specific EM protocols (negative staining, processing of the } gastrointestinal contents, etc....) to visualize these pathogens (ranging } from viruses to bacteria and protozoans). We have a Jeol 1010 EM and perform } the routine EM preparations designed to examine tissues. } } In the interest of bring quality healthcare to our people, we will be very } much obliged, } } ronnie matias } }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
We perform about 1000 exams/yr for viruses using EM. Below are some refs (check out especially 2, 5, 9, 11, 14). Also, at the bottom is a synopsis I give to our medical students; maybe it will be useful.
As for enteric viruses, EM is the best way to look for them. Many of them do not grow in tissue culture, and those that can be coaxed, do so under special conditions that are not employed in the routine culture lab. Biochemical identification (e.g., immunological kits, PCR, Western Blots) require a specific reagent to recognize the virus and if you don't use the right reagent, you will miss viruses (e.g., if you run a test for rotavirus, you will miss adenovirus, etc.) Furthermore, there aren't biochemical reagents for all viruses. With EM, you can see a wide variety of viruses by doing a negative stain of an aqueous extract of stool.
Good luck, Sara
REFERENCES ON VIRUS IDENTIFICATION BY ELECTRON MICROSCOPY
1. Cegielski JP, Msengi AE, Miller SE. 1994. Enteric viruses associated with HIV infection in Tanzanian children with chronic diarrhea. Ped AIDS HIV Inf 5:296-299.
2. Doane FW, Anderson N. 1986. Electron Microscopy in Diagnostic Virology: A Practical Guide and Atlas, Cambridge University Press, New York.
3. Lew JF, Glass, RI, Petric M, LeBaron CW, Hammond GW, Miller SE, Robinson C, Boutilier J, Riepenhoff-Talty M, Payne CM, Franklin R, Oshiro LS, and Jaqua M-J. 1990. Six year retrospective surveillance of gastroenteritis viruses identified at ten electron microscopy centers in the United States and Canada Pediatr Infect Dis J 9:709-714.
4. Mclean DM, Wong KK. 1984. Same Day Diagnosis of Human Virus Infections, CRC Press, Boca Raton, FL.
5. Miller SE. 1986. Detection and identification of viruses by electron microscopy. J Electron Microsc Tech 4:265-301.
5. Miller SE, Howell DN. 1988. Viral infections in the acquired immunodeficiency syndrome. J Electron Microsc Tech 8:41-78.
7. Miller SE. 1988. Diagnostic virology by electron microscopy. ASM News 54:475-481.
8. Miller SE. 1989. Electron microscopy in rapid viral diagnosis. EMSA Bull 19:53-59.
9. Hayat MA, Miller SE. 1990. Negative Staining: Applications and Methods. McGraw-Hill, New York.
10. Miller SE. 1991. Evaluation of electron microscopic information available from clinical samples. In de la Maza LM, Peterson EM (eds.), Medical Virology, Vol 10. Irvine, CA. pp 21-54.
11. Miller SE. 1995. Diagnosis of viral infection by electron microscopy. In Lennette EH, Lennette DA, Lennette ET (eds), Diagnostic Procedures for Viral, Rickettsial and Chlamydial Infections, American Public Health Assoc, Washington, DC. pp 35-76.
12. Oshiro LS, Miller SE. 1992. Application of electron microscopy to the diagnosis of viral infections. In Lennette EH (ed), Laboratory Diagnosis of Viral Infections. Marcel Dekker, New York. pp 45-68.
13. Palmer E, Martin ML. 1982. An Atlas of Mammalian Viruses, CRC Press, Boca Raton, FL.
14. Palmer E, Martin ML. 1988. Electron Microscopy in Viral Diagnosis, CRC Press, Boca Raton, FL.
15. Murphy FA, Fauquet CM, Bishop DHL, Ghabrial SA, Jarvis AW, GP Martelli, Mayo MA, Summers MD. 1995. Virus Taxonomy. Sixth Report of the International Committee on Taxonomy of Viruses. Springer-Verlag, New York.
2, 5, 9, 11-14 = Methods and micrographs 2, 14 = Excellent atlases 7, 8 = Brief synopses of technique and basis of virus ID 15 = Good general reference, but don't memorize!! Lists all viruses, not just human.
TWO WAYS TO LOOK AT VIRUSES BY EM: Negative staining Used with liquid samples Shows whole virus (size, surface structure important) Thin sectioning Used with cells and tissues Shows a slice of an infected cell Shows relationship of viruses to cell organelles (internal structure and cell organelle association important) Location within the cell is a clue to nucleic acid type DNA viruses are usually seen in the nucleus (some exceptions). RNA viruses are usually seen in the cytoplasm (some exceptions). Exceptions: Naked viruses get out of the cell by lysis; if the cell is very sick, there may be virions everywhere, e.g., adenovirus (DNA). Poxviruses are DNA viruses, but are constructed in the cytoplasm. Hepatitis B core particles (DNA) may be in the nucleus and cytoplasm. Dane particles (complete viruses) are only cytoplasmic. Some paramyxovirus (RNA) nucleocapsids, not the complete virion, can be seen in the nucleus (e.g., measlesvirus). Enveloped viruses are associated with cell membranes; the type of cell membrane (plasma membrane, nuclear membrane, endoplasmic reticulum, vacuoles) can be a clue to virus identification.
SUMMARY OF VIRUS TYPES (based on morphology): Naked viruses All human naked viruses are icosahedral (spherical). There are 3 size ranges from 20-35 nm, 45-55 nm, 70-90 nm. Examples: Small: parvovirus, enterovirus, rhinovirus Medium: polyomavirus, papillomavirus Large: rotavirus/reovirus, adenovirus Enveloped viruses All have a lipoprotein membrane on the outside of the nucleocapsid. Most derive this layer by budding through cell membranes; this pliable layer makes the virion pleomorphic. Poxviruses synthesize their shell de novo; sometimes they pick up an extra covering by budding. They are not pleomorphic, but different varieties may be more ovoid, others, brick-shaped. Most range in size from 40-400 nm. (Filoviruses are 80 nm by up to 1400 nm.) The envelope has proteins on the outside that may look like spikes or fuzz (e.g., influenza virus, coronavirus). Sometimes the spikes are too short to be distinguished by EM, making the virion appear smooth (e.g., herpesviruses, rubella virus). The nucleocapsid shape can be a. icosahedral, like the naked viruses; however, some are morphologically nondescript as seen by EM; b. filamentous, like a "slinky." c. complex (poxvirus cores look like dumbbells with lateral bodies) d. morphologically nondescript (just dense without any particular shape). Enveloped DNA viruses (except pox) construct their nucleocapsids in the nucleus. Enveloped RNA viruses construct their nucleocapsids in the cytoplasm; (measles nucleocapsids can sometimes be seen in the nucleus).
On Sat, 26 Feb 2000, Ronald R. Matias wrote:
} Date: Sat, 26 Feb 2000 08:19:49 +0800 } From: Ronald R. Matias {rrm-at-skyinet.net} } To: Microscopy-at-sparc5.microscopy.com } Subject: Need help on EM negative staining } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Hello and Help! } } We are a hospital based research lab. starting to use the electron } microscope to identify or confirm pathogens that cause gastrointestinal } disturbances. We would like to know if someone out there could recommend } appropriate specific EM protocols (negative staining, processing of the } gastrointestinal contents, etc....) to visualize these pathogens (ranging } from viruses to bacteria and protozoans). We have a Jeol 1010 EM and perform } the routine EM preparations designed to examine tissues. } } In the interest of bring quality healthcare to our people, we will be very } much obliged, } } ronnie matias } }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
The University of Kentucky invites applications for a Research Associate in the area of electron microscopy. The Electron Microscopy Facility houses two SEMs, two TEMs one AFM and extensive sample preparation equipment that are used by undergraduate and graduate students in support of research and education. The primary responsibility of the Research Associate will be to oversee the JEOL 2010F Field Emission TEM and auxiliary equipment, including an Oxford energy dispersive x-ray spectrometer (EDS), Gatan electron energy loss imaging filter and acquisition hardware and software. S/he will be responsible for coordinating and administering maintenance, access, training and use of the equipment by internal and external users. S/he will also be encouraged to develop sponsored research programs and collaborate with university faculty members. The qualified candidate will have a Ph.D. and practical experience in analytical electron microscopy and demonstrate good communication skills. The salary will be commensurate with qualifications and experience. The University of Kentucky offers comprehensive insurance and benefits packages and is an equal opportunity employer. Please send applications to:
The University of Kentucky invites applications for a Research Associate in the area of electron microscopy. The Electron Microscopy Facility houses two SEMs, two TEMs one AFM and extensive sample preparation equipment that are used by undergraduate and graduate students in support of research and education. The primary responsibility of the Research Associate will be to oversee the JEOL 2010F Field Emission TEM and auxiliary equipment, including an Oxford energy dispersive x-ray spectrometer (EDS), Gatan electron energy loss imaging filter and acquisition hardware and software. S/he will be responsible for coordinating and administering maintenance, access, training and use of the equipment by internal and external users. S/he will also be encouraged to develop sponsored research programs and collaborate with university faculty members. The qualified candidate will have a Ph.D. and practical experience in analytical electron microscopy and demonstrate good communication skills. The salary will be commensurate with qualifications and experience. The University of Kentucky offers comprehensive insurance and benefits packages and is an equal opportunity employer. Please send applications to:
I don't know if you have received an answer since you posted this message, but if you'd like, we could have a look at your images and try to help you. What we need is of course the image(s) and a description of what you actually want to measure. Please be as specific as you can. Please send the material to my email address below.
Thanks.
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Exchange Administrator Sent: Wednesday, February 23, 2000 8:35 PM To: Michael Bode
Dear all, I am doing the image analysis of SEM micrographs. The micrographs contain both dense (light) and air cell (dark) areas. But two areas are not discrete. I'm having a difficulty of quantifying those separated fractions. I wonder if anyone has any ideas of how to solve the problem. Looking forward for your suggestions. Regards, Supanee
Occassionally someone sends a question to the listserver and there is no response. It doesn't happen often so I thought I would send this question again.
Several weeks ago, I was given some inner membranes from E. coli to be run up to Epon and sectioned for TEM. I treated the sample like tissue culture and pelleted the membranes. Fixed in 3% glut in PBS and post-fixed in 1% OsO4. Routine dehydration in alcohols. Embedded in EMbed 812 with BDMA in Beem Capsules. Left in oven 48 hours. When I tried to section, the tissue was too soft and appeared to not be infiltrated well. Put back into the oven for 2-3 more days. No difference. Thinking I had a neural meltdown while processing the samples, I tried again on fresh samples, paying particular attention to details. I also used new bottles of embedding media. Same results. The part of the block without the tissue is its usual hardness. The part of the block with the tissue (the tip of the beem capsule) is soft and tacky.
Has anyone experienced anything like this? Any suggestions on how I can get the sample into epon?
Thanks in advance for any ideas and/or suggestions.
George Lawton Chief Electron Microscopist Molecular and Cellular Imaging Facility UT Southwestern Medical Center at Dallas 5323 Harry Hines Blvd. Dallas, Tx 75390-9039 Phone: 214-648-7291 eMail: George.Lawton-at-email.swmed.edu
If you were working with a pellet in the bottom of a tube or BEEm cap, it is possible that you were not getting good exchange of reagents at the tip of the tube during dehydration and embedding. Or is you were centifuging in the final resin mixture before putting into the oven it is possible that your sample was packed so tightly that there was not enough resin in it to polymerize properly.
At 12:48 PM 02/28/2000 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Gregory W. Erdos, Ph.D. Ph. 352-392-1295 Assistant Director, Biotechnology Program PO Box 110580 Fax: 352-846-0251 University of Florida Gainesville, FL 32611
We have an old Leitz Orthoplan microscope and are looking for the Ultrapak incident light illuminator (housing with fixed ring mirror and interchangeable UO objectives of 4X to 60X primary magnification). These components have not been produced in many years. If anyone has this equipment, we would be pleased to pay for it and for the shipping. Thank you very much!
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Dear Doug,
We have some experience in using micro-PIXE (proton induced X-ray emission) and the so-called nuclear microprobe in toxicological research. It is more sensitive than EDS and WDS, is very easy to quantify (standardless technique). In principle you can have detection limits of the order of 1 ppm in point analysis mode. Specimens can be of any thickness and their actual thickness is assessed by the simultaneous use of the BS technique (proton backscatering spectrometry). What in my opinion makes it really attractive for this purpose is its very good scanning capability. Scan sizes are from ca. 2.5 mm x 2.5 mm down to the beam resolution, which is of the order of few micrometer (can be down to 1 micrometer or even less, but in most of our uses few micrometers is good enough). The sample preparation is similar as for EDS, using cryotechniques to avoid elemental losses and redistribution. We have so far done studies on Ni, Mo, Cd. The detection limits for As are better than for Mo and Cd. Of course, this technique cannot indicate the state of oxidation, you can really treat it as a "more sensitive EDS".
If you want more information, I can refer you to our earlier work and perhaps you might be interested in contacting me.
Best regards
xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx Dr Wojciech J. Przybylowicz Materials Research Group National Accelerator Centre PO Box 72 Faure 7131 South Africa E-mail: przybylowicz-at-nac.ac.za Fax: +27-21-8433543 Phone: +27-21-8433820 http://www.nac.ac.za/IBA/ http://www.nac.ac.za/materials/ xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx
I seem to recall a discussion of software that was available to keep track of printing jobs submitted to a network printer for billing purposes? An argument here for not installing a networked dyesub printer for digital images is who is going to be responsible for watching who's using it and how many prints they're making. We have had some problems like this with the Epson 850 which is on the network and somebody has ran through two color cartridges in a short time. Thanks for any advice.
We are looking for a program (preferably Macintosh-based) that will automatically measure the diameter of spheres that have been imaged in a TEM.
Typically, the spheres measure about 3-5 mm on the TEM negative and often are touching or slightly overlapped. I know that NIH Image should do this, but I have not been able to get reproduceable data using this program.
Someone told me about a program called "Bolero", written by some Spanish university researchers but it is an $8-10K program. Too rich for our tastes.
Any suggestions?
We do appreciate your suggestions.
John
#################################################################### John J. Bozzola, Ph.D., Director Micro-Imaging and Analysis Center 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
I agree with Greg: sounds like an infiltration problem. Your best bet is to encase the bugs in agar. One way is to fix briefly (2-5 min) in glut, then pellet and let sit for a while (1 hr). This glues them together.
Don't resuspend them in the glut or it will fix the walls of the bugs so that they won't stick together.
Cut off the bottom of the tube if plastic or scrape out the cells if tube is glass, onto Parafilm and drain with wedge of filter paper. Divide into 1 cubic mm (or smaller) portions and cover with molter agar. Cut away any excess. Wash. Osmicate and embed as you would a piece of tissue.
Other folks have suspended bugs in molter agar and pelleted them through the agar. You have to do this before the agar cools. Then cut off the end with the cells.
Don't fix the agar/cells in glut; it will cross-link the agar so that the subsequent solutions can't get in. The glut left in the bugs after they're fixed and before they're encased is OK and will wash out.
Hope this helps.
Sara
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
In a message dated 2/28/00 11:44:37 PM, bozzola-at-siu.edu writes:
} Typically, the spheres measure about 3-5 mm on the TEM negative and } often are touching or slightly overlapped. I know that NIH Image } should do this, but I have not been able to get reproduceable data } using this program.
If none of the spheres have been cut off in the section so that you have stereological corrections to make for the missing parts, and you aren't having problems with thresholding, etc., then I don't see what your problem is. Use a watershed to separate the touching features, and use the maximum feret's diameter to estimate the sphere diameter, not the area (which may be reduced due to overaps). Or you can use the maximum values of the euclidean distance map (the ultimate eroded point) as the sphere radii and not even bother with the watershed.
Hello, Is there a web site where I can find more information about quantitative techniques in microscopy on SEM's? In particular the XPP correction procedure?
Many thanks Rachel
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You may want to try Triad Scientific Inc. in Edison, NJ. , triadsci-at-webspan.net or (800) 867-6690. When I was there last month to purchase a vacuum oven, a whole truck lot of optical microscopes and parts (Leitz, etc.) was being unloaded. Most were still in the original boxes. J. Roy Nelson Material Testing Laboratory Pennington, NJ mtl-at-njcc.com
Margie Bryant wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Date: Feb.28, 2000 } } Hello, } } We have an old Leitz Orthoplan microscope and are } looking for the Ultrapak incident light illuminator (housing with fixed } ring mirror and interchangeable UO objectives of 4X to 60X primary } magnification). These components have not been produced in many years. } If anyone has this equipment, we would be pleased to pay for it and for } the shipping. Thank you very much! } } Sincerely, } } Margie Bryant } } From: Margie } Bryant {mbryant-at-com1.med.usf.edu
I've found the same problem sometimes with Epon/Araldite in Eppendorf tubes. I think the pellet is so tightly bound to the bottom of the tube by the time 100% resin infiltration is reached (especially if you centrifuge along the way) that the resin doesn't have good access to the cells or material at the 'bottom'. I now lift (ease gently) the pellet off the bottom of the tube with a toothpick to ensure that the media are totally surrounding the pellet. Another choice is to cut away all the gooey resin and hope that the material at the 'top' of the pellet is well infiltrated and polymerized.
Regards, Marilyn
George Lawton wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Occassionally someone sends a question to the listserver and there is no response. It doesn't happen often so I thought I would send this question again. } } Several weeks ago, I was given some inner membranes from E. coli to be run up to Epon and sectioned for TEM. I treated the sample like tissue culture and pelleted the membranes. Fixed in } 3% glut in PBS and post-fixed in 1% OsO4. Routine dehydration in alcohols. Embedded in } EMbed 812 with BDMA in Beem Capsules. Left in oven 48 hours. When I tried to section, the tissue was too soft and appeared to not be infiltrated well. Put back into the oven for 2-3 more days. No difference. Thinking I had a neural meltdown while processing the samples, I tried again on fresh samples, paying particular attention to details. I also used new bottles of embedding media. Same results. } The part of the block without the tissue is its usual hardness. The part of the block with the tissue } (the tip of the beem capsule) is soft and tacky. } } Has anyone experienced anything like this? Any suggestions on how I can get the sample into epon? } } Thanks in advance for any ideas and/or suggestions. } } George Lawton } Chief Electron Microscopist } Molecular and Cellular Imaging Facility } UT Southwestern Medical Center at Dallas } 5323 Harry Hines Blvd. } Dallas, Tx 75390-9039 } Phone: 214-648-7291 } eMail: George.Lawton-at-email.swmed.edu
Two things come to mind. First, if processed in a microfuge tube, loosen the pellet from the tube and make sure all fluids and epoxy mixtures come into contact with all surfaces (as already recommended). Second, I always make it a practice to preheat the BEEM capsules and labels in the embedding oven all day or overnight before use to drive off any moisture. This really seems to help in our humid environment!
Good luck and aloha, Tina
Sunny and 80F but with mosquitoes.
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Rick: What you want to do is fairly easy if you are using Windows NT. It will maintain a log of the user name, number of prints, date, name of print job, etc. I posted instructions to this group in 1997. These instructions were published in Microscopy and Microanalysis Vol.3, number 6, p. 569. If you don't have access to this, let me know and I will send the directions. Stanley L. Flegler, Acting Director Center for Advanced Microscopy Michigan State University
Question: Currently studying for a project on HREM of semiconductors, namely GaN; I would like to know where I can obtain some good information on this subject. My lecturer does not have much insight in this area, I need to study its structure, and I am currently using the Cerius(2) software package to simulate images of GaN.
The Centre for Microscopy & Microanalysis (CMM) at The University of Queensland will demonstrate its web-based interactive Virtual Electron Microscopy service this Thursday (March 2, 2000) between 11am and 1pm. (Queensland Time; GMT +10)
CyberSTEM (WWW accessible Scanning and Transmission Electron Microscopy) is a service that became officially permanent on the web after Centre Directors approved funding last week.
URL: http://www.uq.edu.au/nanoworld/online.html
The CMM's videoconference service - begun in 1995 - has evolved into a webcam-based service that allows unlimited live connections to microscopy sessions conducted at the Centre. Visitors can interact with staff during organised sessions via telephone, by chat programs or via email. (Please note: Live interaction is only available during specifically organised sessions - direct all other inquiries to the address/email noted in the signature below.)
Though primarily intended to service paying customers who cannot physically attend sessions, video streaming from two of their microscopes (more later they hope) will be regularly accessible by the general public.
Sincerely,
Duncan
-- ************************************************************ Duncan Waddell (BSc) Senior Scientific Officer Centre for Microscopy and Microanalysis The University of Queensland, St. Lucia, Qld, 4072 Australia Telephone: +61-7-3365-4216 Facsimile: +61-7-3365-4422 WWW: http://www.uq.edu.au/nanoworld/nanohome.html ************************************************************ Any opinion expressed is that of the writer, and not necessarily that of CMM or of the University. ************************************************************
Dear George, In all my experience working up bacteria and bacterial fragments, I have always, and by the way this is the simplest, filtered all suspended cells on Millipore filters and processed the samples on the filters. This yielded excellent results. The fragments, cells stayed on the filters and never detached. Filter your suspended fragment solution using a 0.2um filter unit system attached to a vacuum system. The aspirator of a faucet works great. Rinse in buffer while on the filter system. Then remove the filter and gently place in a plastic or glass petri dish. Again, gently drop fixative over the sample enough to cover your filter and carefully slice it into your appropriate sizes and process from there. You will have success. If you have further questions, just give me a call.
Blessings, Maria
Maria Fazio-Zanakis Bioimaging and Molecular Histology Aventis Pharmaceuticals 1-908-231-3357 Fax: 1-908-231-3962 Email: Maria.Fazio-Zanakis-at-Aventis.com
-----Original Message----- } From: George Lawton [mailto:George.Lawton-at-email.swmed.edu] Sent: Monday, February 28, 2000 1:49 PM To: microscopy-at-sparc5.microscopy.com
Occassionally someone sends a question to the listserver and there is no response. It doesn't happen often so I thought I would send this question again.
Several weeks ago, I was given some inner membranes from E. coli to be run up to Epon and sectioned for TEM. I treated the sample like tissue culture and pelleted the membranes. Fixed in 3% glut in PBS and post-fixed in 1% OsO4. Routine dehydration in alcohols. Embedded in EMbed 812 with BDMA in Beem Capsules. Left in oven 48 hours. When I tried to section, the tissue was too soft and appeared to not be infiltrated well. Put back into the oven for 2-3 more days. No difference. Thinking I had a neural meltdown while processing the samples, I tried again on fresh samples, paying particular attention to details. I also used new bottles of embedding media. Same results. The part of the block without the tissue is its usual hardness. The part of the block with the tissue (the tip of the beem capsule) is soft and tacky.
Has anyone experienced anything like this? Any suggestions on how I can get the sample into epon?
Thanks in advance for any ideas and/or suggestions.
George Lawton Chief Electron Microscopist Molecular and Cellular Imaging Facility UT Southwestern Medical Center at Dallas 5323 Harry Hines Blvd. Dallas, Tx 75390-9039 Phone: 214-648-7291 eMail: George.Lawton-at-email.swmed.edu
Can anyone help me? I am doing research on the plant Achillea millefolium (Yarrow)and I need to find out the ploidy level of 2 seed sources. I am looking for information on companies/institutions that will do this work for me and a rough estimate of the cost. Many thanks, Fiona Russell.
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I recently tried to help out Dr. M. Rohde who was embedding in Spurr's formulation and attempting to locate an antigen.
I said that Spurr's formulation has no place in immunocytochemistry for all the reasons I listed then. This is an oversimplification (I am always harassed for time when trying to answer questions completely.) Let me tell you that I, myself, worked with a graduate student who used Spurr's medium, etched for one hour in sodium metaperiodate, and then got wonderful Au label. The reason for this success was that she had an antigen that was so sturdy and such plentiful supply that it was a candidate for the prozone effect. Her same exact protocol (actually mine) when tried by us in our laboratory was a spectacular failure with our antigen which we soon discovered was scarce and delicate and needed amplification up to the point where I wondered whether we were actually increasing the thickness of the section by amplification methods! Sort of like painting a wall until the room gets smaller! At any rate - yes, Spurr's can be used for immunocytochemistry. However, if at first you don't succeed, don't try again. Go to another epoxy, and then to an acrylic. I should not make absolute statements as I did. I did not mean to discourage those who successfully use Spurr's to abandon it. Besides, compared to what there is to be known about immunocytochemistry, my amount of knowledge can be dropped into a thimble with room left to stuff an entire thumb in after it.
I always reserve the right to be wrong!
Hildy Crowley Sr. Electron Microscopy Specialist Dep. of Biol. Sciences University of Denver Denver, CO
I remember seeing posts over the years regarding web-based equipment sign-up systems, but didn't save any of them. Can anyone direct me to an archived discussion thread, or have current suggestions on a very simple and easy way to set this up? I want to put this under a dept. web page on our intranet only, so security is not a major issue. All I want is a simple calendar that users can view and sign up for time slots. No retribution if time not used, but must require user's name so that they can be tracked down for questioning if needed. I have no experience in HTML programming but am not afraid to learn.
Thanks as always, Karen
-- Karen S. Zaruba kszaruba-at-mmm.com 3M Company, St. Paul, MN
Any suggestions on a really good video printer to include with an SEM purchase? We've had trouble over the years with the standard roll printers getting paper jams, which required sending out for servicing (i.e. lots of down time). We're willing to sacrifice some speed for something reliable. Are there "fast" ( { 1 min. per print) video printers that use the cut sheet paper?
Thanks, Karen
P.S. vendors please respond by E-mail (not phone) or dare to face my wrath. -- Karen S. Zaruba kszaruba-at-mmm.com 3M Company, St. Paul, MN
We have 2 Tracor Northern/Noran TN 5500 EDX systems that need a new home. We are looking for a possible trade for comparable priced equipment/software. If anyone is interested, please contact me directly.
Harry Ekstrom Honeywell Materials Laboratory Phoenix, AZ 602-231-2744
Sorry for the inconvenience, but we mistakenly scheduled our Tripod Polisher¨ Workshop over Memorial Day Weekend. While I'm sure that you would all love to spend your holiday weekend here with us, we felt it necessary to move the workshop to the following week. The workshop is now scheduled for June 2- 3, 2000 in San Clemente, CA.
For those of you who have already registered, you should have already been contacted and given the option of re-booking for the new date or canceling your reservation. Again, I apologize for the error. Certainly, if anyone wishes to come and visit us over the Memorial Day Weekend, we will welcome you with open arms - but you'll have to wait until the following weekend for the workshop!
David Henriks Vice President TEL: 800-728-2233 (toll free in the USA) South Bay Technology, Inc. +1-949-492-2600 1120 Via Callejon FAX: +1-949-492-1499 San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com